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University of Alabama at BirminghamBrain Tumor SPOREG. Yancey Gillespie, Ph.D., Program Director For more information on this specific SPORE's institution, please visit: http://www.braintumor.uab.edu
Overall Summary - "Molecular Therapeutics for Anaplastic Gliomas"Patients diagnosed with anaplastic gliomas face a dismal prognosis as overall survival has remained essentially unchanged despite more than 50 years of extensive basic and clinical science research. The principal goal of the UAB Brain Tumor SPORE is to have a positive impact on this unacceptable situation by translating the laboratory-based efforts of four multi-disciplinary groups of scientists into novel, yet practical, Phase I clinical interventions that address the needs for more effective treatments. Previously, our SPORE investigators (5 projects) successfully translated their basic findings into 2 Phase I interventional clinical trials, a glioma tissue analysis that defined the loss of a particular growth regulatory mechanism in gliomas and a genetic epidemiology survey that defined the significant association of one of two HLA phenotypes with either indolent or rapid glioma progression. The themes of this revised competitive renewal SPORE application include: Anti-angiogenesis, Oncolytic Virus therapy, Antibody-mediated Therapy, Apoptosis, and Autophagy. Four translational research projects are proposed, each of which is designed to initiate one or more interventional clinical trials within a 3- to 5-year time frame. These include: 1) Inhibition of Malignant Glioma Growth by Recombinant Kringle 5 (rK5) which will optimize the anti-angiogenic and pro-apoptotic effects of a recombinant peptide on tumor endothelial and human glioma cells when used in conjunction with radiotherapy; 2) Optimized Chimeric HSV for Anaplastic Glioma Therapy will develop a clinical application for a chimeric herpes simplex virus that has been engineered to be safe while significantly enhancing its replicative properties and overall ability to infect and kill glioma cells; 3) Death Receptor Antibody Therapy for Anaplastic Gliomas will employ combinations of conventional radiotherapy and/or chemotherapy with a UAB-developed, humanized anti-DR5 monoclonal antibody administered either intravenously or by convection enhanced delivery to induce apoptosis in tumor-associated endothelium and glioma cells; and 4) Lysosomotropic Therapy of Anaplastic Gliomas will characterize blood-brain barrier permeable chloroquine and fluoroquinolone analogues that induce glioma cell autophagy. These projects will be supported by 5 Cores: 1) Administrative, 2) Brain Tumor Tissue, 3) Clinical Trials, 4) Biostatistics/Bioinformatics, and 5) Brain Tumor Animal Models. The Career Development Program recruited 2 new investigators to brain tumor translational research and will continue to do so. We will continue the very successful Developmental Research Program that supported 15 investigators, 9 of whom were new to brain tumor research. Three of the proposed projects in this renewal were originally Developmental Projects. The University, School of Medicine and Comprehensive Cancer Center strongly support this SPORE application. We will continue our multiple, active collaborations with other Brain Tumor SPOREs and will foster new interactions with other SPORE programs locally and nationally.
