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Spector, Neil Lee

Positions:

Sandra Coates Associate Professor

Medicine, Medical Oncology
School of Medicine

Associate Professor of Medicine

Medicine, Medical Oncology
School of Medicine

Associate Professor of Pharmacology & Cancer Biology

Pharmacology & Cancer Biology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

M.D. 1982

M.D. — University of Medicine and Dentistry of New Jersey

Intern, Medicine

University of Texas at Dallas

First Year Neurology Resident, Neurology

University of Texas at Dallas

Medical Resident, Medicine

University of Texas at Dallas

News:

Dr. Neil Spector: Latest advances in cancer care

September 30, 2015 — KTEP-El Paso’s “Science Studio"

Researchers turn to canine clinical trials to advance cancer therapies

April 01, 2016 — JAMA: The Journal of the American Medical Association

Grants:

Postdoctoral training in genomic medicine research

Administered By
Duke Center for Applied Genomics and Precision Medicine
AwardedBy
National Institutes of Health
Role
Co-Mentor
Start Date
June 14, 2017
End Date
May 31, 2022

Regional Oncolytic Poliovirus Immunotherapy for Breast Cancer

Administered By
Surgery, Surgical Sciences
AwardedBy
Department of Defense
Role
Co Investigator
Start Date
August 01, 2016
End Date
July 31, 2021

Role of ErbB Receptor Signaling in Regulating Normal and Leukemic Stem Cell Fate

Administered By
Medicine, Hematological Malignancies
AwardedBy
National Institutes of Health
Role
Mentor
Start Date
September 09, 2014
End Date
August 31, 2019

Developing a HER3 Vaccine to Prevent Resistance to Endocrine Therapy

Administered By
Surgery, Surgical Sciences
AwardedBy
Department of Defense
Role
Co Investigator
Start Date
September 30, 2012
End Date
September 29, 2018

Oncogenic Signaling Networks

Administered By
Surgery, Surgical Sciences
AwardedBy
Department of Defense
Role
Co Investigator
Start Date
September 30, 2012
End Date
September 29, 2018

Immunolight Renewal

Administered By
Biomedical Engineering
AwardedBy
Immunolight, LLC
Role
Co-Principal Investigator
Start Date
February 01, 2014
End Date
December 31, 2017

Circumventing Therapeutic Resistance and the Emergence of Disseminated Breast Cancer Cells

Administered By
Chemistry
AwardedBy
United States Army Medical Research Acquisition Activity
Role
Co-Principal Investigator
Start Date
July 01, 2013
End Date
June 30, 2017

Circumventing Therapeutic Resistance and the Emergency of Disseminated Breast Cancer Cells through Non-Invasive Optical

Administered By
Duke Cancer Institute
AwardedBy
United States Army Medical Research Acquisition Activity
Role
Principal Investigator
Start Date
July 01, 2013
End Date
June 30, 2017

Kinase Target of Diverse Cell Surface Receptors in Cancer Invasion and Metastasis

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
July 21, 2011
End Date
April 30, 2017

HIF-1 driven therapeutic resistance mechanisms in chest wall recurrences of inflammatory breast cancer (IBC)

Administered By
Radiation Oncology
AwardedBy
Inflammatory Breast Cancer Research Foundation
Role
Collaborator
Start Date
December 01, 2013
End Date
December 31, 2016

The Role of HER2 Signaling Networks in Early Stage Breast Cancer Initiation and Resistance to Tamoxifen Prevention

Administered By
Medicine, Medical Oncology
AwardedBy
UT MD Anderson Cancer Center
Role
Co Investigator
Start Date
September 25, 2009
End Date
September 24, 2015

Does Cytoplasmic PELP1 Predict Resistance To Tamoxifen Chemoprevention?

Administered By
Medicine, Medical Oncology
AwardedBy
National Institutes of Health
Role
Co-Mentor
Start Date
September 01, 2010
End Date
August 31, 2015

Molecular mechanisms of chemoresistance in breast cancer

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
May 01, 2010
End Date
April 30, 2012

Small Animal PET / CT Molecular Imaging

Administered By
Radiology, Nuclear Medicine
AwardedBy
National Institutes of Health
Role
Major User
Start Date
April 01, 2011
End Date
March 31, 2012

ACOSOG Competing Renewal

Administered By
Duke Clinical Research Institute
AwardedBy
National Institutes of Health
Role
Committee Chair
Start Date
May 01, 2008
End Date
November 30, 2011

Quantitative and Qualitative Differential Expression Proteomics

Administered By
Institutes and Centers
AwardedBy
National Institutes of Health
Role
Major User
Start Date
July 15, 2010
End Date
July 14, 2011

Role of XIAP in Therapeutic Resistance in Inflammatory Breast Cancer

Administered By
Surgery
AwardedBy
Dept. of the Army -- USAMRAA
Role
Consultant
Start Date
July 01, 2008
End Date
August 31, 2010
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Publications:

A Fluorescent Hsp90 Probe Demonstrates the Unique Association between Extracellular Hsp90 and Malignancy in Vivo.

Extracellular expression of heat shock protein 90 (eHsp90) by tumor cells is correlated with malignancy. Development of small molecule probes that can detect eHsp90 in vivo may therefore have utility in the early detection of malignancy. We synthesized a cell impermeable far-red fluorophore-tagged Hsp90 inhibitor to target eHsp90 in vivo. High resolution confocal and lattice light sheet microscopy show that probe-bound eHsp90 accumulates in punctate structures on the plasma membrane of breast tumor cells and is actively internalized. The extent of internalization correlates with tumor cell aggressiveness, and this process can be induced in benign cells by overexpressing p110HER2. Whole body cryoslicing, imaging, and histology of flank and spontaneous tumor-bearing mice strongly suggests that eHsp90 expression and internalization is a phenomenon unique to tumor cells in vivo and may provide an "Achilles heel" for the early diagnosis of metastatic disease and targeted drug delivery.

Authors
Crowe, LB; Hughes, PF; Alcorta, DA; Osada, T; Smith, AP; Totzke, J; Loiselle, DR; Lutz, ID; Gargesha, M; Roy, D; Roques, J; Darr, D; Lyerly, HK; Spector, NL; Haystead, TAJ
MLA Citation
Crowe, LB, Hughes, PF, Alcorta, DA, Osada, T, Smith, AP, Totzke, J, Loiselle, DR, Lutz, ID, Gargesha, M, Roy, D, Roques, J, Darr, D, Lyerly, HK, Spector, NL, and Haystead, TAJ. "A Fluorescent Hsp90 Probe Demonstrates the Unique Association between Extracellular Hsp90 and Malignancy in Vivo." ACS chemical biology 12.4 (April 2017): 1047-1055.
PMID
28103010
Source
epmc
Published In
ACS Chemical Biology
Volume
12
Issue
4
Publish Date
2017
Start Page
1047
End Page
1055
DOI
10.1021/acschembio.7b00006

Clonal Evolutionary Analysis during HER2 Blockade in HER2-Positive Inflammatory Breast Cancer: A Phase II Open-Label Clinical Trial of Afatinib +/- Vinorelbine.

Inflammatory breast cancer (IBC) is a rare, aggressive form of breast cancer associated with HER2 amplification, with high risk of metastasis and an estimated median survival of 2.9 y. We performed an open-label, single-arm phase II clinical trial (ClinicalTrials.gov NCT01325428) to investigate the efficacy and safety of afatinib, an irreversible ErbB family inhibitor, alone and in combination with vinorelbine in patients with HER2-positive IBC. This trial included prospectively planned exome analysis before and after afatinib monotherapy.HER2-positive IBC patients received afatinib 40 mg daily until progression, and thereafter afatinib 40 mg daily and intravenous vinorelbine 25 mg/m2 weekly. The primary endpoint was clinical benefit; secondary endpoints were objective response (OR), duration of OR, and progression-free survival (PFS). Of 26 patients treated with afatinib monotherapy, clinical benefit was achieved in 9 patients (35%), 0 of 7 trastuzumab-treated patients and 9 of 19 trastuzumab-naïve patients. Following disease progression, 10 patients received afatinib plus vinorelbine, and clinical benefit was achieved in 2 of 4 trastuzumab-treated and 0 of 6 trastuzumab-naïve patients. All patients had treatment-related adverse events (AEs). Whole-exome sequencing of tumour biopsies taken before treatment and following disease progression on afatinib monotherapy was performed to assess the mutational landscape of IBC and evolutionary trajectories during therapy. Compared to a cohort of The Cancer Genome Atlas (TCGA) patients with HER2-positive non-IBC, HER2-positive IBC patients had significantly higher mutational and neoantigenic burden, more frequent gain-of-function TP53 mutations and a recurrent 11q13.5 amplification overlapping PAK1. Planned exploratory analysis revealed that trastuzumab-naïve patients with tumours harbouring somatic activation of PI3K/Akt signalling had significantly shorter PFS compared to those without (p = 0.03). High genomic concordance between biopsies taken before and following afatinib resistance was observed with stable clonal structures in non-responding tumours, and evidence of branched evolution in 8 of 9 tumours analysed. Recruitment to the trial was terminated early following the LUX-Breast 1 trial, which showed that afatinib combined with vinorelbine had similar PFS and OR rates to trastuzumab plus vinorelbine but shorter overall survival (OS), and was less tolerable. The main limitations of this study are that the results should be interpreted with caution given the relatively small patient cohort and the potential for tumour sampling bias between pre- and post-treatment tumour biopsies.Afatinib, with or without vinorelbine, showed activity in trastuzumab-naïve HER2-positive IBC patients in a planned subgroup analysis. HER2-positive IBC is characterized by frequent TP53 gain-of-function mutations and a high mutational burden. The high mutational load associated with HER2-positive IBC suggests a potential role for checkpoint inhibitor therapy in this disease.ClinicalTrials.gov NCT01325428.

Authors
Goh, G; Schmid, R; Guiver, K; Arpornwirat, W; Chitapanarux, I; Ganju, V; Im, S-A; Kim, S-B; Dechaphunkul, A; Maneechavakajorn, J; Spector, N; Yau, T; Afrit, M; Ahmed, SB; Johnston, SR; Gibson, N; Uttenreuther-Fischer, M; Herrero, J; Swanton, C
MLA Citation
Goh, G, Schmid, R, Guiver, K, Arpornwirat, W, Chitapanarux, I, Ganju, V, Im, S-A, Kim, S-B, Dechaphunkul, A, Maneechavakajorn, J, Spector, N, Yau, T, Afrit, M, Ahmed, SB, Johnston, SR, Gibson, N, Uttenreuther-Fischer, M, Herrero, J, and Swanton, C. "Clonal Evolutionary Analysis during HER2 Blockade in HER2-Positive Inflammatory Breast Cancer: A Phase II Open-Label Clinical Trial of Afatinib +/- Vinorelbine." PLoS medicine 13.12 (December 6, 2016): e1002136-.
PMID
27923043
Source
epmc
Published In
PLoS medicine
Volume
13
Issue
12
Publish Date
2016
Start Page
e1002136
DOI
10.1371/journal.pmed.1002136

A role for the androgen receptor in the treatment of male breast cancer.

Male breast cancer (BC) is relatively rare, making up less than 1% of all breast cancer cases in the United States. Treatment guidelines for male BC are derived from studies on the treatment of female BC, and are based molecular and clinical characteristics, such as hormone receptor positivity. For female estrogen receptor positive (ER+) breast cancers, the standard of care includes three classes of endocrine therapies: selective estrogen receptor modulators, aromatase inhibitors, and pure anti-estrogens. In contrast to female ER+ breast cancers, there is less known about the optimal treatment for male ER+ BC. Furthermore, in contrast to ER, less is known about the role of the androgen receptor (AR) in male and female BC. We report here the treatment of a 28-year-old man with metastatic AR+, ER+ breast cancer otherwise refractory to chemotherapy, who has had a durable clinical response to hormonal suppression with the combination of aromatase inhibition (Letrozole) in conjunction with a GnRH agonist (Leuprolide).

Authors
Zhu, J; Davis, CT; Silberman, S; Spector, N; Zhang, T
MLA Citation
Zhu, J, Davis, CT, Silberman, S, Spector, N, and Zhang, T. "A role for the androgen receptor in the treatment of male breast cancer." Critical reviews in oncology/hematology 98 (February 2016): 358-363. (Review)
PMID
26669267
Source
epmc
Published In
Critical Reviews in Oncology/Hematology
Volume
98
Publish Date
2016
Start Page
358
End Page
363
DOI
10.1016/j.critrevonc.2015.11.013

X-Ray Psoralen Activated Cancer Therapy (X-PACT).

This work investigates X-PACT (X-ray Psoralen Activated Cancer Therapy): a new approach for the treatment of solid cancer. X-PACT utilizes psoralen, a potent anti-cancer therapeutic with current application to proliferative disease and extracorporeal photopheresis (ECP) of cutaneous T Cell Lymphoma. An immunogenic role for light-activated psoralen has been reported, contributing to long-term clinical responses. Psoralen therapies have to-date been limited to superficial or extracorporeal scenarios due to the requirement for psoralen activation by UVA light, which has limited penetration in tissue. X-PACT solves this challenge by activating psoralen with UV light emitted from novel non-tethered phosphors (co-incubated with psoralen) that absorb x-rays and re-radiate (phosphoresce) at UV wavelengths. The efficacy of X-PACT was evaluated in both in-vitro and in-vivo settings. In-vitro studies utilized breast (4T1), glioma (CT2A) and sarcoma (KP-B) cell lines. Cells were exposed to X-PACT treatments where the concentrations of drug (psoralen and phosphor) and radiation parameters (energy, dose, and dose rate) were varied. Efficacy was evaluated primarily using flow cell cytometry in combination with complimentary assays, and the in-vivo mouse study. In an in-vitro study, we show that X-PACT induces significant tumor cell apoptosis and cytotoxicity, unlike psoralen or phosphor alone (p<0.0001). We also show that apoptosis increases as doses of phosphor, psoralen, or radiation increase. Finally, in an in-vivo pilot study of BALBc mice with syngeneic 4T1 tumors, we show that the rate of tumor growth is slower with X-PACT than with saline or AMT + X-ray (p<0.0001). Overall these studies demonstrate a potential therapeutic effect for X-PACT, and provide a foundation and rationale for future studies. In summary, X-PACT represents a novel treatment approach in which well-tolerated low doses of x-ray radiation are delivered to a specific tumor site to generate UVA light which in-turn unleashes both short- and potentially long-term antitumor activity of photo-active therapeutics like psoralen.

Authors
Oldham, M; Yoon, P; Fathi, Z; Beyer, WF; Adamson, J; Liu, L; Alcorta, D; Xia, W; Osada, T; Liu, C; Yang, XY; Dodd, RD; Herndon, JE; Meng, B; Kirsch, DG; Lyerly, HK; Dewhirst, MW; Fecci, P; Walder, H; Spector, NL
MLA Citation
Oldham, M, Yoon, P, Fathi, Z, Beyer, WF, Adamson, J, Liu, L, Alcorta, D, Xia, W, Osada, T, Liu, C, Yang, XY, Dodd, RD, Herndon, JE, Meng, B, Kirsch, DG, Lyerly, HK, Dewhirst, MW, Fecci, P, Walder, H, and Spector, NL. "X-Ray Psoralen Activated Cancer Therapy (X-PACT)." PloS one 11.9 (January 2016): e0162078-.
Website
http://hdl.handle.net/10161/13034
PMID
27583569
Source
epmc
Published In
PloS one
Volume
11
Issue
9
Publish Date
2016
Start Page
e0162078
DOI
10.1371/journal.pone.0162078

CHAMBER: A Regional Performance Improvement CME Initiative for Breast Cancer Health Care Providers.

CHAMBER was a regional educational initiative for providers of care to patients with HER2+ breast cancer. The study goals were to (1) enhance testing for HER2/neu overexpression in patients with invasive breast cancer; (2) increase the appropriate use of targeted therapy for patients with HER2+ breast cancer; and (3) enhance patients' coping ability. This Performance Improvement Continuing Medical Education (PI-CME) initiative included clinical practice assessment, educational activities, and reassessment. Chart review revealed a high rate of HER2 testing (98%) before and after education. Targeted therapy for patients with HER2+ breast cancer declined after the program (from 96% to 61%), perhaps attributable to an increase in awareness of medical reasons to avoid use of targeted therapy. Assessment for patients' emotional coping ability increased after education (from 55% to 76%; P=.01). Rates of testing for HER2 amplification and assessment of emotional well-being after education were consistent with ASCO Quality Oncology Practice Initiative benchmark values. Documentation of actions to address emotional problems remained an area for improvement.

Authors
Sutton, LM; Geradts, J; Hamilton, EP; Havlin, KA; Kimmick, GG; Marcom, PK; Spector, NL; Watson, M; Rabin, DU; Bruno, TO; Noe, A; Miller, S; Subramaniam, C; Layton, S; Grichnik, K
MLA Citation
Sutton, LM, Geradts, J, Hamilton, EP, Havlin, KA, Kimmick, GG, Marcom, PK, Spector, NL, Watson, M, Rabin, DU, Bruno, TO, Noe, A, Miller, S, Subramaniam, C, Layton, S, and Grichnik, K. "CHAMBER: A Regional Performance Improvement CME Initiative for Breast Cancer Health Care Providers." J Natl Compr Canc Netw 13.8 (August 2015): 1005-1011.
PMID
26285246
Source
pubmed
Published In
Journal of the National Comprehensive Cancer Network : JNCCN
Volume
13
Issue
8
Publish Date
2015
Start Page
1005
End Page
1011

Lapatinib Plasma and Tumor Concentrations and Effects on HER Receptor Phosphorylation in Tumor.

The paradigm shift in cancer treatment from cytotoxic drugs to tumor targeted therapies poses new challenges, including optimization of dose and schedule based on a biologically effective dose, rather than the historical maximum tolerated dose. Optimal dosing is currently determined using concentrations of tyrosine kinase inhibitors in plasma as a surrogate for tumor concentrations. To examine this plasma-tumor relationship, we explored the association between lapatinib levels in tumor and plasma in mice and humans, and those effects on phosphorylation of human epidermal growth factor receptors (HER) in human tumors.Mice bearing BT474 HER2+ human breast cancer xenografts were dosed once or twice daily (BID) with lapatinib. Drug concentrations were measured in blood, tumor, liver, and kidney. In a randomized phase I clinical trial, 28 treatment-naïve female patients with early stage HER2+ breast cancer received lapatinib 1000 or 1500 mg once daily (QD) or 500 mg BID before evaluating steady-state lapatinib levels in plasma and tumor.In mice, lapatinib levels were 4-fold higher in tumor than blood with a 4-fold longer half-life. Tumor concentrations exceeded the in vitro IC90 (~ 900 nM or 500 ng/mL) for inhibition of HER2 phosphorylation throughout the 12-hour dosing interval. In patients, tumor levels were 6- and 10-fold higher with QD and BID dosing, respectively, compared to plasma trough levels. The relationship between tumor and plasma concentration was complex, indicating multiple determinants. HER receptor phosphorylation varied depending upon lapatinib tumor concentrations, suggestive of changes in the repertoire of HER homo- and heterodimers.Plasma lapatinib concentrations underestimated tumor drug levels, suggesting that optimal dosing should be focused on the site of action to avoid to inappropriate dose escalation. Larger clinical trials are required to determine optimal dose and schedule to achieve tumor concentrations that maximally inhibit HER receptors.NCT00359190.

Authors
Spector, NL; Robertson, FC; Bacus, S; Blackwell, K; Smith, DA; Glenn, K; Cartee, L; Harris, J; Kimbrough, CL; Gittelman, M; Avisar, E; Beitsch, P; Koch, KM
MLA Citation
Spector, NL, Robertson, FC, Bacus, S, Blackwell, K, Smith, DA, Glenn, K, Cartee, L, Harris, J, Kimbrough, CL, Gittelman, M, Avisar, E, Beitsch, P, and Koch, KM. "Lapatinib Plasma and Tumor Concentrations and Effects on HER Receptor Phosphorylation in Tumor." PloS one 10.11 (January 2015): e0142845-.
PMID
26571496
Source
epmc
Published In
PloS one
Volume
10
Issue
11
Publish Date
2015
Start Page
e0142845
DOI
10.1371/journal.pone.0142845

Identification of an allosteric small-molecule inhibitor selective for the inducible form of heat shock protein 70

©2014 Elsevier Ltd. All rights reserved. Summary Inducible Hsp70 (Hsp70i) is overexpressed in a wide spectrum of human tumors, and its expression correlates with metastasis, poor outcomes, and resistance to chemotherapy in patients. Identification of small-molecule inhibitors selective for Hsp70i could provide new therapeutic tools for cancer treatment. In this work, we used fluorescence-linked enzyme chemoproteomic strategy (FLECS) to identify HS-72, an allosteric inhibitor selective for Hsp70i. HS-72 displays the hallmarks of Hsp70 inhibition in cells, promoting substrate protein degradation and growth inhibition. Importantly, HS-72 is selective for Hsp70i over the closely related constitutively active Hsc70. Studies with purified protein show HS-72 acts as an allosteric inhibitor, reducing ATP affinity. In vivo HS-72 is well-tolerated, showing bioavailability and efficacy, inhibiting tumor growth and promoting survival in a HER2+ model of breast cancer. The HS-72 scaffold is amenable to resynthesis and iteration, suggesting an ideal starting point for a new generation of anticancer therapeutics targeting Hsp70i.

