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: 10285093
• 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/10285093
• 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/antagonist; major outer membrane protein; metaplastic cell transformation; mimetics; mutant; neoplastic; neoplastic cell; novel strategies; overexpression; precision medicine; prevent; refractory cancer; resistance mechanism; response; 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 食管癌和恶性胸膜间皮瘤是治疗的差异 反复侮辱和石棉的反复侮辱中的3000个体细胞突变。 肿瘤细胞通过新的突变和旁路途径绕过靶标。 靶向米孔室途径代表了一种实质上改进的方法，因为它们是 致癌驱动蛋白和途径突变的下游。 最近，我们已经表明，巴雷特的食管细胞慢性肿瘤诱发了胆汁盐诱导 通过称为“少数MOMP的机制”（线粒体外膜）的恶性转化 透化）MOMP由B细胞淋巴瘤-2（BCl-2）蛋白质定期 Bcl-2同源3（BH3）结构域的促抗凋亡蛋白和抗凋亡蛋白。 将凋亡机械的固有途径激活到水平而不是窒息的水平，从而导致细胞死亡。 促进基因组不稳定性，细胞转化和肿瘤发生。 Barrett的细胞通过抗磷灰石蛋白的上调MCL-1来抵抗凋亡 MCL-1，少数MOMP从致命的mitchondrial激活转变为弗兰克腹部和肿瘤细胞 死亡。 通过确定相关的抗吞噬蛋白的抗性。 Mimetics，靶向那些特定的蛋白质。 过表达Bcl-2蛋白的鼠细胞中的BH3模拟物。 概括肿瘤对治疗的反应。 识别直接从患者肿瘤耐药的蛋白质，然后基于精确的靶向靶向 Bcl-2蛋白提供了一种克服治疗难治性癌症的治疗策略。 我们的目标是破坏线粒体平衡，使癌变能够直接阻止凋亡 靶向负责抗性的蛋白质。 用DBP-PDO模型将食管癌癌症癌症癌症的反应。 通过阻止补偿性抗寄生虫蛋白的少数MOMP向凋亡 DBP-PDO模型是在肿瘤活检中七天内临床可行的生物测定法 研究将使我们能够避免由By By By By By By By By By By By By By By By By By By By By By By By By By By By By By By By By By By Cancer contable机制和突变。 靶向下游线粒体电阻机制。 MOMP朝向弗兰克凋亡，从而使这些细胞容易受到标准疗法的影响。