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Hanks, Brent A.

Overview:

My lab is interested in elucidating the molecular and cellular mechanisms involved in tumor-mediated immune suppression and cancer immunotherapy resistance. Our overriding hypothesis is that tumor cells and/or their associated stromal elements elicit soluble factors that tolerize local dendritic cell populations and/or recruit other immunosuppressive cell populations to the tumor bed; thereby, interfering with the generation of an effective anti-tumor immune response. This work has both basic and translational significance in that it is capable of providing 1. novel pharmacological targets for enhancing anti-tumor immunity and 2.  much needed biomarkers for guiding the management of cancer patients with immunotherapies.  We perform these investigations utilizing both transgenic murine models as well as clinical specimens derived from cancer patients undergoing immunotherapy.  We focus these studies on melanoma, non-small cell lung cancer, pancreatic cancer, and colon cancer. 

We currently have the following ongoing projects in our lab:
1.  Investigating mechanisms by which developing cancers alter the metabolism of local dendritic cells thereby hijacking this antigen-presenting cell population to generate an immunotolerant tumor microenvironment. 
2.  Identifying soluble factors expressed by cancers which manipulate local dendritic cell function to drive regulatory T cell  differentiation within the tumor microenvironment as well as any potential oncogenic signaling pathways driving this process.
3.  Characterizing mechanisms of innate and adaptive resistance mechanisms to checkpoint inhibitor therapies.
4.  Examining the role of the tumor stroma in interfering with immunotherapy efficacy.
5.  Design and development of novel dendritic cell-based vaccine strategies

Positions:

Assistant Professor of Medicine

Medicine, Medical Oncology
School of Medicine

Member of the Duke Cancer Institute

Duke Cancer Institute
School of Medicine

Education:

Ph.D. 2004

Ph.D. — Baylor College of Medicine

M.D. 2006

M.D. — Baylor College of Medicine

Internship And Residency, Internal Medicine

Duke University School of Medicine

Fellowship, Hematology/Oncology

Duke University School of Medicine

Grants:

Awakening the dormant tumor: the role of the tumor microenvironment in breast cancer recurrence

Administered By
Pharmacology & Cancer Biology
AwardedBy
National Institutes of Health
Role
Collaborator
Start Date
August 01, 2017
End Date
July 31, 2021

Melanoma-mediated Dendritic Cell Tolerization and Immune Evasion

Administered By
Medicine, Medical Oncology
AwardedBy
National Institutes of Health
Role
Principal Investigator
Start Date
August 04, 2015
End Date
July 31, 2020

Metabolic Reprogramming of Dendritic Cell-based Cancer Vaccines to Enhance Anti-Tumor Immunity

Administered By
Medicine, Medical Oncology
AwardedBy
Alliance for Cancer Gene Therapy
Role
Principal Investigator
Start Date
June 10, 2016
End Date
June 09, 2019

A phase 1 / 2 randomized, blinded, placebo controlled study of Ipilimumab in combination with INCB024360 or placebo in u

Administered By
Duke Cancer Institute
AwardedBy
Incyte Corporation
Role
Principal Investigator
Start Date
October 01, 2013
End Date
September 30, 2018

Investigating Oncogenic Signaling Pathways that Drive Wnt Ligand-mediated Immune Tolerance in Melanoma

Administered By
Medicine, Medical Oncology
AwardedBy
Conquer Cancer Foundation
Role
Principal Investigator
Start Date
July 01, 2017
End Date
June 30, 2018

Merck Sharp & Dohme Corp. Study Agreement

Administered By
Medicine, Medical Oncology
AwardedBy
Merck
Role
Principal Investigator
Start Date
July 01, 2015
End Date
June 30, 2018

Seahorse XFe96 Extracellular Flux Analyzer

Administered By
Pediatrics, Endocrinology
AwardedBy
North Carolina Biotechnology Center
Role
Major User
Start Date
June 07, 2017
End Date
June 06, 2018

Investigation of the FZD8-Fc Wnt Ligand Antagonist and the Fzd Receptor Monoclonal Antibody in Combination with Immune Checkpoint Blockade in Pre-Clinical Melanoma, Non-Small Cell Lung Cancer, and Col

Administered By
Medicine, Medical Oncology
AwardedBy
OncoMed Pharmaceuticals
Role
Principal Investigator
Start Date
October 16, 2015
End Date
April 16, 2018

CITN-07

Administered By
Medicine, Medical Oncology
AwardedBy
Fred Hutchinson Cancer Research Center
Role
Principal Investigator
Start Date
September 01, 2014
End Date
December 31, 2017

CITN-09 A Phase II Study of MK-3475 in Patients with Advanced Merkel Cell Carcinoma (MCC)

Administered By
Medicine, Medical Oncology
AwardedBy
Fred Hutchinson Cancer Research Center
Role
Principal Investigator
Start Date
September 01, 2016
End Date
August 31, 2017

A Phase II, Open-label, Multicenter, Randomized Study of CDX-1401, a Dendritic Cell Targeting NY-ESO-1 Vaccine, in Patients with Malignant Melanoma Pre-Treated with Recombinant CDX-301, a Recombinant

Administered By
Medicine, Medical Oncology
AwardedBy
Fred Hutchinson Cancer Research Center
Role
Principal Investigator
Start Date
September 01, 2015
End Date
August 31, 2017

Phase II Pilot Trial of an Indoleamine 2, 3,diosygenase -1 ( IDO1) Inhibitor (INCBO24360) Plus a Multipeptide Melanoma Vaccine ( MELTAC 12.1 ) in Patients with Advanced Melanoma

Administered By
Medicine, Medical Oncology
AwardedBy
Fred Hutchinson Cancer Research Center
Role
Principal Investigator
Start Date
September 01, 2015
End Date
August 31, 2017

Therapeutic Targeting of the TGF-BSignaling Axis to Modulate the Tumor Immune Microenvironment and Enhance Melanoma Immu

Administered By
Medicine, Medical Oncology
AwardedBy
Melanoma Research Alliance
Role
Principal Investigator
Start Date
November 01, 2013
End Date
January 31, 2017

Investigation of TEW-7197 in Combination with Immune Checkpoint Inhibition in the BRAFV600E-PTEN-/- Melanoma Model

Administered By
Medicine, Medical Oncology
AwardedBy
MedPacto, Inc
Role
Principal Investigator
Start Date
November 07, 2014
End Date
December 31, 2015

A phase II pilot trial of an Indoleamine2,3, dioxygenase-1 (IDO1) Inhibitor (INCB024360) plus a multipeptide melanoma v

Administered By
Duke Cancer Institute
AwardedBy
Fred Hutchinson Cancer Research Center
Role
Principal Investigator
Start Date
September 01, 2013
End Date
August 31, 2014

A phase II, open-label, multicenter, randomized study of CDX-1401 a Dendritic Cell targeting NY-ESO-1 vaccine, in patien

Administered By
Duke Cancer Institute
AwardedBy
Fred Hutchinson Cancer Research Center
Role
Principal Investigator
Start Date
September 01, 2013
End Date
August 31, 2014

Role of Type III TGF-b Receptor in Mediating Immunosuppression During Breast Cancer Progression

Administered By
Medicine, Medical Oncology
AwardedBy
Department of Defense
Role
Principal Investigator
Start Date
September 30, 2010
End Date
October 29, 2013
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Awards:

Alliance for Cancer Gene Therapy Young Investigator Award. Alliance for Cancer Gene Therapy.

Type
National
Awarded By
Alliance for Cancer Gene Therapy
Date
January 01, 2016

Duke Scholar Award . Duke University Health System.

Type
School
Awarded By
Duke University Health System
Date
January 01, 2016

National Finalist Stand-Up To Cancer-Merck Catalyst Award. SU2C.