Authors
Howe, MK; Bodoor, K; Carlson, DA; Hughes, PF; Alwarawrah, Y; Loiselle, DR; Jaeger, AM; Darr, DB; Jordan, JL; Hunter, LM; Molzberger, ET; Gobillot, TA; Thiele, DJ; Brodsky, JL; Spector, NL; Haystead, TAJ
MLA Citation
Howe, MK, Bodoor, K, Carlson, DA, Hughes, PF, Alwarawrah, Y, Loiselle, DR, Jaeger, AM, Darr, DB, Jordan, JL, Hunter, LM, Molzberger, ET, Gobillot, TA, Thiele, DJ, Brodsky, JL, Spector, NL, and Haystead, TAJ. "Identification of an allosteric small-molecule inhibitor selective for the inducible form of heat shock protein 70." Chemistry and Biology 21.12 (December 18, 2014): 1648-1659.
Source
scopus
Published In
Chemistry & Biology
Volume
21
Issue
12
Publish Date
2014
Start Page
1648
End Page
1659
DOI
10.1016/j.chembiol.2014.10.016

Identification of an allosteric small-molecule inhibitor selective for the inducible form of heat shock protein 70.

Inducible Hsp70 (Hsp70i) is overexpressed in a wide spectrum of human tumors, and its expression correlates with metastasis, poor outcomes, and resistance to chemotherapy in patients. Identification of small-molecule inhibitors selective for Hsp70i could provide new therapeutic tools for cancer treatment. In this work, we used fluorescence-linked enzyme chemoproteomic strategy (FLECS) to identify HS-72, an allosteric inhibitor selective for Hsp70i. HS-72 displays the hallmarks of Hsp70 inhibition in cells, promoting substrate protein degradation and growth inhibition. Importantly, HS-72 is selective for Hsp70i over the closely related constitutively active Hsc70. Studies with purified protein show HS-72 acts as an allosteric inhibitor, reducing ATP affinity. In vivo HS-72 is well-tolerated, showing bioavailability and efficacy, inhibiting tumor growth and promoting survival in a HER2+ model of breast cancer. The HS-72 scaffold is amenable to resynthesis and iteration, suggesting an ideal starting point for a new generation of anticancer therapeutics targeting Hsp70i.

Authors
Howe, MK; Bodoor, K; Carlson, DA; Hughes, PF; Alwarawrah, Y; Loiselle, DR; Jaeger, AM; Darr, DB; Jordan, JL; Hunter, LM; Molzberger, ET; Gobillot, TA; Thiele, DJ; Brodsky, JL; Spector, NL; Haystead, TAJ
MLA Citation
Howe, MK, Bodoor, K, Carlson, DA, Hughes, PF, Alwarawrah, Y, Loiselle, DR, Jaeger, AM, Darr, DB, Jordan, JL, Hunter, LM, Molzberger, ET, Gobillot, TA, Thiele, DJ, Brodsky, JL, Spector, NL, and Haystead, TAJ. "Identification of an allosteric small-molecule inhibitor selective for the inducible form of heat shock protein 70." Chemistry & biology 21.12 (December 11, 2014): 1648-1659.
PMID
25500222
Source
epmc
Published In
Chemistry & Biology
Volume
21
Issue
12
Publish Date
2014
Start Page
1648
End Page
1659
DOI
10.1016/j.chembiol.2014.10.016

Pertuzumab protects the achilles' heel of trastuzumab--emtansine.

Trastuzumab emtansine (T-DM1) represents a significant advancement in the treatment of HER2(+) breast cancers. Its clinical efficacy however will be limited by the development of therapeutic resistance. In this report, the HER3 ligand neuregulin is shown to mediate T-DM1 resistance, which was overcome by administration of pertuzumab, a steric inhibitor of HER2 dimerization.

Authors
Gwin, WR; Spector, NL
MLA Citation
Gwin, WR, and Spector, NL. "Pertuzumab protects the achilles' heel of trastuzumab--emtansine." Clin Cancer Res 20.2 (January 15, 2014): 278-280.
PMID
24240115
Source
pubmed
Published In
Clinical cancer research : an official journal of the American Association for Cancer Research
Volume
20
Issue
2
Publish Date
2014
Start Page
278
End Page
280
DOI
10.1158/1078-0432.CCR-13-2626

Photo-activated psoralen binds the ErbB2 catalytic kinase domain, blocking ErbB2 signaling and triggering tumor cell apoptosis.

Photo-activation of psoralen with UVA irradiation, referred to as PUVA, is used in the treatment of proliferative skin disorders. The anti-proliferative effects of PUVA have been largely attributed to psoralen intercalation of DNA, which upon UV treatment, triggers the formation of interstrand DNA crosslinks (ICL) that inhibit transcription and DNA replication. Here, we show that PUVA exerts antitumor effects in models of human breast cancer that overexpress the ErbB2 receptor tyrosine kinase oncogene, through a new mechanism. Independent of ICL formation, the antitumor effects of PUVA in ErbB2+ breast cancer models can instead be mediated through inhibition of ErbB2 activation and signaling. Using a mass spectroscopy-based approach, we show for the first time that photo-activated 8MOP (8-methoxypsoralen) interacts with the ErbB2 catalytic autokinase domain. Furthermore, PUVA can reverse therapeutic resistance to lapatinib and other ErbB2 targeted therapies, including resistance mediated via expression of a phosphorylated, truncated form of ErbB2 (p85(ErbB2)) that is preferentially expressed in tumor cell nuclei. Current ErbB2 targeted therapies, small molecule kinase inhibitors or antibodies, do not block the phosphorylated, activated state of p85(ErbB2). Here we show that PUVA reduced p85(ErbB2) phosphorylation leading to tumor cell apoptosis. Thus, in addition to its effects on DNA and the formation of ICL, PUVA represents a novel ErbB2 targeted therapy for the treatment of ErbB2+ breast cancers, including those that have developed resistance to other ErbB2 targeted therapies.

Authors
Xia, W; Gooden, D; Liu, L; Zhao, S; Soderblom, EJ; Toone, EJ; Beyer, WF; Walder, H; Spector, NL
MLA Citation
Xia, W, Gooden, D, Liu, L, Zhao, S, Soderblom, EJ, Toone, EJ, Beyer, WF, Walder, H, and Spector, NL. "Photo-activated psoralen binds the ErbB2 catalytic kinase domain, blocking ErbB2 signaling and triggering tumor cell apoptosis." PloS one 9.2 (January 2014): e88983-.
PMID
24551203
Source
epmc
Published In
PloS one
Volume
9
Issue
2
Publish Date
2014
Start Page
e88983
DOI
10.1371/journal.pone.0088983

Preclinical activity profile and therapeutic efficacy of the HSP90 inhibitor ganetespib in triple-negative breast cancer.

PURPOSE: Treatment options for patients with triple-negative breast cancer (TNBC) are largely limited to systemic chemotherapies, which have shown disappointing efficacy in the metastatic setting. Here, we undertook a comprehensive evaluation of the activity of ganetespib, a potent inhibitor of HSP90, in this malignancy. EXPERIMENTAL DESIGN: The antitumor and antimetastatic activity of ganetespib was investigated using TNBC cell lines and xenograft models. Combinatorial drug analyses were performed with chemotherapeutic agents and concomitant effects on DNA damage and cell-cycle disruption were assessed in vitro; antitumor efficacy was assessed in vivo. Metabolic and objective tumor responses were evaluated in patients with metastatic TNBC undergoing ganetespib treatment. RESULTS: Ganetespib simultaneously deactivated multiple oncogenic pathways to potently reduce cell viability in TNBC cell lines, and suppressed lung metastases in experimental models. Ganetespib potentiated the cytotoxic activity of doxorubicin via enhanced DNA damage and mitotic arrest, conferring superior efficacy to the doxorubicin-cyclophosphamide regimen in TNBC xenografts. Ganetespib also promoted mitotic catastrophe and apoptosis in combination with taxanes in vitro, and these effects translated to significantly improved combinatorial activity in vivo. Marked tumor shrinkage of metastatic lung and lymphatic lesions were seen in patients on ganetespib monotherapy. CONCLUSION: The preclinical activity profile and clinical evidence of tumor regression suggest that ganetespib offers considerable promise as a new therapeutic candidate to target TNBC.

Authors
Proia, DA; Zhang, C; Sequeira, M; Jimenez, J-P; He, S; Spector, N; Shapiro, GI; Tolaney, S; Nagai, M; Acquaviva, J; Smith, DL; Sang, J; Bates, RC; El-Hariry, I
MLA Citation
Proia, DA, Zhang, C, Sequeira, M, Jimenez, J-P, He, S, Spector, N, Shapiro, GI, Tolaney, S, Nagai, M, Acquaviva, J, Smith, DL, Sang, J, Bates, RC, and El-Hariry, I. "Preclinical activity profile and therapeutic efficacy of the HSP90 inhibitor ganetespib in triple-negative breast cancer." Clinical cancer research : an official journal of the American Association for Cancer Research 20.2 (January 2014): 413-424.
PMID
24173541
Source
epmc
Published In
Clinical cancer research : an official journal of the American Association for Cancer Research
Volume
20
Issue
2
Publish Date
2014
Start Page
413
End Page
424
DOI
10.1158/1078-0432.ccr-13-2166

Optical and radioiodinated tethered Hsp90 inhibitors reveal selective internalization of ectopic Hsp90 in malignant breast tumor cells.

Inhibitors of heat-shock protein 90 (Hsp90) have demonstrated an unusual selectivity for tumor cells despite its ubiquitous expression. This phenomenon has remained unexplained, but could be influenced by ectopically expressed Hsp90 in tumors. In this work, we synthesized Hsp90 inhibitors that can carry optical or radioiodinated probes via a polyethyleneglycol tether. We show that these tethered inhibitors selectively recognize cells expressing ectopic Hsp90 and become internalized. The internalization process is blocked by Hsp90 antibodies, suggesting that active cycling of the protein occurs at the plasma membrane. In mice, we observed exquisite accumulation of the fluor-tethered versions within breast tumors at very sensitive levels. Cell-based assays with the radiolabeled version showed picomolar detection in cells that express ectopic Hsp90. Our findings show that fluor-tethered or radiolabeled inhibitors that target ectopic Hsp90 can be used to detect breast cancer malignancies through noninvasive imaging.

Authors
Barrott, JJ; Hughes, PF; Osada, T; Yang, X-Y; Hartman, ZC; Loiselle, DR; Spector, NL; Neckers, L; Rajaram, N; Hu, F; Ramanujam, N; Vaidyanathan, G; Zalutsky, MR; Lyerly, HK; Haystead, TA
MLA Citation
Barrott, JJ, Hughes, PF, Osada, T, Yang, X-Y, Hartman, ZC, Loiselle, DR, Spector, NL, Neckers, L, Rajaram, N, Hu, F, Ramanujam, N, Vaidyanathan, G, Zalutsky, MR, Lyerly, HK, and Haystead, TA. "Optical and radioiodinated tethered Hsp90 inhibitors reveal selective internalization of ectopic Hsp90 in malignant breast tumor cells." Chem Biol 20.9 (September 19, 2013): 1187-1197.
PMID
24035283
Source
pubmed
Published In
Chemistry and Biology
Volume
20
Issue
9
Publish Date
2013
Start Page
1187
End Page
1197
DOI
10.1016/j.chembiol.2013.08.004

A network of substrates of the E3 ubiquitin ligases MDM2 and HUWE1 control apoptosis independently of p53.

In the intrinsic pathway of apoptosis, cell-damaging signals promote the release of cytochrome c from mitochondria, triggering activation of the Apaf-1 and caspase-9 apoptosome. The ubiquitin E3 ligase MDM2 decreases the stability of the proapoptotic factor p53. We show that it also coordinated apoptotic events in a p53-independent manner by ubiquitylating the apoptosome activator CAS and the ubiquitin E3 ligase HUWE1. HUWE1 ubiquitylates the antiapoptotic factor Mcl-1, and we found that HUWE1 also ubiquitylated PP5 (protein phosphatase 5), which indirectly inhibited apoptosome activation. Breast cancers that are positive for the tyrosine receptor kinase HER2 (human epidermal growth factor receptor 2) tend to be highly aggressive. In HER2-positive breast cancer cells treated with the HER2 tyrosine kinase inhibitor lapatinib, MDM2 was degraded and HUWE1 was stabilized. In contrast, in breast cancer cells that acquired resistance to lapatinib, the abundance of MDM2 was not decreased and HUWE1 was degraded, which inhibited apoptosis, regardless of p53 status. MDM2 inhibition overcame lapatinib resistance in cells with either wild-type or mutant p53 and in xenograft models. These findings demonstrate broader, p53-independent roles for MDM2 and HUWE1 in apoptosis and specifically suggest the potential for therapy directed against MDM2 to overcome lapatinib resistance.

Authors
Kurokawa, M; Kim, J; Geradts, J; Matsuura, K; Liu, L; Ran, X; Xia, W; Ribar, TJ; Henao, R; Dewhirst, MW; Kim, W-J; Lucas, JE; Wang, S; Spector, NL; Kornbluth, S
MLA Citation
Kurokawa, M, Kim, J, Geradts, J, Matsuura, K, Liu, L, Ran, X, Xia, W, Ribar, TJ, Henao, R, Dewhirst, MW, Kim, W-J, Lucas, JE, Wang, S, Spector, NL, and Kornbluth, S. "A network of substrates of the E3 ubiquitin ligases MDM2 and HUWE1 control apoptosis independently of p53. (Published online)" Sci Signal 6.274 (May 7, 2013): ra32-.
Website
http://hdl.handle.net/10161/8398
PMID
23652204
Source
pubmed
Published In
Science Signaling
Volume
6
Issue
274
Publish Date
2013
Start Page
ra32
DOI
10.1126/scisignal.2003741

Correction: phase 1 clinical trial of HER2-specific immunotherapy with concomitant HER2 kinase inhibtion.

Authors
Hamilton, E; Blackwell, K; Hobeika, AC; Clay, TM; Broadwater, G; Ren, XR; Chen, W; Castro, H; Lehmann, F; Spector, N; Wei, J; Osada, T; Lyerly, HK; Morse, MA
MLA Citation
Hamilton, E, Blackwell, K, Hobeika, AC, Clay, TM, Broadwater, G, Ren, XR, Chen, W, Castro, H, Lehmann, F, Spector, N, Wei, J, Osada, T, Lyerly, HK, and Morse, MA. "Correction: phase 1 clinical trial of HER2-specific immunotherapy with concomitant HER2 kinase inhibtion." J Transl Med 11 (2013): 82-.
PMID
23536971
Source
pubmed
Published In
Journal of Translational Medicine
Volume
11
Publish Date
2013
Start Page
82
DOI
10.1186/1479-5876-11-82

An heregulin-EGFR-HER3 autocrine signaling axis can mediate acquired lapatinib resistance in HER2+ breast cancer models.

INTRODUCTION: The human epidermal growth factor receptor 2 (HER2) receptor tyrosine kinase (RTK) oncogene is an attractive therapeutic target for the treatment of HER2-addicted tumors. Although lapatinib, an FDA-approved small-molecule HER2 and epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor (TKI), represents a significant therapeutic advancement in the treatment of HER2+ breast cancers, responses to lapatinib have not been durable. Consequently, elucidation of mechanisms of acquired therapeutic resistance to HER-directed therapies is of critical importance. METHODS: Using a functional protein-pathway activation mapping strategy, along with targeted genomic knockdowns applied to a series of isogenic-matched pairs of lapatinib-sensitive and resistant cell lines, we now report an unexpected mechanism of acquired resistance to lapatinib and similar TKIs. RESULTS: The signaling analysis revealed that whereas HER2 was appropriately inhibited in lapatinib-resistant cells, EGFR tyrosine phosphorylation was incompletely inhibited. Using a targeted molecular knockdown approach to interrogate the causal molecular underpinnings of EGFR-persistent activation, we found that lapatinib-resistant cells were no longer oncogene addicted to HER2-HER3-PI3K signaling, as seen in the parental lapatinib-sensitive cell lines, but instead were dependent on a heregulin (HRG)-driven HER3-EGFR-PI3K-PDK1 signaling axis. Two FDA-approved EGFR TKIs could not overcome HRG-HER3-mediated activation of EGFR, or reverse lapatinib resistance. The ability to overcome EGFR-mediated acquired therapeutic resistance to lapatinib was demonstrated through molecular knockdown of EGFR and treatment with the irreversible pan-HER TKI neratinib, which blocked HRG-dependent phosphorylation of HER3 and EGFR, resulting in apoptosis of resistant cells. In addition, whereas HRG reversed lapatinib-mediated antitumor effects in parental HER2+ breast cancer cells, neratinib was comparatively resistant to the effects of HRG in parental cells. Finally, we showed that HRG expression is an independent negative predictor of clinical outcome in HER2+ breast cancers, providing potential clinical relevance to our findings. CONCLUSIONS: Molecular analysis of acquired therapeutic resistance to lapatinib identified a new resistance mechanism based on incomplete and "leaky" inhibition of EGFR by lapatinib. The selective pressure applied by incomplete inhibition of the EGFR drug target resulted in selection of ligand-driven feedback that sustained EGFR activation in the face of constant exposure to the drug. Inadequate target inhibition driven by a ligand-mediated autocrine feedback loop may represent a broader mechanism of therapeutic resistance to HER TKIs and suggests adopting a different strategy for selecting more effective TKIs to advance into the clinic.

Authors
Xia, W; Petricoin, EF; Zhao, S; Liu, L; Osada, T; Cheng, Q; Wulfkuhle, JD; Gwin, WR; Yang, X; Gallagher, RI; Bacus, S; Lyerly, HK; Spector, NL
MLA Citation
Xia, W, Petricoin, EF, Zhao, S, Liu, L, Osada, T, Cheng, Q, Wulfkuhle, JD, Gwin, WR, Yang, X, Gallagher, RI, Bacus, S, Lyerly, HK, and Spector, NL. "An heregulin-EGFR-HER3 autocrine signaling axis can mediate acquired lapatinib resistance in HER2+ breast cancer models." Breast Cancer Res 15.5 (2013): R85-.
PMID
24044505
Source
pubmed
Published In
Breast Cancer Research
Volume
15
Issue
5
Publish Date
2013
Start Page
R85
DOI
10.1186/bcr3480

Polyclonal HER2-specific antibodies induced by vaccination mediate receptor internalization and degradation in tumor cells.

INTRODUCTION: Sustained HER2 signaling at the cell surface is an oncogenic mechanism in a significant proportion of breast cancers. While clinically effective therapies targeting HER2 such as mAbs and tyrosine kinase inhibitors exist, tumors overexpressing HER2 eventually progress despite treatment. Thus, abrogation of persistent HER2 expression at the plasma membrane to synergize with current approaches may represent a novel therapeutic strategy. METHODS: We generated polyclonal anti-HER2 antibodies (HER2-VIA) by vaccinating mice with an adenovirus expressing human HER2, and assessed their signaling effects in vitro and anti-tumor effects in a xenograft model. In addition, we studied the signaling effects of human HER2-specific antibodies induced by vaccinating breast cancer patients with a HER2 protein vaccine. RESULTS: HER2-VIA bound HER2 at the plasma membrane, initially activating the downstream kinases extracellular signal-regulated protein kinase 1/2 and Akt, but subsequently inducing receptor internalization in clathrin-coated pits in a HER2 kinase-independent manner, followed by ubiquitination and degradation of HER2 into a 130 kDa fragment phosphorylated at tyrosine residues 1,221/1,222 and 1,248. Following vaccination of breast cancer patients with the HER2 protein vaccine, HER2-specific antibodies were detectable and these antibodies bound to cell surface-expressed HER2 and inhibited HER2 signaling through blocking tyrosine 877 phosphorylation of HER2. In contrast to the murine antibodies, human anti-HER2 antibodies induced by protein vaccination did not mediate receptor internalization and degradation. CONCLUSION: These data provide new insight into HER2 trafficking at the plasma membrane and the changes induced by polyclonal HER2-specific antibodies. The reduction of HER2 membrane expression and HER2 signaling by polyclonal antibodies induced by adenoviral HER2 vaccines supports human clinical trials with this strategy for those breast cancer patients with HER2 therapy-resistant disease.

Authors
Ren, X-R; Wei, J; Lei, G; Wang, J; Lu, J; Xia, W; Spector, N; Barak, LS; Clay, TM; Osada, T; Hamilton, E; Blackwell, K; Hobeika, AC; Morse, MA; Lyerly, HK; Chen, W
MLA Citation
Ren, X-R, Wei, J, Lei, G, Wang, J, Lu, J, Xia, W, Spector, N, Barak, LS, Clay, TM, Osada, T, Hamilton, E, Blackwell, K, Hobeika, AC, Morse, MA, Lyerly, HK, and Chen, W. "Polyclonal HER2-specific antibodies induced by vaccination mediate receptor internalization and degradation in tumor cells. (Published online)" Breast Cancer Res 14.3 (June 7, 2012): R89-.
PMID
22676470
Source
pubmed
Published In
Breast Cancer Research
Volume
14
Issue
3
Publish Date
2012
Start Page
R89
DOI
10.1186/bcr3204

Amplification and high-level expression of heat shock protein 90 marks aggressive phenotypes of human epidermal growth factor receptor 2 negative breast cancer.