Inhibition of Anaplastic Glioma Growth by Kringle 5 (K5) (Project 1)Candece L. Gladson, M.D., L. Burt Nabors, M.D. Angiogenesis is necessary for the sustained growth and invasion of malignant tumors, and is a characteristic feature of anaplastic gliomas. Anti-angiogenic therapy has been shown be an important new therapeutic approach for other types of malignant tumors, especially when combined with standard therapy; Thus, we propose to evaluate potential anti-angiogenic approaches for the treatment of anaplastic gliomas. The kringle 5 domain of plasminogen has been shown to promote an anti-angiogenic effect on microvessel endothelial cells (MvEC) propagated in vitro. rK5 is a naturally occurring peptide in the body and initiates its apoptotic effect on endothelial cells that express the stress antigen, glucose-related protein 78 (GRP78). Only endothelial cells involved in the neovascularization process seen in aggressive malignancies express GRP78. Thus, rK5 affords a safe and potentially effective adjunct to chemoradiation in patients with high-grade gliomas. rK5 may abrogate resistance to standard chemo-radiation therapies and improve therapeutic outcome. A careful appraisal of the potential surrogate markers for angiogenesis may allow improved monitoring of these therapies. Our preliminary data indicate that recombinant Kringle 5 (rK5) induces apoptosis of brain MvEC in a dose- and time-dependent manner and that prior irradiation significantly sensitizes brain MvEC to killing by rK5. Further, we present evidence that rK5 inhibits intracerebral glioma growth in syngeneic mouse glioma and glioma xenograft mouse models, most likely though an inhibition of angiogenesis. Our goal is to determine the mechanisms of action of rK5 in combination with radiation as the basis for optimizing this therapeutic approach. A major impediment to rapid and thorough analysis of anti-angiogenic therapies in animal models and in patients with malignant tumors is the lack of generally accepted surrogate markers of angiogenesis. Potential non-invasive surrogate markers of angiogenesis include: (i) total number of circulating endothelial cells (CECs), (ii) total number of circulating putative endothelial progenitor cells (CEPs), (iii) serum levels of bFGF, (iv) serum levels of SDF1α, and (v) Perfusion MRI characteristics of the tumor. However, these markers have not been thoroughly evaluated in terms of their performance in comparison with accepted “gold standard” markers of angiogenesis. We will first examine whether any of the 5 aforementioned markers provide useful surrogate markers of angiogenesis using two intracerebral mouse models of malignant glioma, a syngeneic and a xenograft model, treated with rK5, alone or with irradiation. Secondly, we will examine the effect of rK5 combined with radiation and temozolomide in a Phase I clinical trial of patients with newly diagnosed glioblastoma multiforme. These studies will: (i) likely lead to further clinical trials of rK5 combined with other agents in patients with malignant gliomas and (ii) will indicate the utility of testing surrogate markers of angiogenesis in patients harboring malignant gliomas, which, if validated in a larger trial, would enhance the development and application of anti-angiogenic therapeutic strategies.
OPTIMIZED CHIMERIC HSV FOR ANAPLASTIC GLIOMA THERAPY (PROJECT 2)Kevin A. Cassady, M.D., James M. Markert, M.D., MPH A principal limitation of HSV-1 viral oncolytic therapy is that the deletion of the γ134.5 gene that renders the virus safe for direct inoculation into CNS tumors also eliminates efficient viral replication in the tumor by inhibiting late virus gene expression. These functions, while encoded by a single gene in HSV-1, are independent phenotypes (as documented extensively in the literature). This application examines an alternative method to improve replication of γ134.5 deletion virus (Δγ134.5) in the tumor, by selectively restoring two functions of the γ134.5 gene, expression of late gene expression and replication in the face of IFN-α, without re-establishing viral neurovirulence. To achieve this end, we introduced a gene from a distantly related herpesvirus gene, Human Cytomegalovirus (HCMV), that facilitates late viral protein synthesis but does not contribute to neurotoxicity. The result is a chimeric HCMV / HSV-1 Δγ134.5 oncolytic vector (herein referred to as chimeric HSV) that is capable of wild-type HSV replication in the tumor but with the same safety profile as a Δγ134.5 vector. The chimeric HSV is superior to Δγ134.5 virus in both xenograft and syngeneic brain tumor models. The goal of this application is to advance the best oncolytic HSV into clinical trial. While pre-clinical studies indicate that the chimeric HSV are superior to existing therapy, the time and cost required for evaluation and advancement of a new biologic to clinic are extensive. Therefore, prior to embarking on this path will identify if there are adjuvant therapies that can improve G207 therapy to match the level of efficacy of the chimeric HSV. If combining additional modalities with G207 can match the efficacy of chimeric HSV therapy, we will proceed with the G207/adjuvant therapy into clinical trial. If the chimeric HSV remains superior, we will proceed with this agent for clinical trial.
