Regulation of glioblastoma cells by GABAergic neurons in human organoid-tumor chimeras

项目摘要/摘要 胶质母细胞瘤(GBM，WHO IV级胶质瘤)是最常见和最致命的原发脑肿瘤之一。 对儿童和成人都有影响。GBM与显著增加的死亡率有关，并且没有 尽管之前进行了许多临床试验，但仍有有效的治疗方法可供患者使用。人们相信，一个更好的 了解大脑中的细胞类型和调节基底膜细胞特性的因素可以提供新的 深入了解其病理机制，揭示治疗基底膜患者的新靶点。这样做的主要目标是 R21建议阐明人GABA能神经元在调节细胞增殖和存活中的作用 人脑GBM球体与人诱导物共培养脑内GBM细胞 多能干细胞(IPSC)来源的端脑有机体(有机体-GBM嵌合体)。为了实现这一目标，我们 将使用最先进的神经科学工具和技术，包括人类干细胞来源的大脑 有机物、光遗传学、狂犬病病毒追踪和单细胞RNA测序，以表征 有机-GBM嵌合体中GABA能神经元与肿瘤细胞之间的通讯及其影响 关于肿瘤细胞存活和增殖的沟通。我们的具体假设是GABA能神经元形成 GBM类器官嵌合体中与肿瘤细胞的功能性突触促进肿瘤细胞的增殖或存活 通过GABA介导的肿瘤细胞去极化。检验这一假设的具体目的是(1)确定 GABA能神经元是否与GBM细胞建立功能性突触来调节肿瘤的生长和生存 以及(2)确定跨突触信号蛋白在调节肿瘤生长和生存中的作用。总的来说， 我们期望这项研究将促进我们对细胞和分子机制的理解。 人脑GBM细胞特性的调节，并为研究 在类人脑环境中，人脑肿瘤与人神经元的相互作用。

PROJECT SUMMARY/ABSTRACT Glioblastoma (GBM, WHO Grade IV glioma) is one of the most common and deadliest primary brain tumors that affects both children and adults. GBM is associated with significantly elevated mortality, and there are no effective therapies available for patients despite many previous clinical trials. It is believed that a better understanding of the cell types and factors regulating GBM cell properties in the brain could provide novel insights into the pathology and reveal new therapeutic targets for treating GBM patients. The main goal of this R21 proposal is to elucidate the role of human GABAergic neurons in regulating the proliferation and survival of GBM cells in the brain using an innovative co-culture approach of human GBM spheroids and human induced pluripotent stem cell (iPSC)-derived telencephalic organoids (organoid-GBM chimera). To achieve this goal, we will use the most advanced neuroscience tools and techniques, including human stem cell-derived brain organoids, optogenetics, rabies virus tracing, and single-cell RNA sequencing, to characterize the communication between GABAergic neurons and tumor cells in organoid-GBM chimera and the effects of this communication on tumor cell survival and proliferation. Our specific hypothesis is that GABAergic neurons form functional synapses with tumor cells in GBM-organoid chimeras to promote tumor cell proliferation or survival via GABA-mediated depolarization of tumor cells. The specific aims to test this hypothesis are (1) to determine whether GABAergic neurons establish functional synapses with GBM cells to regulate tumor growth and survival and (2) to determine the role of trans-synaptic signaling proteins in regulating tumor growth and survival. Overall, we expect that this study will advance our understanding of the cellular and molecular mechanisms involved in the regulation of GBM cell properties in the human brain and provide a novel framework for studying the interactions of human brain tumors with human neurons in a human brain-like environment.