INTRODUCTION: Although human epidermal growth factor receptor 2 (HER2) positive or estrogen receptor (ER) positive breast cancers are treated with clinically validated anti-HER2 or anti-estrogen therapies, intrinsic and acquired resistance to these therapies appears in a substantial proportion of breast cancer patients and new therapies are needed. Identification of additional molecular factors, especially those characterized by aggressive behavior and poor prognosis, could prioritize interventional opportunities to improve the diagnosis and treatment of breast cancer. METHODS: We compiled a collection of 4,010 breast tumor gene expression data derived from 23 datasets that have been posted on the National Center for Biotechnology Information (NCBI) Gene Expression Omnibus (GEO) database. We performed a genome-scale survival analysis using Cox-regression survival analyses, and validated using Kaplan-Meier Estimates survival and Cox Proportional-Hazards Regression survival analyses. We conducted a genome-scale analysis of chromosome alteration using 481 breast cancer samples obtained from The Cancer Genome Atlas (TCGA), from which combined expression and copy number data were available. We assessed the correlation between somatic copy number alterations and gene expression using analysis of variance (ANOVA). RESULTS: Increased expression of each of the heat shock protein (HSP) 90 isoforms, as well as HSP transcriptional factor 1 (HSF1), was correlated with poor prognosis in different subtypes of breast cancer. High-level expression of HSP90AA1 and HSP90AB1, two cytoplasmic HSP90 isoforms, was driven by chromosome coding region amplifications and were independent factors that led to death from breast cancer among patients with triple-negative (TNBC) and HER2-/ER+ subtypes, respectively. Furthermore, amplification of HSF1 was correlated with higher HSP90AA1 and HSP90AB1 mRNA expression among the breast cancer cells without amplifications of these two genes. A collection of HSP90AA1, HSP90AB1 and HSF1 amplifications defined a subpopulation of breast cancer with up-regulated HSP90 gene expression, and up-regulated HSP90 expression independently elevated the risk of recurrence of TNBC and poor prognosis of HER2-/ER+ breast cancer. CONCLUSIONS: Up-regulated HSP90 mRNA expression represents a confluence of genomic vulnerability that renders HER2 negative breast cancers more aggressive, resulting in poor prognosis. Targeting breast cancer with up-regulated HSP90 may potentially improve the effectiveness of clinical intervention in this disease.

Authors
Cheng, Q; Chang, JT; Geradts, J; Neckers, LM; Haystead, T; Spector, NL; Lyerly, HK
MLA Citation
Cheng, Q, Chang, JT, Geradts, J, Neckers, LM, Haystead, T, Spector, NL, and Lyerly, HK. "Amplification and high-level expression of heat shock protein 90 marks aggressive phenotypes of human epidermal growth factor receptor 2 negative breast cancer. (Published online)" Breast Cancer Res 14.2 (April 17, 2012): R62-.
PMID
22510516
Source
pubmed
Published In
Breast Cancer Research
Volume
14
Issue
2
Publish Date
2012
Start Page
R62
DOI
10.1186/bcr3168

Phase 1 clinical trial of HER2-specific immunotherapy with concomitant HER2 kinase inhibition [corrected].

BACKGROUND: Patients with HER2-overexpressing metastatic breast cancer, despite initially benefiting from the monoclonal antibody trastuzumab and the EGFR/HER2 tyrosine kinase inhibitor lapatinib, will eventually have progressive disease. HER2-based vaccines induce polyclonal antibody responses against HER2 that demonstrate enhanced anti-tumor activity when combined with lapatinib in murine models. We wished to test the clinical safety, immunogenicity, and activity of a HER2-based cancer vaccine, when combined with lapatinib. METHODS: We immunized women (n = 12) with metastatic, trastuzumab-refractory, HER2-overexpressing breast cancer with dHER2, a recombinant protein consisting of extracellular domain (ECD) and a portion of the intracellular domain (ICD) of HER2 combined with the adjuvant AS15, containing MPL, QS21, CpG and liposome. Lapatinib (1250 mg/day) was administered concurrently. Peripheral blood antibody and T cell responses were measured. RESULTS: This regimen was well tolerated, with no cardiotoxicity. Anti-HER2-specific antibody was induced in all patients whereas HER2-specific T cells were detected in one patient. Preliminary analyses of patient serum demonstrated downstream signaling inhibition in HER2 expressing tumor cells. The median time to progression was 55 days, with the majority of patients progressing prior to induction of peak anti-HER2 immune responses; however, 300-day overall survival was 92% (95% CI: 77-100%). CONCLUSIONS: dHER2 combined with lapatinib was safe and immunogenic with promising long term survival in those with HER2-overexpressing breast cancers refractory to trastuzumab. Further studies to define the anticancer activity of the antibodies induced by HER2 vaccines along with lapatinib are underway. TRIAL REGISTRY: ClinicalTrials.gov NCT00952692.

Authors
Hamilton, E; Blackwell, K; Hobeika, AC; Clay, TM; Broadwater, G; Ren, X-R; Chen, W; Castro, H; Lehmann, F; Spector, N; Wei, J; Osada, T; Lyerly, HK; Morse, MA
MLA Citation
Hamilton, E, Blackwell, K, Hobeika, AC, Clay, TM, Broadwater, G, Ren, X-R, Chen, W, Castro, H, Lehmann, F, Spector, N, Wei, J, Osada, T, Lyerly, HK, and Morse, MA. "Phase 1 clinical trial of HER2-specific immunotherapy with concomitant HER2 kinase inhibition [corrected]. (Published online)" J Transl Med 10 (February 10, 2012): 28-.
PMID
22325452
Source
pubmed
Published In
Journal of Translational Medicine
Volume
10
Publish Date
2012
Start Page
28
DOI
10.1186/1479-5876-10-28

What can we learn from the age- and race/ethnicity- specific rates of inflammatory breast carcinoma?

Inflammatory Breast Carcinoma (IBC), the most aggressive type of breast tumor with unique clinicopathological presentation, is hypothesized to have distinct etiology with a socioeconomic status (SES) component. Using the Surveillance, Epidemiology and End Results (SEER) Program data for 2004-2007, we compare incidence rates of IBC to non-inflammatory locally advanced breast cancer (LABC) among racial/ethnic groups with different SES. The analysis includes women 20-84 years of age. To examine evidence for the distinct etiology of IBC, we analyzed age-distribution patterns of IBC and non-inflammatory LABC, using a mathematical carcinogenesis model. Based on the Collaborative Staging Extension codes, 2,942 incident IBC cases (codes 71 and 73) and 5,721 non-inflammatory LABC cases (codes 40-62) were identified during the four-year study period. Age-adjusted rates of IBC among non-Hispanic White and Hispanic women were similar (2.5/100,000 in both groups). Similar rates were also found in non-inflammatory LABC in these two groups (4.8/100,000 and 4.2/100,000, respectively). In African-American women, the IBC (3.91/100,000) and non-inflammatory LABC (8.47/100,000) rates were greater compared with other ethnic/racial sub-groups. However, the ratio of rates of IBC/non-inflammatory LABC was similar among all the racial/ethnic groups, suggesting that African-American women are susceptible to aggressive breast tumors in general but not specifically to IBC. The mathematical model successfully predicted the observed age-specific rates of both examined breast tumors and revealed distinct patterns. IBC rates increased until age 65 and then slightly decreased, whereas non-inflammatory LABC rates steadily increased throughout the entire age interval. The number of critical transition carcinogenesis stages (m-stages) predicted by the model were 6.3 and 8.5 for IBC and non-inflammatory LABC, respectively, supporting different etiologies of these breast tumors.

Authors
Il'yasova, D; Siamakpour-Reihani, S; Akushevich, I; Akushevich, L; Spector, N; Schildkraut, J
MLA Citation
Il'yasova, D, Siamakpour-Reihani, S, Akushevich, I, Akushevich, L, Spector, N, and Schildkraut, J. "What can we learn from the age- and race/ethnicity- specific rates of inflammatory breast carcinoma?." Breast Cancer Res Treat 130.2 (November 2011): 691-697.
PMID
21850396
Source
pubmed
Published In
Breast Cancer Research and Treatment
Volume
130
Issue
2
Publish Date
2011
Start Page
691
End Page
697
DOI
10.1007/s10549-011-1719-4

Truncated ErbB2 expressed in tumor cell nuclei contributes to acquired therapeutic resistance to ErbB2 kinase inhibitors.

ErbB2 tyrosine kinase inhibitors (TKI) block tyrosine autophosphorylation and activation of the full-length transmembrane ErbB2 receptor (p185(ErbB2)). In addition to p185(ErbB2), truncated forms of ErbB2 exist in breast cancer cell lines and clinical tumors. The contribution of these truncated forms, specifically those expressed in tumor cell nuclei, to the development of therapeutic resistance to ErbB2 TKIs has not been previously shown. Here, we show that expression of a 95-kDa tyrosine phosphorylated form of ErbB2, herein referred to as p95L (lapatinib-induced p95) was increased in ErbB2(+) breast cancer cells treated with potent ErbB2 TKIs (lapatinib, GW2974). Expressed in tumor cell nuclei, tyrosine phosphorylation of p95L was resistant to inhibition by ErbB2 TKIs. Furthermore, the expression of p95L was increased in ErbB2(+) breast cancer models of acquired therapeutic resistance to lapatinib that mimic the clinical setting. Pretreatment with proteasome inhibitors blocked p95L induction in response to ErbB2 TKIs, implicating the role of the proteasome in the regulation of p95L expression. In addition, tyrosine phosphorylated C-terminal fragments of ErbB2, generated by alternate initiation of translation and similar in molecular weight to p95L, were expressed in tumor cell nuclei, where they too were resistant to inhibition by ErbB2 TKIs. When expressed in the nuclei of lapatinib-sensitive ErbB2(+) breast cancer cells, truncated ErbB2 rendered cells resistant to lapatinib-induced apoptosis. Elucidating the function of nuclear, truncated forms of ErbB2, and developing therapeutic strategies to block their expression and/or activation may enhance the clinical efficacy of ErbB2 TKIs.

Authors
Xia, W; Liu, Z; Zong, R; Liu, L; Zhao, S; Bacus, SS; Mao, Y; He, J; Wulfkuhle, JD; Petricoin, EF; Osada, T; Yang, X-Y; Hartman, ZC; Clay, TM; Blackwell, KL; Lyerly, HK; Spector, NL
MLA Citation
Xia, W, Liu, Z, Zong, R, Liu, L, Zhao, S, Bacus, SS, Mao, Y, He, J, Wulfkuhle, JD, Petricoin, EF, Osada, T, Yang, X-Y, Hartman, ZC, Clay, TM, Blackwell, KL, Lyerly, HK, and Spector, NL. "Truncated ErbB2 expressed in tumor cell nuclei contributes to acquired therapeutic resistance to ErbB2 kinase inhibitors." Mol Cancer Ther 10.8 (August 2011): 1367-1374.
PMID
21673090
Source
pubmed
Published In
Molecular cancer therapeutics
Volume
10
Issue
8
Publish Date
2011
Start Page
1367
End Page
1374
DOI
10.1158/1535-7163.MCT-10-0991

Synergism from combined immunologic and pharmacologic inhibition of HER2 in vivo.

The monoclonal antibody trastuzumab and the EGFR/HER2 tyrosine kinase inhibitor lapatinib improve the clinical outcome of patients with HER2-overexpressing breast cancer. However, the majority of metastatic cancers will eventually progress, suggesting the need for other therapies. Because HER2 overexpression persists, we hypothesized that the anti-HER2 immune response induced by cancer vaccines would be an effective strategy for treating trastuzumab- and lapatinib-refractory tumors. Furthermore, we hypothesized that the antibody response could synergize with lapatinib to enhance tumor inhibition. We developed a recombinant adenoviral vector expressing a kinase-inactive HER2 (Ad-HER2-ki) to use as a cancer vaccine. Vaccine-induced polyclonal HER2-specific antiserum was analyzed for receptor internalization and signaling effects alone and in combination with lapatinib. Ad-HER2-ki vaccine-induced potent T cell and antibody responses in mice and the vaccine-induced polyclonal HER2-specific antiserum mediated receptor internalization and degradation much more effectively than trastuzumab. Our in vitro studies demonstrated that HER2 vaccine-induced antibodies effectively caused a decrease in HER2 expression, but when combined with lapatinib caused significant inhibition of HER2 signaling, decreased pERK and pAKT levels and reduced breast tumor cell proliferation. In addition, a known mechanism of resistance to lapatinib, induction of survivin, was inhibited. The combination of Ad-HER2-ki plus lapatinib also showed superior antitumor efficacy in vivo. Based on these results, we feel clinical studies using this approach to target HER2-overexpressing breast cancer, including trastuzumab- and lapatinib-resistant tumors is warranted.

Authors
Morse, MA; Wei, J; Hartman, Z; Xia, W; Ren, X-R; Lei, G; Barry, WT; Osada, T; Hobeika, AC; Peplinski, S; Jiang, H; Devi, GR; Chen, W; Spector, N; Amalfitano, A; Lyerly, HK; Clay, TM
MLA Citation
Morse, MA, Wei, J, Hartman, Z, Xia, W, Ren, X-R, Lei, G, Barry, WT, Osada, T, Hobeika, AC, Peplinski, S, Jiang, H, Devi, GR, Chen, W, Spector, N, Amalfitano, A, Lyerly, HK, and Clay, TM. "Synergism from combined immunologic and pharmacologic inhibition of HER2 in vivo." Int J Cancer 126.12 (June 15, 2010): 2893-2903.
PMID
19856307
Source
pubmed
Published In
International Journal of Cancer
Volume
126
Issue
12
Publish Date
2010
Start Page
2893
End Page
2903
DOI
10.1002/ijc.24995

An adenoviral vaccine encoding full-length inactivated human Her2 exhibits potent immunogenicty and enhanced therapeutic efficacy without oncogenicity.

PURPOSE: Overexpression of the breast cancer oncogene HER2 correlates with poor survival. Current HER2-directed therapies confer limited clinical benefits and most patients experience progressive disease. Because refractory tumors remain strongly HER2+, vaccine approaches targeting HER2 have therapeutic potential, but wild type (wt) HER2 cannot safely be delivered in immunogenic viral vectors because it is a potent oncogene. We designed and tested several HER2 vaccines devoid of oncogenic activity to develop a safe vaccine for clinical use. EXPERIMENTAL DESIGN: We created recombinant adenoviral vectors expressing the extracellular domain of HER2 (Ad-HER2-ECD), ECD plus the transmembrane domain (Ad-HER2-ECD-TM), and full-length HER2 inactivated for kinase function (Ad-HER2-ki), and determined their immunogenicity and antitumor effect in wild type (WT) and HER2-tolerant mice. To assess their safety, we compared their effect on the cellular transcriptome, cell proliferation, anchorage-dependent growth, and transformation potential in vivo. RESULTS: Ad-HER2-ki was the most immunogenic vector in WT animals, retained immunogenicity in HER2-transgenic tolerant animals, and showed strong therapeutic efficacy in treatment models. Despite being highly expressed, HER2-ki protein was not phosphorylated and did not produce an oncogenic gene signature in primary human cells. Moreover, in contrast to HER2-wt, cells overexpressing HER2-ki were less proliferative, displayed less anchorage-independent growth, and were not transformed in vivo. CONCLUSIONS: Vaccination with mutationally inactivated, nononcogenic Ad-HER2-ki results in robust polyclonal immune responses to HER2 in tolerant models, which translates into strong and effective antitumor responses in vivo. Ad-HER2-ki is thus a safe and promising vaccine for evaluation in clinical trials.

Authors
Hartman, ZC; Wei, J; Osada, T; Glass, O; Lei, G; Yang, X-Y; Peplinski, S; Kim, D-W; Xia, W; Spector, N; Marks, J; Barry, W; Hobeika, A; Devi, G; Amalfitano, A; Morse, MA; Lyerly, HK; Clay, TM
MLA Citation
Hartman, ZC, Wei, J, Osada, T, Glass, O, Lei, G, Yang, X-Y, Peplinski, S, Kim, D-W, Xia, W, Spector, N, Marks, J, Barry, W, Hobeika, A, Devi, G, Amalfitano, A, Morse, MA, Lyerly, HK, and Clay, TM. "An adenoviral vaccine encoding full-length inactivated human Her2 exhibits potent immunogenicty and enhanced therapeutic efficacy without oncogenicity." Clin Cancer Res 16.5 (March 1, 2010): 1466-1477.
PMID
20179231
Source
pubmed
Published In
Clinical cancer research : an official journal of the American Association for Cancer Research
Volume
16
Issue
5
Publish Date
2010
Start Page
1466
End Page
1477
DOI
10.1158/1078-0432.CCR-09-2549

Resistance to ErbB2 tyrosine kinase inhibitors in breast cancer is mediated by calcium-dependent activation of RelA.

The widespread clinical use of therapies targeting the ErbB2 receptor tyrosine kinase oncogene represents a significant advance in breast cancer treatment. However, the development of therapeutic resistance represents a dilemma limiting their clinical efficacy, particularly small-molecule tyrosine kinase inhibitors that block ErbB2 autophosphorylation and activation. Here, we show that lapatinib (GW572016), a highly selective, small-molecule inhibitor of the ErbB2 and epidermal growth factor receptor tyrosine kinases, which was recently approved for the treatment of advanced-stage ErbB2(+) breast cancer, unexpectedly triggered a cytoprotective stress response in ErbB2(+) breast cancer cell lines, which was mediated by the calcium-dependent activation of RelA, the prosurvival subunit of NF-kappaB. Abrogation of lapatinib-induced RelA activation using either small interfering RNA constructs or an intracellular calcium chelator enhanced the apoptotic effects of lapatinib in parental ErbB2(+) breast cancer cells and overcame therapeutic resistance to lapatinib in ErbB2(+) breast cancer lines that had been rendered resistant to lapatinib through chronic exposure to the drug, mimicking the clinical setting. In addition, analysis of changes in phospho-RelA expression in sequential clinical biopsies from ErbB2(+) breast cancers treated with lapatinib monotherapy revealed marginally statistically significant differences between responders and nonresponders, which was consistent with our preclinical findings. Elucidating the regulation of RelA by lapatinib in ErbB2(+) breast cancers, and showing its role in the development of therapeutic resistance to lapatinib, identifies another therapeutic target to overcome or prevent the onset of resistance to lapatinib in some women with ErbB2(+) breast cancers.

Authors
Xia, W; Bacus, S; Husain, I; Liu, L; Zhao, S; Liu, Z; Moseley, MA; Thompson, JW; Chen, FL; Koch, KM; Spector, NL
MLA Citation
Xia, W, Bacus, S, Husain, I, Liu, L, Zhao, S, Liu, Z, Moseley, MA, Thompson, JW, Chen, FL, Koch, KM, and Spector, NL. "Resistance to ErbB2 tyrosine kinase inhibitors in breast cancer is mediated by calcium-dependent activation of RelA." Mol Cancer Ther 9.2 (February 2010): 292-299.
PMID
20124457
Source
pubmed
Published In
Molecular cancer therapeutics
Volume
9
Issue
2
Publish Date
2010
Start Page
292
End Page
299
DOI
10.1158/1535-7163.MCT-09-1041

Reply to F. Bellati et al

Authors
Spector, NL; Blackwell, KL
MLA Citation
Spector, NL, and Blackwell, KL. "Reply to F. Bellati et al." Journal of Clinical Oncology 28.21 (2010): e371-.
Source
scival
Published In
Journal of Clinical Oncology
Volume
28
Issue
21
Publish Date
2010
Start Page
e371
DOI
10.1200/JCO.2010.28.8571

Phase II study to evaluate the efficacy and safety of neoadjuvant lapatinib plus paclitaxel in patients with inflammatory breast cancer

Purpose: We conducted a phase II, open-label, multicenter study to evaluate the efficacy, safety, and tolerability of daily lapatinib plus weekly paclitaxel in treatment-naïve patients with inflammatory breast cancer (IBC). Patients and Methods: The primary end point was pathologic complete response (pCR). Secondary end points included combined clinical response rate (based on Response Evaluation Criteria in Solid Tumors (RECIST) criteria and clinically evaluable skin disease criteria). Patients were assigned to either cohort A (human epidermal growth factor receptor 2 [HER2] 2+ or 3+ by immunohistochemistry [IHC] or fluorescent in situ hybridization [FISH] -amplified ± epidermal growth factor receptor [EGFR] expression) or cohort B (HER2-negative/EGFR-positive). A subpopulation of cohort A considered HER2-positive by the current definition of overexpression (3+ by IHC or FISH-amplified) was also analyzed. Patients received lapatinib at 1,500 mg/d for 14 days, then lapatinib at 1,500 mg/d plus weekly paclitaxel (80 mg/m 2) for 12 weeks, followed by surgical resection or additional chemotherapy. Results: Forty-nine women were enrolled (cohort A, n = 42; cohort B, n = 7). Cohort B was terminated because of slow accrual and lack of efficacy observed in IBC patients with HER2-negative/EGFR-positive tumors enrolled onto the parallel study, EGF103009. pCR occurred in 18.2% (95% CI, 5.2% to 40.3%) of cohort A patients. Combined clinical response rate was 78.6% (95% CI, 63.2% to 89.7%) in all cohort A patients and 78.1% (95% CI, 60.0% to 90.7%) in the HER2-positive subset. Common adverse events included diarrhea, rash, alopecia, and nausea (> 50% of patients in both cohorts). The incidence of grade 3 diarrhea was 55%. Conclusion: Lapatinib monotherapy for 14 days followed by lapatinib plus paclitaxel for 12 weeks provided clinical benefit in IBC patients with HER2-overexpressing tumors without unexpected toxicity. © 2010 by American Society of Clinical Oncology.