DEATH RECEPTOR ANTIBODY THERAPY FOR ANAPLASTIC GLIOMAS (PROJECT 3)Donald J. Buchsbaum, Ph.D., John B. Fiveash, M.D. The goal of the current proposal is to enhance the therapeutic potential of TRA-8, an apoptosis-inducing antibody, for anaplastic glioma therapy. This project initially received support from the Brain Tumor SPORE Developmental Research Program in addition to funding by the UAB Breast and Ovarian Cancer SPORE programs and ABC2 Foundation. TRA-8, initially developed by T. Zhou at UAB, binds Death Receptor-5 (DR5, TRAIL-R2) and induces strong apoptosis of many types of cancer cells, including glioma, without hepatocellular toxicity. Daiichi Sankyo, a Japanese pharmaceutical company, has licensed TRA-8, and in collaboration with UAB, developed a humanized version of TRA-8 (CS-1008), which was entered in 2006 in a now completed first Phase I human study for solid tumors at UAB. CS-1008, a humanized monoclonal antibody to Death Receptor-5 expressed on human glioma cells and tumor associated endothelium, induces apoptosis in some glioma cells but not others. Studies in this project will identify a molecular basis for this variability and will define criteria of potential response to therapy. Anti-DR5 was developed at UAB and in Phase I trial of non-glioma cancer patients had a systemic 8-16 day half-life. Clinical testing of CS-1008 should define a more effective therapy for anaplastic gliomas. To develop an effective anti-death receptor strategy for glioma therapy, several fundamental issues have to be addressed: (1) cell surface DR5 expression may not solely associate with TRA-8-induced apoptosis of glioma cells, thus requiring a biomarker for predicting TRA-8 response; (2) both spontaneous and induced resistance of tumor cells to TRA-8-induced apoptosis is a major concern; and (3) effective and safe delivery of TRA-8 to gliomas in situ. We have developed two novel models to study TRA-8-induced apoptosis of primary gliomas: (1) a primary tumor tissue slice culture method to evaluate TRA-8 sensitivity rapidly in patient glioma specimens; and (2) a key signaling mechanism regulating TRA-8 susceptibility in tumor cells. DDX3, a RNA helicase DEAD box protein family member, constitutively binds a non-death domain region of DR5 and recruits apoptosis inhibitory proteins via CARD (caspase recruitment domain) to the DR5 cytoplasmic tail, leading to inhibition of initial death signal transduction. The functional status of protein complexes of DR5/DDX3/cIAP1 in tumor cells may be both a (a) TRA-8 response biomarker and (b) a key target for enhancement of DR5-mediated apoptosis by chemotherapy and radiation therapy. We hypothesize that radiation therapy and chemotherapy will enhance pro-apoptotic effects of TRA-8 in gliomas. We will accomplish our Specific Aims to: (1) further elucidate the mechanisms of TRA-8 resistance and develop a novel biomarker for use in gliomas; (2) develop methodology for ex vivo predictive assays of TRA-8 sensitivity utilizing the Krumdieck tissue slicer and human brain microvascular endothelial cell culture; (3) demonstrate the ability of systemically administered TRA-8, in concert with temozolomide chemotherapy and irradiation, to produce a significant antitumor effect on xenografts established from human glioma tumors in the brain of nude mice; and, (4) design and perform a Phase I trial of CS-1008 with chemoradiation in patients with newly diagnosed GBM.
