Dynamic BH3 Profiling with Patient Derived Organoids of Esophageal Cancer and Mesothelioma Enable Precision-Based Targeting of the Mitochondrial Apoptotic Pathway

使用患者来源的食管癌和间皮瘤类器官进行动态 BH3 分析，实现线粒体凋亡途径的精确靶向

• 批准号: 10459596
• 负责人: Robert Taylor Ripley
• 金额: $ 8万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10459596
• 关键词: Apoptosis; Apoptotic; Asbestos; BCL-2 Protein; BCL2 gene; Barrett Esophagus; Bile Reflux; Biochemical; Biological Assay; Biopsy; Bypass; Carcinogens; Cell Culture Techniques; Cell Death; Cells; Cessation of life; Chronic; Cisplatin; Clinical; Cytotoxic Chemotherapy; Data; Drug Targeting; Environmental Carcinogens; Equilibrium; Esophageal Adenocarcinoma; Exposure to; Family; Genomic Instability; Goals; Induction of Apoptosis; Link; MCL1 gene; Malignant - descriptor; Malignant Neoplasms; Malignant Pleural Mesothelioma; Malignant neoplasm of esophagus; Measures; Mesothelioma; Methods; Minority; Mission; Mitochondria; Modeling; Mus; Mutate; Mutation; Oncogenic; Organoids; Outer Mitochondrial Membrane; Pathway interactions; Patient Care; Patient Selection; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Prediction of Response to Therapy; Protein Family; Proteins; Public Health; Reflux; Research; Resistance; Sampling; Somatic Mutation; Stimulus; Survival Rate; Techniques; Testing; Therapeutic; Translating; Up-Regulation; base; bile salts; cancer cell; carcinogenesis; carcinogenicity; chemotherapy; clinically actionable; combinatorial; driver mutation; improved; indexing; inhibitor; major outer membrane protein; metaplastic cell transformation; mimetics; mutant; neoplastic; neoplastic cell; novel strategies; overexpression; precision medicine; prevent; refractory cancer; resistance mechanism; small hairpin RNA; therapeutic target; therapy resistant; treatment response; tumor; tumor growth; tumor metabolism; tumorigenesis

SUMMARY / ABSTRACT Esophageal cancer and malignant pleural mesothelioma are difficult to treat because they typically harbor over 3000 somatic mutations from the repeated insults of reflux and asbestos. Identifying one mutation or pathway to target is circumvented by the tumor cell through new mutations and bypass pathways. For this reason, targeting the mitochondrial pathways represents a substantially improved approach because they are downstream of oncogenic driver proteins and pathway mutations. Recently, we have shown that chronic exposure of pre-neoplastic, Barrett’s esophageal cells to bile salt induced malignant transformation through a mechanism termed, ‘Minority MOMP (mitochondrial outer membrane permeabilization)’. MOMP is regulated by the B-cell lymphoma-2 (Bcl-2) family of proteins that are divided into pro- and anti-apoptotic proteins that interact at Bcl-2 homology-3 (BH3) domains. Minority MOMP partially activates the intrinsic pathway of the apoptotic machinery to a level not sufficient to result in cell death; rather, it promotes genomic instability, cellular transformation, and tumorigenesis. We noted that in ‘Minority MOMP’, Barrett’s cells resisted apoptosis through the upregulation of the anti-apoptotic protein, Mcl-1. When we targeted Mcl-1, Minority MOMP shifted from the sub-lethal mitochondrial activation to frank apoptosis and tumor cells died. Dynamic BH3 profiling (DBP) provides an assay to measure which anti-apoptotic proteins are responsible for the resistance for each tumor. By determining the relevant anti-apoptotic protein, a class of compounds, BH3 mimetics, target those specific protein. Recently, a biochemical ‘toolkit’ utilized DBP to identify the appropriate BH3 mimetic in murine cells that overexpressed bcl-2 proteins. Patient-derived organoids (PDO) can recapitulate tumor response to therapy. If a comprehensive biochemical toolkit utilizing DBP in PDOs identifies proteins that enable resistance directly from patient tumors, then precision-based targeting of the Bcl-2 proteins provides a therapeutic strategy to overcome treatment-refractory cancers. Our goal is to disrupt the mitochondrial balance that enables carcinogenesis but blocks apoptosis by directly targeting the proteins responsible for resistance. We will establish a biochemical toolkit to predict treatment response in esophageal cancer and mesothelioma by utilizing a DBP-PDO model. Targeting these proteins will shift Minority MOMP toward apoptosis by blocking the compensatory anti-apoptotic proteins. Our hypothesis is that the DBP-PDO model is a clinically actionable bioassay within seven days from tumor biopsy. This current research will allow us to circumvent intractable bypass mechanisms and mutations of cancer cells altogether by targeting the downstream mitochondrial resistance mechanism. This novel strategy is expected to shift Minority MOMP toward frank apoptosis and thus render these cells vulnerable to standard therapies.

摘要/摘要 食道癌和恶性胸膜间皮瘤很难治疗，因为它们通常隐藏着 反流和石棉的反复侵害导致 3000 种体细胞突变，识别一种突变或途径。 肿瘤细胞通过新的突变和旁路途径绕过了靶点。 针对线粒体途径代表了一种显着改进的方法，因为它们 致癌驱动蛋白和途径突变的下游。 最近，我们发现，肿瘤前巴雷特食管细胞长期暴露于胆盐会诱导 通过称为“少数 MOMP（线粒体外膜）”的机制进行恶性转化 MOMP 受 B 细胞淋巴瘤-2 (Bcl-2) 蛋白家族的调节，该蛋白家族分为以下几类： 在 Bcl-2 同源性 3 (BH3) 结构域部分相互作用的促凋亡蛋白和抗凋亡蛋白。 激活细胞凋亡机制的内在途径，其水平不足以导致细胞死亡； 我们在“少数 MOMP”中注意到，促进基因组不稳定、细胞转化和肿瘤发生。 当我们靶向时，Barrett 细胞通过抗凋亡蛋白 Mcl-1 的上调来抵抗细胞凋亡。 Mcl-1，少数 MOMP 从亚致死线粒体激活转变为明显的细胞凋亡和肿瘤细胞 动态 BH3 分析 (DBP) 提供了一种测定方法来测量哪些抗凋亡蛋白负责。 对于每种肿瘤的耐药性，通过确定相关的抗凋亡蛋白，一类化合物，BH3。 最近，一个生化“工具包”利用 DBP 来识别合适的蛋白质。 过度表达 bcl-2 蛋白的小鼠细胞中的 BH3 模拟物可以。 概括肿瘤对治疗的反应 如果在 PDO 中使用 DBP 的综合生化工具包。 识别能够直接从患者肿瘤中产生耐药性的蛋白质，然后精确靶向 Bcl-2 蛋白提供了克服难治性癌症的治疗策略。 我们的目标是通过直接破坏线粒体平衡来促进致癌作用但阻止细胞凋亡 我们将建立一个生化工具包来预测治疗。 通过利用 DBP-PDO 模型来靶向这些蛋白质将产生食管癌和间皮瘤的反应。 通过阻断补偿性抗凋亡蛋白，将少数 MOMP 转向凋亡。我们的假设是。 DBP-PDO 模型是肿瘤活检后 7 天内可进行临床操作的生物测定。 研究将使我们能够通过以下方式完全规避棘手的旁路机制和癌细胞突变： 针对下游线粒体耐药机制，这一新策略有望改变少数群体。 MOMP 导致明显的细胞凋亡，从而使这些细胞容易受到标准疗法的影响。