Authors
Boussen, H; Cristofanilli, M; Zaks, T; DeSilvio, M; Salazar, V; Spector, N
MLA Citation
Boussen, H, Cristofanilli, M, Zaks, T, DeSilvio, M, Salazar, V, and Spector, N. "Phase II study to evaluate the efficacy and safety of neoadjuvant lapatinib plus paclitaxel in patients with inflammatory breast cancer." Journal of Clinical Oncology 28.20 (2010): 3248-3255.
PMID
20530274
Source
scival
Published In
Journal of Clinical Oncology
Volume
28
Issue
20
Publish Date
2010
Start Page
3248
End Page
3255
DOI
10.1200/JCO.2009.21.8594

Understanding the mechanisms behind trastuzumab therapy for human epidermal growth factor receptor 2-positive breast cancer.

PURPOSE: Targeted therapy with the humanized monoclonal antibody trastuzumab has become a mainstay for human epidermal growth factor receptor 2 (HER2) -positive breast cancer (BC). The mechanisms of action of trastuzumab have not been fully elucidated, and data available to date are reviewed here. The impact of the mechanisms of action on clinical benefit also is discussed. METHODS: An extensive literature review of trastuzumab and proposed mechanisms of action was performed. RESULTS: At least five potential extracellular and intracellular antitumor mechanisms of trastuzumab have been identified in the preclinical setting. These include activation of antibody-dependent cellular cytotoxicity, inhibition of extracellular domain cleavage, abrogation of intracellular signaling, reduction of angiogenesis, and decreased DNA repair. These effects lead to tumor cell stasis and/or death. Clinical benefit from trastuzumab-based therapy in both early and advanced BC has been demonstrated. The benefit of trastuzumab use beyond progression has also been shown, which indicates the need for continuous suppression of the HER2 pathway. Targeting both HER2, with various approaches, and other pathways may enhance the clinical benefit observed with trastuzumab and overcome potential resistance. Novel combinations include pertuzumab (a HER2 dimerization inhibitor), lapatinib (a HER1/HER2 tyrosine kinase inhibitor), bevacizumab (an antiangiogenic agent), tanespimycin (a heat shock protein inhibitor), antiestrogen therapies, and an antibody-drug conjugate (trastuzumab-DM1). CONCLUSION: Trastuzumab is the foundation of care for patients with HER2-positive BC. Emerging data from studies of other targeted agents may provide alternative treatment combinations to maximize the clinical benefit from trastuzumab and prevent or delay resistance. The continued development of trastuzumab highlights promising treatment approaches for the future.

Authors
Spector, NL; Blackwell, KL
MLA Citation
Spector, NL, and Blackwell, KL. "Understanding the mechanisms behind trastuzumab therapy for human epidermal growth factor receptor 2-positive breast cancer." J Clin Oncol 27.34 (December 1, 2009): 5838-5847. (Review)
PMID
19884552
Source
pubmed
Published In
Journal of Clinical Oncology
Volume
27
Issue
34
Publish Date
2009
Start Page
5838
End Page
5847
DOI
10.1200/JCO.2009.22.1507

Lapatinib monotherapy in patients with HER2-overexpressing relapsed or refractory inflammatory breast cancer: final results and survival of the expanded HER2+ cohort in EGF103009, a phase II study

Background: Inflammatory breast cancer is an aggressive and biologically distinct form with a higher frequency of HER2 overexpression than other breast cancers. For patients with resistance to conventional anthracycline or taxane and trastuzumab treatment, options are limited. Lapatinib, an oral reversible inhibitor of epidermal growth factor receptor tyrosine kinases, previously had a 50% response rate in a cohort of 30 patients with HER2-overexpressing (HER2+) recurrent or anthracycline-refractory inflammatory breast cancer. We aimed to assess efficacy of lapatinib in an expanded cohort of patients with relapsed or refractory HER2+ disease. Methods: From March, 2005, to September, 2007, 126 patients with relapsed or refractory HER2+ inflammatory breast cancer were treated with lapatinib 1500 mg once daily in a non-randomised, open-label, phase II study. Pretreatment tumour biopsies were done to verify pathological features of inflammatory breast cancer. Skin disease was assessed every 4 weeks, and response in sites of measurable locally advanced or metastatic disease were assessed by response evaluation in solid tumours (RECIST) criteria every 8 weeks. The primary aim was to assess combined objective response rate, by clinically evaluable skin disease criteria and RECIST, if applicable. Analyses were done by intention to treat; patients with missing data were treated as non-responders. This study is registered with ClinicalTrials.gov, number NCT00105950. Findings: Clinical presentation and biomarker analysis showed a tumour molecular profile consistent with inflammatory breast cancer. No patients had complete response. 49 patients (39%; 95% CI 30-48) had partial response. Median progression-free survival was 14·6 weeks (95% CI 12·1-16·0), with median duration of response of 20·9 weeks (12·7-32·1). Likelihood of response to lapatinib was not affected by previous treatment with trastuzumab. 130 (92%) of 141 patients had at least one adverse event; 45 (32%) had serious adverse events, the most common were dyspnoea (eight patients) and pleural effusion (six). Five patients had fatal adverse events that were possibly treatment related. Interpretation: Lapatinib monotherapy is a potentially effective treatment for relapsed or refractory HER2+ inflammatory breast cancer. Funding: GlaxoSmithKline. © 2009 Elsevier Ltd. All rights reserved.

Authors
Kaufman, B; Trudeau, M; Awada, A; Blackwell, K; Bachelot, T; Salazar, V; DeSilvio, M; Westlund, R; Zaks, T; Spector, N; Johnston, S
MLA Citation
Kaufman, B, Trudeau, M, Awada, A, Blackwell, K, Bachelot, T, Salazar, V, DeSilvio, M, Westlund, R, Zaks, T, Spector, N, and Johnston, S. "Lapatinib monotherapy in patients with HER2-overexpressing relapsed or refractory inflammatory breast cancer: final results and survival of the expanded HER2+ cohort in EGF103009, a phase II study." The Lancet Oncology 10.6 (2009): 581-588.
PMID
19394894
Source
scival
Published In
The Lancet Oncology
Volume
10
Issue
6
Publish Date
2009
Start Page
581
End Page
588
DOI
10.1016/S1470-2045(09)70087-7

A phase I and pharmacokinetic study of oral lapatinib administered once or twice daily in patients with solid malignancies

Purpose: This study determined the range of tolerable doses, clinical safety, pharmacokinetics, and preliminary evidence of clinical activity following once or twice daily administration of lapatinib in patients with solid malignancies. Experimental Design: Cancer patients (n = 81) received oral doses of lapatinib ranging from 175 to 1,800 mg once daily or 500 to 900 mg twice daily. Clinical assessments of safety and antitumor activity were recorded and blood was sampled for pharmacokinetic assessments. The effect of a low-fat meal on lapatinib pharmacokinetics was assessed in a subset of patients. Results: Lapatinib was well tolerated, such that dose escalation was limited at 1,800 mg once daily only by pill burden. Twice-daily dosing was implemented to further explore tolerability, and was limited by diarrhea to 500 mg twice daily. The most commonly reported adverse events with once-daily dosing were diarrhea (48%), nausea (40%), rash (40%), and fatigue (38%) and with twice-daily dosing were diarrhea (85%), rash (54%), and nausea (34%). Lapatinib serum concentrations accumulated upon repeated dosing, increasing nearly in proportion with dose, and were significantly increased when dosed with food or administered twice daily. One patient with head and neck cancer achieved a confirmed complete response and 22 patients had stable disease of≥8 weeks including three patients with stable disease of >10 months (renal, lung, and salivary gland cancers). Conclusion: Lapatinib was well tolerated following once and twice daily administration. Systemicexposure to lapatinib was dependent on the dose, duration and frequency of dosing, and prandial state. Clinical activity was observed. © 2009 American Association for Cancer Research.

Authors
III, HAB; Taylor, CW; Jones, SF; Koch, KM; Versola, MJ; Arya, N; Fleming, RA; Smith, DA; Pandite, L; Spector, N; Wilding, G
MLA Citation
III, HAB, Taylor, CW, Jones, SF, Koch, KM, Versola, MJ, Arya, N, Fleming, RA, Smith, DA, Pandite, L, Spector, N, and Wilding, G. "A phase I and pharmacokinetic study of oral lapatinib administered once or twice daily in patients with solid malignancies." Clinical Cancer Research 15.21 (2009): 6702-6708.
PMID
19825948
Source
scival
Published In
Clinical cancer research : an official journal of the American Association for Cancer Research
Volume
15
Issue
21
Publish Date
2009
Start Page
6702
End Page
6708
DOI
10.1158/1078-0432.CCR-09-0369

Acquired resistance to small molecule ErbB2 tyrosine kinase inhibitors.

Breast cancers overexpressing the ErbB2 (HER2) receptor tyrosine kinase oncogene are treated with targeted therapies such as trastuzumab (Herceptin), an anti-ErbB2 antibody, and lapatinib (GW572016/Tykerb), a selective small molecule inhibitor of ErbB2 and epidermal growth factor receptor tyrosine kinases that was recently approved for ErbB2+ breast cancers that progressed on trastuzumab-based therapy. The efficacy of lapatinib as a monotherapy or in combination with chemotherapy, however, is limited by the development of therapeutic resistance that typically occurs within 12 months of starting therapy. In contrast to small molecule inhibitors targeting other receptor tyrosine kinases where resistance has been attributed to mutations within the targeted receptor, ErbB2 mutations have not been commonly found in breast tumors. Instead, acquired resistance to lapatinib seems to be mediated by redundant survival pathways that are activated as a consequence of marked inhibition of ErbB2 kinase activity. For example, inhibition of phosphatidylinositol3 kinase-Akt in lapatinib-treated cells leads to derepression of FOXO3A, a transcription factor that up-regulates estrogen receptor (ER) signaling, resulting in a switch in the regulation of survival factors (e.g., survivin) and cell survival from ErbB2 alone to ER and ErbB2 in resistant cells. In this review, we discuss the effects of lapatinib on signaling networks in ErbB2+ breast cancer cells to elucidate potential mechanisms of therapeutic resistance and strategies to overcome or prevent its development.

Authors
Chen, FL; Xia, W; Spector, NL
MLA Citation
Chen, FL, Xia, W, and Spector, NL. "Acquired resistance to small molecule ErbB2 tyrosine kinase inhibitors." Clin Cancer Res 14.21 (November 1, 2008): 6730-6734. (Review)
PMID
18980964
Source
pubmed
Published In
Clinical cancer research : an official journal of the American Association for Cancer Research
Volume
14
Issue
21
Publish Date
2008
Start Page
6730
End Page
6734
DOI
10.1158/1078-0432.CCR-08-0581

The effect of anti-VEGF therapy on immature myeloid cell and dendritic cells in cancer patients.

Impairment of dendritic cells (DC), the most effective activators of anticancer immune responses, is one mechanism for defective antitumor immunity, but the causes of DC impairment are incompletely understood. We evaluated the association of impaired DC differentiation with angiogenesis-associated molecules D-dimer, vascular endothelial growth factor (VEGF), urokinase plasminogen activator (uPA), and plasminogen activator inhibitor (PAI-1) in peripheral blood from 41 patients with lung, breast, and colorectal carcinoma. Subsequently, we studied the effect of administration of the anti-VEGF antibody (bevacizumab) on DC maturation and function in vivo. Compared with healthy volunteers, cancer patients had a bias toward the immunoregulatory DC2, had deficits in DC maturation after overnight in vitro culture, and had a significant increase in immature myeloid cell progenitors of DC (0.50 +/- 0.31% vs. 0.32 +/- 0.16% of peripheral blood mononuclear cells, respectively, P = 0.011). A positive correlation was found between the percentage of DC2 and PAI-1 (R = 0.50) and between immature myeloid cells and VEGF (R = 0.52). Bevacizumab administration to cancer patients was associated with a decrease in the accumulation of immature progenitor cells (0.39 +/- 0.30% vs. 0.27 +/- 0.24%, P = 0.012) and induced a modest increase in the DC population in peripheral blood (0.47 +/- 0.23% vs. 0.53 +/- 0.30%). Moreover, anti-VEGF antibody treatment enhanced allo-stimulatory capacity of DC and T cell proliferation against recall antigens. These data suggest that DC differentiation is negatively associated with VEGF levels and may be one explanation for impaired anticancer immunity, especially in patients with advanced malignancies.

Authors
Osada, T; Chong, G; Tansik, R; Hong, T; Spector, N; Kumar, R; Hurwitz, HI; Dev, I; Nixon, AB; Lyerly, HK; Clay, T; Morse, MA
MLA Citation
Osada, T, Chong, G, Tansik, R, Hong, T, Spector, N, Kumar, R, Hurwitz, HI, Dev, I, Nixon, AB, Lyerly, HK, Clay, T, and Morse, MA. "The effect of anti-VEGF therapy on immature myeloid cell and dendritic cells in cancer patients." Cancer Immunol Immunother 57.8 (August 2008): 1115-1124.
PMID
18193223
Source
pubmed
Published In
Cancer Immunology, Immunotherapy
Volume
57
Issue
8
Publish Date
2008
Start Page
1115
End Page
1124
DOI
10.1007/s00262-007-0441-x

Phase II study of predictive biomarker profiles for response targeting human epidermal growth factor receptor 2 (HER-2) in advanced inflammatory breast cancer with lapatinib monotherapy

Purpose: Inflammatory breast cancer (IBC) is one of the most aggressive forms of breast cancer. Lapatinib, an oral reversible inhibitor of epidermal growth factor receptor (EGFR) and human EGFR 2 (HER-2), demonstrated clinical activity in four of five IBC patients in phase I trials. We conducted a phase II trial to confirm the sensitivity of IBC to lapatinib, to determine whether response is HER-2 or EGFR dependent, and to elucidate a molecular signature predictive of lapatinib sensitivity. Patients and Methods: Our open-label multicenter phase II trial (EGF103009) assessed clinical activity and safety of lapatinib monotherapy in patients with recurrent or anthracycline-refractory IBC. Patients were assigned to cohorts A (HER-2-overexpressing [HER-2+]) or B(HER-2-/EGFR+) and fresh pretreatment tumor biopsies were collected. Results: Forty-five patients (30 in cohort A; 15 in cohort B) received lapatinib 1,500 mg once daily continuously. Clinical presentation and biomarker analyses demonstrated a tumor molecular signature consistent with IBC. Lapatinib was generally well tolerated, with primarily grade 1/2 skin and GI toxicities. Fifteen patients (50%) in cohort A had clinical responses to lapatinib in skin and/or measurable disease (according to Response Evaluation Criteria in Solid Tumors) compared with one patient in cohort B. Within cohort A, phosphorylated (p) HER-3 and lack of p53 expression predicted for response to lapatinib (P < .05). Tumors coexpressing pHER-2 and pHER-3 were more likely to respond to lapatinib (nine of 10 v four of 14; P = .0045). Prior trastuzumab therapy and loss of phosphate and tensin homolog 10 (PTEN) did not preclude response to lapatinib. Conclusion: Lapatinib is well tolerated with clinical activity in heavily pretreated HER-2+, but not EGFR+/HER-2-, IBC. In this study, coexpression of pHER-2 and pHER-3 in tumors seems to predict for a favorable response to lapatinib. These findings warrant further investigation of lapatinib monotherapy or combination therapy in HER-2+ IBC. © 2008 by American Society of Clinical Oncology.

Authors
Johnston, S; Trudeau, M; Kaufman, B; Boussen, H; Blackwell, K; LoRusso, P; Lombardi, DP; Ahmed, SB; Citrin, DL; DeSilvio, ML; Harris, J; Westlund, RE; Salazar, V; Zaks, TZ; Spector, NL
MLA Citation
Johnston, S, Trudeau, M, Kaufman, B, Boussen, H, Blackwell, K, LoRusso, P, Lombardi, DP, Ahmed, SB, Citrin, DL, DeSilvio, ML, Harris, J, Westlund, RE, Salazar, V, Zaks, TZ, and Spector, NL. "Phase II study of predictive biomarker profiles for response targeting human epidermal growth factor receptor 2 (HER-2) in advanced inflammatory breast cancer with lapatinib monotherapy." Journal of Clinical Oncology 26.7 (2008): 1066-1072.
PMID
18212337
Source
scival
Published In
Journal of Clinical Oncology
Volume
26
Issue
7
Publish Date
2008
Start Page
1066
End Page
1072
DOI
10.1200/JCO.2007.13.9949

Expert roundtable: Emerging questions in ErbB2-positive breast cancer; February 22, 2007

Authors
Spector, N; Pegram, M; Perez, EA; Piccart, M
MLA Citation
Spector, N, Pegram, M, Perez, EA, and Piccart, M. "Expert roundtable: Emerging questions in ErbB2-positive breast cancer; February 22, 2007." Clinical Breast Cancer 8.SUPPL. 3 (2008): S131-S141.
Source
scival
Published In
Clinical Breast Cancer
Volume
8
Issue
SUPPL. 3
Publish Date
2008
Start Page
S131
End Page
S141
DOI
10.3816/CBC.2008.s.009

Treatment of metastatic ErbB2-positive breast cancer: Options after progression on trastuzumab

Although trastuzumab-based therapy has changed the treatment paradigm for ErbB2-positive breast cancers, most patients eventually develop progressive disease. Of particular interest is the issue of disease progression in the central nervous system, a safe haven from high molecular weight antibodies like trastuzumab, which have limited ability to cross the blood-brain barrier. This review will discuss therapeutic options for when disease progression has occurred on trastuzumab-based therapies, including central nervous system progression, continuation of trastuzumab-based therapy, addition of novel targeted therapies, and the use of small-molecule tyrosine kinase inhibitors targeting ErbB receptors.

Authors
Spector, N
MLA Citation
Spector, N. "Treatment of metastatic ErbB2-positive breast cancer: Options after progression on trastuzumab." Clinical Breast Cancer 8.SUPPL. 3 (2008): S94-S99.
Source
scival
Published In
Clinical Breast Cancer
Volume
8
Issue
SUPPL. 3
Publish Date
2008
Start Page
S94
End Page
S99
DOI
10.3816/CBC.2008.s.005

Activation of AMP-activated protein kinase by human EGF receptor 2/EGF receptor tyrosine kinase inhibitor protects cardiac cells.

The human EGF receptor (HER) 2 receptor tyrosine kinase is a survival factor for human cardiomyocytes, and its inhibition may explain the increased incidence of cardiomyopathy associated with the anti-HER2 monoclonal antibody trastuzumab (Genentech, South San Francisco, CA), particularly in patients with prior exposure to cardiotoxic chemotherapies e.g., anthracyclines. Here, we show that GW2974 (HER2/EGF receptor tyrosine kinase inhibitor), but not trastuzumab, activates AMP-activated protein kinase (AMPK), initiating a metabolic stress response in human cardiomyocytes that protects against TNFalpha-induced cell death. GW2974 stimulates calcium dependent fatty acid oxidation in vitro and in the myocardium of GW2974-treated rodents. Calcium chelation or siRNA-targeted AMPK knockdown blocks GW2974 induced fatty acid oxidation. In addition, inhibition of AMPK by a specific inhibitor resulted in increased killing of cardiomyocytes. Elucidating the effects of HER2-targeted therapies on AMPK may predict for risk of cardiomyopathy and provide a novel HER2-targeted strategy designed to protect myocardium from the pro-apoptotic effects of pro-inflammatory cytokines released in response to cardiac injury by chemotherapy or acute ischemia.

Authors
Spector, NL; Yarden, Y; Smith, B; Lyass, L; Trusk, P; Pry, K; Hill, JE; Xia, W; Seger, R; Bacus, SS
MLA Citation
Spector, NL, Yarden, Y, Smith, B, Lyass, L, Trusk, P, Pry, K, Hill, JE, Xia, W, Seger, R, and Bacus, SS. "Activation of AMP-activated protein kinase by human EGF receptor 2/EGF receptor tyrosine kinase inhibitor protects cardiac cells." Proc Natl Acad Sci U S A 104.25 (June 19, 2007): 10607-10612.
PMID
17556544
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
104
Issue
25
Publish Date
2007
Start Page
10607
End Page
10612
DOI
10.1073/pnas.0701286104

Lapatinib antitumor activity is not dependent upon phosphatase and tensin homologue deleted on chromosome 10 in ErbB2-overexpressing breast cancers.

