Molecular determinants of cellular heterogeneity and therapeutic resistance in GBM.

尽管积极的治疗，包括手术，然后是放射/化疗GBM复发，几乎所有 无进展生存期仅10周的患者。目前还没有有效的治疗方法 复发性GBM主要是由于我们对疾病的分子理解不足。胶质瘤干细胞 是肿瘤内异质性、肿瘤扩散、治疗抗性和复发的关键决定因素 治疗后。该项目题为细胞异质性的分子决定因素和治疗 GBM中的耐药性汇集了一个跨学科的专家团队，使用最先进的GBM细胞模型， 单细胞基因组学和创新的基因传递技术，以了解GSC驱动机制 有助于在GBM中产生肿瘤增殖和治疗抗性细胞，目的是定义 新的治疗靶点。本提案的目标1将寻求确定干细胞驱动机制如何 诱导治疗抗性GSC。从机制上讲，我们将测试Oct 4和Sox 2驱动/维持 通过激活TGFBR 2的表达和功能， Oct 4/Sox 2：ELF 3：TGFBR 2轴。作为一项中期战略，剖析推动治疗的分子机制- 我们将联合收割机结合我们经过验证的细胞系统和最先进的10 x Genomics Chromium 管道询问患者来源的神经胶质瘤细胞系的转录组和染色质状态， 单细胞水平的抗性对应物。该提案的目标2将集中于开发新的分子 靶向治疗抗性GBM细胞群体的药剂。我们将继续保持积极势头， 基于miRNA的治疗方法的最新进展，以设计更好地抑制肿瘤的分子方法 生长，防止治疗抗性细胞亚群的出现，并使治疗抗性细胞对 在GBM的人异种移植物模型中的化疗/辐射。我们的初步数据显示，miR-149- 3 p可以抑制8 协同作用同时驱动肿瘤维持和治疗的假定癌基因 阻力本提案的目的2.1将探索miR-149- 3 p对治疗的机制贡献， GSC的抗性表型。目标2.2将研究我们新发现的概念的临床前可翻译性 通过测试新的miRNAs组合，更有效地使因基因突变而失调的致癌网络正常化， GBM中的干细胞驱动机制。完成这项研究将：（i）确定能够 （ii）定义能够转变为治疗抗性状态的细胞群体;（iii） 确定与这些细胞群相关的转录组和染色质变化， 治疗靶向;（iv）提供新的合理的临床前治疗以潜在地治疗复发性GBM。 该项目的完成将为理解和更有效地治疗耐药提供路线图 GBM，从而改善患者预后并挽救生命。

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.