Type
National
Awarded By
SU2C
Date
January 01, 2016

DCI Pilot Research Award. Duke Cancer Institute.

Type
University
Awarded By
Duke Cancer Institute
Date
January 01, 2015

Melanoma Research Alliance Young Investigator Award. Melanoma Research Alliance, DCI, Frank Courtney .

Type
National
Awarded By
Melanoma Research Alliance, DCI, Frank Courtney
Date
January 01, 2013

American Society Clinical Oncology Merit Award. ASCO.

Type
National
Awarded By
ASCO
Date
January 01, 2012

Publications:

Identifying baseline immune-related biomarkers to predict clinical outcome of immunotherapy.

As cancer strikes, individuals vary not only in terms of factors that contribute to its occurrence and development, but as importantly, in their capacity to respond to treatment. While exciting new therapeutic options that mobilize the immune system against cancer have led to breakthroughs for a variety of malignancies, success is limited to a subset of patients. Pre-existing immunological features of both the host and the tumor may contribute to how patients will eventually fare with immunotherapy. A broad understanding of baseline immunity, both in the periphery and in the tumor microenvironment, is needed in order to fully realize the potential of cancer immunotherapy. Such interrogation of the tumor, blood, and host immune parameters prior to treatment is expected to identify biomarkers predictive of clinical outcome as well as to elucidate why some patients fail to respond to immunotherapy. To approach these opportunities for progress, the Society for Immunotherapy of Cancer (SITC) reconvened the Immune Biomarkers Task Force. Comprised of an international multidisciplinary panel of experts, Working Group 4 sought to make recommendations that focus on the complexity of the tumor microenvironment, with its diversity of immune genes, proteins, cells, and pathways naturally present at baseline and in circulation, and novel tools to aid in such broad analyses.

Authors
Gnjatic, S; Bronte, V; Brunet, LR; Butler, MO; Disis, ML; Galon, J; Hakansson, LG; Hanks, BA; Karanikas, V; Khleif, SN; Kirkwood, JM; Miller, LD; Schendel, DJ; Tanneau, I; Wigginton, JM; Butterfield, LH
MLA Citation
Gnjatic, S, Bronte, V, Brunet, LR, Butler, MO, Disis, ML, Galon, J, Hakansson, LG, Hanks, BA, Karanikas, V, Khleif, SN, Kirkwood, JM, Miller, LD, Schendel, DJ, Tanneau, I, Wigginton, JM, and Butterfield, LH. "Identifying baseline immune-related biomarkers to predict clinical outcome of immunotherapy." Journal for immunotherapy of cancer 5 (January 2017): 44-. (Review)
PMID
28515944
Source
epmc
Published In
Journal for ImmunoTherapy of Cancer
Volume
5
Publish Date
2017
Start Page
44
DOI
10.1186/s40425-017-0243-4

Safety and Efficacy of Radiation Therapy in Advanced Melanoma Patients Treated With Ipilimumab.

Ipilimumab and radiation therapy (RT) are standard treatments for advanced melanoma; preclinical models suggest the potential for synergy. However, limited clinical information exists regarding safety and optimal timing of the combination.We reviewed the records of consecutive patients with unresectable stage 3 or 4 melanoma treated with ipilimumab. Patients were categorized as having received RT or not. Differences were estimated between these 2 cohorts.We identified 88 patients treated with ipilimumab. At baseline, the ipilimumab-plus-RT group (n=44) had more unfavorable characteristics. Despite this, overall survival, progression-free survival, and both immune-related and non-immune-related toxicity were not statistically different (P=.67). Patients who received ipilimumab before RT had an increased duration of irradiated tumor response compared with patients receiving ipilimumab after RT (74.7% vs 44.8% at 12 months; P=.01, log-rank test). In addition, patients receiving ablative RT had non-statistically significantly improved median overall survival (19.6 vs 10.2 months), as well as 6-month (95.1% vs 72.7%) and 12-month (79.7% vs 48.5%) survival rates, compared with those treated with conventionally fractionated RT.We found that both ablative and conventionally fractionated RT can be safely administered with ipilimumab without a clinically apparent increase in toxicity. Patients who received ipilimumab before RT had an increased duration of irradiated tumor response.

Authors
Qin, R; Olson, A; Singh, B; Thomas, S; Wolf, S; Bhavsar, NA; Hanks, BA; Salama, JK; Salama, AKS
MLA Citation
Qin, R, Olson, A, Singh, B, Thomas, S, Wolf, S, Bhavsar, NA, Hanks, BA, Salama, JK, and Salama, AKS. "Safety and Efficacy of Radiation Therapy in Advanced Melanoma Patients Treated With Ipilimumab." International journal of radiation oncology, biology, physics 96.1 (September 2016): 72-77.
PMID
27375168
Source
epmc
Published In
International Journal of Radiation Oncology, Biology, Physics
Volume
96
Issue
1
Publish Date
2016
Start Page
72
End Page
77
DOI
10.1016/j.ijrobp.2016.04.017

Isolated recto-sigmoid colitis: a new imaging pattern of ipilimumab-associated colitis.

The purpose of this study is to describe typical CT findings and distinct imaging patterns of ipilimumab-associated colitis in immunotherapeutic treatment of melanoma.This HIPAA-compliant retrospective study included 86 patients with melanoma imaged with CT or PET/CT of the abdomen and pelvis during or shortly after administration of ipilimumab. Twelve of 86 patients (14%) developed symptoms of colitis and underwent CT imaging of the abdomen and pelvis while symptomatic. Two radiologists reviewed CT images to evaluate for the presence of CT findings of colitis including mesenteric vessel engorgement, pericolonic inflammatory change, hyperenhancement of colonic mucosa, colonic wall thickening, fluid-filled colonic distension, pneumoperitoneum, pneumatosis, and diverticulosis in the inflamed segment of colon. One nuclear medicine radiologist reviewed PET images for abnormally increased FDG uptake in the colon. The diagnosis of ipilimumab-associated colitis was made based on clinical presentation, imaging findings, and laboratory data.Common CT findings of ipilimumab-associated colitis included colonic mucosal hyperenhancement (10/12 [83%]), mesenteric vessel engorgement (9/12 [75.0%]), colonic wall thickening (9/12 [75%]), and pericolonic fat stranding (2/12 [16%]). No patient developed pneumatosis or pneumoperitoneum. Diffuse colitis was present in 4/12 (33%) patients. Segmental colitis with associated diverticulosis (was present in 2/12 (17%) patients). A third pattern, isolated recto-sigmoid colitis without diverticulosis, was observed in 6/12 (50%) patients. All patients with colitis demonstrated recto-sigmoid involvement.A third radiologic pattern of ipilimumab-associated colitis was observed in this study: isolated recto-sigmoid colitis without diverticulosis. All patterns of ipilimumab-associated colitis include recto-sigmoid involvement.

Authors
Barina, AR; Bashir, MR; Howard, BA; Hanks, BA; Salama, AK; Jaffe, TA
MLA Citation
Barina, AR, Bashir, MR, Howard, BA, Hanks, BA, Salama, AK, and Jaffe, TA. "Isolated recto-sigmoid colitis: a new imaging pattern of ipilimumab-associated colitis." Abdominal radiology (New York) 41.2 (February 2016): 207-214.
PMID
26867901
Source
epmc
Published In
Abdominal radiology (New York)
Volume
41
Issue
2
Publish Date
2016
Start Page
207
End Page
214
DOI
10.1007/s00261-015-0560-3

Immune evasion pathways and the design of dendritic cell-based cancer vaccines.

