The Experts below are selected from a list of 3534 Experts worldwide ranked by ideXlab platform
Claudia Palena - One of the best experts on this subject based on the ideXlab platform.
-
Phase I Trial of a Modified Vaccinia Ankara Priming Vaccine Followed by a Fowlpox Virus Boosting Vaccine Modified to Express Brachyury and Costimulatory Molecules in Advanced Solid Tumors
The oncologist, 2019Co-Authors: Julie Collins, Renee N. Donahue, Yo-ting Tsai, Michell Manu, Claudia Palena, Margaret E. Gatti-mays, Jennifer L. Marte, Ravi A. Madan, Fatima Karzai, Christopher R. HeeryAbstract:LESSONS LEARNED Modified vaccinia Ankara-Bavarian Nordic (MVA-BN)-Brachyury followed by fowlpox virus-BN-Brachyury was well tolerated upon administration to patients with advanced cancer. Sixty-three percent of patients developed CD4+ and/or CD8+ T-cell responses to brachyury after vaccination. BN-Brachyury vaccine also induced T-cell responses against CEA and MUC1, which are cascade antigens, that is, antigens not encoded in the vaccines. BACKGROUND Brachyury, a transcription factor, plays an integral role in the epithelial-mesenchymal transition, metastasis, and tumor resistance to chemotherapy. It is expressed in many tumor types, and rarely in normal tissues, making it an ideal immunologic target. Bavarian Nordic (BN)-Brachyury consists of vaccination with modified vaccinia Ankara (MVA) priming followed by fowlpox virus (FPV) boosting, each encoding transgenes for brachyury and costimulatory molecules. METHODS Patients with metastatic solid tumors were treated with two monthly doses of MVA-brachyury s.c., 8 × 108 infectious units (IU), followed by FPV-brachyury s.c., 1 × 109 IU, for six monthly doses and then every 3 months for up to 2 years. The primary objective was to determine safety and tolerability. RESULTS Eleven patients were enrolled from March 2018 to July 2018 (one patient was nonevaluable). No dose-limiting toxicities were observed. The most common treatment-related adverse event was grade 1/2 injection-site reaction observed in all patients. Best overall response was stable disease in six patients, and the 6-month progression-free survival rate was 50%. T cells against brachyury and cascade antigens CEA and MUC1 were detected in the majority of patients. CONCLUSION BN-Brachyury vaccine is well tolerated and induces immune responses to brachyury and cascade antigens and demonstrates some evidence of clinical benefit.
-
Loss of the Cyclin-Dependent Kinase Inhibitor 1 in the Context of Brachyury-Mediated Phenotypic Plasticity Drives Tumor Resistance to Immune Attack
Frontiers Media S.A., 2018Co-Authors: Duane H Hamilton, Kristen K. Mccampbell, Claudia PalenaAbstract:The acquisition of mesenchymal features by carcinoma cells is now recognized as a driver of metastasis and tumor resistance to a range of anticancer therapeutics, including chemotherapy, radiation, and certain small-molecule targeted therapies. With the recent successful implementation of immunotherapies for the treatment of various types of cancer, there is growing interest in understanding whether an immunological approach could be effective at eradicating carcinoma cells bearing mesenchymal features. Recent studies, however, demonstrated that carcinoma cells that have acquired mesenchymal features may also exhibit decreased susceptibility to lysis mediated by immune effector cells, including antigen-specific CD8+ T cells, innate natural killer (NK), and lymphokine-activated killer (LAK) cells. Here, we investigated the mechanism involved in the immune resistance of carcinoma cells that express very high levels of the transcription factor brachyury, a molecule previously shown to drive the acquisition of mesenchymal features by carcinoma cells. Our results demonstrate that very high levels of brachyury expression drive the loss of the cyclin-dependent kinase inhibitor 1 (p21CIP1, p21), an event that results in decreased tumor susceptibility to immune-mediated lysis. We show here that reconstitution of p21 expression markedly increases the lysis of brachyury-high tumor cells mediated by antigen-specific CD8+ T cells, NK, and LAK cells, TNF-related apoptosis-inducing ligand, and chemotherapy. Several reports have now demonstrated a role for p21 loss in cancer as an inducer of the epithelial–mesenchymal transition. The results from the present study situate p21 as a central player in many of the aspects of the phenomenon of brachyury-mediated mesenchymalization of carcinomas, including resistance to chemotherapy and immune-mediated cytotoxicity. We also demonstrate here that the defects in tumor cell death described in association with very high levels of brachyury could be alleviated via the use of a WEE1 inhibitor. Several vaccine platforms targeting brachyury have been developed and are undergoing clinical evaluation. These studies provide further rationale for the use of WEE1 inhibition in combination with brachyury-based immunotherapeutic approaches
-
brachyury a vaccine target is overexpressed in triple negative breast cancer
Endocrine-related Cancer, 2016Co-Authors: Duane H Hamilton, Claudia Palena, Mario Roselli, Patrizia Ferroni, Leopoldo Costarelli, F Cavaliere, Mariateresa Taffuri, Fiorella GuadagniAbstract:Patients diagnosed with triple-negative breast cancer (TNBC) have a high rate of tumor metastasis and a poor prognosis. The treatment option for these patients is currently chemotherapy, which results in very low response rates. Strategies that exploit the immune system for the treatment of cancer have now shown the ability to improve survival in several tumor types. Identifying potential targets for immune therapeutic interventions is an important step in developing novel treatments for TNBC. In this study, in silico analysis of publicly available datasets and immunohistochemical analysis of primary and metastatic tumor biopsies from TNBC patients were conducted to evaluate the expression of the transcription factor brachyury, which is a driver of tumor metastasis and resistance and a target for cancer vaccine approaches. Analysis of breast cancer datasets demonstrated a predominant expression of brachyury mRNA in TNBC and in basal vs luminal or HER2 molecular breast cancer subtypes. At the protein level, variable levels of brachyury expression were detected both in primary and metastatic TNBC lesions. A strong association was observed between nuclear brachyury protein expression and the stage of disease, with nuclear brachyury being more predominant in metastatic vs primary tumors. Survival analysis also demonstrated an association between high levels of brachyury in the primary tumor and poor prognosis. Two brachyury-targeting cancer vaccines are currently undergoing clinical evaluation; the data presented here provide rationale for using brachyury-targeting immunotherapy approaches for the treatment of TNBC.