LYSOSOMOTROPIC THERAPY FOR ANAPLASTIC GLIOMAS (PROJECT 4)Kevin A. Roth, M.D., Ph.D., L. Burt Nabors, M.D. More effective, less toxic therapies are desperately needed to improve survival of patients diagnosed with anaplastic gliomas. We have identified, in preliminary studies, a class of compounds operationally termed lysosomotropic agents that have significant glial tumoricidal activity in vitro. Many of these agents, such as chloroquine, quinacrine and quinolone antibiotics, are already FDA approved for other medical conditions facilitating their rapid advancement into human brain tumor patient clinical trials. Successful brain tumor treatment requires both inhibition of survival-promoting and activation of death-inducing molecular pathways. We have been investigating the interrelations between pro-survival autophagy pathways and apoptotic death-associated molecules in the nervous system for almost ten years. Our laboratory was the first to report that chloroquine, a lysosomotropic agent that inhibits lysosome function and promotes autophagic vacuole accumulation, induces neuronal cell death in part through p53-dependent activation of the intrinsic apoptotic death pathway and in part, via non-apoptotic death pathways. We have hypothesized that chloroquine acts to kill cells not simply by inhibiting the pro-survival action of autophagy but also by enhancing lysosomal-dependent pro-death signaling. Chloroquine has recently been found to be a potent adjuvant to standard chemotherapeutic agents in several animal models of cancer and most importantly, in the treatment of human glioblastoma multiforme (GBM) patients. The mechanism(s) by which chloroquine potentiates tumor cell death and the therapeutic potential of other lysosomotropic agents in high grade glial neoplasms are poorly defined. In this application, we focus on the molecular pathways by which chloroquine, quinacrine, and other tumoricidal lysosomotropic agents that we have identified promote malignant glial cell death and will critically test the therapeutic potential of these compounds using in vivo mouse models of GBM prior to direct Phase I/II clinical trials of quinacrine and other FDA approved lysosomotropic agents for treatment of patients with newly diagnosed GBM. In aim one, we will test the hypotheses that chloroquine, quinacrine, and related lysosomotropic agents induce glial tumor cell death through both apoptotic and autophagic death pathways that are at least in part mediated by lysosomal membrane permeabilization and dysregulated cathepsin activity. Aim two will test the in vivo tumoricidal effects of chloroquine, quinacrine, and related compounds, alone and in combination with temozolomide and irradiation treatment, using human GBM explants in nude mice. In Aim three, we will perform a phase I/II human clinical trial of quinacrine, a potent in vitro glial tumoricidal agent with superior blood-brain barrier penetration than chloroquine, in patients with newly diagnosed GBM. In total, these highly integrated studies will extend our previous studies of apoptotic and autophagic death pathways and yield new insights into the clinical utility of lysosomotropic therapy for patients with anaplastic gliomas.
ADMINISTRATIVE CORE (CORE A)G. Yancey Gillespie, Ph.D., James M. Markert, M.D., MPH Administration and coordination of a complex translational program that is closely aligned with a matrix Comprehensive Cancer Center and an active clinical program for patients with brain tumors is essential for timely progression from basic science discovery to clinical trial. Integration of each project in this SPORE with its cores requires a thorough understanding of the roles of each component. A major strength is the capacity of the SPORE Administration to terminate unproductive projects and institute promising ones. The principal role of the Administrative Core is to provide centralized management support, and collaborative research support for all research projects of the Specialized Program of Research Excellence (SPORE) in Neurological Cancer. The Program Director and the Co-Leader will provide scientific and administrative leadership to facilitate expeditious translation of our basic science advances into therapeutic opportunities. Administrative functions include oversight of operations and budgetary issues for all SPORE investigators, Developmental Research and Career Development Awardees. This includes coordination of the organization of the various oversight and advisory committees that will periodically monitor the progress of the individual SPORE projects, specifically the External and Internal Advisory Committees and the Executive Committee and its subcommittees. As we have in the past, full Projects that are not meeting their milestones towards successful translation of their research will be ended and the funds distributed to new projects that show greater promise of translation to an interventional clinical trial. Likewise, projects that successfully translate their basic research to clinical trial(s) will be considered to have achieved the main objective(s) and will be replaced with projects that are just as likely to achieve clinical translation. To ensure a continual source of potential replacement projects, this core will manage the recruitment, selection, oversight and other activities that are central to the Developmental Research and Career Development Programs. This core will coordinate the relocation of the Neuro-Oncology Program to administrative and laboratory space in the Comprehensive Cancer Center space. The SPORE Director and Co-Director will ensure continued integration of the Brain Tumor SPORE with the UAB Comprehensive Cancer Center through periodic meetings with Cancer Center staff. The CCC currently provides fiscal accounting support for SPORE investigators and this will continue.
