Calcium Channels in Glioblastoma

胶质母细胞瘤中的钙通道

基本信息

批准号：
10708091
负责人：
Roger Abounader
金额：
$ 50.87万
依托单位：
UNIVERSITY OF VIRGINIA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-21 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10708091
关键词：
Animal Model Apoptosis Brain CRISPR screen Calcium Calcium Channel Calcium Channel Blockers Calcium Signaling Cell Proliferation Cell physiology Cells Chemotherapy and/or radiation Clinical Clinical Trials Coculture Techniques Combined Modality Therapy Cytosol Data Development Drug Synergism Event Extracellular Space FDA approved Genetic Transcription Genomics Glioblastoma Growth Human Immune Immunocompetent In Vitro Knowledge Life Expectancy Malignant Neoplasms Malignant neoplasm of brain Mediating Mibefradil Modality Molecular Mus Neurons Neurosciences Operative Surgical Procedures Pharmaceutical Preparations Phase Prognosis Proteomics Publications Radiation therapy Recurrence Regimen Research Role Synapses Synaptic plasticity T-Type Calcium Channels Testing Therapeutic Transgenic Organisms Translating Tumor Promotion Work Xenograft Model Xenograft procedure angiogenesis cell growth cell motility chemotherapy clinical translation druggable target improved in vitro Assay in vivo mouse model multidisciplinary neoplastic cell neuro-oncology neurotransmission new combination therapies novel therapeutic intervention novel therapeutics optimal treatments screening standard of care stem cells therapeutic target tool tumor tumor growth tumor microenvironment

项目摘要

ABSTRACT Glioblastoma (GBM) is the most common and most deadly primary malignant brain tumor. Calcium signaling regulates a plethora of cancer-associated molecular and cellular processes including cell proliferation, apoptosis, motility, angiogenesis, differentiation, gene transcription as well as neurotransmission and synaptic plasticity. Calcium influx from the extracellular space to the cytosol is regulated by T-Type calcium channels (TTCC). Our preliminary data show that TTCC are upregulated in GBM cells, stem cells (GSC) and human tumors and that their blockage leads to inhibition of cancer-promoting parameters in tumor cell-intrinsic and microenvironment-dependent manners. Based on these data, we hypothesize that TTCC strongly regulate GBM molecular events and GBM-microenvironment interactions to drive tumor growth, and that targeting TTCC in combination with other modalities is a promising GBM therapy. To test this hypothesis, we propose to investigate the tumor cell-intrinsic and microenvironment-dependent functions, mechanisms of action, and therapeutic targeting of TTCC in GBM. In Aim 1, we will determine the GBM cell-intrinsic role and mechanisms of action of TTCC in new mouse models with intact microenvironment. We will develop new RCAS/Tva and transgenic immune competent TTCC mouse models and use them to study the role of TTCC in an intact GBM microenvironment. We will also use genomic and proteomic screenings and molecular and functional approaches to uncover the mechanisms of action of TTCC in these GBM tumors. In Aim 2, we will uncover the role of tumor microenvironment TTCC in mediating tumor-promoting neuron/GBM interactions. We hypothesize that neuronal TTCC and GBM TTCC cooperate to regulate the tumor-promoting interactions between GBM cells and neurons that were recently discovered. To test this hypothesis, we will use co-cultures and GBM animal models to investigate the role of TTCC in regulating neuron/GBM synaptic formation, calcium influx into tumor cells, and tumor growth and malignancy. In Aim 3, we will develop and test new strategies for the therapeutic targeting of TTCC in GBM. We have a repurposed FDA approved TTCC blocker, mibefradil, that was demonstrated to be safe and possibly effective in a phase I recurrent GBM trial. We will test the effects of mibefradil on the growth of GBM xenografts, syngeneic tumors and RCAS/Tva GBM mice using the standard clinical Stupp Regimen. We will also perform in vitro and in vivo synthetic lethal CRISPR screens to uncover druggable targets and drugs that synergize with mibefradil. We will then test combinations of mibefradil and the synthetic lethal drugs in GBM animal models. Altogether, the findings will generate new knowledge on the functions and mechanisms of action of TTCC in GBM and its microenvironment, develop new tools for the study of TTCC, uncover the role of TTCC in mediating tumor-promoting neuron/GBM interactions, and develop and test new efficacious GBM combination therapies that could be translated into clinical trials.

抽象的胶质母细胞瘤（GBM）是最常见和最致命的原发性恶性脑肿瘤。钙信号传导调节大量与癌症相关的分子和细胞过程，包括细胞增殖、细胞凋亡、运动性、血管生成、分化、基因转录以及神经传递和突触可塑性。钙从细胞外空间流入细胞质受 T 型钙通道 (TTCC) 调节。我们的初步数据显示，TTCC 在 GBM 细胞、干细胞 (GSC) 和人类肿瘤中表达上调，并且它们的阻断导致肿瘤细胞固有的和促进癌症的参数受到抑制微环境依赖的方式。基于这些数据，我们假设 TTCC 强烈监管 GBM 分子事件和 GBM-微环境相互作用驱动肿瘤生长，并且靶向 TTCC 与其他疗法相结合是一种有前途的 GBM 治疗方法。为了检验这个假设，我们建议研究肿瘤细胞固有的和微环境依赖性的功能、机制 TTCC 在 GBM 中的作用和治疗靶向。在目标 1 中，我们将确定 GBM 细胞的内在作用以及 TTCC 在具有完整微环境的新小鼠模型中的作用机制。我们将开发新的RCAS/Tva和转基因免疫能力TTCC小鼠模型，并用它们来研究TTCC的作用在完整的 GBM 微环境中。我们还将使用基因组和蛋白质组筛选以及分子和揭示 TTCC 在这些 GBM 肿瘤中的作用机制的功能方法。在目标 2 中，我们将揭示肿瘤微环境TTCC在介导促癌神经元/GBM中的作用互动。我们假设神经元 TTCC 和 GBM TTCC 合作调节肿瘤促进最近发现的 GBM 细胞和神经元之间的相互作用。为了检验这个假设，我们将使用共培养和 GBM 动物模型研究 TTCC 在调节神经元/GBM 突触中的作用形成、钙流入肿瘤细胞以及肿瘤生长和恶性肿瘤。在目标 3 中，我们将开发和测试 GBM 中 TTCC 治疗靶向的新策略。我们有 FDA 批准的 TTCC 改造用途阻断剂米贝拉地尔 (mibefradil) 在 I 期复发性 GBM 试验中被证明是安全的，并且可能有效。我们将测试米贝拉地尔对 GBM 异种移植物、同基因肿瘤和 RCAS/Tva GBM 小鼠生长的影响使用标准临床 Stupp 方案。我们还将进行体外和体内合成致死性 CRISPR 筛选以发现可成药的靶点和与米贝拉地尔协同作用的药物。然后我们将测试组合 GBM 动物模型中米贝拉地尔和合成致死药物的研究。总而言之，这些发现将产生新的了解 TTCC 在 GBM 及其微环境中的功能和作用机制，开发新的研究 TTCC 的工具，揭示 TTCC 在介导肿瘤促进神经元/GBM 相互作用中的作用，开发和测试新的有效的 GBM 联合疗法，并将其转化为临床试验。