-
nuclear brachyury expression is consistent in chordoma common in germ cell tumors and small cell carcinomas and rare in other carcinomas and sarcomas an immunohistochemical study of 5229 cases
The American Journal of Surgical Pathology, 2015Co-Authors: Markku Miettinen, Christopher R. Heery, Jeffrey Schlom, Zengfeng Wang, Jerzy Lasota, Claudia PalenaAbstract:Brachyury is a transcription factor of the T-box family typically expressed in notochord and chordoma. Some studies report brachyury as highly specific for chordoma, whereas others have concluded that brachyury is expressed in many types of common carcinomas by RT-PCR and immunohistochemistry and could be involved in the epithelial-mesenchymal transition and metastatic process. In this study, we immunohistochemically evaluated 5229 different tumors for nuclear brachyury expression using a new rabbit monoclonal antibody and automated immunostaining (Leica Bond Max). Only nuclear labeling was scored, and antibody dilution of 1:2000 was used. In normal tissues, only rare cells in seminiferous tubules were labeled; all other organs were negative. All chordomas (75/76), except a sarcomatous one, were positive, whereas chondrosarcomas were negative. Among epithelial tumors, positivity was often detected in embryonal carcinoma (74%) and seminoma (45%). Pulmonary small cell carcinoma was often positive (41%), whereas pulmonary and pancreatic adenocarcinomas only rarely showed nuclear brachyury-positivity (3–4%). Common carcinomas such as ductal carcinomas of breast, or adenocarcinomas of the prostate only exceptionally showed nuclear positivity (< 1%). No colorectal, hepatocellular, renal cell, squamous cell, thyroid or urothelial carcinoma, or mesothelioma showed nuclear brachyury-positivity. Among mesenchymal and neuroectodermal tumors, only isolated cases of melanoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, and follicular lymphoma showed nuclear expression. However, as shown previously with lung carcinoma, experiments with lower antibody dilutions (1:200–1:500) showed weak cytoplasmic and nuclear labeling in breast cancers. In addition to chordoma, we show here for the first time that nuclear brachyury expression is prevalent in embryonal carcinoma, seminoma, and small cell carcinoma of the lung but very rare in common carcinomas, sarcomas, and melanoma. With these reservations, we have demonstrated the presence of nuclear brachyury immunoreactivity to be a sensitive and fairly specific marker for chordoma.
-
wee1 inhibition alleviates resistance to immune attack of tumor cells undergoing epithelial mesenchymal transition
Cancer Research, 2014Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Kam Sze Tsang, Claudia PalenaAbstract:Aberrant expression of the T-box transcription factor brachyury in human carcinomas drives the phenomenon of epithelial-mesenchymal transition (EMT), a phenotypic modulation that facilitates tumor dissemination and resistance to conventional therapies, including chemotherapy and radiation. By generating isogenic cancer cell lines with various levels of brachyury expression, we demonstrate that high levels of brachyury also significantly reduce the susceptibility of cancer cells to lysis by both antigen-specific T cells and natural killer cells. Our results indicated that resistance of brachyury-high tumor cells to immune-mediated attack was due to inefficient caspase-dependent apoptosis, manifested as inefficient nuclear lamin degradation in the presence of activated effector caspases. We correlated this phenomenon to loss of cell cycle kinase CDK1, which mediates lamin phosphorylation. In support of a causal connection, pre-treatment of tumor cells with a specific inhibitor of WEE1, a negative regulator kinase of CDK1, could counter the defective apoptosis of tumor cells expressing high levels of brachyury. Thus, our findings suggested that reconstituting CDK1 activity to threshold levels may be sufficient to restore immunosurveillance of mesenchymal-like cancer cells that have escaped previous immune detection or eradication.
Duane H Hamilton - One of the best experts on this subject based on the ideXlab platform.
-
Loss of the Cyclin-Dependent Kinase Inhibitor 1 in the Context of Brachyury-Mediated Phenotypic Plasticity Drives Tumor Resistance to Immune Attack
Frontiers Media S.A., 2018Co-Authors: Duane H Hamilton, Kristen K. Mccampbell, Claudia PalenaAbstract:The acquisition of mesenchymal features by carcinoma cells is now recognized as a driver of metastasis and tumor resistance to a range of anticancer therapeutics, including chemotherapy, radiation, and certain small-molecule targeted therapies. With the recent successful implementation of immunotherapies for the treatment of various types of cancer, there is growing interest in understanding whether an immunological approach could be effective at eradicating carcinoma cells bearing mesenchymal features. Recent studies, however, demonstrated that carcinoma cells that have acquired mesenchymal features may also exhibit decreased susceptibility to lysis mediated by immune effector cells, including antigen-specific CD8+ T cells, innate natural killer (NK), and lymphokine-activated killer (LAK) cells. Here, we investigated the mechanism involved in the immune resistance of carcinoma cells that express very high levels of the transcription factor brachyury, a molecule previously shown to drive the acquisition of mesenchymal features by carcinoma cells. Our results demonstrate that very high levels of brachyury expression drive the loss of the cyclin-dependent kinase inhibitor 1 (p21CIP1, p21), an event that results in decreased tumor susceptibility to immune-mediated lysis. We show here that reconstitution of p21 expression markedly increases the lysis of brachyury-high tumor cells mediated by antigen-specific CD8+ T cells, NK, and LAK cells, TNF-related apoptosis-inducing ligand, and chemotherapy. Several reports have now demonstrated a role for p21 loss in cancer as an inducer of the epithelial–mesenchymal transition. The results from the present study situate p21 as a central player in many of the aspects of the phenomenon of brachyury-mediated mesenchymalization of carcinomas, including resistance to chemotherapy and immune-mediated cytotoxicity. We also demonstrate here that the defects in tumor cell death described in association with very high levels of brachyury could be alleviated via the use of a WEE1 inhibitor. Several vaccine platforms targeting brachyury have been developed and are undergoing clinical evaluation. These studies provide further rationale for the use of WEE1 inhibition in combination with brachyury-based immunotherapeutic approaches