Emerging data is suggesting that the process of dendritic cell (DC) tolerization is an important step in tumorigenesis. Our understanding of the networks within the tumor microenvironment that functionally tolerize DC function is evolving while methods for genetically manipulating DC populations in situ continue to develop. A more intimate understanding of the paracrine signaling pathways which mediate immune evasion by subverting DC function promises to provide novel strategies for improving the clinical efficacy of DC-based cancer vaccines. This will likely require a better understanding of both the antigen expression profile and the immune evasion network of the tumor and its associated stromal tissues.

Authors
Hanks, BA
MLA Citation
Hanks, BA. "Immune evasion pathways and the design of dendritic cell-based cancer vaccines." Discovery medicine 21.114 (February 2016): 135-142. (Review)
Website
http://hdl.handle.net/10161/13295
PMID
27011049
Source
epmc
Published In
Discovery medicine
Volume
21
Issue
114
Publish Date
2016
Start Page
135
End Page
142

Genetic risk analysis of a patient with fulminant autoimmune type 1 diabetes mellitus secondary to combination ipilimumab and nivolumab immunotherapy.

Checkpoint inhibitor immunotherapy is becoming an effective treatment modality for an increasing number of malignancies. As a result, autoinflammatory side-effects are also being observed more commonly in the clinic. We are currently unable to predict which patients will develop more severe toxicities associated with these treatment regimens.We present a patient with stage IV melanoma that developed rapid onset autoimmune type 1 diabetes (T1D) in response to combination ipilimumab and nivolumab immunotherapy. At the time of the patient's presentation with diabetes ketoacidosis, a confirmed anti-GAD antibody seroconversion was noted. Longer-term follow-up of this patient has demonstrated a durable complete response based on PET CT imaging along with a persistently undetectable C-peptide level. Single nucleotide polymorphism gene sequencing and HLA risk allele analysis has revealed the patient to lack any established genetic predisposition to the development of autoimmune T1D.While larger studies are necessary to better understand the role of genetic risk factors for the development of autoimmune toxicities in those patients undergoing checkpoint inhibitor immunotherapy, these results suggest that pre-screening patients for known T1D risk alleles may not be indicated. Additional investigation is needed to determine whether an approach such as T cell receptor clonotypic analysis to identify the presence of autoreactive T cell clones may be an effective approach for predicting which patients are at risk for the development of autoinflammatory toxicities while undergoing checkpoint inhibitor immunotherapy.

Authors
Lowe, JR; Perry, DJ; Salama, AKS; Mathews, CE; Moss, LG; Hanks, BA
MLA Citation
Lowe, JR, Perry, DJ, Salama, AKS, Mathews, CE, Moss, LG, and Hanks, BA. "Genetic risk analysis of a patient with fulminant autoimmune type 1 diabetes mellitus secondary to combination ipilimumab and nivolumab immunotherapy." Journal for immunotherapy of cancer 4 (January 2016): 89-.
Website
http://hdl.handle.net/10161/13294
PMID
28031819
Source
epmc
Published In
Journal for ImmunoTherapy of Cancer
Volume
4
Publish Date
2016
Start Page
89
DOI
10.1186/s40425-016-0196-z

Melanoma-Derived Wnt5a Promotes Local Dendritic-Cell Expression of IDO and Immunotolerance: Opportunities for Pharmacologic Enhancement of Immunotherapy.

The β-catenin signaling pathway has been demonstrated to promote the development of a tolerogenic dendritic cell (DC) population capable of driving regulatory T-cell (Treg) differentiation. Further studies have implicated tolerogenic DCs in promoting carcinogenesis in preclinical models. The molecular mechanisms underlying the establishment of immune tolerance by this DC population are poorly understood, and the methods by which developing cancers can co-opt this pathway to subvert immune surveillance are currently unknown. This work demonstrates that melanoma-derived Wnt5a ligand upregulates the durable expression and activity of the indoleamine 2,3-dioxygenase-1 (IDO) enzyme by local DCs in a manner that depends upon the β-catenin signaling pathway. These data indicate that Wnt5a-conditioned DCs promote the differentiation of Tregs in an IDO-dependent manner, and that this process serves to suppress melanoma immune surveillance. We further show that the genetic silencing of the PORCN membrane-bound O-acyl transferase, which is necessary for melanoma Wnt ligand secretion, enhances antitumor T-cell immunity, and that the pharmacologic inhibition of this enzyme synergistically suppresses melanoma progression when combined with anti-CTLA-4 antibody therapy. Finally, our data suggest that β-catenin signaling activity, based on a target gene expression profile that includes IDO in human sentinel lymph node-derived DCs, is associated with melanoma disease burden and diminished progression-free survival. This work implicates the Wnt-β-catenin signaling pathway as a novel therapeutic target in the melanoma immune microenvironment and demonstrates the potential impact of manipulating DC function as a strategy for optimizing tumor immunotherapy.

Authors
Holtzhausen, A; Zhao, F; Evans, KS; Tsutsui, M; Orabona, C; Tyler, DS; Hanks, BA
MLA Citation
Holtzhausen, A, Zhao, F, Evans, KS, Tsutsui, M, Orabona, C, Tyler, DS, and Hanks, BA. "Melanoma-Derived Wnt5a Promotes Local Dendritic-Cell Expression of IDO and Immunotolerance: Opportunities for Pharmacologic Enhancement of Immunotherapy." Cancer immunology research 3.9 (September 2015): 1082-1095.
Website
http://hdl.handle.net/10161/13296
PMID
26041736
Source
epmc
Published In
Cancer Immunology Research
Volume
3
Issue
9
Publish Date
2015
Start Page
1082
End Page
1095
DOI
10.1158/2326-6066.cir-14-0167

Targeting the Wnt5a-beta-catenin pathway in the melanoma microenvironment to augment checkpoint inhibitor immunotherapy.

Authors
Hanks, BA; Zhao, F; Evans, K; Holtzhausen, A; Tsutsui, M; Tyler, D
MLA Citation
Hanks, BA, Zhao, F, Evans, K, Holtzhausen, A, Tsutsui, M, and Tyler, D. "Targeting the Wnt5a-beta-catenin pathway in the melanoma microenvironment to augment checkpoint inhibitor immunotherapy." May 20, 2015.
Source
wos-lite
Published In
Journal of Clinical Oncology
Volume
33
Issue
15
Publish Date
2015

Rapid complete response of metastatic melanoma in a patient undergoing ipilimumab immunotherapy in the setting of active ulcerative colitis.

While blockade of the cytotoxic T-lymphocyte antigen-4 (CTLA-4) T cell regulatory receptor has become a commonly utilized strategy in the management of advanced melanoma, many questions remain regarding the use of this agent in patient populations with autoimmune disease. We present a case involving the treatment of a patient with stage IV melanoma and ulcerative colitis (UC) with anti-CTLA-4 antibody immunotherapy. Upon initial treatment, the patient developed grade III colitis requiring tumor necrosis factor-alpha (TNF-α) blocking antibody therapy, however re-treatment with anti-CTLA-4 antibody following a total colectomy resulted in a rapid complete response accompanied by the development of a tracheobronchitis, a previously described extra-intestinal manifestation of UC. This case contributes to the evolving literature on the use of checkpoint inhibitors in patients also suffering from autoimmune disease, supports future clinical trials investigating the use of these agents in patients with autoimmune diseases, and suggests that an understanding of the specific molecular pathways involved in a patient's autoimmune pathology may provide insight into the development of more effective novel combinatorial immunotherapeutic strategies.

