Mechanistic biomarkers to enable Bcl2 inhibitor therapies for neuroblastoma

机制生物标志物使 Bcl2 抑制剂能够治疗神经母细胞瘤

• 批准号: 10356118
• 负责人: MICHAEL D HOGARTY
• 金额: $ 39.37万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-02-18 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10356118
• 关键词: Adopted; Adult; Apoptosis; Apoptotic; BCL-2 Protein; BH3 Domain; BIM Bcl-2-binding protein; Biological Assay; Biological Markers; Cancer Etiology; Cancer Patient; Cell Line; Cellular Stress; Cessation of life; Chronic; Clinic; Clinical; Clinical Trials Design; Co-Immunoprecipitations; Combined Modality Therapy; Data; Death Domain; Dependence; Development; Diagnosis; Diagnostic tests; Environment; Exposure to; Family; Family member; Formalin; Genomics; Goals; Growth; Hematologic Neoplasms; Heterogeneity; In Situ; In Vitro; Infrastructure; Invaded; Knowledge; Ligation; Lymphocyte; MAP Kinase Gene; MCL1 gene; MEKs; Malignant Neoplasms; Measures; Mediating; Mitochondria; Mitogen-Activated Protein Kinase Inhibitor; Modeling; Mutation; Myeloid Leukemia; Neuroblastoma; Outcome; Paraffin Embedding; Pathway interactions; Patients; Pediatric Neoplasm; Pharmaceutical Preparations; Play; Primary Neoplasm; Process; Protein Family; Proteins; Public Health; Relapse; Research; Resistance; Role; Signal Transduction; Site; Slide; Solid Neoplasm; Stress; Tertiary Protein Structure; Therapeutic; Validation; Work; base; cancer care; cancer cell; cell growth; clinical care; clinical efficacy; clinically translatable; cytochrome c; diagnostic assay; diagnostic tool; drug efficacy; genomic predictors; improved; in vivo; in-vitro diagnostics; inhibitor; inhibitor therapy; innovation; insight; mortality; neoplastic cell; novel diagnostics; patient derived xenograft model; patient subsets; precision medicine; predictive marker; predictive test; protein protein interaction; resistance mechanism; response; survival prediction; synergism; tumor

Project Summary Drugs that inhibit Bcl2-family survival proteins promise to change the landscape of cancer care. Cancers activate stress signals in the form of BH3-domain proteins like Bim as they escape cellular growth constraints and invade hostile environments. To remain viable, tumors use survival proteins like Bcl2, Bclx, Bclw and Mcl1 to sequester these BH3 proteins through specific protein-protein interactions (PPIs). This blocks their apoptotic signal but also renders such tumors continually dependent on this function. Drugs that competitively displace BH3 proteins from the survival protein sequestering them unleash a potent apoptotic signal. Indeed, venetoclax is a Bcl2 inhibitor that has demonstrated striking clinical efficacy, garnering FDA-approval for the treatment of chronic lymphocytic and adult myelogenous leukemias.These tumor types do not have Bcl2-activatingmutations but are empirically defined to be dependent on Bcl2 for their survival. In contrast, many solid tumors have heterogeneity in which survival protein they use to block BH3 signals, and the absence of biomarkers that predict sensitivity to this emerging drug class remains a barrier. Our objectives are to identify predictive biomarkers and develop diagnostic tools to leverage Bcl2-family inhibitors for clinical use. We created a national infrastructure to generate patient-derivedxenograftand cell line models of the lethal childhood tumor, neuroblastoma, and utilized innovative functional assays to define the Bcl2-family protein they depend on for survival. We discovered that neuroblastomas have endogenously activated Bim neutralized through a PPI with a single dominant survival protein. This sequesters Bim's apoptotic activity, but also encodes a continual dependency, and defines a mechanistic biomarker defining the survival protein required for viability. An unanticipated finding is that this survival dependency is highly cancer cell intrinsic and stable: consistent in patient-matched tumors from primary and metastatic sites, and at diagnosis and relapse. In tumors with Bim bound by Bcl2 (Bim:Bcl2 PPI), Bcl2 inhibitors like venetoclax are highly active in vitro and in vivo. In tumors with Bim bound by Mcl1 (Bim:Mcl1 PPI), Bcl2 inhibitors have no activity. Further, we find that all neuroblastomas with MAPK pathway mutations are in the Bim:Mcl1 class. Surprisingly, Mcl1 inhibitors also have no activity for this subset, despite displacing Bim from Mcl1. In these tumors, Bim is re-sequestered by Bclx, and all are exquisitely sensitive to combined Mcl1/Bclx inhibition. This demonstrates the robustness of our predictive Bim PPI biomarker that we will exploit to identify all survival dependency classes in neuroblastoma. We will define the extent to which the biomarker remains a stable intrinsic tumor feature that predicts selective vulnerability to Bcl2-family inhibitors. We also leverage therapeutic MAPK inhibitors to antagonize Mcl1 dependency. Importantly, we seek to develop in vitro diagnostic tools to identify predictive BimPPIs using proximity-ligation assays, and to credential genomic MAPK biomarkers to define Mcl1 dependence. Collectively, our work applies precision medicine approaches to assign Bcl2-family inhibitors for clinical use, and inform clinical trial designs, including predicting rational combination therapies.