-
brachyury a vaccine target is overexpressed in triple negative breast cancer
Endocrine-related Cancer, 2016Co-Authors: Duane H Hamilton, Claudia Palena, Mario Roselli, Patrizia Ferroni, Leopoldo Costarelli, F Cavaliere, Mariateresa Taffuri, Fiorella GuadagniAbstract:Patients diagnosed with triple-negative breast cancer (TNBC) have a high rate of tumor metastasis and a poor prognosis. The treatment option for these patients is currently chemotherapy, which results in very low response rates. Strategies that exploit the immune system for the treatment of cancer have now shown the ability to improve survival in several tumor types. Identifying potential targets for immune therapeutic interventions is an important step in developing novel treatments for TNBC. In this study, in silico analysis of publicly available datasets and immunohistochemical analysis of primary and metastatic tumor biopsies from TNBC patients were conducted to evaluate the expression of the transcription factor brachyury, which is a driver of tumor metastasis and resistance and a target for cancer vaccine approaches. Analysis of breast cancer datasets demonstrated a predominant expression of brachyury mRNA in TNBC and in basal vs luminal or HER2 molecular breast cancer subtypes. At the protein level, variable levels of brachyury expression were detected both in primary and metastatic TNBC lesions. A strong association was observed between nuclear brachyury protein expression and the stage of disease, with nuclear brachyury being more predominant in metastatic vs primary tumors. Survival analysis also demonstrated an association between high levels of brachyury in the primary tumor and poor prognosis. Two brachyury-targeting cancer vaccines are currently undergoing clinical evaluation; the data presented here provide rationale for using brachyury-targeting immunotherapy approaches for the treatment of TNBC.
-
wee1 inhibition alleviates resistance to immune attack of tumor cells undergoing epithelial mesenchymal transition
Cancer Research, 2014Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Kam Sze Tsang, Claudia PalenaAbstract:Aberrant expression of the T-box transcription factor brachyury in human carcinomas drives the phenomenon of epithelial-mesenchymal transition (EMT), a phenotypic modulation that facilitates tumor dissemination and resistance to conventional therapies, including chemotherapy and radiation. By generating isogenic cancer cell lines with various levels of brachyury expression, we demonstrate that high levels of brachyury also significantly reduce the susceptibility of cancer cells to lysis by both antigen-specific T cells and natural killer cells. Our results indicated that resistance of brachyury-high tumor cells to immune-mediated attack was due to inefficient caspase-dependent apoptosis, manifested as inefficient nuclear lamin degradation in the presence of activated effector caspases. We correlated this phenomenon to loss of cell cycle kinase CDK1, which mediates lamin phosphorylation. In support of a causal connection, pre-treatment of tumor cells with a specific inhibitor of WEE1, a negative regulator kinase of CDK1, could counter the defective apoptosis of tumor cells expressing high levels of brachyury. Thus, our findings suggested that reconstituting CDK1 activity to threshold levels may be sufficient to restore immunosurveillance of mesenchymal-like cancer cells that have escaped previous immune detection or eradication.
-
immunological targeting of tumor cells undergoing an epithelial mesenchymal transition via a recombinant brachyury yeast vaccine
Oncotarget, 2013Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Jeffrey Schlom, Mary T Litzinger, Alessandra Jales, James W Hodge, Andressa Ardiani, David Apelian, Claudia PalenaAbstract:// Duane H. Hamilton 1,* , Mary T. Litzinger 1,* , Alessandra Jales 1 , Bruce Huang 1 , Romaine I. Fernando 1 , James W. Hodge 1 , Andressa Ardiani 1 , David Apelian 2 , Jeffrey Schlom 1,* , and Claudia Palena 1,* 1 Laboratory of Tumor Immunology and Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 2 GlobeImmune Inc., Louisville, Colorado * These authors contributed equally to the work Correspondence: Claudia Palena, email: // Keywords : epithelial-mesenchymal transition, brachyury, T-box transcription factor, cancer vaccine, T-cell immunotherapy Received : August 16, 2013 Accepted : September 24, 2013 Published : September 26, 2013 Abstract The embryonic T-box transcription factor brachyury is aberrantly expressed in a range of human tumors. Previous studies have demonstrated that brachyury is a driver of the epithelial-mesenchymal transition (EMT), a process associated with cancer progression. Brachyury expression in human tumor cells enhances tumor invasiveness in vitro and metastasis in vivo , and induces resistance to various conventional therapeutics including chemotherapy and radiation. These characteristics, and the selective expression of brachyury for a range of human tumor types vs. normal adult tissues, make brachyury an attractive tumor target. Due to its intracellular localization and the “undruggable” character of transcription factors, available options to target brachyury are currently limited. Here we report on the development and characterization of an immunological platform for the efficient targeting of brachyury-positive tumors consisting of a heat-killed, recombinant Saccharomyces cerevisiae (yeast)–brachyury vector-based vaccine (designated as GI-6301) that expresses the full-length human brachyury protein. We demonstrate that human dendritic cells treated with recombinant yeast-brachyury can activate and expand brachyury-specific CD4 + and CD8 + T cells in vitro that, in turn, can effectively lyse human tumor cells expressing the brachyury protein. Vaccination of mice with recombinant yeast-brachyury is also shown here to elicit brachyury-specific CD4 + and CD8 + T-cell responses, and to induce anti-tumor immunity in the absence of toxicity. Based on these results, a Phase I clinical trial of GI-6301 is currently ongoing in patients with advanced tumors; to our knowledge, this is the first vaccine platform aimed at targeting a driver of tumor EMT that has successfully reached the clinical stage.