Authors
Bostwick, AD; Salama, AK; Hanks, BA
MLA Citation
Bostwick, AD, Salama, AK, and Hanks, BA. "Rapid complete response of metastatic melanoma in a patient undergoing ipilimumab immunotherapy in the setting of active ulcerative colitis." Journal for immunotherapy of cancer 3 (January 2015): 19-.
Website
http://hdl.handle.net/10161/11661
PMID
25992290
Source
epmc
Published In
Journal for ImmunoTherapy of Cancer
Volume
3
Publish Date
2015
Start Page
19
DOI
10.1186/s40425-015-0064-2

Immunotherapy following regional chemotherapy treatment of advanced extremity melanoma.

Following regional chemotherapy (RC) for melanoma, approximately 75 % of patients will progress. The role of immunotherapy after RC has not been well established.A prospective, single-institution database of 243 patients with in-transit melanoma (1995-2013) was queried for patients who had progression of disease after RC with melphalan and subsequently received systemic immunotherapy. Fifteen patients received IL-2 only, 12 received ipilimumab only, and 6 received IL-2 followed by ipilimumab. Fisher's exact test was used to determine if there was a difference in number of complete responders after immunotherapy.With IL-2 alone, all patients progressed. After ipilimumab alone, three patients had a complete response and nine had progressive disease. Six additional patients received IL-2 first then ipilimumab. All six progressed on IL-2 but three went on to have a complete response to ipilimumab while three progressed. The use of ipilimumab at any time in patients who progressed after RC was associated with higher rate of complete response compared to use of IL-2 alone (33 vs. 0 %; p = 0.021).Patients with progression after regional therapy for melanoma may benefit from immunologic therapy. In this group of patients, immune checkpoint blockade with ipilimumab has a higher complete response rate than T cell stimulation with IL-2, with no complete responders in the IL-2 only group. Furthermore, the complete response rate for ipilimumab in our cohort is higher than reported response rates in the literature for ipilimumab alone, suggesting that the effects of immunotherapy may be bolstered by previous regional treatment.

Authors
Jiang, BS; Beasley, GM; Speicher, PJ; Mosca, PJ; Morse, MA; Hanks, B; Salama, A; Tyler, DS
MLA Citation
Jiang, BS, Beasley, GM, Speicher, PJ, Mosca, PJ, Morse, MA, Hanks, B, Salama, A, and Tyler, DS. "Immunotherapy following regional chemotherapy treatment of advanced extremity melanoma." Annals of surgical oncology 21.8 (August 2014): 2525-2531.
PMID
24700302
Source
epmc
Published In
Annals of Surgical Oncology
Volume
21
Issue
8
Publish Date
2014
Start Page
2525
End Page
2531
DOI
10.1245/s10434-014-3671-0

Combinatorial TGF-beta signaling blockade and anti-CTLA-4 antibody immunotherapy in a murine BRAF(V600E)-PTEN-/-transgenic model of melanoma

Authors
Hanks, BA; Holtzhausen, A; Evans, K; Held, M; Blobe, GC
MLA Citation
Hanks, BA, Holtzhausen, A, Evans, K, Held, M, and Blobe, GC. "Combinatorial TGF-beta signaling blockade and anti-CTLA-4 antibody immunotherapy in a murine BRAF(V600E)-PTEN-/-transgenic model of melanoma." May 20, 2014.
Source
wos-lite
Published In
Journal of Clinical Oncology
Volume
32
Issue
15
Publish Date
2014

Immunotherapy following regional chemotherapy treatment of advanced extremity melanoma

Purpose. Following regional chemotherapy (RC) for melanoma, approximately 75 % of patients will progress. The role of immunotherapy after RC has not been well established. Methods. A prospective, single-institution database of 243 patients with in-transit melanoma (1995-2013) was queried for patients who had progression of disease after RC with melphalan and subsequently received systemic immunotherapy. Fifteen patients received IL-2 only, 12 received ipilimumab only, and 6 received IL-2 followed by ipilimumab. Fisher's exact test was used to determine if there was a difference in number of complete responders after immunotherapy. Results. With IL-2 alone, all patients progressed. After ipilimumab alone, three patients had a complete response and nine had progressive disease. Six additional patients received IL-2 first then ipilimumab. All six progressed on IL-2 but three went on to have a complete response to ipilimumab while three progressed. The use of ipilimumab at any time in patients who progressed after RC was associated with higher rate of complete response compared to use of IL-2 alone (33 vs. 0 %; p = 0.021). Conclusions. Patients with progression after regional therapy for melanoma may benefit from immunologic therapy. In this group of patients, immune checkpoint blockade with ipilimumab has a higher complete response rate than T cell stimulation with IL-2, with no complete responders in the IL-2 only group. Furthermore, the complete response rate for ipilimumab in our cohort is higher than reported response rates in the literature for ipilimumab alone, suggesting that the effects of immunotherapy may be bolstered by previous regional treatment. © 2014 Society of Surgical Oncology.

Authors
Jiang, BS; Beasley, GM; Speicher, PJ; Mosca, PJ; Morse, MA; Hanks, B; Salama, A; Tyler, DS
MLA Citation
Jiang, BS, Beasley, GM, Speicher, PJ, Mosca, PJ, Morse, MA, Hanks, B, Salama, A, and Tyler, DS. "Immunotherapy following regional chemotherapy treatment of advanced extremity melanoma." Annals of Surgical Oncology 21.8 (January 1, 2014): 2525-2531.
Source
scopus
Published In
Annals of Surgical Oncology
Volume
21
Issue
8
Publish Date
2014
Start Page
2525
End Page
2531
DOI
10.1245/s10434-014-3671-0

Early Carcinogenesis Involves the Establishment of Immune Privilege via Intrinsic and Extrinsic Regulation of Indoleamine 2,3-dioxygenase-1: Translational Implications in Cancer Immunotherapy.

Although prolonged genetic pressure has been conjectured to be necessary for the eventual development of tumor immune evasion mechanisms, recent work is demonstrating that early genetic mutations are capable of moonlighting as both intrinsic and extrinsic modulators of the tumor immune microenvironment. The indoleamine 2,3-dioxygenase-1 (IDO) immunoregulatory enzyme is emerging as a key player in tumor-mediated immune tolerance. While loss of the tumor suppressor, BIN-1, and the over-expression of cyclooxygenase-2 have been implicated in intrinsic regulation of IDO, recent findings have demonstrated the loss of TβRIII and the upregulation of Wnt5a by developing cancers to play a role in the extrinsic control of IDO activity by local dendritic cell populations residing within tumor and tumor-draining lymph node tissues. Together, these genetic changes are capable of modulating paracrine signaling pathways in the early stages of carcinogenesis to establish a site of immune privilege by promoting the differentiation and activation of local regulatory T cells. Additional investigation of these immune evasion pathways promises to provide opportunities for the development of novel strategies to synergistically enhance the efficacy of the evolving class of T cell-targeted "checkpoint" inhibitors.

Authors
Holtzhausen, A; Zhao, F; Evans, KS; Hanks, BA
MLA Citation
Holtzhausen, A, Zhao, F, Evans, KS, and Hanks, BA. "Early Carcinogenesis Involves the Establishment of Immune Privilege via Intrinsic and Extrinsic Regulation of Indoleamine 2,3-dioxygenase-1: Translational Implications in Cancer Immunotherapy." Frontiers in immunology 5 (January 2014): 438-. (Review)
Website
http://hdl.handle.net/10161/11662
PMID
25339948
Source
epmc
Published In
Frontiers in Immunology
Volume
5
Publish Date
2014
Start Page
438
DOI
10.3389/fimmu.2014.00438

Type III TGF-β receptor downregulation generates an immunotolerant tumor microenvironment.

