Organoid Acquired Resistance

类器官获得性抗性

• 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
• 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

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.

项目摘要 该U54应用程序3的项目3的总体目标是基于器官的获取机制的定义 对非小细胞肺癌（NSCLC）的靶向治疗的抗性（AR）。 NSCLC员工的治疗 针对EGFR，ALK，KRAS G12C，ROS1和PD-1/PD-L1途径的许多疗法。然而， 尽管有初步反应，但这些分子靶向的疗法均未治愈，几乎所有患者 最终屈服于进行性疾病。在继电器中显着获得的阻力功能，例如 吉非替尼或osimertinib治疗后的EGFR T790M或C797S突变；但是，已知的AR机制 对于接受靶向治疗的患者，无法确定相当一部分的患者。进一步加剧了这一点 困境是肿瘤内在与肿瘤微环境（TME）影响的不同贡献。这 特别是TME包含细胞外基质（ECM），成纤维细胞，血管和免疫成分 在生命中影响AR的发展和表现。 TME依赖性和非依赖性的研究 由于明显缺乏体外人类实验系统，AR的机制进一步阻碍了AR的机制 在患者活检中对TME的整体概括。 我们的应用利用Sourav Bandyopadhyay和Calvin Kuo组的协同优势 在NSCLC中，有机体和蛋白质组学技术解决了EGFR和KRAS G12C AR的机制。我们 从获得的原发性肿瘤活检中进行晚期蛋白质组学方法和类器官培养 阻力（来自项目1）以及人类免疫中生长的患者衍生源（PDX）肿瘤 重构小鼠（来自项目2）。我们的器官是“仅上皮”或新型的空气界面（ALI） 与肿瘤浸润的成纤维细胞和免疫的类器官培养 子集（T，B，NK，髓样）。 AIM 1利用NSCLC Organoid CRISPR屏幕的结果确定是否是否 osimertinib敏感性局部可以在临床活检/PDX AR器官中克服，延迟或改变获得的耐药性， 用TME机制强调命中。 AIM 2使用我们的质谱平台来表征总数 蛋白质组和磷酸 - 蛋白质组的治疗，残留和获得的耐药器官的变化以及 PDX，同时通过遗传，机械和 药理学研究。最后，项目1已确定MIF/CD74/CD44和CD47/SIRPA肿瘤 - 纤维细胞 - 在AR期间更新的巨噬细胞途径，然后在AIM 3中使用ALI类型探索该途径 来自AR临床活检和PDX肿瘤。总体而言，我们提出了一种全面的翻译方法 利用完整的器官和蛋白质组学专业知识，可以直接研究NSCLC的活检和PDX 具有对靶向治疗的耐药性的患者，可发展改善治疗方法。