-
the embryonic transcription factor brachyury blocks cell cycle progression and mediates tumor resistance to conventional antitumor therapies
Cell Death and Disease, 2013Co-Authors: Bruce Huang, Duane H Hamilton, Romaine I Fernando, Mary T Litzinger, James W Hodge, J R Cohen, Claudia PalenaAbstract:The T-box transcription factor Brachyury, a molecule frequently detected in human cancers but seldom found in normal adult tissue, has recently been characterized as a driver of the epithelial-to-mesenchymal switch of human carcinomas. In the current investigation, we present data demonstrating that in two different human lung carcinoma models expression of Brachyury strongly correlates with increased in vitro resistance to cytotoxic therapies, such as chemotherapy and radiation. We also demonstrate that chemotherapy treatment in vitro selects for tumor cells with high levels of Brachyury and that the degree of resistance to therapy correlates with the level of Brachyury expression. In vitro and in vivo, human lung carcinoma cells with higher levels of Brachyury divide at slower rates than those with lower levels of Brachyury, a phenomenon associated with marked downregulation of cyclin D1, phosphorylated Rb and CDKN1A (p21). Chromatin immunoprecipitation and luciferase reporter assays revealed that Brachyury binds to a half T-box consensus site located within the promoter region of the p21 gene, indicating a potential mechanism for the observed therapeutic resistance associated with Brachyury expression. Finally, we demonstrate that in vivo treatment of tumor xenografts with chemotherapy results in the selective growth of resistant tumors characterized by high levels of Brachyury expression. Altogether, these results suggest that Brachyury expression may attenuate cell cycle progression, enabling tumor cells to become less susceptible to chemotherapy and radiation in human carcinomas.
Romaine I Fernando - One of the best experts on this subject based on the ideXlab platform.
-
wee1 inhibition alleviates resistance to immune attack of tumor cells undergoing epithelial mesenchymal transition
Cancer Research, 2014Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Kam Sze Tsang, Claudia PalenaAbstract:Aberrant expression of the T-box transcription factor brachyury in human carcinomas drives the phenomenon of epithelial-mesenchymal transition (EMT), a phenotypic modulation that facilitates tumor dissemination and resistance to conventional therapies, including chemotherapy and radiation. By generating isogenic cancer cell lines with various levels of brachyury expression, we demonstrate that high levels of brachyury also significantly reduce the susceptibility of cancer cells to lysis by both antigen-specific T cells and natural killer cells. Our results indicated that resistance of brachyury-high tumor cells to immune-mediated attack was due to inefficient caspase-dependent apoptosis, manifested as inefficient nuclear lamin degradation in the presence of activated effector caspases. We correlated this phenomenon to loss of cell cycle kinase CDK1, which mediates lamin phosphorylation. In support of a causal connection, pre-treatment of tumor cells with a specific inhibitor of WEE1, a negative regulator kinase of CDK1, could counter the defective apoptosis of tumor cells expressing high levels of brachyury. Thus, our findings suggested that reconstituting CDK1 activity to threshold levels may be sufficient to restore immunosurveillance of mesenchymal-like cancer cells that have escaped previous immune detection or eradication.
-
immunological targeting of tumor cells undergoing an epithelial mesenchymal transition via a recombinant brachyury yeast vaccine
Oncotarget, 2013Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Jeffrey Schlom, Mary T Litzinger, Alessandra Jales, James W Hodge, Andressa Ardiani, David Apelian, Claudia PalenaAbstract:// Duane H. Hamilton 1,* , Mary T. Litzinger 1,* , Alessandra Jales 1 , Bruce Huang 1 , Romaine I. Fernando 1 , James W. Hodge 1 , Andressa Ardiani 1 , David Apelian 2 , Jeffrey Schlom 1,* , and Claudia Palena 1,* 1 Laboratory of Tumor Immunology and Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 2 GlobeImmune Inc., Louisville, Colorado * These authors contributed equally to the work Correspondence: Claudia Palena, email: // Keywords : epithelial-mesenchymal transition, brachyury, T-box transcription factor, cancer vaccine, T-cell immunotherapy Received : August 16, 2013 Accepted : September 24, 2013 Published : September 26, 2013 Abstract The embryonic T-box transcription factor brachyury is aberrantly expressed in a range of human tumors. Previous studies have demonstrated that brachyury is a driver of the epithelial-mesenchymal transition (EMT), a process associated with cancer progression. Brachyury expression in human tumor cells enhances tumor invasiveness in vitro and metastasis in vivo , and induces resistance to various conventional therapeutics including chemotherapy and radiation. These characteristics, and the selective expression of brachyury for a range of human tumor types vs. normal adult tissues, make brachyury an attractive tumor target. Due to its intracellular localization and the “undruggable” character of transcription factors, available options to target brachyury are currently limited. Here we report on the development and characterization of an immunological platform for the efficient targeting of brachyury-positive tumors consisting of a heat-killed, recombinant Saccharomyces cerevisiae (yeast)–brachyury vector-based vaccine (designated as GI-6301) that expresses the full-length human brachyury protein. We demonstrate that human dendritic cells treated with recombinant yeast-brachyury can activate and expand brachyury-specific CD4 + and CD8 + T cells in vitro that, in turn, can effectively lyse human tumor cells expressing the brachyury protein. Vaccination of mice with recombinant yeast-brachyury is also shown here to elicit brachyury-specific CD4 + and CD8 + T-cell responses, and to induce anti-tumor immunity in the absence of toxicity. Based on these results, a Phase I clinical trial of GI-6301 is currently ongoing in patients with advanced tumors; to our knowledge, this is the first vaccine platform aimed at targeting a driver of tumor EMT that has successfully reached the clinical stage.