Cancers subvert the host immune system to facilitate disease progression. These evolved immunosuppressive mechanisms are also implicated in circumventing immunotherapeutic strategies. Emerging data indicate that local tumor-associated DC populations exhibit tolerogenic features by promoting Treg development; however, the mechanisms by which tumors manipulate DC and Treg function in the tumor microenvironment remain unclear. Type III TGF-β receptor (TGFBR3) and its shed extracellular domain (sTGFBR3) regulate TGF-β signaling and maintain epithelial homeostasis, with loss of TGFBR3 expression promoting progression early in breast cancer development. Using murine models of breast cancer and melanoma, we elucidated a tumor immunoevasion mechanism whereby loss of tumor-expressed TGFBR3/sTGFBR3 enhanced TGF-β signaling within locoregional DC populations and upregulated both the immunoregulatory enzyme indoleamine 2,3-dioxygenase (IDO) in plasmacytoid DCs and the CCL22 chemokine in myeloid DCs. Alterations in these DC populations mediated Treg infiltration and the suppression of antitumor immunity. Our findings provide mechanistic support for using TGF-β inhibitors to enhance the efficacy of tumor immunotherapy, indicate that sTGFBR3 levels could serve as a predictive immunotherapy biomarker, and expand the mechanisms by which TGFBR3 suppresses cancer progression to include effects on the tumor immune microenvironment.

Authors
Hanks, BA; Holtzhausen, A; Evans, KS; Jamieson, R; Gimpel, P; Campbell, OM; Hector-Greene, M; Sun, L; Tewari, A; George, A; Starr, M; Nixon, A; Augustine, C; Beasley, G; Tyler, DS; Osada, T; Morse, MA; Ling, L; Lyerly, HK; Blobe, GC
MLA Citation
Hanks, BA, Holtzhausen, A, Evans, KS, Jamieson, R, Gimpel, P, Campbell, OM, Hector-Greene, M, Sun, L, Tewari, A, George, A, Starr, M, Nixon, A, Augustine, C, Beasley, G, Tyler, DS, Osada, T, Morse, MA, Ling, L, Lyerly, HK, and Blobe, GC. "Type III TGF-β receptor downregulation generates an immunotolerant tumor microenvironment." J Clin Invest 123.9 (September 2013): 3925-3940.
Website
http://hdl.handle.net/10161/13297
PMID
23925295
Source
pubmed
Published In
Journal of Clinical Investigation
Volume
123
Issue
9
Publish Date
2013
Start Page
3925
End Page
3940
DOI
10.1172/JCI65745

Improved time to progression for transarterial chemoembolization compared with transarterial embolization for patients with unresectable hepatocellular carcinoma.

BACKGROUND: Embolizing branches of the hepatic artery lengthens survival for patients with unresectable hepatocellular carcinoma (HCC), but the benefit of combining chemotherapy with the embolizing particles remains controversial. METHODS: A retrospective review was undertaken of sequential patients with advanced HCC undergoing embolization in the past 10 years at 2 neighboring institutions and with 2 years of follow-up data. TACE was generally performed with doxorubicin plus mitomycin C. RESULTS: One hundred twenty-four patients were included; 77 received TACE and 47 received TAE. On multivariable analysis stratified by institution, type of embolization and CLIP score significantly predicted PFS and time to progression (TTP), whereas CLIP score and AFP independently predicted overall survival (OS). TACE significantly prolonged PFS and TTP (P = .0004 and P = .001, respectively), but not OS (P = .83). CONCLUSIONS: The addition of chemotherapy to TAE prolongs PFS and TTP. Future efforts should focus on adjunctive therapies after the embolization to increase survival.

Authors
Morse, MA; Hanks, BA; Suhocki, P; Doan, PL; Liu, EA; Frost, P; Bernard, SA; Tsai, A; Moore, DT; O'Neil, BH
MLA Citation
Morse, MA, Hanks, BA, Suhocki, P, Doan, PL, Liu, EA, Frost, P, Bernard, SA, Tsai, A, Moore, DT, and O'Neil, BH. "Improved time to progression for transarterial chemoembolization compared with transarterial embolization for patients with unresectable hepatocellular carcinoma." Clin Colorectal Cancer 11.3 (September 2012): 185-190.
PMID
22280845
Source
pubmed
Published In
Clinical colorectal cancer
Volume
11
Issue
3
Publish Date
2012
Start Page
185
End Page
190
DOI
10.1016/j.clcc.2011.11.003

Effect of the loss of the type III TGF beta receptor during tumor progression on tumor microenvironment: Preclinical development of TGF beta inhibition and TGF beta-related biomarkers to enhance immunotherapy efficacy.

Authors
Hanks, BA; Holtzhausen, A; Gimpel, P; Jamieson, R; Campbell, OM; Sun, L; Augustine, CK; Tyler, DS; Osada, T; Morse, M; Ling, LE; Lyerly, HK; Blobe, GC
MLA Citation
Hanks, BA, Holtzhausen, A, Gimpel, P, Jamieson, R, Campbell, OM, Sun, L, Augustine, CK, Tyler, DS, Osada, T, Morse, M, Ling, LE, Lyerly, HK, and Blobe, GC. "Effect of the loss of the type III TGF beta receptor during tumor progression on tumor microenvironment: Preclinical development of TGF beta inhibition and TGF beta-related biomarkers to enhance immunotherapy efficacy." May 20, 2012.
Source
wos-lite
Published In
Journal of Clinical Oncology
Volume
30
Issue
15
Publish Date
2012

Role of the type III TGF-β receptor in modulating antitumor immunity during breast cancer progression.

10540 Background: We have shown that breast cancers downregulate the expression of the type III TGF-β receptor (TβRIII) tumor suppressor during tumor progression. Previous work has shown TβRIII to undergo ectodomain shedding, enabling the sequestration of the soluble TGF-β ligand and the inhibition of the TGF-β signaling pathway.  The TGF-β cytokine inhibits dendritic cell (DC)-dependent antigen presentation.  We hypothesize that the downregulation of TβRIII during breast tumor development permits enhanced TGF-β signaling within the local DCs of the tumor microenvironment allowing the tumor to evade the host immune response.Our data suggest that the tumor suppressor properties of TβRIII in the 4T1 murine metastatic breast cancer model are diminished in immunosuppressed hosts. Indeed, loss of TβRIII allows for the progression of more immunogenic Her2/neu-expressing 4T1 tumors and suppresses Her2/neu-specific T cell responses. Flow cytometry and rt-PCR studies indicate that breast tumors which lack TβRIII expression exhibit reduced numbers of infiltrating CD8+ T cells and increased regulatoy T cells (Tregs), findings which are supported by human microarray data. In addition, DCs within TβRIIIlo tumors and their draining lymph nodes (TDLNs) express enhanced levels of the indoleamine 2,3-dioxygenase (IDO) enzyme as well as the CCL22 chemokine, which correlates with expanded local Treg populations. Studies have shown 4T1-RIII conditioned media to inhibit TGF-β signaling within DCs and to suppress the TGF-β-mediated inhibition of DC maturation. Our work is showing that DCs within the TDLNs of TβRIIIhi tumors exhibit a more mature phenotype.The increased TGF-β signaling capacity of DCs residing in TβRIIIlo tumors may allow for increased local CCL22 expression; thereby promoting Treg recruitment and allowing for CTLA-4-mediated IDO upregulation by local DCs.This pathway represents a novel mechanism for evading the host anti-tumor immune response, supports the targeting of TGF-β as a strategy to enhance the efficacy of immunotherapeutic approaches for solid tumors, and suggests that serum levels of soluble TβRIII may represent a useful biomarker for immunotherapy.

