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 食道癌和恶性胸膜间皮瘤很难治疗，因为它们通常藏在 反射X和石棉的反复事件发生了3000个体细胞突变。识别一个突变或途径 通过新的突变和旁路途径，肿瘤细胞绕过靶标。为此原因， 针对线粒体途径代表了一种大大改进的方法，因为它们是 致癌驱动蛋白和途径突变的下游。 最近，我们表明，巴雷特的食管诱导的疏松前塑性细胞长期暴露于 通过称为“少数MOMP（线粒体外膜）的机制”的恶性转化 透化）’。 MOMP受B细胞淋巴瘤-2（BCL-2）蛋白质的调节，分为 在Bcl-2同源性3（BH3）结构域相互作用的促凋亡蛋白和抗凋亡蛋白。少数MOMP部分 将凋亡机制的固有途径激活到不足以导致细胞死亡的水平。相反，它 促进基因组不稳定性，细胞转化和肿瘤发生。我们注意到，在“少数MOMP”中， Barrett的细胞通过上调抗凋亡蛋白MCL-1的上调抵抗凋亡。当我们目标时 MCL-1，少数MOMP从致命的线粒体激活转变为弗兰克凋亡和肿瘤细胞 死了。动态BH3分析（DBP）提供了评估，以测量哪些抗凋亡蛋白是负责的 对于每个肿瘤的抗性。通过确定相关的抗凋亡蛋白，一类化合物BH3 模拟物，靶向那些特定的蛋白质。最近，生化的“工具包”利用DBP来识别适当的 BH3模拟于过表达Bcl-2蛋白的鼠细胞中。患者衍生的类器官（PDO）可以 概括肿瘤对治疗的反应。如果在PDOS中使用DBP的全面生化工具包 鉴定蛋白质直接从患者肿瘤中抗性，然后基于精确的靶向 Bcl-2蛋白提供了克服治疗难治性癌症的治疗策略。 我们的目标是破坏能够实现癌变但直接阻止凋亡的线粒体平衡 靶向负责抗性的蛋白质。我们将建立一个生化工具包，以预测治疗 通过使用DBP-PDO模型，食管癌和间皮瘤的反应。针对这些蛋白质将 通过阻断补偿性抗凋亡蛋白，将少数MOMP转移到凋亡中。我们的假设是 DBP-PDO模型在肿瘤活检的7天内是临床上可起作用的生物测定法。此电流 研究将使我们能够完全避免棘手的旁路机制和癌细胞的突变 靶向下游线粒体电阻机制。这种新颖的策略有望改变少数 MOMP朝着弗兰克凋亡，从而使这些细胞容易受到标准疗法的影响。