Credentialing next-generation human glioma models for precision therapeutics

摘要 尽管在EGFR驱动的肺癌方面取得了显着成功，但精准治疗在EGFR驱动的肺癌方面却失败了 神经胶质瘤，最常见和致命的原发性脑肿瘤。EGFR治疗失败的原因 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， 神经胶质瘤，包括肿瘤内异质性，专门针对侵袭性脑肿瘤细胞设计的药物 位于血脑屏障（BBB）后且完整，以及适应性耐药。在这里，我们将开发 并在分子上证明EGFR驱动的新型人脑胶质瘤模型可用于临床前开发， 基于EGFR酪氨酸激酶抑制剂（TKI）的治疗。该提案的基础来自于 Furnari实验室开发了一种新的平台（iGBM），用于使用CRISPR构建神经胶质瘤模型。 基因组编辑，并建立了肿瘤内遗传异质性作为一个共生的驱动因素， 肿瘤发生米勒实验室在小分子实验疗法方面拥有丰富的经验， 基因工程神经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，多重抑制剂珠与质谱联用，以评估胶质瘤 kinome envelope研究表明，动态kinome重编程有助于靶向耐药， 在神经胶质瘤模型中。他现在在伯明翰的亚拉巴马大学，当地的合作者已经 在生物学上忠实的人类患者源性异种移植物（PDX）模型方面拥有丰富的经验。的 O 'Rourke实验室是开发复杂的胶质母细胞瘤类器官（GBO）模型的先驱， 忠实地概括了分子和细胞异质性人类肿瘤的生物学。在这个多PI 项目，我们将联合收割机我们的专业知识，以解决以下目标：（1）开发新的基因 由最常见的EGFR细胞外结构域突变驱动的工程化人类模型。然后我们将 生物学和分子学证明这些模型对遗传匹配的PDX和GBO， 基因组学、表观基因组学、转录组学和激酶组蛋白质组学，并使用 一组EGFR TKI，包括一种设计用于特异性靶向完整肿瘤细胞后面的侵袭性胶质瘤细胞的药物。 BBB. (2)通过生物学、分子学和EGFR鉴定异质性EGFR突变体iGBM模型 TKI治疗分析。因此，我们将开发具有定义的驱动突变的人类模型， 对PDX/GBO进行临床前药物开发是有用的。模型将用于开发未来 合理的联合治疗，对抗耐药性，增强EGFR TKI疗效。这项工作将 因此有助于实现神经肿瘤学中精确治疗的未满足需求。

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.