Trastuzumab antitumor activity in ErbB2-overexpressing breast cancers seems to be dependent upon the presence of phosphatase and tensin homologue deleted on chromosome 10 (PTEN), a phosphatase that dampens phosphatidylinositol 3-kinase-Akt signaling. Consequently, PTEN deficiency, which occurs in 50% of breast cancers, predicts for resistance to trastuzumab monotherapy. Here, we show that lapatinib, a small-molecule inhibitor of ErbB1 and ErbB2 tyrosine kinases, exerts its antitumor activity in a PTEN-independent manner. Steady-state phosphorylated ErbB2 (p-ErbB2) and p-Akt (S473) protein levels were inhibited within 30 min following lapatinib but not in response to trastuzumab in BT474 and Au565 cells (two ErbB2-overexpressing breast cancer cell lines that are sensitive to the proapoptotic effects of lapatinib). Whereas trastuzumab reportedly inhibits SRC phosphorylation (Y416), which in turn reduced SRC-ErbB2 protein interactions, lapatinib had no effect on either variable. To assess the potential functional role that PTEN might play in lapatinib antitumor activity, we selectively knocked down PTEN in BT474 and Au565 cells using small interfering RNA transfection. Loss of PTEN did not affect induction of tumor cell apoptosis by lapatinib in either cell line. In addition, lapatinib inhibited Akt phosphorylation in MDA-MB-468 cells, an ErbB1-expressing/ErbB2 non-overexpressing breast cancer line, despite their PTEN-null status. Moreover, patients with ErbB2-overexpressing inflammatory breast cancers responded to lapatinib monotherapy regardless of PTEN status. Thus, lapatinib seems to exert its antitumor activity in ErbB2-overexpressing breast cancers in a PTEN-independent manner. These data emphasize the importance of assessing PTEN status in tumors when selecting ErbB2-targeted therapies in patients with breast cancer.

Authors
Xia, W; Husain, I; Liu, L; Bacus, S; Saini, S; Spohn, J; Pry, K; Westlund, R; Stein, SH; Spector, NL
MLA Citation
Xia, W, Husain, I, Liu, L, Bacus, S, Saini, S, Spohn, J, Pry, K, Westlund, R, Stein, SH, and Spector, NL. "Lapatinib antitumor activity is not dependent upon phosphatase and tensin homologue deleted on chromosome 10 in ErbB2-overexpressing breast cancers." Cancer Res 67.3 (February 1, 2007): 1170-1175.
PMID
17283152
Source
pubmed
Published In
Cancer Research
Volume
67
Issue
3
Publish Date
2007
Start Page
1170
End Page
1175
DOI
10.1158/0008-5472.CAN-06-2101

HER2 therapy. Small molecule HER-2 tyrosine kinase inhibitors

Overexpression of the human epidermal growth factor receptor (HER)-2 oncogenic receptor tyrosine kinase, which occurs in 25% of breast cancers, portends poor clinical outcome and consequently represents an attractive target for therapeutic intervention. Small molecule tyrosine kinase inhibitors that compete with ATP binding at the cytoplasmic catalytic kinase domain of HER-2 block autophosphorylation and activation of HER-2, resulting in inhibition of downstream proliferation and survival signals. These agents have exhibited clinical activity in patients with HER-2 overexpressing breast cancers. Here we review the development of HER-2 tyrosine kinase inhibitors, their mechanisms of action, their biological and clinical activities, their safety profile, and combination strategies including conventional cytotoxics and other targeted agents. © 2007 BioMed Central Ltd.

Authors
Spector, N; Xia, W; El-Hariry, I; Yarden, Y; Bacus, S
MLA Citation
Spector, N, Xia, W, El-Hariry, I, Yarden, Y, and Bacus, S. "HER2 therapy. Small molecule HER-2 tyrosine kinase inhibitors." Breast Cancer Research 9.2 (2007).
Source
scival
Published In
Breast Cancer Research
Volume
9
Issue
2
Publish Date
2007
DOI
10.1186/bcr1652

A reciprocal tensin-3-cten switch mediates EGF-driven mammary cell migration

Cell migration driven by the epidermal growth factor receptor (EGFR) propels morphogenesis and involves reorganization of the actin cytoskeleton. Although de novo transcription precedes migration, transcript identity remains largely unknown. Through their actin-binding domains, tensins link the cytoskeleton to integrin-based adhesion sites. Here we report that EGF downregulates tensin-3 expression, and concomitantly upregulates cten, a tensin family member that lacks the actin-binding domain. Knockdown of cten or tensin-3, respectively, impairs or enhances mammary cell migration. Furthermore, cten displaces tensin-3 from the cytoplasmic tail of integrin β1, thereby instigating actin fibre disassembly. In invasive breast cancer, cten expression correlates not only with high EGFR and HER2, but also with metastasis to lymph nodes. Moreover, treatment of inflammatory breast cancer patients with an EGFR/HER2 dual-specificity kinase inhibitor significantly downregulated cten expression. In conclusion, a transcriptional tensin-3-cten switch may contribute to the metastasis of mammary cancer.

Authors
Katz, M; Amit, I; Citri, A; Shay, T; Carvalho, S; Lavi, S; Milanezi, F; Lyass, L; Amariglio, N; Jacob-Hirsch, J; Ben-Chetrit, N; Tarcic, G; Lindzen, M; Avraham, R; Liao, Y-C; Trusk, P; Lyass, A; Rechavi, G; Spector, NL; Lo, SH; Schmitt, F; Bacus, SS; Yarden, Y
MLA Citation
Katz, M, Amit, I, Citri, A, Shay, T, Carvalho, S, Lavi, S, Milanezi, F, Lyass, L, Amariglio, N, Jacob-Hirsch, J, Ben-Chetrit, N, Tarcic, G, Lindzen, M, Avraham, R, Liao, Y-C, Trusk, P, Lyass, A, Rechavi, G, Spector, NL, Lo, SH, Schmitt, F, Bacus, SS, and Yarden, Y. "A reciprocal tensin-3-cten switch mediates EGF-driven mammary cell migration." Nature Cell Biology 9.8 (2007): 961-969.
PMID
17643115
Source
scival
Published In
Nature Cell Biology
Volume
9
Issue
8
Publish Date
2007
Start Page
961
End Page
969
DOI
10.1038/ncb1622

A model of acquired autoresistance to a potent ErbB2 tyrosine kinase inhibitor and a therapeutic strategy to prevent its onset in breast cancer.

The development of acquired resistance to ErbB2 tyrosine kinase inhibitors limits the clinical efficacy of this class of cancer therapeutics. Little is known about the mechanism(s) of acquired resistance to these agents. Here we establish a model of acquired resistance to N-{3-chloro-4-[(3-fluorobenzyl) oxy]phenyl}-6-[5-({[2 (methylsulfonyl)ethyl]amino}methyl)-2-furyl]-4-quinazolinamine (lapatinib), an inhibitor of ErbB2 and ErbB1 tyrosine kinases by chronically exposing lapatinib-sensitive ErbB2-overexpressing breast cancer cells to lapatinib, simulating the clinic where lapatinib is administered on a daily chronic basis. Analysis of baseline gene expression in acquired lapatinib-resistant and parental cells indicates estrogen receptor (ER) signaling involvement in the development of resistance. Using gene interference, we confirm that acquired resistance to lapatinib is mediated by a switch in cell survival dependence and regulation of a key antiapoptotic mediator from ErbB2 alone to codependence upon ER and ErbB2 rather than loss of ErbB2 expression or insensitivity of ErbB2 signaling to lapatinib. Increased ER signaling in response to lapatinib is enhanced by the activation of factors facilitating the transcriptional activity of ER, notably FOXO3a and caveolin-1. Importantly, we confirm that lapatinib induces ER signaling in tumor biopsies from patients with ErbB2-overexpressing breast cancers receiving lapatinib therapy. These findings provided the rationale for preventing the development of acquired resistance by simultaneously inhibiting both ER and ErbB2 signaling pathways. Establishing clinically relevant models of acquired resistance to ErbB2 kinase inhibitors will enhance therapeutic strategies to improve clinical outcomes for patients with ErbB2-overexpressing breast cancers.

Authors
Xia, W; Bacus, S; Hegde, P; Husain, I; Strum, J; Liu, L; Paulazzo, G; Lyass, L; Trusk, P; Hill, J; Harris, J; Spector, NL
MLA Citation
Xia, W, Bacus, S, Hegde, P, Husain, I, Strum, J, Liu, L, Paulazzo, G, Lyass, L, Trusk, P, Hill, J, Harris, J, and Spector, NL. "A model of acquired autoresistance to a potent ErbB2 tyrosine kinase inhibitor and a therapeutic strategy to prevent its onset in breast cancer." Proc Natl Acad Sci U S A 103.20 (May 16, 2006): 7795-7800.
PMID
16682622
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
103
Issue
20
Publish Date
2006
Start Page
7795
End Page
7800
DOI
10.1073/pnas.0602468103

Regulation of survivin by ErbB2 signaling: therapeutic implications for ErbB2-overexpressing breast cancers.

In breast cancer, overexpression of ErbB2 or aberrant regulation of survivin, a member of the inhibitor of apoptosis family, is associated with resistance to chemo/hormone therapy and predicts for a poor clinical outcome. A functional link between the two predictive factors has not been previously shown. Here, using genetic and pharmacologic approaches to block ErbB2 signaling, we show that ErbB2 regulates survivin protein expression in ErbB2-overexpressing breast cancer cells. Selective knockdown of ErbB2 using small interfering RNA markedly reduced survivin protein, resulting in apoptosis of ErbB2-overexpressing breast cancer cell lines such as BT474. Alternatively, inhibition of ErbB2 signaling using lapatinib (GW572016), a reversible small-molecule inhibitor of ErbB1/ErbB2 tyrosine kinases, at pharmacologically relevant concentrations, leads to marked inhibition of survivin protein with subsequent apoptosis. The effect of lapatinib on survivin seems to be predominantly posttranslational, mediated by ubiquitin-proteosome degradation as lactacystin, a proteosome inhibitor, reverses these effects. Furthermore, lapatinib down-regulated the expression of His-tagged survivin, which was under the transcriptional control of a heterologous promoter, providing additional evidence supporting a posttranslational mechanism of regulation. In contrast, trastuzumab and gefitinib failed to down-regulate survivin in ErbB2-overexpressing breast cancer cells. Importantly, the clinical relevance of these findings was illustrated in patients with ErbB2-overexpressing breast cancer whose clinical response to lapatinib was associated with marked inhibition of survivin in their tumors. These findings shed new light on the mechanism by which ErbB2 overexpression protects against apoptotic stimuli in breast cancer and identifies therapeutic interventions to improve clinical outcomes in these aggressive tumors.

Authors
Xia, W; Bisi, J; Strum, J; Liu, L; Carrick, K; Graham, KM; Treece, AL; Hardwicke, MA; Dush, M; Liao, Q; Westlund, RE; Zhao, S; Bacus, S; Spector, NL
MLA Citation
Xia, W, Bisi, J, Strum, J, Liu, L, Carrick, K, Graham, KM, Treece, AL, Hardwicke, MA, Dush, M, Liao, Q, Westlund, RE, Zhao, S, Bacus, S, and Spector, NL. "Regulation of survivin by ErbB2 signaling: therapeutic implications for ErbB2-overexpressing breast cancers." Cancer Res 66.3 (February 1, 2006): 1640-1647.
PMID
16452223
Source
pubmed
Published In
Cancer Research
Volume
66
Issue
3
Publish Date
2006
Start Page
1640
End Page
1647
DOI
10.1158/0008-5472.CAN-05-2000

Delivery of a healthy baby after first-trimester maternal exposure to lapatinib

We report the case of a woman who conceived while being treated on a phase I clincal trial with lapatinib, a dual inhibitor of epidermal growth factor receptor (EGFR) and HER2/neu, for metastatic breast cancer. Despite approximately 11 weeks of exposure to lapatinib in the first and second trimesters, the pregnancy was uncomplicated and resulted in the delivery of a healthy baby. Although concomitant cancer and pregnancy is relatively rare, the increasing use of biologic agents among fertile women - sometimes for as long as a year in the adjuvant setting - increases the probability that some women will conceive while taking a growth factor pathway inhibitor. As with systemic chemotherapy given during pregnancy, there exists the potential for teratogenicity or fetal demise from exposure of the developing embryo to inhibitors of EGFR and HER2/neu. Despite the positive outcome of this case, continued caution is warranted with the use of EGFR and HER2/neu inhibitors in pregnancy.

Authors
Kelly, H; Graham, M; Humes, E; Dorflinger, LJ; Boggess, KA; O'Neil, BH; Harris, J; Spector, NL; Dees, EC
MLA Citation
Kelly, H, Graham, M, Humes, E, Dorflinger, LJ, Boggess, KA, O'Neil, BH, Harris, J, Spector, NL, and Dees, EC. "Delivery of a healthy baby after first-trimester maternal exposure to lapatinib." Clinical Breast Cancer 7.4 (2006): 339-341.
Source
scival
Published In
Clinical Breast Cancer
Volume
7
Issue
4
Publish Date
2006
Start Page
339
End Page
341
DOI
10.3816/CBC.2006.n.048

Rational development of targeted cancer therapies using biomarkers

As the complex network of signalling pathways involved in regulating tumor cell growth and survival is unraveled, tractable targets for therapeutic drug development have been identified. Questions regarding the best approach towards developing these targeted therapies remain [eg, what is the best strategy for (i) selecting doses, (ii) identifying target patient populations for clinical trials, and (iii) designing combination therapies based on scientific rationale]. Since these agents exert biological and clinical effects that are generally distinct from traditional cytotoxic agents, a different paradigm for their development has been suggested. Here we will discuss how incorporation of biomaker analysis in early-phase clinical trials can provide valuable information to guide further development of targeted therapies.

Authors
Bacus, S; Yarden, Y; Xia, W; Spector, NL
MLA Citation
Bacus, S, Yarden, Y, Xia, W, and Spector, NL. "Rational development of targeted cancer therapies using biomarkers." Laboratory Medicine 37.8 (2006): 482-489.
Source
scival
Published In
Laboratory medicine
Volume
37
Issue
8
Publish Date
2006
Start Page
482
End Page
489
DOI
10.1309/C0YY-YW2L-55EQ-90CN

Combining lapatinib (GW572016), a small molecule inhibitor of ErbB1 and ErbB2 tyrosine kinases, with therapeutic anti-ErbB2 antibodies enhances apoptosis of ErbB2-overexpressing breast cancer cells.

Antibodies and small molecule tyrosine kinase inhibitors targeting ErbB2 exhibit distinct, noncross resistant mechanisms of action. Here, apoptosis of ErbB2-overexpressing breast cancer cells was enhanced by combining lapatinib, an inhibitor of ErbB1 and ErbB2 tyrosine kinases, with anti-ErbB2 antibodies, including (i) trastuzumab, a humanized monoclonal antibody, and (ii) pAb, rabbit polyclonal antisera generated by vaccination with a human ErbB2 fusion protein. Treating ErbB2-overexpressing breast cancer cell lines with a relatively low concentration of lapatinib alone resulted in a minimal increase in tumor cell apoptosis with an associated decrease in steady-state protein levels of p-ErbB2, p-Akt, p-Erk1/2, and notably survivin, compared to baseline. Exposure to pAb alone reduced total ErbB2 protein, disrupting ErbB3 transactivation, leading to a marked inhibition of p-Akt; however, survivin protein levels remained unchanged and apoptosis only increased slightly. Treatment with trastuzumab alone had relatively little effect on survivin and apoptosis was unaffected. Combining lapatinib with either pAb or trastuzumab markedly downregulated survivin protein and enhanced tumor cell apoptosis. The association between the inhibition of survivin and enhanced apoptosis following the combination of ErbB2-targeted therapies provides a biological effect in order to identify therapeutic strategies that promote tumor cell apoptosis and might improve clinical response.

Authors
Xia, W; Gerard, CM; Liu, L; Baudson, NM; Ory, TL; Spector, NL
MLA Citation
Xia, W, Gerard, CM, Liu, L, Baudson, NM, Ory, TL, and Spector, NL. "Combining lapatinib (GW572016), a small molecule inhibitor of ErbB1 and ErbB2 tyrosine kinases, with therapeutic anti-ErbB2 antibodies enhances apoptosis of ErbB2-overexpressing breast cancer cells." Oncogene 24.41 (September 15, 2005): 6213-6221.
PMID
16091755
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
24
Issue
41
Publish Date
2005
Start Page
6213
End Page
6221
DOI
10.1038/sj.onc.1208774

Phase I safety, pharmacokinetics, and clinical activity study of lapatinib (GW572016), a reversible dual inhibitor of epidermal growth factor receptor tyrosine kinases, in heavily pretreated patients with metastatic carcinomas.

PURPOSE: This study (EGF10004) assessed the safety/tolerability, pharmacokinetics, and clinical activity of daily oral dosing with lapatinib (GW572016) in patients with ErbB1-expressing and/or ErbB2-overexpressing advanced-stage refractory solid tumors. PATIENTS AND METHODS: Heavily pretreated patients with ErbB1-expressing and/or ErbB2-overexpressing metastatic cancers were randomly assigned to one of five dose cohorts of lapatinib administered once daily. Pharmacokinetic samples were obtained on days 1 and 20. Clinical response was assessed every 8 weeks. RESULTS: Sixty-seven patients with metastatic solid tumors were treated with lapatinib. The most frequently reported drug-related adverse events were diarrhea (42%) and rash (31%). No grade 4 drug-related adverse events were reported. Five grade 3 drug-related toxicities (gastrointestinal events and rash) were experienced by four patients. Drug-related interstitial pneumonitis or cardiac dysfunction associated with other ErbB-targeted therapies was not reported. Four patients with trastuzumab-resistant metastatic breast cancer-two of whom were classified as having inflammatory breast cancer-had partial responses (PRs). Twenty-four patients with various other carcinomas experienced stable disease, of whom 10 received lapatinib for > or = 6 months. The relationships between lapatinib dose or serum concentration and clinical response could not be adequately characterized due to the limited response data. The incidence of diarrhea increased with increasing dose, whereas the incidence of rash was not related to dose. CONCLUSION: Lapatinib was well tolerated at doses ranging from 500 to 1,600 mg once daily. Clinical activity was observed in heavily pretreated patients with ErbB1-expressing and/or ErbB2-overexpressing metastatic cancers, including four PRs in patients with trastuzumab-resistant breast cancers and prolonged stable disease in 10 patients.

Authors
Burris, HA; Hurwitz, HI; Dees, EC; Dowlati, A; Blackwell, KL; O'Neil, B; Marcom, PK; Ellis, MJ; Overmoyer, B; Jones, SF; Harris, JL; Smith, DA; Koch, KM; Stead, A; Mangum, S; Spector, NL
MLA Citation
Burris, HA, Hurwitz, HI, Dees, EC, Dowlati, A, Blackwell, KL, O'Neil, B, Marcom, PK, Ellis, MJ, Overmoyer, B, Jones, SF, Harris, JL, Smith, DA, Koch, KM, Stead, A, Mangum, S, and Spector, NL. "Phase I safety, pharmacokinetics, and clinical activity study of lapatinib (GW572016), a reversible dual inhibitor of epidermal growth factor receptor tyrosine kinases, in heavily pretreated patients with metastatic carcinomas." J Clin Oncol 23.23 (August 10, 2005): 5305-5313.
PMID
15955900
Source
pubmed
Published In
Journal of Clinical Oncology
Volume
23
Issue
23
Publish Date
2005
Start Page
5305
End Page
5313
DOI
10.1200/JCO.2005.16.584

Study of the biologic effects of lapatinib, a reversible inhibitor of ErbB1 and ErbB2 tyrosine kinases, on tumor growth and survival pathways in patients with advanced malignancies.

PURPOSE: This was a pilot study to assess the biologic effects of lapatinib on various tumor growth/survival pathways in patients with advanced ErbB1 and/or ErbB2-overexpressing solid malignancies. PATIENTS AND METHODS: Heavily pretreated patients with metastatic cancers overexpressing ErbB2 and/or expressing ErbB1 were randomly assigned to one of five dose cohorts of lapatinib (GW572016) administered orally once daily continuously. The biologic effects of lapatinib on tumor growth and survival pathways were assessed in tumor biopsies obtained before and after 21 days of therapy. Clinical response was determined at 8 weeks. RESULTS: Sequential tumor biopsies from 33 patients were examined. Partial responses occurred in four patients with breast cancer, and disease stabilization occurred in 11 others with various malignancies. Responders exhibited variable levels of inhibition of p-ErbB1, p-ErbB2, p-Erk1/2, p-Akt, cyclin D1, and transforming growth factor alpha. Even some nonresponders demonstrated varying degrees of biomarker inhibition. Increased tumor cell apoptosis (TUNEL) occurred in patients with evidence of tumor regression but not in nonresponders (progressive disease). Clinical response was associated with a pretreatment TUNEL score > 0 and increased pretreatment expression of ErbB2, p-ErbB2, Erk1/2, p-Erk1/2, insulin-like growth factor receptor-1, p70 S6 kinase, and transforming growth factor alpha compared with nonresponders. CONCLUSION: Lapatinib exhibited preliminary evidence of biologic and clinical activity in ErbB1 and/or ErbB2-overexpressing tumors. However, the limited sample size of this study and the variability of the biologic endpoints suggest that further work is needed to prioritize biomarkers for disease-directed studies, and underscores the need for improved trial design strategies in early clinical studies of targeted agents.

