Targeted Inhibition in Leukemia

白血病的靶向抑制

• 批准号: 10212981
• 负责人: Jose A Cancelas
• 金额: $ 46.56万
• 依托单位: CINCINNATI CHILDRENS HOSP MED CTR
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-08 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10212981
• 关键词: Acute Myelocytic Leukemia; Adult; Allosteric Site; Apoptosis; Applications Grants; B-Cell Acute Lymphoblastic Leukemia; B-cell precursor acute lymphoblastic leukemia cell; Binding; Binding Sites; Biochemical Genetics; Biological Markers; Biophysics; Breast; Cancer Model; Cell Death; Cells; Cessation of life; Chemoresistance; Childhood; Childhood Acute Lymphocytic Leukemia; Childhood Acute Myeloid Leukemia; Clinic; Data; Development; Disease; Dose; Drug Binding Site; Drug Kinetics; Drug Targeting; Exhibits; Genes; Genetic; Glioblastoma; Goals; Growth; Guanine; Guanine Nucleotide Exchange Factors; Hematopoiesis; Human; Immunophenotyping; Immunotherapy; In Vitro; Intervention; Leukemia Acute Lymphoblastic Chemotherapy; Leukemic Cell; MAP Kinase Gene; Malignant Neoplasms; Malignant neoplasm of lung; Malignant neoplasm of prostate; Mediator of activation protein; Metallothionein; Methodology; Modeling; Molecular; Molecular Conformation; Molecular Target; Monomeric GTP-Binding Proteins; Neoplasms; Oncogenes; Oncogenic; Oncology; Pathway interactions; Patients; Ph+ ALL; Pharmaceutical Preparations; Pharmacodynamics; Phase I Clinical Trials; Philadelphia; Prognosis; Property; Proteins; Proto-Oncogene Proteins c-abl; Public Health; Relapse; Research; Resistance; Role; Salvage Therapy; Signal Pathway; Signal Transduction; Site; Site-Directed Mutagenesis; Small GTPase Activators; Solid Neoplasm; Structure; Testing; Therapeutic; Toxic effect; Translating; Transplantation; Tyrosine Kinase Inhibitor; Xenograft procedure; addiction; analog; base; bcr-abl Fusion Proteins; cancer stem cell; chemical group; chemotherapy; combinatorial; cytotoxic; drug efficacy; genetic approach; human disease; human model; in vitro Assay; in vivo; inhibitor/antagonist; leukemia; leukemic stem cell; malignant breast neoplasm; malignant stomach neoplasm; mouse model; non-oncogenic; novel; novel therapeutics; overexpression; patient derived xenograft model; preclinical safety; prevent; refractory cancer; repository; small molecule; small molecule inhibitor; stem; stem cells; therapeutically effective; tumor