Authors
Hanks, BA; Campbell, OM; Lee, JD; Morse, M; Clay, TM; Lyerly, HK; Blobe, GC
MLA Citation
Hanks, BA, Campbell, OM, Lee, JD, Morse, M, Clay, TM, Lyerly, HK, and Blobe, GC. "Role of the type III TGF-β receptor in modulating antitumor immunity during breast cancer progression." Journal of clinical oncology : official journal of the American Society of Clinical Oncology 29.15_suppl (May 2011): 10540-.
PMID
28021827
Source
epmc
Published In
Journal of Clinical Oncology
Volume
29
Issue
15_suppl
Publish Date
2011
Start Page
10540

Pharmacological inhibition of TGFβ as a strategy to augment the antitumor immune response.

There is considerable evidence suggesting that a variety of malignancies utilize the TGFβ cytokine to evade immune surveillance mechanisms to facilitate tumor growth and metastatic progression. The recently developed large- and small-molecule TGFβ inhibitors have demonstrated antitumor efficacy in several preclinical tumor models. Further investigation has revealed these agents to be critically dependent upon the host's immune system, suggesting that the inhibition of TGFβ may overcome the immunosuppressive tumor microenvironment and, ultimately, augment the antitumor immune response. These findings strongly support combining this strategy with other immunotherapeutic approaches for the treatment of metastatic cancer. This review discusses the immunoregulatory and antitumor properties of these pharmacological inhibitors of TGFβ signaling as either independent agents or in combination with various immunotherapeutic strategies, their potential side effects, as well as additional avenues of research that may be necessary for their eventual clinical application.

Authors
Hanks, BA; Morse, MA
MLA Citation
Hanks, BA, and Morse, MA. "Pharmacological inhibition of TGFβ as a strategy to augment the antitumor immune response." Curr Opin Investig Drugs 11.12 (December 2010): 1342-1353. (Review)
PMID
21154116
Source
pubmed
Published In
Current Opinion in Investigational Drugs
Volume
11
Issue
12
Publish Date
2010
Start Page
1342
End Page
1353

Role of the type III TGF-b receptor in mediating immunosuppression during breast cancer progression

Authors
Hanks, BA; Lee, JD; Morse, M; Clay, TM; Lyerly, HK; Blobe, AC
MLA Citation
Hanks, BA, Lee, JD, Morse, M, Clay, TM, Lyerly, HK, and Blobe, AC. "Role of the type III TGF-b receptor in mediating immunosuppression during breast cancer progression." May 20, 2010.
Source
wos-lite
Published In
Journal of Clinical Oncology
Volume
28
Issue
15
Publish Date
2010

The efficacy and tolerability of transarterial chemo-embolization (TACE) compared with transarterial embolization (TAE) for patients with unresectable hepatocellular carcinoma (HCC)

Authors
Hanks, BA; Suhocki, PV; DeLong, DM; Doan, PL; Liu, E; Tsai, AL; Burke, CT; Bernard, SA; O'Neil, BH; Morse, MA
MLA Citation
Hanks, BA, Suhocki, PV, DeLong, DM, Doan, PL, Liu, E, Tsai, AL, Burke, CT, Bernard, SA, O'Neil, BH, and Morse, MA. "The efficacy and tolerability of transarterial chemo-embolization (TACE) compared with transarterial embolization (TAE) for patients with unresectable hepatocellular carcinoma (HCC)." JOURNAL OF CLINICAL ONCOLOGY 26.15 (May 20, 2008).
Source
wos-lite
Published In
Journal of Clinical Oncology
Volume
26
Issue
15
Publish Date
2008

The efficacy and tolerability of transarterial chemo-embolization (TACE) compared with transarterial embolization (TAE) for patients with unresectable hepatocellular carcinoma (HCC).

4595 Background: Radiologic procedures that involve embolizing branches of the hepatic artery lengthen survival for patients with unresectable HCC, but the benefit of administering intrahepatic arterial chemotherapy during the embolization procedure is uncertain.A retrospective review of all patients with HCC and undergoing an embolization in the last 10 years and with 2 years of follow-up data was undertaken at two institutions to compare the survival and TTP of patients receiving TACE compared with TAE. Prognostic factors analyzed included age, gender, race, largest tumor size, number of tumors, macrovascular invasion, AFP, Child's class, and CLIP score.122 pts with the following characteristics were included in the analysis: 71% Caucasian, 22% African American, 52% HCV+, 13% HBV+, 66% with cirrhosis, Child's A/B/C 70/25/5%, CLIP Score 0,1/2/>3 52/26/21%, 72% had either 1 or 2 tumors, 14% macrovascular invasion, 16% portal vein branch thrombosis, 20% extrahepatic disease. The procedure (TACE (51%, the majority receiving doxorubicin plus mitomycin C) and TAE (49%).was completed in one session in 87%. A subsequent embolization was performed in 37% for progression or new disease. The embolizing material was ethiodised oil in 39%, polyacrylamide/gelatin in 43%, and polyvinyl alcohol in 41%. The procedures were equally well tolerated. Although 37% of patients experienced an adverse event, readmission to the hospital was required in only 10%. One patient experienced fulminant hepatic failure. The median survival for TAE was 19.6 mo (95% CI 10.6-28.4) compared with 12.9mo (95% CI 7.0-19.5) for TACE (p=0.066). The TTP for TAE was 3.2mo (95% CI 2.1-6.0) compared with 4.1 mo (95% CI 2.1-5.6 for TACE (p=0.79). On multivariate analysis, the trend towards a difference in survival for TAE was eliminated and only CLIP score, AFP, and largest tumor remained as prognostic factors.This retropsective review suggests that adding chemotherapy to embolization does not improve survival compared with bland embolization in pts with HCC. Future efforts should focus on adjunctive therapies following the embolization to prevent progression or new primary tumors. No significant financial relationships to disclose.

Authors
Hanks, BA; Suhocki, PV; DeLong, DM; Doan, PL; Liu, E; Tsai, AL; Burke, CT; Bernard, SA; O'Neil, BH; Morse, MA
MLA Citation
Hanks, BA, Suhocki, PV, DeLong, DM, Doan, PL, Liu, E, Tsai, AL, Burke, CT, Bernard, SA, O'Neil, BH, and Morse, MA. "The efficacy and tolerability of transarterial chemo-embolization (TACE) compared with transarterial embolization (TAE) for patients with unresectable hepatocellular carcinoma (HCC)." Journal of clinical oncology : official journal of the American Society of Clinical Oncology 26.15_suppl (May 2008): 4595-.
PMID
27948705
Source
epmc
Published In
Journal of Clinical Oncology
Volume
26
Issue
15_suppl
Publish Date
2008
Start Page
4595

Enhanced activation of human dendritic cells by inducible CD40 and Toll-like receptor-4 ligation.

Despite the potency of dendritic cells (DC) as antigen-presenting cells for priming adaptive immunity, DC-based cancer vaccines have been largely insufficient to effectively reduce tumor burden or prevent tumor progression in most patients. To enhance DC-based vaccines, we used the combination of a synthetic ligand-inducible CD40 receptor (iCD40) along with Toll-like receptor-4 (TLR-4) ligation in human monocyte-derived DCs. The iCD40 receptor permits targeted, reversible activation of CD40 in vivo, potentially bypassing the essential role of CD4(+) T cells for activation of DCs. As a rigorous preclinical study of this approach, we evaluated key parameters of DC activation and function. Whereas neither iCD40 nor TLR-4 signaling alone led to high levels of interleukin (IL)-12p70 and IL-6, using iCD40 in combination with lipopolysaccharide (LPS) or monophosphoryl lipid A led to strongly synergistic production of both. Furthermore, this approach led to high expression of DC maturation markers, epitope-specific CTL and T helper 1 responses, as well as DC migration in vitro and in vivo. Moreover, use of iCD40-modified and LPS-stimulated DCs led to targeted expansion of autologous T cells against tumor-associated antigens, including prostate-specific membrane antigen, and elimination of preestablished tumors, supporting this technology as a potent strategy for DC-based cancer immunotherapy.