项目摘要 抑制Bcl 2家族生存蛋白的药物有望改变癌症治疗的前景。癌症 激活BH 3结构域蛋白质形式的应激信号，如Bim，因为它们逃避细胞生长限制 并入侵敌对环境。为了保持活力，肿瘤使用存活蛋白，如Bcl 2，Bclx，Bclw和Mcl 1 通过特定的蛋白质-蛋白质相互作用（PPI）来隔离这些BH 3蛋白质。这可以阻止它们的凋亡 信号，但也使这种肿瘤持续依赖于这种功能。竞争性取代 BH 3蛋白从生存蛋白中分离出来，释放出一种强有力的凋亡信号。的确，venetoclax 是一种Bcl 2抑制剂，已证明具有显著的临床疗效，获得FDA批准用于治疗 慢性淋巴细胞白血病和成人骨髓性白血病。这些肿瘤类型没有Bcl 2激活突变 但根据经验被定义为依赖Bcl 2存活。相比之下，许多实体瘤具有 他们使用生存蛋白来阻断BH 3信号的异质性，以及缺乏预测BH 3信号的生物标志物。 对这类新兴毒品的敏感性仍然是一个障碍。我们的目标是确定预测性生物标志物， 开发诊断工具，以利用Bcl 2家族抑制剂用于临床。我们建立了国家基础设施， 产生致命的儿童肿瘤，神经母细胞瘤的患者来源的异种移植物和细胞系模型， 创新的功能测定，以确定Bcl 2家族蛋白，他们依赖于生存。我们发现 神经母细胞瘤通过PPI中和内源性激活的Bim， 蛋白这隔离了Bim的凋亡活性，但也编码了一种持续的依赖性，并定义了一种依赖性。 定义生存力所需的存活蛋白的机械生物标志物。一个意想不到的发现是， 生存依赖性是高度癌细胞内在的和稳定的：在原发性肿瘤患者匹配的肿瘤中一致 和转移部位，以及在诊断和复发时。在Bim与Bcl 2结合的肿瘤中（Bim：Bcl 2 PPI），Bcl 2 抑制剂如维奈托克在体外和体内都具有高活性。在Bim与Mcl 1结合的肿瘤中（Bim：Mcl 1 PPI）， bcl 2抑制剂没有活性。此外，我们发现所有有MAPK通路突变的神经母细胞瘤都是在神经母细胞瘤中发生的。 Bim：Mcl 1类。令人惊讶的是，Mcl 1抑制剂对该亚组也没有活性，尽管从 Mcl 1.在这些肿瘤中，Bim被Bclx重新隔离，并且所有这些肿瘤都对Mcl 1/Bclx组合非常敏感 抑制作用这证明了我们的预测Bim PPI生物标志物的稳健性，我们将利用它来识别 神经母细胞瘤的所有生存依赖分类。我们将定义生物标志物在多大程度上仍然是 稳定的内在肿瘤特征预测对Bcl 2家族抑制剂的选择性脆弱性。我们还利用 治疗性MAPK抑制剂拮抗Mcl 1依赖性。重要的是，我们寻求开发体外诊断 使用邻位连接测定法鉴定预测性BimPPI的工具，以及鉴定基因组MAPK生物标志物的工具 定义Mcl 1依赖性。总的来说，我们的工作应用精确医学方法来分配Bcl 2家族 抑制剂的临床应用，并告知临床试验设计，包括预测合理的联合治疗。