BRAIN TUMOR TISSUE CORE (CORE B)Cheryl A. Palmer, M.D., G. Yancey Gillespie, Ph.D. Properly collected, highly annotated glioma tumor tissues and matched normal blood DNA specimens of high quality are essential to ensuring the quality of data derived from studies conducted with these clinical materials. Advanced molecular analyses of human tissues at the gene, mRNA and protein levels will facilitate systems biology-based data interpretation. Moreover, extensive immunohistopathologic and genetic analysis experience and expertise of Core Investigators offer a unique resource for all SPORE investigators. The Brain Tumor Tissue (BTT) Core Facility will assist each SPORE Project Leader to investigate the role of various molecules (proteins, mRNA, DNA) in brain tumor proliferation, invasion and angiogenesis. The BTT Core Facility will help confirm and validate the role of various therapeutic agents by providing a repository for human malignant glioma tissues and cells as well as non-neoplastic brain tissue. The primary objective will be to obtain and archive fresh tissue samples from brain tumor patients operated at University Hospital (UAB) and The Children’s Hospital of Alabama (TCHA) for primary or recurrent disease as well as brain tissue from patients operated for intractable epilepsy, trauma or other non-malignant disorders. All tissues collected will be excess tissue not needed for diagnostic purposes and all records will be encoded so that patient confidentiality is assiduously preserved. The BTT Core Facility will validate its collection/cryopreservation procedures by performing routine DNA/RNA extractions and PCR/RT-PCR analyses for specific marker (STR) and standard housekeeping genes. Quality of archived tissues and cells will be monitored by routine histochemical/immunohistochemical studies. Tissue collection, archiving, use and results is fully integrated with a computerized database maintained by the Biostatistics/Bioinformatics Core Facility. This database is updated regularly with basic demographic and treatment response data on all brain tumor (or non-neoplastic brain tissue donor) patients treated and/or followed at UAB or TCHA. All information collected and entered into the computerized database using encoding techniques to ensure patient confidentiality will be reviewed by a health professional. A secondary objective will be to prepare explant cultures of brain tumors and cultures of normal astrocytes and microvascular endothelial cells for Project Investigators. Thirdly, personnel in the Core Facility will routinely perform molecular biology and histologic studies. Each tumor will be probed by Low Density Array for expression of genes of special significance to each project. Tumor tissue will be probed by immunohistochemical and fluorescence-in-situ hybridization methods, based on specific genes of interest currently investigated by program project members. Newly diagnosed and recurrent high grade glioma xenografts will be established by the Brain Tumor Animal Models Core. Finally, the BTT Core Facility staff will work with each Project member to utilize our database for correlative analyses based on extent of evaluable treatment response and gene expression
CLINICAL TRIALS CORE (CORE C)L. Burt Nabors, M.D., James M. Markert, M.D., MPH The overall purpose of the Clinical Core is to provide the infrastructure and expertise to aid the investigators of the SPORE into translating their laboratory-based findings into clinical protocols. The Specific Aims for this Core are: 1) to provide for optimal support of investigators in this project to undertake clinical research involving patients with CNS malignancies, 2) to provide patients with CNS malignancies and referring physicians access to clinical research treatments developed by investigators of this project, and 3) to facilitate the introduction of protocols derived from the translational research of this SPORE to a national cooperative consortium of brain tumor clinical centers. The Core will be directed by Dr. L. Burton Nabors, who will provide overall leadership and administration of the Core and will oversee its interactions with the Cancer Center, the General Clinical Research Center, and other institutional components that can support the Core’s operations. He will be aided in this regard by Dr. James M. Markert, Core Co-Leader, the Clinical Core Committee, whose membership include the Clinical Co-Investigators of each project, the other Core Directors, and members with expertise in radiation oncology and neuropsychology. This committee will meet monthly to review the status of translational progress in each project, and will assist Dr. Nabors in the writing and implementation of clinical protocols resulting from this translational research. Four interventional clinical trials are proposed in this SPORE application and Inter-SPORE trials and other clinically related protocols will be instituted during the course of this research. Each protocol combines patient therapy with biologic correlates. A Clinical Trials core, operated by experienced and knowledgeable professionals, is essential for rapid and effective translation of basic research findings as mandated by the SPORE Program. This Core will support each of the 4 projects and other funded SPORE research. All clinical protocols are initially discussed in the Brain Tumor Working Group and approved protocols are forwarded to the Comprehensive Cancer Center's Clinical Trials Review Committee where the scientific, and clinical basis for the protocol, protocol methodologies, consent form(s) and other meritorious components are assessed as well as the need for nursing and biostatistical support. The CTRC submits the protocol to the IRB as a Cancer Trial. The Comprehensive Cancer Center will monitor the trial through its Data Safety Monitoring Committee, established by a NIH-approved DSMB plan.