ABSTRACT We have identified the first in vivo and in vitro small molecule inhibitor of Vav3, a signaling hub that is overex- pressed in many cancers and an activator of the small GTPase Rac. Rac is a major mediator of oncogenic and non-oncogenic addiction in cancer stem/progenitor cells. At low dose, our inhibitor eliminates TKI-resistance in vivo, prolongs the survival of a mouse model of pre-B-ALL, and eradicates cancer stem cell propagation in a model of mouse serial transplantation. It induces apoptosis of primary pediatric Philadelphia-positive (Ph+) and Ph-like B-ALL and chemotherapy-resistant RAM immunophenotype primary pediatric AML cells. It specifically targets leukemic cells while sparing normal hematopoiesis in vivo and shows no toxicity. In addition, it is active in oncogenic Ras xenografts mouse models of human solid tumors. Given this broad activity and the wide in- volvement of Vav3 and Rac in human disease, it is likely that our inhibitor will be efficacious in several human cancers resistant to current therapies. Even though we focus on Ph+ B-cell acute lymphoblastic leukemia (Ph+ B-ALL) as a simpler cancer model to validate the mechanism of action of our inhibitor, we will test its efficacy in models of pediatric TKI-resistant B- ALL and AML. Despite the introduction of ABL tyrosine kinase inhibitor (TKI) therapy and more recently highly- toxic immunotherapies, Ph+ B-ALL and AML remain poor prognosis diseases, especially in adults, as a result of frequent relapse and resistance to current therapies. The long-term goal of this grant application is a multidrug approach consisting of a TKI and our drug or an optimized derivative as a new therapy for ALL and AML without the toxicity associated with current salvage therapy approaches. We postulate that multitarget approaches in ALL and AML are necessary to prevent resistance to single-agent TKI therapy. Based on our preliminary data, we hypothesize that our drug increases death of leukemia initiating and propa- gating cells and overcomes TKI-resistance by targeting Vav3. The goal of the proposed research is to (1) validate the Vav3/Rac signaling axis as our drug’s target using biochemical and genetic approaches and determine in vivo implications; (2) to identify our drug’s binding site on Vav3 using biophysical, structural, and genetic ap- proaches, and validate the site using site-directed mutagenesis. Finally, (3) we will take advantage of the CCHMC Oncology Leukemia/Solid Tumor Repository to test our drug’s efficacy in PDX models of chemotherapy- resistant pediatric ALL and AML alone and in combination with existing TKI approaches and validate metallothi- onein as a biomarker. If successful, we would like to see our drug, or a more potent analog, move into pre-clinical safety analysis and potentially into a Phase I clinical trial in ALL and AML resistant to TKI therapies. Allosteric targeting of the Vav3 autoinhibited conformation as proposed here could be generalized to other ‘undruggable’ protein-protein interfaces.!

抽象的 我们已经确定了VAV3的第一个体内和体外小分子抑制剂，这是一个过度的信号枢纽 被压在许多癌症和小型GTPase RAC的激活剂中。 RAC是致癌和 癌症茎/祖细胞中的非结构成瘾。低剂量，我们的抑制剂消除了TKI抗性 体内，延长了b-b-all小鼠模型的存活，而放射性癌症干细胞在A中的传播 小鼠系列移植的模型。它诱导原发性小儿费城阳性（pH+）和 类似pH的B-ALL和耐化疗的RAM免疫表型原发性小儿AML细胞。特别是 靶向白血病细胞，同时在体内保留正常的造血症，并且表现出无毒性。另外，它是活跃的 在致癌性RAS异种移植物中，人类实体瘤的小鼠模型。考虑到这项广泛的活动和广泛的活动 VAV3和RAC在人类疾病中的体积，我们的抑制剂可能会在几个人类中有效 癌症对当前疗法有抵抗力。 即使我们专注于pH+ B细胞急性淋巴细胞白血病（pH+ b-all）作为更简单的癌症模型 验证抑制剂的作用机理，我们将在小儿耐药B-的模型中测试其有效 全部和AML。尽管引入了ABL酪氨酸激酶抑制剂（TKI）疗法，并且最近高度高度 有毒免疫疗法，pH+ B-all和AML仍然是不良的预后疾病，尤其是在成年人中，由于 经常缓解当前疗法。该赠款申请的长期目标是多饮 由TKI和我们的药物或优化衍生物作为所有人的新疗法和AML的方法 与当前的打捞方法相关的毒性。我们假设多白毒在 所有和AML都是防止对单药TKI治疗的耐药性所必需的。 基于我们的初步数据，我们假设我们的药物增加了性白血病的死亡，并提高了预言 通过靶向VAV3，门控单元和克服TKI抗性。拟议研究的目的是验证 VAV3/RAC信号轴是使用生化和遗传方法作为我们药物的靶标的 体内含义； （2）使用生物物理，结构和遗传性ap-识别我们药物在VAV3上的结合位点 使用定点诱变来探测并验证该地点。最后，（3）我们将利用 CCHMC肿瘤学白血病/实体肿瘤存储库，以测试我们在化学疗法模型中的药物效率 - 抗性小儿全部和AML，并与现有的TKI方法结合使用并验证金属丝 Onein作为生物标志物。如果成功的话，我们希望看到我们的药物或更有潜在的类似物，进入临床前 安全分析，并有可能在全体I期临床试验中进行，并且对TKI疗法具有抗性。变构 靶向VAV3自动抑制的构象，如这里所提出的，可以推广到其他“不可能” 蛋白质蛋白接口。！