Authors
Lapteva, N; Seethammagari, MR; Hanks, BA; Jiang, J; Levitt, JM; Slawin, KM; Spencer, DM
MLA Citation
Lapteva, N, Seethammagari, MR, Hanks, BA, Jiang, J, Levitt, JM, Slawin, KM, and Spencer, DM. "Enhanced activation of human dendritic cells by inducible CD40 and Toll-like receptor-4 ligation." Cancer Res 67.21 (November 1, 2007): 10528-10537.
PMID
17974997
Source
pubmed
Published In
Cancer Research
Volume
67
Issue
21
Publish Date
2007
Start Page
10528
End Page
10537
DOI
10.1158/0008-5472.CAN-07-0833

Re-engineered CD40 receptor enables potent pharmacological activation of dendritic-cell cancer vaccines in vivo.

Modest clinical outcomes of dendritic-cell (DC) vaccine trials call for the refinement of DC vaccine design. Although many potential antigens have been identified, development of methods to enhance antigen presentation by DCs has lagged. We have engineered a potent, drug-inducible CD40 (iCD40) receptor that permits temporally controlled, lymphoid-localized, DC-specific activation. iCD40 is comprised of a membrane-localized cytoplasmic domain of CD40 fused to drug-binding domains. This allows it to respond to a lipid-permeable, high-affinity dimerizer drug while circumventing ectodomain-dependent negative-feedback mechanisms. These modifications permit prolonged activation of iCD40-expressing DCs in vivo, resulting in more potent CD8(+) T-cell effector responses, including the eradication of previously established solid tumors, relative to activation of DCs ex vivo (P < 0.01), typical of most clinical DC protocols. In addition, iCD40-mediated DC activation exceeded that achieved by stimulating the full-length, endogenous CD40 receptor both in vitro and in vivo. Because iCD40 is insulated from the extracellular environment and can be activated within the context of an immunological synapse, iCD40-expressing DCs have a prolonged lifespan and should lead to more potent vaccines, perhaps even in immune-compromised patients.

Authors
Hanks, BA; Jiang, J; Singh, RAK; Song, W; Barry, M; Huls, MH; Slawin, KM; Spencer, DM
MLA Citation
Hanks, BA, Jiang, J, Singh, RAK, Song, W, Barry, M, Huls, MH, Slawin, KM, and Spencer, DM. "Re-engineered CD40 receptor enables potent pharmacological activation of dendritic-cell cancer vaccines in vivo." Nat Med 11.2 (February 2005): 130-137.
PMID
15665830
Source
pubmed
Published In
Nature Medicine
Volume
11
Issue
2
Publish Date
2005
Start Page
130
End Page
137
DOI
10.1038/nm1183

Template-based docking of a prolactin receptor proline-rich motif octapeptide to FKBP12: implications for cytokine receptor signaling.

A conserved proline-rich motif (PRM) in the cytoplasmic domain of cytokine receptors has been suggested to be a signaling switch regulated by the action of the FK506 binding protein (FKBP) family of peptidylprolyl isomerases (O'Neal KD, Yu-Lee LY, Shearer WT, 1995, Ann NY Acad Sci 766:282-284). We have docked the prolactin receptor PRM (Ile1-Phe2-Pro3-Pro4-Val5-Pro6-Gly7-Pro8) to the ligand binding site of FKBP12. The procedure involved conformational search restricted by NMR restraints (O'Neal KD et al., 1996, Biochem J 315:833-844), energy minimization of the octapeptide conformers so obtained, template-based docking of a selected conformer to FKBP12, and energy refinement of the resulting complex. The template used was the crystal structure of a cyclic FK506-peptide hybrid bound to FKBP12. Val5-Pro6 of the PRM was taken to be the biologically relevant Xaa-Pro bond. The docked conformer is stabilized by two intramolecular hydrogen bonds, N7H7-->O4 and N2H2-->O8, and two intermolecular ones, Ile56; N-H-->O = C:Pro6 and Tyr82:O-H-->O = C:Gly7. This conformer features a Type I beta-turn and has extensive hydrophobic contacts with the FKBP12 binding surface. The observed interactions support the hypothesis that FKBP12 catalyzes cis-trans isomerization in the PRM when it is part of the longer cytoplasmic domain of a cytokine receptor, and suggest a significant role for the PRM in signal transduction.

Authors
Soman, KV; Hanks, BA; Tien, H; Chari, MV; O'Neal, KD; Morrisett, JD
MLA Citation
Soman, KV, Hanks, BA, Tien, H, Chari, MV, O'Neal, KD, and Morrisett, JD. "Template-based docking of a prolactin receptor proline-rich motif octapeptide to FKBP12: implications for cytokine receptor signaling." Protein science : a publication of the Protein Society 6.5 (May 1997): 999-1008.
PMID
9144770
Source
epmc
Published In
Protein Science
Volume
6
Issue
5
Publish Date
1997
Start Page
999
End Page
1008
DOI
10.1002/pro.5560060505

Role of conserved residues within the carboxy phosphate domain of carbamoyl phosphate synthetase.

Carbamoyl phosphate synthetase (CPS) catalyzes the formation of carbamoyl phosphate from glutamine, bicarbonate, and 2 mol of MgATP. The heterodimeric protein is composed of a small amidotransferase subunit and a larger synthetase subunit. The synthetase subunit contains a large tandem repeat for each of the nucleotides used in the overall synthesis of carbamoyl phosphate. A working model for the three-dimensional fold of the carboxy phosphate domain of CPS was constructed on the basis of amino acid sequence alignments and the X-ray crystal structure coordinates for biotin carboxylase and D-alanine:D-alanine ligase. This model was used to select ten residues within the carboxy phosphate domain of CPS for modification and subsequent characterization of the kinetic constants for the mutant proteins. Residues R82, R129, R169, D207, E215, N283, and Q285 were changed to alanine residues; residues E299 and R303 to glutamine; and residue N301 to aspartate. No significant changes in the catalytic constants were observed upon mutation of either R82 or D207, and thus these residues appear to be nonessential for binding and/or catalytic activity. The Michaelis constant for ATP was most affected by modification of residues R129, R169, Q285, and N301. The binding of bicarbonate was most affected by the mutagenesis of residues E215, E299, N301, and R303. The mutation of residues E215, N283, E299, N301, and R303 resulted in proteins which were unable to synthesize carbamoyl phosphate at a significant rate. All of the mutations, with the exception of the N301D mutant, primarily affected the enzyme by altering the step for the phosphorylation of bicarbonate. However, mutation of N301 to aspartic acid also disrupted the catalytic step involved in the phosphorylation of carbamate. These results are consistent with a role for the N-terminal half of the synthetase subunit of CPS that is primarily directed at the initial phosphorylation of bicarbonate by the first ATP utilized in the overall synthesis of carbamoyl phosphate. The active site structure appears to be very similar to the ones previously determined for D-alanine:D-alanine ligase and biotin carboxylase.

Authors
Stapleton, MA; Javid-Majd, F; Harmon, MF; Hanks, BA; Grahmann, JL; Mullins, LS; Raushel, FM
MLA Citation
Stapleton, MA, Javid-Majd, F, Harmon, MF, Hanks, BA, Grahmann, JL, Mullins, LS, and Raushel, FM. "Role of conserved residues within the carboxy phosphate domain of carbamoyl phosphate synthetase." Biochemistry 35.45 (November 1996): 14352-14361.
PMID
8916922
Source
epmc
Published In
Biochemistry
Volume
35
Issue
45
Publish Date
1996
Start Page
14352
End Page
14361
DOI
10.1021/bi961183y

Comparison of the functional differences for the homologous residues within the carboxy phosphate and carbamate domains of carbamoyl phosphate synthetase.