BIOSTATISTICS AND BIOINFORMATICS CORE (CORE D)Sreelatha Meleth, Ph.D., Alan Cantor, Ph.D. The primary objective of the Biostatistics/Bioinformatics Core Facility is to provide centralized statistical services and collaborative research support for the research projects of the Specialized Program of Research Excellence (SPORE) for Neurological Cancer. The Biostatistics/Bioinformatics Core Facility will serve as the focal point from which SPORE investigators, developmental research and career development awardees draw statistical expertise for the design, management and analysis of their research projects. Biostatistics/bioinformatics services supported by SPORE funds are in addition to those currently offered through the Biostatistics Unit of the Comprehensive Cancer Center and support special analytical and non-hypothesis driven research activities (database, genetics statistics, website). These special activities enhance the biostatistics/bioinformatics services that are needed by SPORE investigators for correlative analyses of translational research findings. The specific aims of the Biostatistics/Bioinformatics Core component are to:
BRAIN TUMOR ANIMAL MODELS CORE (CORE E)G. Yancey Gillespie, Ph.D. This core facility will assist each SPORE Project Leader to test, in relevant animal models of brain tumors, preclinical safety and efficacy of novel therapies designed to achieve an improved anti-glioma effect. Animal models we will employ are likely to identify both toxicity and efficacy of various therapeutic modalities proposed to treat patients with malignant brain tumors. The BTAM Core will centralize SPORE animal experimentation, standardizing expert surgical and animal handling techniques and minimizing chances for trivial interferences that could confound comparative analyses. Tumor volume, tumor mass and survival statistics will be collected where appropriate. Normal and tumor tissues will be collected and submitted to each investigator or will be processed in this core for gene expression or histopathologic analyses. Project 1 will use both inducible and transplantable syngeneic gliomas in immunocompetent mice to test the capacity of recombinant Kringle 5 peptide of plasminogen XVIII to elicit an effective anti-angiogenic and anti-glioma response. Project 2 will evaluate the capacity of a genetically engineered chimeric herpes simplex virus, type 1 to produce oncolysis of human gliomas growing as xenografts in the brains of nude mice. Pre-IND safety studies in HSV-sensitive New-World owl monkeys (Aotus spp.) will be conducted to define any unanticipated toxicities to primate brain. Project 3 will use human glioma xenografts to determine the mechanism(s) of apoptosis of gloma cells and tumor-associated endothelial cells induced with a monoclonal antibody to DR-5 death receptor. Safety and efficacy will be defined for intracranial human gliomas in nude mice. Project 4 will compare safety and efficacy of quinacrine and fluoroquinolones to that of chloroquine all of which induce autophagy in human intracranial glioma xenografts in nude mice. These lysosomotrophic agents enhance glioma cell sensitivity to temozolomide chemotherapy and radiation therapy. The BTAM Core will support preclinical animal studies by Developmental Research and Career Development awardees. The BTAM Core is an essential component to the process of translating novel therapies from the laboratory to clinical application. Animal testing, performed in a highly standardized fashion by trained, skilled and experienced professionals, is a prerequisite for FDA approval to initiate-IRB approved clinical trials in humans. Moreover, our brain tumor models replicate, in most ways, the biology and physiology of high grade gliomas in patients and as such can be predictive of the likelihood of success or failure of novel therapies. The BTAM Core will conduct all NMR imaging studies of tumor-bearing mice in these preclinical evaluations in the 9.4T Small Animal NMR Facility and will conduct 4.7T NMR imaging and spectroscopic studies in nonhuman primate (Aotus) monkeys for Project 2. Finally, the Core will continue to evaluate serially passaged human glioma xenografts as well as specific transgenic models for suitability for preclinical toxicity and efficacy analyses for each modality proposed by the individual Projects within this SPORE. SPORE Investigators at UABGillespie, G. Yancey, Ph.D., Surgery/Neurosurgery |
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