-
the embryonic transcription factor brachyury blocks cell cycle progression and mediates tumor resistance to conventional antitumor therapies
Cell Death and Disease, 2013Co-Authors: Bruce Huang, Duane H Hamilton, Romaine I Fernando, Mary T Litzinger, James W Hodge, J R Cohen, Claudia PalenaAbstract:The T-box transcription factor Brachyury, a molecule frequently detected in human cancers but seldom found in normal adult tissue, has recently been characterized as a driver of the epithelial-to-mesenchymal switch of human carcinomas. In the current investigation, we present data demonstrating that in two different human lung carcinoma models expression of Brachyury strongly correlates with increased in vitro resistance to cytotoxic therapies, such as chemotherapy and radiation. We also demonstrate that chemotherapy treatment in vitro selects for tumor cells with high levels of Brachyury and that the degree of resistance to therapy correlates with the level of Brachyury expression. In vitro and in vivo, human lung carcinoma cells with higher levels of Brachyury divide at slower rates than those with lower levels of Brachyury, a phenomenon associated with marked downregulation of cyclin D1, phosphorylated Rb and CDKN1A (p21). Chromatin immunoprecipitation and luciferase reporter assays revealed that Brachyury binds to a half T-box consensus site located within the promoter region of the p21 gene, indicating a potential mechanism for the observed therapeutic resistance associated with Brachyury expression. Finally, we demonstrate that in vivo treatment of tumor xenografts with chemotherapy results in the selective growth of resistant tumors characterized by high levels of Brachyury expression. Altogether, these results suggest that Brachyury expression may attenuate cell cycle progression, enabling tumor cells to become less susceptible to chemotherapy and radiation in human carcinomas.
-
abstract a64 brachyury mediated epithelial mesenchymal transition of human carcinoma cells is associated with an increased resistance to immune mediated attack
Cancer Research, 2011Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Kwongyok Tsang, Jeffrey Schlom, Claudia PalenaAbstract:The epithelial-mesenchymal transition (EMT) has recently been recognized as a relevant process during the progression of carcinomas. Epithelial tumor cells undergoing EMT have been shown to acquire a mesenchymal-like phenotype, cell motility, and invasiveness, in vivo metastatic propensity and stem cell-like properties, including the ability to resist cell death initiated by traditional cancer treatment modalities such as chemotherapy and radiation. The development of therapeutic interventions aimed at interfering with EMT is therefore emerging as a rational approach for the prevention of cancer metastasis and alleviation of therapeutic resistance. Promising results recently obtained in preclinical and clinical studies with cancer vaccines may lead to their use for the treatment of various types of cancer; the concept of utilizing a vaccine to specifically target essential regulators of EMT is yet to be exploited in the field of tumor immunology. Recently, we have demonstrated that Brachyury, a T-box transcription factor essential for embryonic development, plays a central role in the induction of EMT in human carcinoma cells. In contrast to other drivers of EMT, Brachyury is highly expressed in various types of human tumors, while its expression in most normal adult tissues is undetectable. Moreover, Brachyury-specific T cells capable of lysing human tumor cell lines in an antigen-specific, HLA-restricted manner can be expanded from the peripheral blood of cancer patients, making Brachyury an attractive target for a vaccination strategy designed to specifically target cells undergoing an EMT. In light of the documented resistance of mesenchymal-like tumor cells to chemotherapy and radiation, we have thoroughly investigated the susceptibility of tumor cells undergoing EMT to immune-mediated attack. Utilizing several epithelial human tumor cell lines stably transfected to express various levels of Brachyury, we have observed that tumor cells with low to moderate amounts of Brachyury can be efficiently lysed by various immune-mediated mechanisms, while very high levels of Brachyury expression correlate with increased resistance to lysis mediated by natural killer (NK) cells, lymphokine-activated killer (LAK) cells, and antigen-specific CD8+ T cells. It has been suggested that EMT is a reversible process, whereby cells can undergo a mesenchymal-epithelial transition (MET). By stably inhibiting the expression of Brachyury in various mesenchymal-like human tumor cell lines, we have also demonstrated that induction of MET restores susceptibility of tumor cells to immune-mediated attack. Currently, the mechanisms responsible for the acquisition of increased resistance to immune-mediated killing by cells undergoing an EMT are being investigated. The results from these studies will be fundamental in designing strategies aimed at rendering metastatic carcinoma cells more susceptible not only to chemotherapy and radiation, but also to immunotherapeutic interventions against the tumor. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr A64.
-
the human t box mesodermal transcription factor brachyury is a candidate target for t cell mediated cancer immunotherapy
Clinical Cancer Research, 2007Co-Authors: Claudia Palena, Romaine I Fernando, Mary T Litzinger, D E Polev, Kwong Y Tsang, L L Krukovskaya, Ancha Baranova, A P Kozlov, Jeffrey SchlomAbstract:Purpose: Identification of tumor antigens is essential in advancing immune-based therapeutic interventions in cancer. Particularly attractive targets are those molecules that are selectively expressed by malignant cells and that are also essential for tumor progression. Experimental Design and Results: We have used a computer-based differential display analysis tool for mining of expressed sequence tag clusters in the human Unigene database and identified Brachyury as a novel tumor antigen. Brachyury, a member of the T-box transcription factor family, is a key player in mesoderm specification during embryonic development. Moreover, transcription factors that control mesoderm have been implicated in the epithelial-mesenchymal transition (EMT), which has been postulated to be a key step during tumor progression to metastasis. Reverse transcription-PCR analysis validated the in silico predictions and showed Brachyury expression in tumors of the small intestine, stomach, kidney, bladder, uterus, ovary, and testis, as well as in cell lines derived from lung, colon, and prostate carcinomas, but not in the vast majority of the normal tissues tested. An HLA-A0201 epitope of human Brachyury was identified that was able to expand T lymphocytes from blood of cancer patients and normal donors with the ability to lyse Brachyury-expressing tumor cells. Conclusions: To our knowledge, this is the first demonstration that (a) a T-box transcription factor and (b) a molecule implicated in mesodermal development, i.e., EMT, can be a potential target for human T-cell–mediated cancer immunotherapy.
Jeffrey Schlom - One of the best experts on this subject based on the ideXlab platform.