Authors
Spector, NL; Xia, W; Burris, H; Hurwitz, H; Dees, EC; Dowlati, A; O'Neil, B; Overmoyer, B; Marcom, PK; Blackwell, KL; Smith, DA; Koch, KM; Stead, A; Mangum, S; Ellis, MJ; Liu, L; Man, AK; Bremer, TM; Harris, J; Bacus, S
MLA Citation
Spector, NL, Xia, W, Burris, H, Hurwitz, H, Dees, EC, Dowlati, A, O'Neil, B, Overmoyer, B, Marcom, PK, Blackwell, KL, Smith, DA, Koch, KM, Stead, A, Mangum, S, Ellis, MJ, Liu, L, Man, AK, Bremer, TM, Harris, J, and Bacus, S. "Study of the biologic effects of lapatinib, a reversible inhibitor of ErbB1 and ErbB2 tyrosine kinases, on tumor growth and survival pathways in patients with advanced malignancies." J Clin Oncol 23.11 (April 10, 2005): 2502-2512.
PMID
15684311
Source
pubmed
Published In
Journal of Clinical Oncology
Volume
23
Issue
11
Publish Date
2005
Start Page
2502
End Page
2512
DOI
10.1200/JCO.2005.12.157

Phase I pharmacokinetic studies evaluating single and multiple doses of oral GW572016, a dual EGFR-ErbB2 inhibitor, in healthy subjects

GW572016 is a dual EGFR-ErbB2 inhibitor that has promise as an anticancer agent. Two phase I studies were conducted to determine the safety, tolerability and pharmacokinetics of single and multiple doses given to healthy subjects. The single dose study evaluated two groups of eight subjects in an ascending dose, 4-way cross-over, while the multiple dose study evaluated twenty-seven healthy volunteers in an ascending dose, double-blind, randomized, placebo-controlled, staggered parallel design. No serious adverse events were seen in either study. The most common adverse events for subjects receiving GW572016 were headache, diarrhea, rash, cold symptoms, gastrointestinal symptoms, and elevated LFTs, which were similar between treatment and placebo groups. Absorption of single doses of GW572016 was slightly delayed, with median t lag of 15 minutes (range 0-90 minutes) and achieved peak serum concentrations at a median of three hours (range 1.5-6 hours) post-dose. Serum concentrations after multiple doses of GW572016 demonstrated no significant accumulation at the 25 mg dose, and approximately 50% accumulation at the 100 mg and 175 mg doses, achieving steady state in six to seven days. A modest time-dependent increase in serum concentrations also was detected with multiple doses of GW572016. Single and multiple oral doses of GW572016 were well tolerated in healthy subjects, and resulted in dose-related systemic exposure of GW572016.

Authors
Bence, AK; Anderson, EB; Halepota, MA; Doukas, MA; DeSimone, PA; Davis, GA; Smith, DA; Koch, KM; Stead, AG; Mangum, S; Bowen, CJ; Spector, NL; Hsieh, S; Adams, VR
MLA Citation
Bence, AK, Anderson, EB, Halepota, MA, Doukas, MA, DeSimone, PA, Davis, GA, Smith, DA, Koch, KM, Stead, AG, Mangum, S, Bowen, CJ, Spector, NL, Hsieh, S, and Adams, VR. "Phase I pharmacokinetic studies evaluating single and multiple doses of oral GW572016, a dual EGFR-ErbB2 inhibitor, in healthy subjects." Investigational New Drugs 23.1 (2005): 39-49.
PMID
15528979
Source
scival
Published In
Investigational New Drugs
Volume
23
Issue
1
Publish Date
2005
Start Page
39
End Page
49
DOI
10.1023/B:DRUG.0000047104.45929.ea

A Phase I trial of preoperative eniluracil plus 5-fluorouracil and radiation for locally advanced or unresectable adenocarcinoma of the rectum and colon.

PURPOSE: Eniluracil, an effective inactivator of dihydropyrimidine dehydrogenase, allows for oral dosing of 5-fluorouracil (5-FU), which avoids the morbidity of continuous infusion 5-FU. We addressed the safety of oral eniluracil and 5-FU combined with preoperative radiotherapy and determined the recommended Phase II dose and dose-limiting toxicity in patients with locally advanced rectal and colon cancer. METHODS AND MATERIALS: Patients with TNM Stage II or III rectal cancer and residual or recurrent colon cancer received eniluracil (starting at 6.0 mg/m(2) every 12 h) and 5-FU (starting at 0.6 mg/m(2) every 12 h). Eniluracil and 5-FU were given with a 5-week course of preoperative radiotherapy of 4500 cGy, with a possible 540-cGy boost. Surgery was performed approximately 4 weeks after completion of chemoradiotherapy. RESULTS: Twenty-two patients were enrolled; 1 patient was withdrawn owing to noncompliance. Chemotherapy was completed in all patients; radiotherapy was completed in 20 patients. The recommended Phase II dose of eniluracil and 5-FU was 8 mg/m(2) every 12 h and 0.8 mg/m(2) every 12 h, respectively. Diarrhea was the dose-limiting toxicity. Eleven of the 17 patients with primary rectal cancer underwent a sphincter-sparing procedure. One patient had a pathologic complete response. CONCLUSION: Preoperative chemoradiotherapy with oral eniluracil and 5-FU is feasible and well tolerated. Additional investigation is warranted.

Authors
Czito, BG; Hong, TJ; Cohen, DP; Tyler, DS; Lee, CG; Anscher, MS; Ludwig, KA; Seigler, HF; Mantyh, C; Morse, MA; Lockhart, AC; Petros, WP; Honeycutt, W; Spector, NL; Ertel, PJ; Mangum, SG; Hurwitz, HI
MLA Citation
Czito, BG, Hong, TJ, Cohen, DP, Tyler, DS, Lee, CG, Anscher, MS, Ludwig, KA, Seigler, HF, Mantyh, C, Morse, MA, Lockhart, AC, Petros, WP, Honeycutt, W, Spector, NL, Ertel, PJ, Mangum, SG, and Hurwitz, HI. "A Phase I trial of preoperative eniluracil plus 5-fluorouracil and radiation for locally advanced or unresectable adenocarcinoma of the rectum and colon." Int J Radiat Oncol Biol Phys 58.3 (March 1, 2004): 779-785.
PMID
14967434
Source
pubmed
Published In
International Journal of Radiation Oncology, Biology, Physics
Volume
58
Issue
3
Publish Date
2004
Start Page
779
End Page
785
DOI
10.1016/S0360-3016(03)01567-0

Truncated ErbB2 receptor (p95ErbB2) is regulated by heregulin through heterodimer formation with ErbB3 yet remains sensitive to the dual EGFR/ErbB2 kinase inhibitor GW572016.

The expression of the NH2 terminally truncated ErbB2 receptor (p95ErbB2) in breast cancer correlates with metastatic disease progression compared with the expression of full-length p185ErbB2. We now show that heregulin (HRG), but not EGF, stimulates p95ErbB2 phosphorylation in BT474 breast cancer cells. Furthermore, phospho-p95ErbB2 forms heterodimers with ErbB3, but not EGFR, while p185ErbB2 heterodimerizes with both EGFR and ErbB3. The predilection of p95ErbB2 to heterodimerize with ErbB3 provides an explanation for its regulation by HRG, an ErbB3 ligand. GW572016, a reversible small molecule inhibitor of EGFR and ErbB2 tyrosine kinases, inhibits baseline p95ErbB2 phosphorylation in BT474 cells and tumor xenografts. Inhibition of p95ErbB2, p185ErbB2, and EGFR phosphorylation by GW572016 resulted in the inhibition of downstream phospho-Erk1/2, phospho-AKT, and cyclin D steady-state protein levels. Increased phosphorylation of p95ErbB2 and AKT in response to HRG was abrogated to varying degrees by GW572016. In contrast, trastuzumab did not inhibit p95ErbB2 phosphorylation or the expression of downstream phospho-Erk1/2, phospho-AKT, or cyclin D. It is tempting to speculate that trastuzumab resistance may be mediated in part by the selection of p95ErbB2-expressing breast cancer cells capable of exerting potent growth and prosurvival signals through p95ErbB2-ErbB3 heterodimers. Thus, p95ErbB2 represents a target for therapeutic intervention, and one that is sensitive to GW572016 therapy.

Authors
Xia, W; Liu, L-H; Ho, P; Spector, NL
MLA Citation
Xia, W, Liu, L-H, Ho, P, and Spector, NL. "Truncated ErbB2 receptor (p95ErbB2) is regulated by heregulin through heterodimer formation with ErbB3 yet remains sensitive to the dual EGFR/ErbB2 kinase inhibitor GW572016." Oncogene 23.3 (January 22, 2004): 646-653.
PMID
14737100
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
23
Issue
3
Publish Date
2004
Start Page
646
End Page
653
DOI
10.1038/sj.onc.1207166

Concurrent exposure to heat shock and H7 synergizes to trigger breast cancer cell apoptosis while sparing normal cells.

Most cancer therapies, including chemotherapy, kill tumor cells by inducing apoptosis. Consequently, the propensity of tumor cells to evade apoptotic signals contributes to therapeutic resistance. Here we show that breast cancer cells exhibiting a highly resistant phenotype undergo apoptosis when exposed to concurrent heat shock and H7, a potent serine/threonine kinase inhibitor. The anti-tumor effects of this combination are synergistic as neither treatment alone adversely affects breast cancer cell growth/survival. In contrast, non-malignant breast epithelial and hematopoietic progenitor cells are resistant to this combination therapy, thereby excluding non-specific cytotoxicity as the cause of tumor cell apoptosis. Heat or other cell stresses, including chemotherapy, preferentially enhance heat shock protein (hsp) synthesis, which serves to protect cells from potentially lethal consequences of heat shock stimuli. Ectopic overexpression of hsps in breast cancer cells protects against chemotherapy-induced apoptosis. Furthermore, increased hsps in primary breast cancers correlates with resistance to therapy and decreased survival. Stress-induced hsp synthesis is mediated by heat shock transcription factor 1 (HSF1). To simulate hsp overexpressing primary breast cancers, a number of breast cancer cell lines were transfected with HSF1d202-316, a constitutively activated form of HSF1 that leads to baseline overexpression of hsps in the absence of stress. Importantly, HSF1d202-316 transfected breast cancer cells undergo apoptosis following concurrent heat shock and H7. In light of its tumor selective activity against breast cancer cells that exhibit a highly resistant phenotype, concurrent H7 and heat shock warrants further investigation as a potential cancer therapy.

Authors
Xia, W; Hardy, L; Liu, L; Zhao, S; Goodman, M; Voellmy, R; Spector, NL
MLA Citation
Xia, W, Hardy, L, Liu, L, Zhao, S, Goodman, M, Voellmy, R, and Spector, NL. "Concurrent exposure to heat shock and H7 synergizes to trigger breast cancer cell apoptosis while sparing normal cells." Breast Cancer Res Treat 77.3 (February 2003): 233-243.
PMID
12602923
Source
pubmed
Published In
Breast Cancer Research and Treatment
Volume
77
Issue
3
Publish Date
2003
Start Page
233
End Page
243

Anti-tumor activity of GW572016: a dual tyrosine kinase inhibitor blocks EGF activation of EGFR/erbB2 and downstream Erk1/2 and AKT pathways.

Dual EGFR/erbB2 inhibition is an attractive therapeutic strategy for epithelial tumors, as ligand-induced erbB2/EGFR heterodimerization triggers potent proliferative and survival signals. Here we show that a small molecule, GW572016, potently inhibits both EGFR and erbB2 tyrosine kinases leading to growth arrest and/or apoptosis in EGFR and erbB2-dependent tumor cell lines. GW572016 markedly reduced tyrosine phosphorylation of EGFR and erbB2, and inhibited activation of Erk1/2 and AKT, downstream effectors of proliferation and cell survival, respectively. Complete inhibition of activated AKT in erbB2 overexpressing cells correlated with a 23-fold increase in apoptosis compared with vehicle controls. EGF, often elevated in cancer patients, did not reverse the inhibitory effects of GW572016. These observations were reproduced in vivo, where GW572016 treatment inhibited activation of EGFR, erbB2, Erk1/2 and AKT in human tumor xenografts. Erk1/2 and AKT represent potential biomarkers to assess the clinical activity of GW572016. Inhibition of activated AKT in EGFR or erbB2-dependent tumors by GW572016 may lead to tumor regressions when used as a monotherapy, or may enhance the anti-tumor activity of chemotherapeutics, since constitutive activation of AKT has been linked to chemo-resistance.

Authors
Xia, W; Mullin, RJ; Keith, BR; Liu, L-H; Ma, H; Rusnak, DW; Owens, G; Alligood, KJ; Spector, NL
MLA Citation
Xia, W, Mullin, RJ, Keith, BR, Liu, L-H, Ma, H, Rusnak, DW, Owens, G, Alligood, KJ, and Spector, NL. "Anti-tumor activity of GW572016: a dual tyrosine kinase inhibitor blocks EGF activation of EGFR/erbB2 and downstream Erk1/2 and AKT pathways." Oncogene 21.41 (September 12, 2002): 6255-6263.
PMID
12214266
Source
pubmed
Published In
Oncogene: Including Oncogene Reviews
Volume
21
Issue
41
Publish Date
2002
Start Page
6255
End Page
6263
DOI
10.1038/sj.onc.1205794

8 Role of immunohistochemical expression of AKT protein in breast carcinoma

Authors
Smith, BL; Altomare, D; Spector, NL; Bacus, SS
MLA Citation
Smith, BL, Altomare, D, Spector, NL, and Bacus, SS. "8 Role of immunohistochemical expression of AKT protein in breast carcinoma." Handbook of Immunohistochemistry and in Situ Hybridization of Human Carcinomas 1 (2002): 307-319.
Source
scival
Published In
Handbook of Immunohistochemistry and in Situ Hybridization of Human Carcinomas
Volume
1
Publish Date
2002
Start Page
307
End Page
319
DOI
10.1016/S1874-5784(04)80042-4

Tumor selective G2/M cell cycle arrest and apoptosis of epithelial and hematological malignancies by BBL22, a benzazepine.

Two distinct benzodiazepine binding sites have been identified, (i) a central site restricted to brain and (ii) a ubiquitously expressed mitochondrial binding site, the so-called peripheral-type benzodiazepine receptor (PBR). In this paper, we show that a benzazepine referred to as BBL22 (2-amino 9-chloro-7-(2-fluorophenyl)-5H-pyrimidol[5,4-d][2]benzazepine), which is classified as a PBR ligand based on structure, induces arrest in G(2)/M phase of the cell cycle in human tumor cell lines of both epithelial and hematopoietic cellular origin. After G(2)/M arrest, several tumor types, notably prostate and certain breast cancer lines exhibited significant apoptosis. Ideally, cancer therapies should selectively target tumor cells while sparing normal cell counterparts. BBL22 exhibited such selectivity, as it did not affect the growth and survival of nonmalignant breast and prostate epithelial lines. Moreover, BBL22 demonstrated structural requirements for this selective antitumor activity as 11 structurally related PBR ligands, including high-affinity ligands Ro5-4864 and PK11195, failed to induce tumor cell growth arrest or apoptosis. The in vivo antitumor activity of BBL22 was examined in a human xenograft model of androgen-independent prostate cancer where BBL22 significantly reduced the growth of PC3 prostate tumors without eliciting overt toxicity. Identification of BBL22 represents a tumor selective therapeutic strategy for a variety of human tumors.

Authors
Xia, W; Spector, S; Hardy, L; Zhao, S; Saluk, A; Alemane, L; Spector, NL
MLA Citation
Xia, W, Spector, S, Hardy, L, Zhao, S, Saluk, A, Alemane, L, and Spector, NL. "Tumor selective G2/M cell cycle arrest and apoptosis of epithelial and hematological malignancies by BBL22, a benzazepine." Proc Natl Acad Sci U S A 97.13 (June 20, 2000): 7494-7499.
PMID
10861014
Source
pubmed
Published In
Proceedings of the National Academy of Sciences of USA
Volume
97
Issue
13
Publish Date
2000
Start Page
7494
End Page
7499

Sensitization of tumor cells to fas killing through overexpression of heat-shock transcription factor 1.

Activation of the heat-shock or stress response is generally considered a cytoprotective response to heat or other proteotoxic stresses. In mammalian cells, stress-induced transcription of heat-shock genes is regulated by heat-shock transcription factor 1 (HSF1). We now show that activation of the Fas death receptor transactivates HSF1 in HeLa cells, a Fas-expressing cervical carcinoma line. Whereas HSF1 is constitutively expressed in a non-DNA-binding, transcriptionally inactive state, activation of Fas leads to enhanced transcription of a heat-shock reporter gene. The effects of Fas on heat-shock-gene transcription do not appear to be a consequence of cell death as they (1) precede apoptotic changes and (2) are not abrogated by YVAD-CMK, an inhibitor of Fas apoptosis that acts by blocking downstream effector proteases. Despite expressing Fas, HeLa cells are relatively insensitive to Fas-mediated killing, indicating that Fas expression alone, although necessary, is not sufficient for apoptosis. By overexpressing a constitutively activated form of HSF1, we sensitize HeLa cells to Fas-mediated killing. These findings shed new light on the interaction between two of the most evolutionarily conserved cell programs in nature, the Fas death pathway and the heat-shock response. Strategies designed to upregulate HSF1 in tumor cells, either through pharmacologic or gene-therapy approaches will hopefully provide a means with which to sensitize tumors to the killing effects of cancer therapies operating through the Fas receptor.

Authors
Xia, W; Voellmy, R; Spector, NL
MLA Citation
Xia, W, Voellmy, R, and Spector, NL. "Sensitization of tumor cells to fas killing through overexpression of heat-shock transcription factor 1." J Cell Physiol 183.3 (June 2000): 425-431.
PMID
10797318
Source
pubmed
Published In
Journal of Cellular Physiology
Volume
183
Issue
3
Publish Date
2000
Start Page
425
End Page
431
DOI
10.1002/(SICI)1097-4652(200006)183:3<425::AID-JCP16>3.0.CO;2-M

Sensitization of tumor cells to Fas killing through overexpression of heat-shock transcription factor 1

Activation of the heat-shock or stress response is generally considered a cytoprotective response to heat or other proteotoxic stresses. In mammalian cells, stress-induced transcription of heat-shock genes is regulated by heat- shock transcription factor 1 (HSF1). We now show that activation of the Fas death receptor transactivates HSF1 in HeLa cells, a Fas-expressing cervical carcinoma line. Whereas HSF1 is constitutively expressed in a non-DNA- binding, transcriptionally inactive state, activation of Fas leads to enhanced transcription of a heat-shock reporter gene. The effects of Fas on heat-shock-gene transcription do not appear to be a consequence of cell death as they (1) precede apoptotic changes and (2) are not abrogated by YVAD-CMK, an inhibitor of Fas apoptosis that acts by blocking downstream effector proteases. Despite expressing Fas, HeLa cells are relatively insensitive to Fas-mediated killing, indicating that Fas expression alone, although necessary, is not sufficient for apoptosis. By overexpressing a constitutively activated form of HSF1, we sensitize HeLa cells to as-mediated killing. These findings shed new light on the interaction between two of the most evolutionarily conserved cell programs in nature, the Fas death pathway and the heat-shock response. Strategies designed to upregulate HSF1 in tumor cells, either through pharmacologic or gene-therapy approaches will hopefully provide a means with which to sensitize tumors to the killing effects of cancer therapies operating through the Fas receptor. (C) 2000 Wiley-Liss, Inc.

Authors
Xia, W; Voellmy, R; Spector, NL
MLA Citation
Xia, W, Voellmy, R, and Spector, NL. "Sensitization of tumor cells to Fas killing through overexpression of heat-shock transcription factor 1." Journal of Cellular Physiology 183.3 (May 17, 2000): 425-431.
Source
scopus
Published In
Journal of Cellular Physiology
Volume
183
Issue
3
Publish Date
2000
Start Page
425
End Page
431
DOI
10.1002/(SICI)1097-4652(200006)183:3<425::AID-JCP16>3.0.CO;2-M

Truncation of Sp1 transcription factor by myeloblastin in undifferentiated HL60 cells.

