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

基本信息

批准号：
10558649
负责人：
MICHAEL D HOGARTY
金额：
$ 38.71万
依托单位：
CHILDREN'S HOSP OF PHILADELPHIA
依托单位国家：
美国
项目类别：
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/10558649
关键词：
Adopted Adult Apoptosis Apoptotic BCL-2 Protein BH3 Domain BIM Bcl-2-binding protein Binding 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 Credentialing 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 Testing Therapeutic Validation Work cancer care cancer cell cell growth clinical care clinical efficacy clinical translation 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

项目摘要

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BCL2家庭生存蛋白的药物有望改变癌症护理的景观。癌症以BH3核蛋白蛋白（如BIM）的形式激活应力信号，因为它们逃脱了细胞生长约束并入侵敌对的环境。为了保持生存，肿瘤使用BCL2，BCLX，BCLW和MCL1等生存蛋白通过特定的蛋白质蛋白相互作用（PPI）隔离这些BH3蛋白。这阻止了他们的凋亡信号但也使这些肿瘤不断取决于此功能。竞争性取代的药物来自生存蛋白的BH3蛋白将其隔离为有效的凋亡信号。确实，venetoclax 是一种表现出惊人的临床功效的BCL2抑制剂，获得了FDA批准以治疗慢性淋巴细胞和成年粒细胞性白血病。但从经验上定义为依赖Bcl2的生存。相反，许多实体瘤有它们用来阻止BH3信号的生存蛋白的异质性，并且没有预测的生物标志物对这一新兴药物类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和开发诊断工具来利用BCL2家庭抑制剂供临床使用。我们创建了一个国家基础设施产生致命儿童肿瘤，神经母细胞瘤的患者 - 衍生品和细胞系模型，并使用创新的功能测定法来定义其依赖生存的Bcl2家族蛋白。我们发现了这一点神经母细胞瘤通过具有单个显性生存的PPI进行了内源激活的BIM激活的BIM 蛋白质。这隔离了BIM的凋亡活动，但也编码了连续的依赖性，并定义了机械生物标志物定义生存力所需的生存蛋白。一个意外的发现是生存依赖性是高度癌细胞固有且稳定的：在原发性的患者匹配肿瘤中一致和转移部位，以及诊断和复发。在由BCL2（BIM：BCL2 PPI）结合的BIM的肿瘤中，BCl2 诸如Venetoclax之类的抑制剂在体外和体内高度活跃。在由MCL1结合的BIM（BIM：MCL1 PPI）的肿瘤中， BCL2抑制剂没有活性。此外，我们发现所有带有MAPK途径突变的神经母细胞瘤都在 BIM：MCL1类。令人惊讶的是，尽管MCL1抑制剂对此子集也没有活性，尽管 MCL1。在这些肿瘤中，BIM由BCLX重新延迟，所有这些都对MCL1/BCLX的组合非常敏感抑制。这证明了我们的预测性BIM PPI生物标志物的鲁棒性，我们将利用这些生物标志物来识别神经母细胞瘤的所有生存依赖类别。我们将定义生物标志物保持在多大程度上稳定的内在肿瘤特征可预测对Bcl2家庭抑制剂的选择性脆弱性。我们也利用治疗性MAPK抑制剂可拮抗Mcl1依赖性。重要的是，我们试图开发体外诊断使用接近结合测定和凭证MAPK生物标志物识别预测性BIMPPI的工具定义MCL1依赖性。总的来说，我们的工作采用精确医学方法来分配Bcl2家庭用于临床使用的抑制剂，并为临床试验设计提供信息，包括预测合理组合疗法。