-
nuclear brachyury expression is consistent in chordoma common in germ cell tumors and small cell carcinomas and rare in other carcinomas and sarcomas an immunohistochemical study of 5229 cases
The American Journal of Surgical Pathology, 2015Co-Authors: Markku Miettinen, Christopher R. Heery, Jeffrey Schlom, Zengfeng Wang, Jerzy Lasota, Claudia PalenaAbstract:Brachyury is a transcription factor of the T-box family typically expressed in notochord and chordoma. Some studies report brachyury as highly specific for chordoma, whereas others have concluded that brachyury is expressed in many types of common carcinomas by RT-PCR and immunohistochemistry and could be involved in the epithelial-mesenchymal transition and metastatic process. In this study, we immunohistochemically evaluated 5229 different tumors for nuclear brachyury expression using a new rabbit monoclonal antibody and automated immunostaining (Leica Bond Max). Only nuclear labeling was scored, and antibody dilution of 1:2000 was used. In normal tissues, only rare cells in seminiferous tubules were labeled; all other organs were negative. All chordomas (75/76), except a sarcomatous one, were positive, whereas chondrosarcomas were negative. Among epithelial tumors, positivity was often detected in embryonal carcinoma (74%) and seminoma (45%). Pulmonary small cell carcinoma was often positive (41%), whereas pulmonary and pancreatic adenocarcinomas only rarely showed nuclear brachyury-positivity (3–4%). Common carcinomas such as ductal carcinomas of breast, or adenocarcinomas of the prostate only exceptionally showed nuclear positivity (< 1%). No colorectal, hepatocellular, renal cell, squamous cell, thyroid or urothelial carcinoma, or mesothelioma showed nuclear brachyury-positivity. Among mesenchymal and neuroectodermal tumors, only isolated cases of melanoma, malignant peripheral nerve sheath tumor, rhabdomyosarcoma, synovial sarcoma, and follicular lymphoma showed nuclear expression. However, as shown previously with lung carcinoma, experiments with lower antibody dilutions (1:200–1:500) showed weak cytoplasmic and nuclear labeling in breast cancers. In addition to chordoma, we show here for the first time that nuclear brachyury expression is prevalent in embryonal carcinoma, seminoma, and small cell carcinoma of the lung but very rare in common carcinomas, sarcomas, and melanoma. With these reservations, we have demonstrated the presence of nuclear brachyury immunoreactivity to be a sensitive and fairly specific marker for chordoma.
-
immunological targeting of tumor cells undergoing an epithelial mesenchymal transition via a recombinant brachyury yeast vaccine
Oncotarget, 2013Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Jeffrey Schlom, Mary T Litzinger, Alessandra Jales, James W Hodge, Andressa Ardiani, David Apelian, Claudia PalenaAbstract:// Duane H. Hamilton 1,* , Mary T. Litzinger 1,* , Alessandra Jales 1 , Bruce Huang 1 , Romaine I. Fernando 1 , James W. Hodge 1 , Andressa Ardiani 1 , David Apelian 2 , Jeffrey Schlom 1,* , and Claudia Palena 1,* 1 Laboratory of Tumor Immunology and Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 2 GlobeImmune Inc., Louisville, Colorado * These authors contributed equally to the work Correspondence: Claudia Palena, email: // Keywords : epithelial-mesenchymal transition, brachyury, T-box transcription factor, cancer vaccine, T-cell immunotherapy Received : August 16, 2013 Accepted : September 24, 2013 Published : September 26, 2013 Abstract The embryonic T-box transcription factor brachyury is aberrantly expressed in a range of human tumors. Previous studies have demonstrated that brachyury is a driver of the epithelial-mesenchymal transition (EMT), a process associated with cancer progression. Brachyury expression in human tumor cells enhances tumor invasiveness in vitro and metastasis in vivo , and induces resistance to various conventional therapeutics including chemotherapy and radiation. These characteristics, and the selective expression of brachyury for a range of human tumor types vs. normal adult tissues, make brachyury an attractive tumor target. Due to its intracellular localization and the “undruggable” character of transcription factors, available options to target brachyury are currently limited. Here we report on the development and characterization of an immunological platform for the efficient targeting of brachyury-positive tumors consisting of a heat-killed, recombinant Saccharomyces cerevisiae (yeast)–brachyury vector-based vaccine (designated as GI-6301) that expresses the full-length human brachyury protein. We demonstrate that human dendritic cells treated with recombinant yeast-brachyury can activate and expand brachyury-specific CD4 + and CD8 + T cells in vitro that, in turn, can effectively lyse human tumor cells expressing the brachyury protein. Vaccination of mice with recombinant yeast-brachyury is also shown here to elicit brachyury-specific CD4 + and CD8 + T-cell responses, and to induce anti-tumor immunity in the absence of toxicity. Based on these results, a Phase I clinical trial of GI-6301 is currently ongoing in patients with advanced tumors; to our knowledge, this is the first vaccine platform aimed at targeting a driver of tumor EMT that has successfully reached the clinical stage.
