Calcium Channels in Glioblastoma

胶质母细胞瘤中的钙通道

• 批准号: 10583656
• 负责人: Roger Abounader
• 金额: $ 54.12万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 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/10583656
• 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; Differentiated Gene; Event; Extracellular Space; FDA approved; Genetic Transcription; Genomics; Glioblastoma; Growth; Human; Immune; Immunocompetent; In Vitro; Knowledge; Lead; 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; Work; Xenograft Model; Xenograft procedure; angiogenesis; base; cell growth; cell motility; clinically translatable; 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; standard of care; stem cells; therapeutic target; tool; tumor; tumor growth; tumor microenvironment

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，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，凋亡， 运动，血管生成，分化，基因转录以及神经传递和突触可塑性。 从细胞外空间到细胞质的钙涌入受T型钙通道（TTCC）调节。我们的 初步数据表明，TTCC在GBM细胞，干细胞（GSC）和人类肿瘤中被上调，并且 它们的阻塞导致抑制肿瘤细胞中癌症促进参数的抑制和 微环境依赖于举止。基于这些数据，我们假设TTCC强烈调节 GBM分子事件和GBM微环境相互作用以驱动肿瘤的生长，并 将TTCC与其他模式结合使用是一种有希望的GBM疗法。为了检验这一假设， 我们建议研究肿瘤细胞中的和微环境依赖性功能，机制 TTCC在GBM中的作用和治疗靶向。在AIM 1中，我们将确定GBM细胞中的角色 TTCC在具有完整微环境的新小鼠模型中的作用机理。我们将发展 新的RCAS/TVA和转基因免疫有能力的TTCC鼠标模型，并使用它们来研究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学筛选以及分子以及 揭示TTCC在这些GBM肿瘤中的作用机理的功能方法。在AIM 2中，我们将 发现肿瘤微环境TTCC在介导肿瘤神经元/GBM中的作用 互动。我们假设神经元TCCC和GBM TTCC合作以调节肿瘤促进 GBM细胞与最近发现的神经元之间的相互作用。为了检验这一假设，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，钙进入肿瘤细胞以及肿瘤的生长和恶性肿瘤。在AIM 3中，我们将开发和测试 GBM中TTCC治疗靶向的新策略。我们有一个重新利用的FDA批准的TTCC Mibefradil的阻滞剂被证明是安全且可能在I期复发性GBM试验中有效的。我们 将测试Mibefradil对GBM异种移植物，合成肿瘤和RCAS/TVA GBM小鼠的生长的影响 使用标准临床Stupp方案。我们还将在体外和体内合成致死CRISPR 筛选与Mibefradil协同作用的可吸毒靶标和药物。然后我们将测试组合 Mibefradil和GBM动物模型中的合成致命药物的摄入。总共，发现将产生新的 关于TTCC在GBM及其微环境中的作用功能和机制的知识，发展了新 研究TTCC的工具，发现TTCC在介导肿瘤神经元/GBM相互作用中的作用， 并开发和测试可以转化为临床试验的新有效的GBM组合疗法。