When HL60 cells are exposed to 1,25-dihydroxyvitamin D3 (1,25D3), they undergo changes approximating the phenotype of the monocyte. Little is known, however, about the regulation and the mechanisms of this transition. It was previously noted that DNA binding by the Sp1 transcription factor in nuclear extracts of HL60 cells is profoundly altered when these cells are induced to differentiate by 1,25D3. In the present study, we show that in untreated HL60 cells only a truncated, approximately 30-kDa Sp1 fragment, encompassing the C-terminal region, binds to the GC element-containing DNA. Full-length 105-kDa Sp1 protein cannot be detected in these cells, although reverse transriptase-polymerase chain reaction reveals the presence of both 5' and 3' ends of Sp1 mRNA. Following treatment with 10(7) M 1,25D3 for 96 hr or in cells made resistant to 1,25D3 or to 1-beta-D-arabinocytosine, the Sp1 protein can be demonstrated. After an exposure to purified myeloblastin, a serine protease, purified recombinant Sp1 protein and extracts of 1,25D3-treated cells show a pattern of DNA binding similar to the pattern seen using extracts of untreated HL60 cells, indicating that the Sp1 protein is a target for myeloblastin. Because myeloblastin is present in naive HL60 cells and is downregulated during their differentiation, inhibition of proteolysis of these transcription factors seems to provide a mechanism through which differentiating HL60 cells can acquire a new repertoire of gene expression, perhaps for the maintenance of the differentiated phenotype.

Authors
Rao, J; Zhang, F; Donnelly, RJ; Spector, NL; Studzinski, GP
MLA Citation
Rao, J, Zhang, F, Donnelly, RJ, Spector, NL, and Studzinski, GP. "Truncation of Sp1 transcription factor by myeloblastin in undifferentiated HL60 cells." Journal of cellular physiology 175.2 (May 1998): 121-128.
PMID
9525470
Source
epmc
Published In
Journal of Cellular Physiology
Volume
175
Issue
2
Publish Date
1998
Start Page
121
End Page
128
DOI
10.1002/(sici)1097-4652(199805)175:2<121::aid-jcp1>3.0.co;2-q

Analysis of the value of empiric vancomycin administration in febrile neutropenia occurring after autologous peripheral blood stem cell transplants

We conducted a retrospective review of 125 patients undergoing high-dose therapy and stem cell rescue in order to evaluate the incidence of documented infection and the utility of the administration of vancomycin empirically. All patients received prophylactic oral quinolone therapy. Because neutropenia in this setting is relatively brief, 21 patients never manifested fever, and no patient died of infection. Of the remaining 104 patients, positive blood cultures were obtained in only 10, nine with a gram stain positive and one with a gram stain negative organism. Sixty-two patients without any evidence of gram positive infection received vancomycin according to the existing algorithm for care of neutropenic fevers. In this population of patients, empiric administration of vancomycin for neutropenic fevers without culture documentation appears to be unnecessary, could be discontinued safely and at substantial cost savings, and might slow the appearance of vancomycin-resistant organisms.

Authors
Koya, R; Andersen, J; Fernandez, H; Goodman, M; Spector, N; Smith, R; Hanlon, J; Cassileth, PA
MLA Citation
Koya, R, Andersen, J, Fernandez, H, Goodman, M, Spector, N, Smith, R, Hanlon, J, and Cassileth, PA. "Analysis of the value of empiric vancomycin administration in febrile neutropenia occurring after autologous peripheral blood stem cell transplants." Bone Marrow Transplantation 21.9 (1998): 923-926.
PMID
9613785
Source
scival
Published In
Bone Marrow Transplantation
Volume
21
Issue
9
Publish Date
1998
Start Page
923
End Page
926

Truncation of sp1 transcription factor by myeloblastin in undifferentiated HL60 cells

When HL60 cells are exposed to 1,25-dihydroxyvitamin D3 (1,25D3), they undergo changes approximating the phenotype of the monocyte. Little is known, however, about the regulation and the mechanisms of this transition. It was previously noted that DNA binding by the Sp1 transcription factor in nuclear extracts of HL60 cells is profoundly altered when these cells are induced to differentiate by 1,25D3. In the present study, we show that in untreated HL60 cells only a truncated, approximately 30-kDa Sp1 fragment, encompassing the C-terminal region, binds to the GC element-containing DNA. Full-length 105-kDa Sp1 protein cannot be detected in these cells, although reverse transriptase-polymerase chain reaction reveals the presence of both 5' and 3' ends of Sp1 mRNA. Following treatment with 10-7 M 1,25D3 for 96 hr or in cells made resistant to 1,25D3 or to 1-β-D-arabinocytosine, the Sp1 protein can be demonstrated. After an exposure to purified myeloblastin, a serine protease, purified recombinant Sp1 protein and extracts of 1,25D3-treated cells show a pattern of DNA binding similar to the pattern seen using extracts of untreated HL60 cells, indicating that the Sp1 protein is a target for myeloblastin. Because myeloblastin is present in naive HL60 cells and is downregulated during their differentiation, inhibition of proteolysis of these transcription factors seems to provide a mechanism through which differentiating HL60 cells can acquire a new repertoire of gene expression, perhaps for the maintenance of the differentiated phenotype.

Authors
Rao, J; Zhang, F; Donnelly, RJ; Spector, NL; Studzinski, GP
MLA Citation
Rao, J, Zhang, F, Donnelly, RJ, Spector, NL, and Studzinski, GP. "Truncation of sp1 transcription factor by myeloblastin in undifferentiated HL60 cells." Journal of Cellular Physiology 175.2 (1998): 121-128.
Source
scival
Published In
Journal of Cellular Physiology
Volume
175
Issue
2
Publish Date
1998
Start Page
121
End Page
128
DOI
10.1002/(SICI)1097-4652(199805)175:2<121::AID-JCP1>3.0.CO;2-Q

Interleukin-2 therapy for advanced chronic myeloid leukemia

Chronic myeloid leukemia (CML) is usually treated with either alpha-interferon or hydrea. Median survival is 6 years. Eventually, in most CML patients, the disease evolves to blast phase with clinical and morphologic characteristics of an acute leukemia. This phase is commonly associated with systemic symptoms and the appearance of new cytogenetic abnormalities. Therapy for this phase is of limited value, resulting in a mean survival of 4 months. We describe four consecutive patients seen at our clinic with advanced stage CML (three blast, one accelerated phase) who were treated with interleukin-2 (Proleukin). The mean survival in these patients was 22 months (range 9-35 months) and two are still alive 25 and 35 months after the start of therapy. One patient had a complete cytogenetic response and another a partial response. Toxicity was minimal and no patient had to discontinue therapy because of it.

Authors
Goodman, M; Spector, N; Rodrigues, G; Cassileth, PA
MLA Citation
Goodman, M, Spector, N, Rodrigues, G, and Cassileth, PA. "Interleukin-2 therapy for advanced chronic myeloid leukemia." Leukemia 12.11 (1998): 1682-1684.
PMID
9823941
Source
scival
Published In
Leukemia
Volume
12
Issue
11
Publish Date
1998
Start Page
1682
End Page
1684

Activation signals regulate heat shock transcription factor 1 in human B lymphocytes.

We previously showed that the ability of human B lymphocytes to elicit a cytoprotective heat shock response when confronted by heat or other stresses was dependent upon the state of cell activation. This was unexpected, considering the highly conserved nature of the heat shock response and the widely held belief that all nonmutated mature cells were capable of eliciting a heat shock response when stressed. To elucidate the mechanism by which activation primes B cells to respond to stresses, we examined heat shock transcription factor 1 (hHSF1) in B cells since this factor appears to be solely responsible for stress-induced transcription of heat shock genes in human cells. In the current report, we show that hHSF1-DNA binding complexes are undetectable in extracts of unactivated B cells. In fact, hHSF1 protein is not constitutively expressed in unactivated B cells, nor is its synthesis stress-inducible. However, following activation, hHSF1 can be found in either a transcriptionally active or an inactive state, depending upon whether the cell has been stressed or not. Thus, activation pathways play an important role in enabling B cells to survive and function properly in the context of physiologic stresses by regulating hHSF1.

Authors
Hardy, L; Goodman, M; Vasquez, A; Chauhan, D; Anderson, KC; Voellmy, R; Spector, NL
MLA Citation
Hardy, L, Goodman, M, Vasquez, A, Chauhan, D, Anderson, KC, Voellmy, R, and Spector, NL. "Activation signals regulate heat shock transcription factor 1 in human B lymphocytes." Journal of cellular physiology 170.3 (March 1997): 235-240.
PMID
9066779
Source
epmc
Published In
Journal of Cellular Physiology
Volume
170
Issue
3
Publish Date
1997
Start Page
235
End Page
240
DOI
10.1002/(sici)1097-4652(199703)170:3<235::aid-jcp3>3.0.co;2-p

Activation signals regulate heat shock transcription factor 1 in human B lymphocytes

We previously showed that the ability of human B lymphocytes to elicit a cytoprotective heat shock response when confronted by heat or other stresses was dependent upon the state of cell activation. This was unexpected, considering the highly conserved nature of the heat shock response and the widely held belief that all nonmutated mature cells were capable of eliciting a heat shock response when stressed. To elucidate the mechanism by which activation primes B cells to respond to stresses, we examined heat shock transcription factor 1 (hHSF1) in B cells since this factor appears to be solely responsible for stress-induced transcription of heat shock genes in human cells. in the current report, we show that hHSF1-DNA binding complexes are undetectable in extracts of unactivated B cells. In fact, hHSF1 protein is not constitutively expressed in unactivated B cells, nor is its synthesis stress-inducible. However, following activation, hHSF1 can be found in either a transcriptionally active or an inactive state, depending upon whether the cell has been stressed or not. Thus, activation pathways play an important role in enabling B cells to survive and function properly in the context of physiologic stresses by regulating hHSF1.

Authors
Hardy, L; Goodman, M; Vasquez, A; Chauhan, D; Anderson, KC; Voellmy, R; Spector, NL
MLA Citation
Hardy, L, Goodman, M, Vasquez, A, Chauhan, D, Anderson, KC, Voellmy, R, and Spector, NL. "Activation signals regulate heat shock transcription factor 1 in human B lymphocytes." Journal of Cellular Physiology 170.3 (1997): 235-240.
Source
scival
Published In
Journal of Cellular Physiology
Volume
170
Issue
3
Publish Date
1997
Start Page
235
End Page
240
DOI
10.1002/(SICI)1097-4652(199703)170:3<235::AID-JCP3>3.0.CO;2-P

28-kDa mammalian heat shock protein, a novel substrate of a growth regulatory protease involved in differentiation of human leukemia cells

Because of their differentiating effects in neoplastic cells in vitro, the use of retinoids in the treatment of various malignant and premalignant conditions is under investigation. To date, signal transduction pathways involved in retinoid-induced differentiation remain poorly understood. Differentiation of HL-60 cells by all-trans-retinoic acid (tRA) is directly mediated by down-regulation of the serine protease myeloblastin (mbn). In this report, we investigate the possibility that the 28-kDa heat shock protein (hsp28), previously linked to differentiation of normal and neoplastic cells including HL-60, may be regulated by mbn. Using NB4 promyelocytic leukemic cells as a differentiative model, we show that tRA induces initial suppression and subsequent up-regulation of hsp28 protein, mirroring tRA-induced changes in mbn protein. The progressive reduction in hsp28 mRNA levels in response to tRA suggests that changes in hsp28 protein levels might be posttranscriptionally mediated, raising the possibility that hsp28 may be targeted by mbn. To address this, we developed an assay using purified mbn and recombinant hsp28 and now show that hsp28 is hydrolyzed by mbn but not its homologue, human neutrophil elastase. Moreover, mbn does not indiscriminately hydrolyze other proteins. Identifying hsp28 as a substrate of mbn strongly suggests that hsp28 may be a key component of the tRA signaling pathway involved in regulating cell differentiation.

Authors
Spector, NL; Hardy, L; Ryan, C; Jr, WHM; Humes, JL; Nadler, LM; Luedke, E
MLA Citation
Spector, NL, Hardy, L, Ryan, C, Jr, WHM, Humes, JL, Nadler, LM, and Luedke, E. "28-kDa mammalian heat shock protein, a novel substrate of a growth regulatory protease involved in differentiation of human leukemia cells." Journal of Biological Chemistry 270.3 (1995): 1003-1006.
PMID
7836350
Source
scival
Published In
Journal of Biological Chemistry
Volume
270
Issue
3
Publish Date
1995
Start Page
1003
End Page
1006
DOI
10.1074/jbc.270.3.1003

A phase II study of continuous infusion recombinant human granulocyte-macrophage colony-stimulating factor as an adjunct to autologous bone marrow transplantation for patients with non-Hodgkin's lymphoma in first remission

Recombinant human granulocyte-macrophage colony-stimulating factor (rhGM-CSF) clearly hastens myeloid recovery in patients with relapsed hematologic malignancies undergoing autologous bone marrow transplantation (ABMT). In efforts to further improve neutrophil engraftment and shorten hospital stay in ABMT patients, rhGM-CSF was administered by a potentially more potent route (continuous infusion) to non-Hodgkin's lymphoma (NHL) patients with better BM reserve (first remission). Time to myeloid engraftment was compared with that of NHL patients treated in first remission at our institution on a similar ABMT protocol but without growth factor support (controls). Median neutrophil engraftment (absolute neutrophil count, 500 cells/μL) in first remission patients treated with rhGM-CSF was 14 days, compared with 22 days in controls (P = .0001). Hospital stays were also significantly reduced for rhGM-CSF patients (P = .0003). Platelet engraftment did not differ between the two groups. Persistent fever and generalized serositis were the primary toxicities. rhGM-CSF, delivered by this route, was efficacious but more toxic than 2-hour rhGM-CSF infusions previously reported by other investigators. Future alterations in both dose and schedule may retain comparable efficacy yet diminish toxicity. © 1994 by The American Society of Hematology.

Authors
O'Day, SJ; Rabinowe, SN; Neuberg, D; Freedman, AS; Soiffer, RJ; Spector, NA; Robertson, MJ; Anderson, K; Whelan, M; Pesek, K; Ritz, J; Nadler, LM
MLA Citation
O'Day, SJ, Rabinowe, SN, Neuberg, D, Freedman, AS, Soiffer, RJ, Spector, NA, Robertson, MJ, Anderson, K, Whelan, M, Pesek, K, Ritz, J, and Nadler, LM. "A phase II study of continuous infusion recombinant human granulocyte-macrophage colony-stimulating factor as an adjunct to autologous bone marrow transplantation for patients with non-Hodgkin's lymphoma in first remission." Blood 83.9 (1994): 2707-2714.
PMID
8167349
Source
scival
Published In
Blood
Volume
83
Issue
9
Publish Date
1994
Start Page
2707
End Page
2714

Regulation of the 28 kDa heat shock protein by retinoic acid during differentiation of human leukemic HL-60 cells

Dysregulation of hematopoietic cellular differentiation contributes to leukemogenesis. Unfortunately, relatively little is known about how cell differentiation is regulated. Considering that heat shock proteins (hsp) and specifically the small hsps have been increasingly linked to growth regulation, we sought to determine whether the mammalian small hsp (hsp28) is a growth-regulatory candidate during hematopoietic cell differentiation. Because of its effects on cell growth and differentiation and its increasing clinical use as a differentiating agent, we examined the effect of retinoic acid (RA) on hsp28 during differentiation of the human leukemic HL-60 cell line. Although hsp28 was constitutively expressed at low levels in untreated HL-60 cells, steady state hsp28 protein increased transiently, concomitant with the onset of G1 cell cycle arrest. Furthermore, hsp28 phosphorylation transiently increased within one hour following treatment with RA. Interestingly, in contrast to other differentiating agents the induction of hsp28 by RA was post-transcriptionally mediated with hsp28 protein and mRNA being discordantly regulated. These observations underscore the complex regulation of hsp28 by RA during granulocytic differentiation of human leukemic cells and indicate hsp28 as an intermediary in the pathway through which retinoids exert their growth and differentiative effects.

Authors
Spector, NL
MLA Citation
Spector, NL. "Regulation of the 28 kDa heat shock protein by retinoic acid during differentiation of human leukemic HL-60 cells." FEBS Letters 337.2 (1994): 184-188.
Source
scival
Published In
FEBS Letters
Volume
337
Issue
2
Publish Date
1994
Start Page
184
End Page
188
DOI
10.1016/0014-5793(94)80270-X

Anti-B4-blocked ricin: A phase I trial of 7-day continuous infusion in patients with B-cell neoplasms

Purpose: This phase I trial was undertaken to determine the maximum-tolerated dose (MTD) and dose-limiting toxicities (DLTs) of the B-cell-restricted immunotoxin anti-B4-blocked ricin (anti-B4-bR) when it is administered by 7-day continuous infusion. Patients and Methods: Thirty-four patients with relapsed and refractory B-cell neoplasms (26 non-Hodgkin's lymphoma [NHL], four chronic lymphocytic leukemia [CLL], four acute lymphoblastic leukemia [ALL]) received 7-day continuous infusion anti-B4-bR. Successive cohorts of at least three patients were treated at doses of 10 to 70 μg/kg/d for 7 days with the dose increased by 10 μg/kg/d for each cohort. The initial three cohorts of patients (10, 20, and 30 μg/kg/d × 7 days) also received a bolus infusion of 20 μg/kg before beginning the continuous infusion. Results: The MTD was reached at 50 μg/kg/d × 7 days. The DLTs were National Cancer Institute Common Toxicity Criteria (NCI CTC) grade IV reversible increases in AST and ALT, and grade IV decreases in platelet counts. Adverse reactions included fevers, nausea, headaches, myalgias, hypoalbuminemia, dyspnea, edema, and capillary leak syndrome. Potentially therapeutic serum levels of anti-B4-bR could be sustained for 4 days in patients treated at the MTD. Two complete responses (CRs), three partial responses (PRs), and 11 transient responses (TRs) were observed. Conclusion: Anti-B4-bR can be administered safely by 7-day continuous infusion with tolerable, reversible toxicities to patients with relapsed B-cell neoplasms. Although occasional responses were seen, future trials will use anti-B4-bR in patients with lower tumor burdens to circumvent the obstacle of immunotoxin delivery to bulk disease. © 1993 by American Society of Clinical Oncology.

Authors
Grossbard, ML; Lambert, JM; Goldmacher, VS; Spector, NL; Kinsella, J; Coral, LEF; Taylor, JA; Blättler, WA; Epstein, CL; Nadler, LM
MLA Citation
Grossbard, ML, Lambert, JM, Goldmacher, VS, Spector, NL, Kinsella, J, Coral, LEF, Taylor, JA, Blättler, WA, Epstein, CL, and Nadler, LM. "Anti-B4-blocked ricin: A phase I trial of 7-day continuous infusion in patients with B-cell neoplasms." Journal of Clinical Oncology 11.4 (1993): 726-737.
PMID
7683045
Source
scival
Published In
Journal of Clinical Oncology
Volume
11
Issue
4
Publish Date
1993
Start Page
726
End Page
737

Monoclonal antibody-purged autologous bone marrow transplantation in adults with acute lymphoblastic leukemia at high risk of relapse

The prognosis for adults with B lineage ALL who have relapsed after an initial remission is poor. High-dose chemoradiotherapy followed by autologous BMT can induce prolonged clinical remissions in some children with recurrent ALL. In this study, we evaluated the efficacy of autologous BMT in adults. Autologous marrow was treated in vitro with J5 and J2 monoclonal antibodies (CD10/CD9) plus rabbit complement to purge residual ALL cells. Twenty-two adults with B lineage ALL were treated with high-dose chemoradiotherapy followed by infusion of J2/J5 purged autologous BM. The median age was 28 years (range 18-54 years). Twenty-one of 22 patients had experienced at least one relapse prior to BMT. All patients achieved complete hematologic engraftment. Disease-free survival (DFS) in this cohort of patients was 20%, with all survivors alive and free of disease between 2.5 and 7.5 years post-BMT. Age at the time of BMT was an important prognostic factor, with patients < 28 years old faring much better than older individuals (DFS, 45% vs 0%, p = 0.01). Our experience suggests that high-dose chemoradiotherapy followed by infusion of J2/J5 purged autologous marrow is as efficacious in young adults as it is in children and is a reasonable alternative for patients who lack HLA-matched donors. Results in older adults are poor, however, and demonstrate the need for more effective transplant strategies in these individuals.