-
abstract a64 brachyury mediated epithelial mesenchymal transition of human carcinoma cells is associated with an increased resistance to immune mediated attack
Cancer Research, 2011Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Kwongyok Tsang, Jeffrey Schlom, Claudia PalenaAbstract:The epithelial-mesenchymal transition (EMT) has recently been recognized as a relevant process during the progression of carcinomas. Epithelial tumor cells undergoing EMT have been shown to acquire a mesenchymal-like phenotype, cell motility, and invasiveness, in vivo metastatic propensity and stem cell-like properties, including the ability to resist cell death initiated by traditional cancer treatment modalities such as chemotherapy and radiation. The development of therapeutic interventions aimed at interfering with EMT is therefore emerging as a rational approach for the prevention of cancer metastasis and alleviation of therapeutic resistance. Promising results recently obtained in preclinical and clinical studies with cancer vaccines may lead to their use for the treatment of various types of cancer; the concept of utilizing a vaccine to specifically target essential regulators of EMT is yet to be exploited in the field of tumor immunology. Recently, we have demonstrated that Brachyury, a T-box transcription factor essential for embryonic development, plays a central role in the induction of EMT in human carcinoma cells. In contrast to other drivers of EMT, Brachyury is highly expressed in various types of human tumors, while its expression in most normal adult tissues is undetectable. Moreover, Brachyury-specific T cells capable of lysing human tumor cell lines in an antigen-specific, HLA-restricted manner can be expanded from the peripheral blood of cancer patients, making Brachyury an attractive target for a vaccination strategy designed to specifically target cells undergoing an EMT. In light of the documented resistance of mesenchymal-like tumor cells to chemotherapy and radiation, we have thoroughly investigated the susceptibility of tumor cells undergoing EMT to immune-mediated attack. Utilizing several epithelial human tumor cell lines stably transfected to express various levels of Brachyury, we have observed that tumor cells with low to moderate amounts of Brachyury can be efficiently lysed by various immune-mediated mechanisms, while very high levels of Brachyury expression correlate with increased resistance to lysis mediated by natural killer (NK) cells, lymphokine-activated killer (LAK) cells, and antigen-specific CD8+ T cells. It has been suggested that EMT is a reversible process, whereby cells can undergo a mesenchymal-epithelial transition (MET). By stably inhibiting the expression of Brachyury in various mesenchymal-like human tumor cell lines, we have also demonstrated that induction of MET restores susceptibility of tumor cells to immune-mediated attack. Currently, the mechanisms responsible for the acquisition of increased resistance to immune-mediated killing by cells undergoing an EMT are being investigated. The results from these studies will be fundamental in designing strategies aimed at rendering metastatic carcinoma cells more susceptible not only to chemotherapy and radiation, but also to immunotherapeutic interventions against the tumor. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr A64.
-
the human t box mesodermal transcription factor brachyury is a candidate target for t cell mediated cancer immunotherapy
Clinical Cancer Research, 2007Co-Authors: Claudia Palena, Romaine I Fernando, Mary T Litzinger, D E Polev, Kwong Y Tsang, L L Krukovskaya, Ancha Baranova, A P Kozlov, Jeffrey SchlomAbstract:Purpose: Identification of tumor antigens is essential in advancing immune-based therapeutic interventions in cancer. Particularly attractive targets are those molecules that are selectively expressed by malignant cells and that are also essential for tumor progression. Experimental Design and Results: We have used a computer-based differential display analysis tool for mining of expressed sequence tag clusters in the human Unigene database and identified Brachyury as a novel tumor antigen. Brachyury, a member of the T-box transcription factor family, is a key player in mesoderm specification during embryonic development. Moreover, transcription factors that control mesoderm have been implicated in the epithelial-mesenchymal transition (EMT), which has been postulated to be a key step during tumor progression to metastasis. Reverse transcription-PCR analysis validated the in silico predictions and showed Brachyury expression in tumors of the small intestine, stomach, kidney, bladder, uterus, ovary, and testis, as well as in cell lines derived from lung, colon, and prostate carcinomas, but not in the vast majority of the normal tissues tested. An HLA-A0201 epitope of human Brachyury was identified that was able to expand T lymphocytes from blood of cancer patients and normal donors with the ability to lyse Brachyury-expressing tumor cells. Conclusions: To our knowledge, this is the first demonstration that (a) a T-box transcription factor and (b) a molecule implicated in mesodermal development, i.e., EMT, can be a potential target for human T-cell–mediated cancer immunotherapy.
Bruce Huang - One of the best experts on this subject based on the ideXlab platform.
-
wee1 inhibition alleviates resistance to immune attack of tumor cells undergoing epithelial mesenchymal transition
Cancer Research, 2014Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Kam Sze Tsang, Claudia PalenaAbstract:Aberrant expression of the T-box transcription factor brachyury in human carcinomas drives the phenomenon of epithelial-mesenchymal transition (EMT), a phenotypic modulation that facilitates tumor dissemination and resistance to conventional therapies, including chemotherapy and radiation. By generating isogenic cancer cell lines with various levels of brachyury expression, we demonstrate that high levels of brachyury also significantly reduce the susceptibility of cancer cells to lysis by both antigen-specific T cells and natural killer cells. Our results indicated that resistance of brachyury-high tumor cells to immune-mediated attack was due to inefficient caspase-dependent apoptosis, manifested as inefficient nuclear lamin degradation in the presence of activated effector caspases. We correlated this phenomenon to loss of cell cycle kinase CDK1, which mediates lamin phosphorylation. In support of a causal connection, pre-treatment of tumor cells with a specific inhibitor of WEE1, a negative regulator kinase of CDK1, could counter the defective apoptosis of tumor cells expressing high levels of brachyury. Thus, our findings suggested that reconstituting CDK1 activity to threshold levels may be sufficient to restore immunosurveillance of mesenchymal-like cancer cells that have escaped previous immune detection or eradication.
