Molecular determinants of cellular heterogeneity and therapeutic resistance in GBM.

尽管积极的治疗包括手术，然后放/化疗，但几乎所有的GBM都复发了 无进展生存期仅为10周的患者。目前还没有经过验证的治疗方法。 复发的GBM在很大程度上是由于我们对这种疾病的分子理解不足。胶质瘤干细胞(GSCs) 是肿瘤内异质性、肿瘤扩散、治疗抵抗和复发的关键决定因素。 在治疗之后。该项目题为细胞异质性的分子决定因素和治疗 GBM中的阻力汇集了一个使用最先进的GBM细胞模型的跨学科专家团队， 单细胞基因组学和创新的基因传递技术，以了解GSC的驱动机制 有助于在GBM中产生肿瘤增殖和治疗耐药细胞，目标是确定 新的治疗靶点。这项提案的目标1将寻求确定干细胞驱动机制如何 诱导治疗耐药的GSCs。从机制上讲，我们将测试Oct4和Sox2驱动/维护的假设 GSCs中通过激活TGFBR2的表达和功能而产生的耐药表型 10月4日/SOX2：ELF3：TGFBR2轴。作为一种中期策略来剖析推动治疗的分子机制- 在GBM中，我们将结合我们经过验证的细胞系统和最先进的10倍基因组铬 询问患者来源的胶质瘤细胞系的转录组和染色质状态的管道-他们的治疗- 单细胞水平上的抗性对应体。这项提议的目标2将集中在开发新的分子 靶向治疗耐药的GBM细胞群体的药物。我们将以我们的积极势头为基础 基于miRNA的治疗设计更好地抑制肿瘤的分子方法的最新进展 生长，防止出现耐药细胞亚群，并使耐药细胞对 人肾小球基底膜异种移植模型的化疗/放射治疗我们的初步数据显示miR-149-3P可以抑制8 可能的癌基因同时协调作用驱动肿瘤的维持和治疗 抵抗。该提案的目标2.1将探索miR-149-3P对治疗的机制贡献- GSCs的耐药表型。Aim 2.2将研究我们新发现的概念的临床前可译性 通过测试miRNAs的新组合来更有效地正常化由 基底膜中的干细胞驱动机制。这项研究的完成将：(I)确定能够 (2)确定能够过渡到耐药状态的细胞群体；(3) 确定与这些细胞群体相关的转录和染色质变化 治疗靶向；(Iv)提供新的合理的临床前治疗方法，以潜在地治疗复发的GBM。 该项目的完成将为理解和更有效地治疗耐药性提供路线图 因此，改善了患者的预后并挽救了生命。

Despite aggressive therapy consisting of surgery followed by radio/chemotherapy GBM recurs in almost all patients with a progression-free survival of only 10 weeks. Currently there are no proven therapies to treat recurrent GBM largely due to our inadequate molecular understanding of the disease. Glioma stem-cells (GSCs) are critical determinants of intra-tumor heterogeneity, tumor propagation, therapeutic resistance, and recurrence following treatment. This project entitled Molecular determinants of cellular heterogeneity and therapeutic resistance in GBM brings together a cross-disciplinary team of experts using state-of-the art GBM cell models, single-cell genomics, and innovative gene-delivery technology to understand how GSC-driving mechanisms contribute to the generation of tumor-propagating and therapy resistant cells in GBM with the goal of defining novel therapeutic targets. Aim 1 of this proposal will seek to determine how stem-cell driving mechanisms induce therapy resistant GSCs. Mechanistically, we will test the hypothesis that Oct4 and Sox2 drive/maintain a therapy-resistant phenotype in GSCs by activating TGFBR2 expression and function via an Oct4/Sox2:ELF3:TGFBR2 axis. As a mid-term strategy to dissect the molecular mechanism driving therapy- resistance in GBM we will combine our validated cell systems and the state-of-the-art 10x Genomics Chromium pipeline to interrogate the transcriptome and chromatin state of patient-derived glioma cell lines their therapy- resistant counterparts at the single cell level. Aim 2 of this proposal will focus on developing novel molecular agents that target therapy-resistant GBM cell populations. We will build on the positive momentum of our recent novel developments in miRNA-based therapeutics to design molecular approaches to better inhibit tumor growth, prevent emergence of therapy-resistant cell subpopulations, and sensitize therapy-resistant cells to chemo/radiation in human xenograft models of GBM. Our preliminary data shows that miR-149-3p can inhibit 8 putative oncogenes simultaneously whose coordinate action drive tumor maintenance and therapeutic resistance. Aim 2.1 of this proposal will explore the mechanistic contribution of miR-149-3p to the therapy- resistant phenotype of GSCs. Aim 2.2 will investigate the pre-clinical translatability of our new-found concepts by testing novel combinations of miRNAs to more effectively normalize oncogenic networks dysregulated by stem-cell driving mechanisms in GBM. Completion of this study will: (i) Define the cell sub-populations capable of tumor propagation; (ii) define cell populations capable of transitioning to a therapy-resistant state; (iii) determine transcriptomic and chromatin changes associated with these cell populations that are amenable to therapeutic targeting; (iv) provide novel rational pre-clinical therapeutics to potentially treat recurrent GBM. Completion of this project will provide a roadmap to both understanding and more effectively treating resistant GBM, hence improving patient outcomes and saving lives.