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

项目总结/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性脑肿瘤之一， 影响儿童和成人。GBM与显著升高的死亡率相关， 尽管之前进行了许多临床试验，但患者仍可以获得有效的治疗方法。据信，一个更好的 了解脑中GBM细胞特性的细胞类型和调节因子可以提供新的 深入了解病理学，并揭示治疗GBM患者的新治疗靶点。这个的主要目标 R21的建议是阐明人GABA能神经元在调节细胞增殖和存活中的作用， 使用人GBM球状体和人诱导的GBM细胞的创新共培养方法， 多能干细胞（iPSC）衍生的端脑类器官（类器官-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，包括人类干细胞衍生的大脑 类器官，光遗传学，狂犬病病毒追踪和单细胞RNA测序，以表征 类器官-GBM嵌合体中GABA能神经元和肿瘤细胞之间的通讯及其作用 肿瘤细胞的存活和增殖。我们的假设是GABA能神经元 GBM-类器官嵌合体中与肿瘤细胞的功能性突触以促进肿瘤细胞增殖或存活 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以调节肿瘤生长和存活 和（2）确定跨突触信号传导蛋白在调节肿瘤生长和存活中的作用。总的来说， 我们希望这项研究将促进我们对参与的细胞和分子机制的理解， GBM细胞特性在人脑中的调节，并提供了一个新的框架， 在类似人脑的环境中，人脑肿瘤与人类神经元的相互作用。

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.