-
immunological targeting of tumor cells undergoing an epithelial mesenchymal transition via a recombinant brachyury yeast vaccine
Oncotarget, 2013Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Jeffrey Schlom, Mary T Litzinger, Alessandra Jales, James W Hodge, Andressa Ardiani, David Apelian, Claudia PalenaAbstract:// Duane H. Hamilton 1,* , Mary T. Litzinger 1,* , Alessandra Jales 1 , Bruce Huang 1 , Romaine I. Fernando 1 , James W. Hodge 1 , Andressa Ardiani 1 , David Apelian 2 , Jeffrey Schlom 1,* , and Claudia Palena 1,* 1 Laboratory of Tumor Immunology and Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, Maryland 2 GlobeImmune Inc., Louisville, Colorado * These authors contributed equally to the work Correspondence: Claudia Palena, email: // Keywords : epithelial-mesenchymal transition, brachyury, T-box transcription factor, cancer vaccine, T-cell immunotherapy Received : August 16, 2013 Accepted : September 24, 2013 Published : September 26, 2013 Abstract The embryonic T-box transcription factor brachyury is aberrantly expressed in a range of human tumors. Previous studies have demonstrated that brachyury is a driver of the epithelial-mesenchymal transition (EMT), a process associated with cancer progression. Brachyury expression in human tumor cells enhances tumor invasiveness in vitro and metastasis in vivo , and induces resistance to various conventional therapeutics including chemotherapy and radiation. These characteristics, and the selective expression of brachyury for a range of human tumor types vs. normal adult tissues, make brachyury an attractive tumor target. Due to its intracellular localization and the “undruggable” character of transcription factors, available options to target brachyury are currently limited. Here we report on the development and characterization of an immunological platform for the efficient targeting of brachyury-positive tumors consisting of a heat-killed, recombinant Saccharomyces cerevisiae (yeast)–brachyury vector-based vaccine (designated as GI-6301) that expresses the full-length human brachyury protein. We demonstrate that human dendritic cells treated with recombinant yeast-brachyury can activate and expand brachyury-specific CD4 + and CD8 + T cells in vitro that, in turn, can effectively lyse human tumor cells expressing the brachyury protein. Vaccination of mice with recombinant yeast-brachyury is also shown here to elicit brachyury-specific CD4 + and CD8 + T-cell responses, and to induce anti-tumor immunity in the absence of toxicity. Based on these results, a Phase I clinical trial of GI-6301 is currently ongoing in patients with advanced tumors; to our knowledge, this is the first vaccine platform aimed at targeting a driver of tumor EMT that has successfully reached the clinical stage.
-
the embryonic transcription factor brachyury blocks cell cycle progression and mediates tumor resistance to conventional antitumor therapies
Cell Death and Disease, 2013Co-Authors: Bruce Huang, Duane H Hamilton, Romaine I Fernando, Mary T Litzinger, James W Hodge, J R Cohen, Claudia PalenaAbstract:The T-box transcription factor Brachyury, a molecule frequently detected in human cancers but seldom found in normal adult tissue, has recently been characterized as a driver of the epithelial-to-mesenchymal switch of human carcinomas. In the current investigation, we present data demonstrating that in two different human lung carcinoma models expression of Brachyury strongly correlates with increased in vitro resistance to cytotoxic therapies, such as chemotherapy and radiation. We also demonstrate that chemotherapy treatment in vitro selects for tumor cells with high levels of Brachyury and that the degree of resistance to therapy correlates with the level of Brachyury expression. In vitro and in vivo, human lung carcinoma cells with higher levels of Brachyury divide at slower rates than those with lower levels of Brachyury, a phenomenon associated with marked downregulation of cyclin D1, phosphorylated Rb and CDKN1A (p21). Chromatin immunoprecipitation and luciferase reporter assays revealed that Brachyury binds to a half T-box consensus site located within the promoter region of the p21 gene, indicating a potential mechanism for the observed therapeutic resistance associated with Brachyury expression. Finally, we demonstrate that in vivo treatment of tumor xenografts with chemotherapy results in the selective growth of resistant tumors characterized by high levels of Brachyury expression. Altogether, these results suggest that Brachyury expression may attenuate cell cycle progression, enabling tumor cells to become less susceptible to chemotherapy and radiation in human carcinomas.
-
abstract a64 brachyury mediated epithelial mesenchymal transition of human carcinoma cells is associated with an increased resistance to immune mediated attack
Cancer Research, 2011Co-Authors: Duane H Hamilton, Romaine I Fernando, Bruce Huang, Kwongyok Tsang, Jeffrey Schlom, Claudia PalenaAbstract:The epithelial-mesenchymal transition (EMT) has recently been recognized as a relevant process during the progression of carcinomas. Epithelial tumor cells undergoing EMT have been shown to acquire a mesenchymal-like phenotype, cell motility, and invasiveness, in vivo metastatic propensity and stem cell-like properties, including the ability to resist cell death initiated by traditional cancer treatment modalities such as chemotherapy and radiation. The development of therapeutic interventions aimed at interfering with EMT is therefore emerging as a rational approach for the prevention of cancer metastasis and alleviation of therapeutic resistance. Promising results recently obtained in preclinical and clinical studies with cancer vaccines may lead to their use for the treatment of various types of cancer; the concept of utilizing a vaccine to specifically target essential regulators of EMT is yet to be exploited in the field of tumor immunology. Recently, we have demonstrated that Brachyury, a T-box transcription factor essential for embryonic development, plays a central role in the induction of EMT in human carcinoma cells. In contrast to other drivers of EMT, Brachyury is highly expressed in various types of human tumors, while its expression in most normal adult tissues is undetectable. Moreover, Brachyury-specific T cells capable of lysing human tumor cell lines in an antigen-specific, HLA-restricted manner can be expanded from the peripheral blood of cancer patients, making Brachyury an attractive target for a vaccination strategy designed to specifically target cells undergoing an EMT. In light of the documented resistance of mesenchymal-like tumor cells to chemotherapy and radiation, we have thoroughly investigated the susceptibility of tumor cells undergoing EMT to immune-mediated attack. Utilizing several epithelial human tumor cell lines stably transfected to express various levels of Brachyury, we have observed that tumor cells with low to moderate amounts of Brachyury can be efficiently lysed by various immune-mediated mechanisms, while very high levels of Brachyury expression correlate with increased resistance to lysis mediated by natural killer (NK) cells, lymphokine-activated killer (LAK) cells, and antigen-specific CD8+ T cells. It has been suggested that EMT is a reversible process, whereby cells can undergo a mesenchymal-epithelial transition (MET). By stably inhibiting the expression of Brachyury in various mesenchymal-like human tumor cell lines, we have also demonstrated that induction of MET restores susceptibility of tumor cells to immune-mediated attack. Currently, the mechanisms responsible for the acquisition of increased resistance to immune-mediated killing by cells undergoing an EMT are being investigated. The results from these studies will be fundamental in designing strategies aimed at rendering metastatic carcinoma cells more susceptible not only to chemotherapy and radiation, but also to immunotherapeutic interventions against the tumor. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the Second AACR International Conference on Frontiers in Basic Cancer Research; 2011 Sep 14-18; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2011;71(18 Suppl):Abstract nr A64.
