Organoid Acquired Resistance

类器官获得性抗性

基本信息

批准号：
10517262
负责人：
Sourav Bandyopadhyay
金额：
$ 28.33万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-30 至 2027-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10517262
关键词：
Acute Address Air Alveolar Antibodies Antibody-drug conjugates Bioinformatics Biology Biopsy Blood Vessels CD44 gene CD47 gene CRISPR screen Cancer Patient Cell Communication Cells Clinical Clinical Treatment Clustered Regularly Interspaced Short Palindromic Repeats Coculture Techniques Development Drug Interactions Drug resistance Endothelium Engineering Epidermal Growth Factor Receptor Epithelial Exhibits Extracellular Matrix Fibroblasts Gefitinib Genetic Glucuronic Acids Glycosaminoglycans Goals Grant Histology Human Immune In Vitro KRAS2 gene Lethal Genes Liquid substance Lung Malignant Neoplasms Mass Spectrum Analysis Modality Modeling Molecular Mus Mutation Myelogenous Nature Non-Small-Cell Lung Carcinoma Organoids PD-1/PD-L1 PDL1 pathway PTPNS1 gene Pathway interactions Patients Penetrance Phagocytes Phagocytosis Pharmaceutical Preparations Pharmacology Study Primary Neoplasm Production Progressive Disease Proteins Proteoglycan Proteome Proteomics ROS1 gene Refractory Disease Regulation Relapse Residual state Resistance Role Sampling Series Signal Transduction System Technology Testing Therapeutic Treatment Efficacy acquired drug resistance advanced disease anticancer treatment base cell type cohesion drug sensitivity improved in vivo inhibitor lead candidate macrophage molecular targeted therapies mutant neoplastic novel patient derived xenograft model phosphoproteomics preservation programs prospective reconstitution resistance mechanism response single-cell RNA sequencing small molecule targeted agent targeted treatment therapy resistant translational approach tumor tumor microenvironment tumor xenograft virtual

项目摘要

PROJECT SUMMARY The overall goal of Project 3 of this U54 application is the organoid-based definition of mechanisms of acquired resistance (AR) to targeted therapy in non-small cell lung cancer (NSCLC). The treatment of NSCLC employs numerous therapies directed against the EGFR, ALK, KRAS G12C, ROS1 and PD-1/PD-L1 pathways. However, despite initial responses, these molecularly-targeted therapies are not curative, and virtually all patients eventually succumb to progressive disease. Acquired resistance features prominently in relapse, such as with EGFR T790M or C797S mutations following gefitinib or osimertinib treatment; however, known AR mechanisms cannot be identified for a significant proportion of patients receiving targeted therapy. Further compounding this dilemma are the distinct contributions of tumor-intrinsic versus tumor microenvironmental (TME) influences. The TME in particular contains extracellular matrix (ECM), fibroblast, vascular and immune components that can vitally impact the development and manifestations of AR. The study of TME-dependent and -independent mechanisms of AR have been further hindered by a distinct lack of in vitro human experimental systems allowing holistic recapitulation of the TME in patient biopsies. Our application leverages the synergistic strengths of the Sourav Bandyopadhyay and Calvin Kuo groups in organoid and proteomics technology to address mechanisms of EGFR and KRAS G12C AR in NSCLC. We conduct advanced proteomic approaches and organoid culture both from primary tumor biopsies at acquired resistance (from Project 1) as well as from patient-derived xenograft (PDX) tumors grown in human immune reconstituted mice (from Project 2). Our organoids are either “epithelial-only” or novel air-liquid interface (ALI) organoid cultures that co-preserve tumor epithelium en bloc with tumor-infiltrating fibroblasts and immune subsets (T, B, NK, myeloid). Aim 1 leverages the results of a NSCLC organoid CRISPR screen to determine if osimertinib-sensitizing loci can overcome, delay or alter acquired resistance in clinical biopsy/PDX AR organoids, emphasizing hits with TME mechanisms. Aim 2 uses our mass spectrometry platforms to characterize total proteome and phospho-proteomic changes in treatment-naive, residual and acquired resistance organoids and PDX, while pursuing lead candidate TROP2 and new target nominations by genetic, mechanistic and pharmacologic studies. Lastly, Project 1 has identified MIF/CD74/CD44 and CD47/SIRPa tumor-fibroblast- macrophage pathways upregulated during AR which are then functionally explored in Aim 3 using ALI organoids from AR clinical biopsies and PDX tumors. Overall, we propose a comprehensive, translational approach that exploits complementary organoid and proteomics expertise to directly study biopsies and PDX from NSCLC patients with acquired resistance to targeted therapy, towards development of improved therapeutic approaches.

项目概要该 U54 应用程序项目 3 的总体目标是基于类器官的获得性机制定义非小细胞肺癌（NSCLC）的靶向治疗耐药（AR）。许多疗法针对 EGFR、ALK、KRAS G12C、ROS1 和 PD-1/PD-L1 通路。尽管有初步反应，这些分子靶向疗法并不能治愈，而且几乎所有患者最终死于进行性疾病，获得性耐药在复发时具有显着特征，例如然而，吉非替尼或奥希替尼治疗后的 EGFR T790M 或 C797S 突变；已知的 AR 机制对于接受靶向治疗的很大一部分患者来说，无法确定这一点，这进一步加剧了这种情况。困境是肿瘤内在影响与肿瘤微环境（TME）影响的独特贡献。 TME 特别含有细胞外基质 (ECM)、成纤维细胞、血管和免疫成分，可以 TME 依赖性和非依赖性的研究对 AR 的发展和表现产生重大影响。由于明显缺乏体外人体实验系统，AR 的机制进一步受到阻碍。患者活检中 TME 的整体再现。我们的应用程序利用了 Sourav Bandyopadhyay 和 Calvin Kuo 团队的协同优势我们致力于研究类器官和蛋白质组学技术，以解决 NSCLC 中 EGFR 和 KRAS G12C AR 的机制。从原发性肿瘤活检中进行先进的蛋白质组学方法和类器官培养耐药性（来自项目 1）以及在人体免疫中生长的患者来源的异种移植 (PDX) 肿瘤重组小鼠（来自项目 2）。我们的类器官要么是“仅上皮细胞”，要么是新型气液界面 (ALI)。类器官培养物共同保存肿瘤上皮与肿瘤浸润成纤维细胞和免疫细胞目标 1 利用 NSCLC 类器官 CRISPR 筛选的结果来确定是否存在子集（T、B、NK、骨髓）。奥希替尼敏感位点可以克服、延迟或改变临床活检/PDX AR 类器官中的获得性耐药性， Aim 2 使用我们的质谱平台来强调 TME 机制的命中。初治、残留和获得性耐药类器官中的蛋白质组和磷酸化蛋白质组变化 PDX，同时通过遗传、机制和方法寻求主要候选 TROP2 和新靶点提名最后，项目 1 确定了 MIF/CD74/CD44 和 CD47/SIRPa 肿瘤-成纤维细胞- AR 期间巨噬细胞通路上调，然后使用 ALI 类器官在 Aim 3 中对其进行功能探索总的来说，我们提出了一种全面的转化方法，利用互补的类器官和蛋白质组学专业知识直接研究 NSCLC 的活检和 PDX 对靶向治疗产生耐药性的患者，寻求开发改进的治疗方法。