Authors
Soiffer, RJ; Roy, DC; Gonin, R; Murray, C; Anderson, KC; Freedman, AS; Rabinowe, SN; Robertson, MJ; Spector, N; Pesek, K; Mauch, P; Nadler, LM; Ritz, J
MLA Citation
Soiffer, RJ, Roy, DC, Gonin, R, Murray, C, Anderson, KC, Freedman, AS, Rabinowe, SN, Robertson, MJ, Spector, N, Pesek, K, Mauch, P, Nadler, LM, and Ritz, J. "Monoclonal antibody-purged autologous bone marrow transplantation in adults with acute lymphoblastic leukemia at high risk of relapse." Bone Marrow Transplantation 12.3 (1993): 243-251.
PMID
8241984
Source
scival
Published In
Bone Marrow Transplantation
Volume
12
Issue
3
Publish Date
1993
Start Page
243
End Page
251

Monoclonal antibody-purged bone marrow transplantation therapy for multiple myeloma

Forty patients with plasma cell dyscrasias underwent high-dose chemoradiotherapy and either anti-B-cell monoclonal antibody (MoAb)-treated autologous, anti-T-cell MoAb-treated HLA-matched sibling allogeneic or syngeneic bone marrow transplantation (BMT). The majority of patients had advanced Durie-Salmon stage myeloma at diagnosis, all were pretreated with chemotherapy, and 17 had received prior radiotherapy. At the time of BMT, all patients demonstrated good performance status with Karnofsky score of 80% or greater and had less than 10% marrow tumor cells; 34 patients had residual monoclonal marrow plasma cells and 38 patients had paraprotein. Following high-dose chemoradiotherapy, there were 18 complete responses (CR), 18 partial responses, one non-responder, and three toxic deaths. Granulocytes greater than 500/μL and untransfused platelets greater than 20,000/μL were noted at a median of 23 (range, 12 to 46) and 25 (range, 10 to 175) days posttransplant (PT), respectively, in 24 of the 26 patients who underwent autografting. In the 14 patients who received allogeneic or syngeneic grafts, granulocytes greater than 500/μL and untransfused platelets greater than 20,000/μL were noted at a median of 19 (range, 12 to 24) and 16 (range, 5 to 32) days PT, respectively. With 24 months median follow-up for survival after autologous BMT, 16 of 26 patients are alive free from progression at 2+ to 55+ months PT; of these, 5 patients remain in CR at 6+ to 55+ months PT. With 24 months median follow-up for survival after allogeneic and syngeneic BMT, 8 of 14 patients are alive free from progression at 8+ to 34+ months PT; of these, 5 patients remain in CR at 8+ to 34+ months PT. This therapy has achieved high response rates and prolonged progression-free survival in some patients and proven to have acceptable toxicity. However, relapses post-BMT, coupled with slow engraftment post-BMT in heavily pretreated patients, suggest that such treatment strategies should be used earlier in the disease course. To define the role of BMT in the treatment of myeloma, its efficacy should be compared with that of conventional chemotherapy in a randomized trial. © 1993 by The American Society of Hematology.

Authors
Anderson, KC; Andersen, J; Soiffer, R; Freedman, AS; Rabinowe, SN; Robertson, MJ; Spector, N; Blake, K; Murray, C; Freeman, A; Coral, F; Marcus, KC; Mauch, P; Nadler, LM; Ritz, J
MLA Citation
Anderson, KC, Andersen, J, Soiffer, R, Freedman, AS, Rabinowe, SN, Robertson, MJ, Spector, N, Blake, K, Murray, C, Freeman, A, Coral, F, Marcus, KC, Mauch, P, Nadler, LM, and Ritz, J. "Monoclonal antibody-purged bone marrow transplantation therapy for multiple myeloma." Blood 82.8 (1993): 2568-2576.
PMID
8400304
Source
scival
Published In
Blood
Volume
82
Issue
8
Publish Date
1993
Start Page
2568
End Page
2576

Autologous and allogeneic bone marrow transplantation for poor prognosis patients with B-cell chronic lymphocytic leukemia

Twenty patients with poor prognosis B-cell chronic lymphocytic leukemia (B-CLL) underwent uniform high-dose chemoradiotherapy followed by rescue with multiple monoclonal antibody-purged autologous bone marrow (BM) (12 patients) or T-cell-depleted allogeneic BM from HLA-identical siblings (8 patients) in a pilot study to assess the feasibility of BM transplantation (BMT) in this disease. All had poor prognosis disease by either staging, BM pattern, tumor doubling time criteria, or cytogenetics. All patients achieved remission criteria (defined as ≤2 cm adenopathy, absence of splenomegaly, ≤20% of the intertrabecular space involved on BM biopsy) before BMT. Despite the use of fludarabine, a median of three treatment regimens were required to achieve BMT eligibility. After BMT, all patients achieved complete hematologic engraftment. Toxicities were not significantly different between autologous versus allogeneic BMT. Two toxic deaths were observed. Of 19 evaluable patients, 17 clinical complete clinical remissions (89%) were observed, with 2 patients (1 allogeneic and 1 autologous) exhibiting persistent BM disease. Complete clinical remissions were documented at the phenotypic and molecular level for the majority of patients in whom dual fluorescence for CD5 and CD20 (15 of 15; 100%) and Ig gene rearrangements (11 of 14; 79%) were performed. Although long-term follow-up is needed to assess any potential impact on the disease-free and overall survival of these patients, this study shows the feasibility of using high-dose chemoradiotherapy and BMT in patients with poor prognosis B-CLL. © 1993 by The American Society of Hematology.

Authors
Rabinowe, SN; Soiffer, RJ; Gribben, JG; Daley, H; Freedman, AS; Daley, J; Pesek, K; Neuberg, D; Pinkus, G; Leavitt, PR; Spector, NA; Grossbard, ML; Anderson, K; Robertson, MJ; Mauch, P; Chayt-Marcus, K; Ritz, J; Nadler, LM
MLA Citation
Rabinowe, SN, Soiffer, RJ, Gribben, JG, Daley, H, Freedman, AS, Daley, J, Pesek, K, Neuberg, D, Pinkus, G, Leavitt, PR, Spector, NA, Grossbard, ML, Anderson, K, Robertson, MJ, Mauch, P, Chayt-Marcus, K, Ritz, J, and Nadler, LM. "Autologous and allogeneic bone marrow transplantation for poor prognosis patients with B-cell chronic lymphocytic leukemia." Blood 82.4 (1993): 1366-1376.
PMID
7688995
Source
scival
Published In
Blood
Volume
82
Issue
4
Publish Date
1993
Start Page
1366
End Page
1376

Heat shock protein is a unique marker of growth arrest during macrophage differentiation of HL-60 cells

Prior to morphologic and functional maturation, terminally differentiating hematopoietic cells first exit the cell cycle and undergo growth arrest. Relatively little is known about which molecules regulate differentiation-induced growth arrest. In the present report, we sought to determine whether the mammalian low molecular weight heat shock protein (hsp28) was a candidate growth-regulatory molecule during human hematopoiesis. To this end, hsp28 protein expression was examined during phorbol ester (PMA)-induced macrophage differentiation of the human HL-60 promyelocytic leukemic cell line. Whereas hsp28 was constitutively expressed at relatively low levels in an unphosphorylated state, hsp28 was rapidly phosphorylated within 4 hr following PMA-induced differentiation, preceding increased hsp28 protein levels at 24-48 h. In contrast to other differentiative agents, hsp28 steady state mRNA and protein were regulated concordantly in response to macrophage differentiation. More importantly, these changes were transient, and occurred concomitant with the down-regulation of cellular proliferation and the onset of G1 phase cell cycle arrest. In total, these observations implicate hsp28 as an intermediary in the myelomonocytic differentiative pathway of promyelocytic leukemic cells, and will shed light on the events regulating this process. ©1993 Wiley-Liss, Inc.

Authors
Spector, NL; Ryan, C; Samson, W; Levine, H; Nadler, LM; Arrigo, A-P
MLA Citation
Spector, NL, Ryan, C, Samson, W, Levine, H, Nadler, LM, and Arrigo, A-P. "Heat shock protein is a unique marker of growth arrest during macrophage differentiation of HL-60 cells." Journal of Cellular Physiology 156.3 (1993): 619-625.
PMID
8360264
Source
scival
Published In
Journal of Cellular Physiology
Volume
156
Issue
3
Publish Date
1993
Start Page
619
End Page
625

Detection by polymerase chain reaction of residual cells with the bcl-2 translocation is associated with increased risk of relapse after autologous bone marrow transplantation for B-cell lymphoma

Although molecular biologic techniques can now detect minimal numbers of residual cancer cells in patients in complete clinical remission, the clinical significance of minimal residual disease has never been conclusively established. If the detection of minimal residual disease predicts which patients will relapse, then therapy could be altered based upon the detection of these cells. The t(14;18) can be detected by polymerase chain reaction (PCR) amplification in 50% of patients with B-cell non-Hodgkin's lymphoma and allows detection of one lymphoma cell in up to 1 million normal cells. To determine the clinical significance of the detection of minimal residual lymphoma cells in the bone marrow (BM) PCR amplification was used to detect the presence of residual lymphoma cells after autologous BM transplantation (ABMT) in serial BM samples from 134 patients with B-cell lymphoma in whom a bcl-2 translocation could be detected. PCR analysis was performed on a total of 542 BM samples obtained while these patients were in complete remission. Disease-free survival was markedly increased in patients with no PCR-detectable lymphoma cells in the marrow compared with those in whom residual lymphoma cells were detected (P < .00001), and the presence of detectable lymphoma cells was associated with a 48-fold increase in the risk of relapse. Of the 77 patients (57%) with no PCR-detectable lymphoma cells in their most recent BM sample, none have relapsed. In contrast, all 33 patients (25%) who have relapsed had PCR-detectable lymphoma cells detected in their BM before clinical relapse occurred. In 19 patients (14%), residual lymphoma cells in the BM were detected early following transplantation and subsequently were no longer detectable, although these patients received no further therapy. In these patients, residual lymphoma cells may already have been irreversibly damaged by the high-dose therapy or an endogenous immune mechanism may be capable of eliminating residual lymphoma cells in some patients. Therefore, although the detection of minimal residual disease by PCR following ABMT in patients with lymphoma identifies those patients at high risk of relapse, the presence of residual minimal disease early after transplantation may not be associated with poor prognosis in a small subset of patients. Confirmatory studies will be required to determine more definitively the role of minimal disease detection to identify which patients require additional therapy. © 1993 by The American Society of Hematology.

Authors
Gribben, JG; Neuberg, D; Freedman, AS; Gimmi, CD; Pesek, KW; Barber, M; Saporito, L; Woo, SD; Coral, F; Spector, N; Rabinowe, SN; Grossbard, ML; Ritz, J; Nadler, LM
MLA Citation
Gribben, JG, Neuberg, D, Freedman, AS, Gimmi, CD, Pesek, KW, Barber, M, Saporito, L, Woo, SD, Coral, F, Spector, N, Rabinowe, SN, Grossbard, ML, Ritz, J, and Nadler, LM. "Detection by polymerase chain reaction of residual cells with the bcl-2 translocation is associated with increased risk of relapse after autologous bone marrow transplantation for B-cell lymphoma." Blood 81.12 (1993): 3449-3457.
PMID
8507880
Source
scival
Published In
Blood
Volume
81
Issue
12
Publish Date
1993
Start Page
3449
End Page
3457

Autologous bone marrow transplantation in poor-prognosis intermediate-grade and high-grade B-cell non-Hodgkin's lymphoma in first remission: A pilot study

Purpose: Using high-dose therapy and outologous bone marrow transplantation (ABMT) to overcome cellular resistance and eradicate minimal disease, we initiated a pilot study during first remission in patients with non-Hodgkin's lymphoma (NHL) to examine whether the long-term disease-free survival (DPS) rate can be improved for patients with poor-prognosis intermediate/ high-grade NHL. Patients and Methods: Twenty-six patients with advanced-stage diffuse intermediate/high-grade B-cell NHL (including 16 patients with diffuse small cleaved-cell [DSC]) were selected at presentation by histologic and clinical characteristics to have less than a 25% probability of long-term DFS with conventional treatment. After induction chemotherapy, 16 patients were in complete remission (CR) and 10 were in a minimal disease state. Patients were then treated with high-dose cyclophosphamide, total-body irradiation (TBI), and anti-B-cell monoclonal antibody-purged ABMT. Results: Following ABMT, no acute in-hospital treatment deaths occurred, and engraftment of granulocytes and platelets was significantly faster than for patients undergoing ABMT who were in second or subsequent remission. Of 26 patients, 21 remain in CR maintained without continued therapy, three relapsed in sites of prior nodal disease (4.8, 5.4, and 28 months post-ABMT), and two died in remission. The DPS rate is estimated to be 85% at 28 months and thereafter. The median followup period for the 21 patients who are alive and disease-free is 32 months. Conclusion: This pilot study suggests that consolidation of first remission with ABMT may improve the longterm DFS rate for diffuse intermediate/high-grade NHL patients at high risk for relapse.

Authors
Freedman, AS; Takvorian, T; Neuberg, D; Mauch, P; Rabinowe, SN; Anderson, KC; Soiffer, RJ; Spector, N; Grossbard, M; Robertson, MJ; Blake, K; Coral, F; Canellos, GP; Ritz, J; Nadler, LM
MLA Citation
Freedman, AS, Takvorian, T, Neuberg, D, Mauch, P, Rabinowe, SN, Anderson, KC, Soiffer, RJ, Spector, N, Grossbard, M, Robertson, MJ, Blake, K, Coral, F, Canellos, GP, Ritz, J, and Nadler, LM. "Autologous bone marrow transplantation in poor-prognosis intermediate-grade and high-grade B-cell non-Hodgkin's lymphoma in first remission: A pilot study." Journal of Clinical Oncology 11.5 (1993): 931-936.
PMID
8487057
Source
scival
Published In
Journal of Clinical Oncology
Volume
11
Issue
5
Publish Date
1993
Start Page
931
End Page
936

Serotherapy of B-cell neoplasms with anti-B4-blocked ricin: a phase I trial of daily bolus infusion.

Anti-B4-blocked Ricin (Anti-B4-bR) is an immunotoxin comprised of the anti-B4 monoclonal antibody (MoAb) and the protein toxin "blocked ricin." The anti-B4 MoAb is directed against the B-lineage-restricted CD19 antigen expressed on more than 95% of normal and neoplastic B cells. Blocked ricin is an altered ricin derivative that has its nonspecific binding eliminated by chemically blocking the galactose binding domains of the B chain. In vitro cytotoxicity studies demonstrate that the IC37 of Anti-B4-bR is 2 x 10(-11) mol/L compared with 4 x 10(-12) mol/L for native ricin. A phase I dose escalation clinical trial was conducted in 25 patients with refractory B-cell malignancies. Anti-B4-bR was administered by daily 1-hour bolus infusion for 5 consecutive days at doses ranging from 1 microgram/kg/d to 60 micrograms/kg/d. Serum levels above 1 nmol/L were achieved transiently in the majority of patients treated at the maximum tolerated dose of 50 micrograms/kg/d for 5 days for a total dose of 250 micrograms/kg. The dose-limiting toxicity was defined by transient, reversible grade 3 elevations in hepatic transaminases, without impaired hepatic synthetic function. Minor toxicities included transient hypoalbuminemia, thrombocytopenia, and fevers. Human antimouse antibody and human anti-ricin antibody were detected in nine patients. One complete response, two partial responses, and eight mixed or transient responses were observed. These results show the in vitro and in vivo cytotoxicity of Anti-B4-bR and indicate that this immunotoxin can be administered as a daily bolus infusion for 5 days with tolerable, reversible toxicity.

Authors
Grossbard, ML; Freedman, AS; Ritz, J; Coral, F; Goldmacher, VS; Eliseo, L; Spector, N; Dear, K; Lambert, JM; Blättler, WA
MLA Citation
Grossbard, ML, Freedman, AS, Ritz, J, Coral, F, Goldmacher, VS, Eliseo, L, Spector, N, Dear, K, Lambert, JM, and Blättler, WA. "Serotherapy of B-cell neoplasms with anti-B4-blocked ricin: a phase I trial of daily bolus infusion." Blood 79.3 (February 1992): 576-585.
PMID
1370636
Source
epmc
Published In
Blood
Volume
79
Issue
3
Publish Date
1992
Start Page
576
End Page
585

Prevention of graft-versus-host disease by selective depletion of CD6-positive T Lymphocytes from Donor Bone Marrow

Purpose: Acute and chronic graft-versus-host disease (GVHD) continues to be the major causes of morbidity and mortality after allogeneic bone marrow transplantation (BMT). In this study, we have evaluated the clinical effects of selective in vitro T-cell depletion of donor allogeneic bone marrow by using a single monoclonal antibody ([MoAb] anti-T12, CD6) and rabbit complement. This antibody recognizes mature T cells, but not other cellular elements such as natural-killer (NK) cells, B cells, and myeloid precursors. Patients and Methods: From August 1983 to April 1991, 112 consecutive adult patients with hematologic malignancies underwent BMT with bone marrow from HLA-identical sibling donors. Marrow was harvested and depleted of mature T lymphocytes ex vivo by the use of three rounds of incubation with an anti-T12 antibody and rabbit complement. The preparative regimen consisted of cyclophosphamide and fractionated total body irradiation (TBI) in 108 patients. No patients received prophylactic immune suppression post-BMT. Purgation by anti-T12 was used as the only method for the prevention of GVHD. Results: Twenty patients (18%) developed acute GVHD (grade 2 to 4); only eight patients developed chronic GVHD. The incidence of GVHD did not increase signifi-cantly with age. Only three of 112 patients (2.7%) exhibited acute graft failure. One patient developed late graft failure that was associated with cytomegolovirus (CMV) infection. Within the subset of 50 patients who had not previously undergone unsuccessful conventional therapy (acute leukemia in first remission or chronic myelogenous leukemia [CML] in stable phase), we estimated by the Kaplan-Meier method that the probability of disease-free survival was 50% at 3 years post-BMT, with a median follow-up of 44 months. The treatment-related mortality rate in this group was only 14% and was independent of patient age. Conclusions: We conclude that selective in vitro T-cell depletion with an anti-T12 monoclonal antibody effectively reduces the incidence of both acute and chronic GVHD after allogeneic BMT without compromising engraftment. Moreover, depletion of CD6-positive cells from donor marrow obviates the need to administer immune suppressive medications to the majority of patients. This approach reduces the morbidity and mortality of allogeneic BMT and permits the BMT of older patients. © 1992 by American Society of Clinical Oncology.

Authors
Soiffer, RJ; Murray, C; Mauch, P; Anderson, KC; Freedman, AS; Rabinowe, SN; Takvorian, T; Robertson, MJ; Spector, N; Gonin, R; Miller, KB; Rudders, RA; Freeman, A; Blake, K; Coral, F; Nodler, LM; Ritz, J
MLA Citation
Soiffer, RJ, Murray, C, Mauch, P, Anderson, KC, Freedman, AS, Rabinowe, SN, Takvorian, T, Robertson, MJ, Spector, N, Gonin, R, Miller, KB, Rudders, RA, Freeman, A, Blake, K, Coral, F, Nodler, LM, and Ritz, J. "Prevention of graft-versus-host disease by selective depletion of CD6-positive T Lymphocytes from Donor Bone Marrow." Journal of Clinical Oncology 10.7 (1992): 1191-1200.
PMID
1607923
Source
scival
Published In
Journal of Clinical Oncology
Volume
10
Issue
7
Publish Date
1992
Start Page
1191
End Page
1200

Growth arrest of human B lymphocytes is accompanied by induction of the low molecular weight mammalian heat shock protein (Hsp28)

A large number of protein and molecular markers have been identified that delineate the early stages of human B cell activation and proliferation. In contrast, few if any molecules are transiently expressed precisely as activated B cells stop proliferating and undergo growth arrest. We demonstrate that the low molecular weight heat shock protein (hsp28) exhibits unique induction kinetics that specifically demarcates this interval. After mitogenic activation of unstimulated splenic B cells, hsp28 protein and phosphorylation transiently increase coinciding precisely with the peak of cellular proliferation and the onset of growth arrest. Although most neoplastic B cells constitutively express hsp28, three cell lines were identified that were hsp28-. No differences in phenotype or growth kinetics were detected between hsp28+ and hsp28- neoplastic B cells demonstrating that hsp28 expression is not essential for cell growth. However, when treated with phorbol ester or heat shock, these hsp28- cell lines synthesize hsp28 followed by the onset growth arrest. The consistency with which hsp28 induction transiently delineates the interval from peak proliferation to the onset of growth arrest suggests hsp28 itself is likely to be involved in regulating this process.

Authors
Spector, NL; Samson, W; Ryan, C; Gribben, J; Urba, W; Welch, WJ; Nadler, LM
MLA Citation
Spector, NL, Samson, W, Ryan, C, Gribben, J, Urba, W, Welch, WJ, and Nadler, LM. "Growth arrest of human B lymphocytes is accompanied by induction of the low molecular weight mammalian heat shock protein (Hsp28)." Journal of Immunology 148.6 (1992): 1668-1673.
PMID
1541812
Source
scival
Published In
Journal of immunology (Baltimore, Md. : 1950)
Volume
148
Issue
6
Publish Date
1992
Start Page
1668
End Page
1673

Activation primes human B lymphocytes to respond to heat shock

Crosslinkage of the B cell antigen receptor by anti-μ beads or SAC results in the selective induction of hsp70. We have observed that activated cells, having enhanced expression of hsp70, survive lethal stimuli much better than their unactivated counterparts. These results are in accordance with the proposal that hsp70 is essential for cells to survive lethal environmental stresses. Moreover, the activation event itself primes B cells thereby enabling them to increase the expression of both hsp70 mRNA and protein. This is the first demonstration that triggering of B cells via crosslinkage of sIg is accompanied by the induction of thermotolerance without the need for a prior sublethal heat treatment.

Authors
Spector, NL; Freedman, AS; Freeman, G; Segil, J; Whitman, JF; Welch, WJ; Nadler, LM
MLA Citation
Spector, NL, Freedman, AS, Freeman, G, Segil, J, Whitman, JF, Welch, WJ, and Nadler, LM. "Activation primes human B lymphocytes to respond to heat shock." Journal of Experimental Medicine 170.5 (1989): 1763-1768.
PMID
2809511
Source
scival
Published In
Journal of Experimental Medicine
Volume
170
Issue
5
Publish Date
1989
Start Page
1763
End Page
1768
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