Carbamoyl phosphate synthetase (CPS) from Escherichia coli catalyzes the formation of carbamoyl phosphate from two molecules of MgATP, bicarbonate, and glutamine. It has been previously shown that the amino- and carboxy-terminal halves of the large subunit of this protein are homologous. A working model for the active site structure of the carboxy-terminal domain of the large subunit of CPS was constructed based upon amino acid sequence alignments and the previously determined three-dimensional structures of two mechanistically related proteins, biotin carboxylase and D-alanine:D-alanine ligase. The model was tested by mutation of ten amino acid residues predicted to be important for binding and/or catalysis. The mutated residues were as follows: R571, R675, R715, D753, E761, N827, Q829, E841, N843, and R845. The mutant proteins were expressed, purified to homogeneity and the catalytic properties determined for a variety of assay formats. The mutants E761A, E841Q, N843D, and R845Q were diminished in their ability to synthesize carbamoyl phosphate. The R715A, Q829A, and R675A mutants displayed elevated Michaelis constants for MgADP in the partial back reaction. The mutants E761A, N827A, E841Q, N843D, and R845Q showed significant increases in the Michaelis constants for either bicarbonate or carbamoyl phosphate. No significant alterations were noted upon mutation of either R571 or D753 to an alanine residue and thus these amino acids do not appear essential for structure or catalytic activity. These results have been utilized to further support the proposal that the C-terminal half of the large subunit of CPS is primarily responsible for the phosphorylation of the carbamate intermediate during the final formation of carbamoyl phosphate. The measured effects on the catalyic activities displayed by these mutations were found to be comparable to the previously determined effects after mutation of the homologous residues located on the N-terminal half of CPS and also for those residues mutated within D-alanine:D-alanine ligase [Shi, Y., & Walsh, C.T. (1995) Biochemistry 34, 2768-2776].

Authors
Javid-Majd, F; Stapleton, MA; Harmon, MF; Hanks, BA; Mullins, LS; Raushel, FM
MLA Citation
Javid-Majd, F, Stapleton, MA, Harmon, MF, Hanks, BA, Mullins, LS, and Raushel, FM. "Comparison of the functional differences for the homologous residues within the carboxy phosphate and carbamate domains of carbamoyl phosphate synthetase." Biochemistry 35.45 (November 1996): 14362-14369.
PMID
8916923
Source
epmc
Published In
Biochemistry
Volume
35
Issue
45
Publish Date
1996
Start Page
14362
End Page
14369
DOI
10.1021/bi961184q

Tumor-mediated Metabolic Re-Programing of Dendritic Cells as a Fundamental Mechanism of Immune Tolerance and Immunotherapy Resistance.

Authors
Zhao, F; Evans, KS; Xiao, C; Hanks, BA
MLA Citation
Zhao, F, Evans, KS, Xiao, C, and Hanks, BA. "Tumor-mediated Metabolic Re-Programing of Dendritic Cells as a Fundamental Mechanism of Immune Tolerance and Immunotherapy Resistance." 2016 Keystone Symposium: Immunometabolism. February 25, 2016 - February 28, 2016. Banff, Alberta, Canada.
Source
manual

The Influence of the Tumor Microenvironment on Checkpoint Inhibitor Efficacy: Lessons Learned from Targeting the TGF-β Signaling Pathway.

Authors
Hanks, BA
MLA Citation
Hanks, BA. "The Influence of the Tumor Microenvironment on Checkpoint Inhibitor Efficacy: Lessons Learned from Targeting the TGF-β Signaling Pathway." 2016 Melanoma Research Alliance Annual Meeting. February 25, 2016 - February 26, 2016. Washington, DC.
Source
manual

Targeting the TGF-β Signaling Pathway to Augment the Efficacy of Immunotherapy Checkpoint Inhibitors in Melanoma

Authors
Hanks, BA
MLA Citation
Hanks, BA. "Targeting the TGF-β Signaling Pathway to Augment the Efficacy of Immunotherapy Checkpoint Inhibitors in Melanoma." 2nd Annual Summit on Melanoma. September 18, 2015 - September 20, 2015. Pasadena, California.
Source
manual

A Phase II Randomized Study of CDX-1401, a Dendritic Cell Targeting NY-ESO-1 Vaccine, in Patients with Malignant Melanoma Pre-Treated with Recombinant CDX-301, a Recombinant Human Flt3 Ligand

Authors
Bhardwaj, N; Ernstoff, M; Curti, B; Hanks, BA; Albertini, M; Luke, J; Yellin, M; Keler, T; Davis, T; Vitale, L; Crocker, A; Friedlander, P; Morishima, C; Cheever, M; Fling, S
MLA Citation
Bhardwaj, N, Ernstoff, M, Curti, B, Hanks, BA, Albertini, M, Luke, J, Yellin, M, Keler, T, Davis, T, Vitale, L, Crocker, A, Friedlander, P, Morishima, C, Cheever, M, and Fling, S. "A Phase II Randomized Study of CDX-1401, a Dendritic Cell Targeting NY-ESO-1 Vaccine, in Patients with Malignant Melanoma Pre-Treated with Recombinant CDX-301, a Recombinant Human Flt3 Ligand." 2016 American Society of Clinical Oncology Annual Meeting. June 3, 2016 - June 7, 2016. Chicago, IL.
Source
manual

The Wnt5a-β-catenin Pathway Triggers a Metabolic Switch That Drives Indoleamine 2,3-dioxygenase Activity and Dendritic Cell Tolerization in the Melanoma Microenvironment.

Authors
Zhao, F; Evans, K; Xiao, C; Holtzhausen, A; Hanks, BA
MLA Citation
Zhao, F, Evans, K, Xiao, C, Holtzhausen, A, and Hanks, BA. "The Wnt5a-β-catenin Pathway Triggers a Metabolic Switch That Drives Indoleamine 2,3-dioxygenase Activity and Dendritic Cell Tolerization in the Melanoma Microenvironment." Society for Immunotherapy of Cancer 2016 Annual Meeting. November 9, 2016 - November 13, 2016. National Harbor, Washington, DC. BioMed Central.
Website
http://hdl.handle.net/10161/14012
Source
manual
Published In
Journal for ImmunoTherapy of Cancer
Volume
82
Issue
4(suppl 1)
Start Page
8
End Page
9

Utilizing Pre-Clinical Melanoma Models to Design Rational Combinatorial Immunotherapy Regimens : Lessons Learned from Targeting the TGF-beta Signaling Pathway.

Authors
Hanks, BA
MLA Citation
Hanks, BA. "Utilizing Pre-Clinical Melanoma Models to Design Rational Combinatorial Immunotherapy Regimens : Lessons Learned from Targeting the TGF-beta Signaling Pathway." 2017 Melanoma Research Alliance Annual Meeting. February 13, 2017 - February 15, 2017. Washington, DC.
Source
manual
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Research Areas:

  • Biomarkers, Pharmacological
  • Cell Line, Tumor
  • Chemokine CCL22
  • Combined Modality Therapy
  • Dendritic Cells
  • Disease-Free Survival
  • Down-Regulation
  • Female
  • Humans
  • Indoleamine-Pyrrole 2,3,-Dioxygenase
  • Lymphocyte Activation
  • Mammary Neoplasms, Experimental
  • Melanoma
  • Melanoma, Experimental
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Molecular Targeted Therapy
  • Neoplasm Staging
  • Neoplasm Transplantation
  • Neoplasms
  • Prognosis
  • Proteoglycans
  • Receptors, Transforming Growth Factor beta
  • Signal Transduction
  • Transforming Growth Factor beta
  • Tumor Escape
  • Tumor Microenvironment