Credentialing next-generation human glioma models for precision therapeutics

摘要 尽管在EGFR驱动的肺癌上取得了显著的成功，但精准治疗在EGFR驱动的肺癌上却失败了 胶质瘤是最常见和最致命的原发脑瘤。EGFR治疗在这方面失败的原因 临床背景包括缺乏忠实地概括EGFR驱动的生物学的临床前模型 胶质瘤，包括肿瘤内的异质性，专为侵袭性脑肿瘤细胞设计的药物 位于血脑屏障(BBB)后方且完好，并具有适应性耐药。在这里，我们将发展 和分子凭证新的、EGFR驱动的人脑胶质瘤模型用于临床前研究 以EGFR酪氨酸激酶抑制剂(TKI)为基础的治疗。该提案的基础来自于 Furnari Lab，他开发了一个新的平台(IGBM)，用于使用CRISPR设计脑胶质瘤模型 基因组编辑，并已建立了肿瘤内遗传异质性作为共生驱动因素 肿瘤发生学。米勒实验室在小分子实验疗法方面拥有丰富的经验 基因工程胶质瘤模型和下一代测序。他还使用了一种新的化学物质 蛋白质组学方法--多重抑制物微球与质谱仪联用--评估脑胶质瘤 并表明动态的动态组重新编程有助于靶向耐药 在神经胶质瘤模型中。他现在在阿拉巴马大学伯明翰分校，当地的合作者在那里 在生物学上忠实的人类患者来源的异种移植(PDX)模型方面有丰富的经验。这个 O‘Rourke实验室是开发复杂的胶质母细胞瘤器质性(GBO)模型的先驱，该模型 忠实地概括了分子和细胞异质性人类肿瘤的生物学。在此多PI中 在这个项目中，我们将结合我们的专业知识来解决以下目标：(1)开发新的基因 由最常见的EGFR胞外区突变驱动的工程化人类模型。到时候我们会的 从生物学和分子水平上证明这些模型可以对抗基因匹配的PDX和GBO 基因组学、表观基因组学、转录组学和动态组蛋白质组学，并使用 一组EGFR TKI，包括一个专门针对完整背后的侵袭性胶质瘤细胞的小组 BBB。(2)从生物学、分子和EGFR三个方面对异种EGFR突变IGBM模型进行验证 TKI治疗性侧写。因此，我们将开发具有定义的驱动程序突变的人类模型，这将是 PDX/GBO的有用辅助药物用于临床前药物开发。模式将被用来发展未来 合理的联合治疗，对抗耐药，提高EGFR TKI疗效。这项工作将 因此，有助于实现神经肿瘤学中尚未满足的精确治疗需求。

ABSTRACT Despite notable success in EGFR-driven lung cancer, precision therapeutics have failed in EGFR-driven gliomas, the most common and deadly primary brain tumors. Reasons for failure of EGFR therapies in this clinical context include the lack of preclinical models that faithfully recapitulate the biology of EGFR-driven gliomas, including intra-tumor heterogeneity, drugs specifically designed to target invasive brain tumor cells located behind and intact blood-brain barrier (BBB), and adaptive drug resistance. Here we will develop and molecularly credential novel, EGFR-driven human glioma models for use in preclinical development of EGFR tyrosine kinase inhibitor (TKI)-based therapies. The foundation of the proposal comes from the Furnari Lab, who developed a novel platform (iGBM) for engineering glioma models using CRISPR genome editing and has established intra-tumor genetic heterogeneity as a symbiotic driver of tumorigenesis. The Miller Lab has extensive experience in small molecule experimental therapeutics using genetically engineered gliomas model and next-generation sequencing. He also used a novel chemical proteomics method, multiplex inhibitor beads coupled with mass spectrometry, to assess the glioma kinome en masse and showed that dynamic kinome reprogramming contributes to targeted drug resistance in glioma models. He is now at the University of Alabama at Birmingham, where local collaborators have extensive experience with biologically faithful human patient-derived xenograft (PDX) models. The O’Rourke Lab is a pioneer in development of sophisticated glioblastoma organoid (GBO) models that faithfully recapitulate the biology of molecularly and cellularly heterogeneous human tumors. In this Multi-PI project, we will combine our expertise to address the following Aims: (1) To develop novel genetically engineered human models driven by the most common EGFR extracellular domain mutations. We will then biologically and molecularly credential these models against genetically-matched PDX and GBO using genomics, epigenomics, transcriptomics, and kinome proteomics, and therapeutically challenge them using a panel of EGFR TKI, including one designed to specifically target invasive glioma cells behind the intact BBB. (2) To credential heterogeneous EGFR mutant iGBM models via biological, molecular, and EGFR TKI therapeutic profiling. We will thus develop human models with defined driver mutations that will be useful adjuncts to PDX/GBO for preclinical drug development. Models will be used to develop future rational combination therapies that combat drug resistance and enhance EGFR TKI efficacy. This work will therefore help realize the unmet need of precision therapeutics in neuro-oncology.