Targeting PTPN11 dependent Hematologic Malignancies

靶向 PTPN11 依赖性血液恶性肿瘤

• 批准号: 10057374
• 负责人: BILL H CHANG
• 金额: $ 35.23万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057374
• 关键词: Acute Myelocytic Leukemia; Adaptor Signaling Protein; Address; Animal Model; Biochemical; Biochemical Pathway; Biological Assay; Biological Models; Bone Marrow Cells; CRISPR/Cas technology; Cell Line; Cell Survival; Cell model; Cells; Chemicals; Chemoresistance; Clinical; Clinical Data; Combined Modality Therapy; Complex; Data; Dependence; Development; Disease; Drug Combinations; Evaluation; Foundations; Goals; Hematologic Neoplasms; Hematology; Hematopoietic; Immunodeficient Mouse; In Vitro; Juvenile Myelomonocytic Leukemia; Knowledge; Leukemic Cell; MAP Kinase Gene; MAPK Signaling Pathway Pathway; MEKs; Malignant Neoplasms; Modeling; Molecular; Mus; Mutation; Pathogenesis; Pathway interactions; Patients; Phosphotransferases; Play; Prognosis; Protein Tyrosine Kinase; Protein Tyrosine Phosphatase; Proteins; RNA Interference; Role; Sampling; Signal Transduction; Solid Neoplasm; Tertiary Protein Structure; Testing; Therapeutic; Transgenic Organisms; Treatment Protocols; Validation; Xenograft procedure; acute myeloid leukemia cell; base; cell killing; clinical development; effective therapy; experience; improved; in vivo; inhibitor/antagonist; leukemia; mouse model; mutant; neoplastic cell; novel; pre-clinical; protein complex; protein phosphatase inhibitor-2; response; small molecule inhibitor; targeted treatment; therapeutic target

Activating mutations in Protein Tyrosine Phosphatase, non-receptor type 11 (PTPN11) are seen in approximately 5-10% percent of patients with acute myeloid leukemia (AML) that independently confers a poor prognosis, and is the leukemic driver in 35% of patients with the rare chemoresistant juvenile myelomonocytic leukemia (JMML). To date, targeting PTPN11 for clinical benefit has not been well established. Our preliminary data using high-throughput functional assays have identified that PTPN11 activity is dependent on an upstream tyrosine kinase, TNK2, and that leukemic cells with activating PTPN11 mutants are sensitive to TNK2 inhibition. The long-term goal of this proposal is to identify novel combinations of targeted therapies using PTPN11/TNK2 dependent leukemias as a model. The immediate goals are to comprehensively understand the molecular mechanisms of PTPN11 activation, and to define therapeutic regimens for treatment based on the central hypothesis that PTPN11-dependent leukemias are directly activated by TNK2. To accomplish these goals, several key AIMS will need to be addressed: 1) Develop a detailed understanding of the biochemical pathways interacting with activated PTPN11. We will use molecular and biochemical studies to dissect the interactions of proteins within PTPN11/TNK2 and downstream pathways through heterologous model systems, PTPN11 mutant AML cell lines, murine bone marrow cells with mutant PTPTN11, and primary mutant PTPN11 AML samples. Further, RNAi and CRISPR technology will be utilized to selectively inhibit expression of candidate proteins to identify critical interactions necessary for PTPN11/TNK2 activity within hematopoietic model systems. These studies will establish the molecular and biochemical foundation necessary for defining PTPN11/TNK2 activation, and help guide novel targeted approaches. 2) Test combination pathway inhibition to overcome escape from monotherapy. We will use our experience and expertise with small molecule inhibitors to identify synergistic combinations of inhibitors to TNK2, PTPN11, modulators of PTPN11/TNK2 activation, and downstream effectors in vitro using cell lines and primary patient samples. Molecular and biochemical studies will be performed on samples that overcome therapy to identify escape mechanisms. These studies will define the use of combination therapy for treatment of PTPN11-driven leukemias and provide critical pre-clinical data for further development of these combinations. 3) Test combination therapies in the animal model. We will test the above combinations in vivo using mouse models harboring PTPN11 mutations, as well as primary patient leukemia samples with PTPN11 mutations xenografted into immunodeficient mice. These studies will build the preclinical foundation for combination of targeted therapies for eventual use in patients with PTPN11-driven leukemias.

蛋白酪氨酸磷酸酶，非受体11型（PTPN 11）的激活突变见于 大约5%-10%的急性髓细胞白血病（AML）患者独立地产生不良的 预后，并且是35%的罕见化疗耐药的青少年骨髓单核细胞白血病患者的白血病驱动因素。 白血病（JMML）。迄今为止，靶向PTPN 11的临床获益尚未得到很好的确立。我们 使用高通量功能测定的初步数据已经确定PTPN 11活性依赖于 上游酪氨酸激酶TNK 2，并且具有活化PTPN 11突变体的白血病细胞对 TNK 2抑制。该提案的长期目标是确定靶向药物的新组合， 使用PTPN 11/TNK 2依赖性白血病作为模型的治疗。近期目标是 全面了解PTPN 11激活的分子机制，并定义 基于PTPN 11依赖性白血病的中心假设的治疗方案 直接被TNK 2激活。为了实现这些目标，需要解决几个关键的AIMS： 详细了解与活化的PTPN 11相互作用的生化途径。我们将 使用分子和生物化学研究来剖析PTPN 11/TNK 2内蛋白质的相互作用， 通过异源模型系统的下游通路，PTPN 11突变AML细胞系，鼠骨 具有突变型PTPTN 11的骨髓细胞和原代突变型PTPN 11 AML样品。此外，RNA干扰和 CRISPR技术将用于选择性抑制候选蛋白的表达，以识别 造血模型系统中PTPN 11/TNK 2活性所必需的关键相互作用。这些研究 将建立定义PTPN 11/TNK 2激活所需的分子和生物化学基础， 帮助指导新的有针对性的方法。2)测试组合途径抑制以克服逃避 单一疗法。我们将利用我们在小分子抑制剂方面的经验和专业知识， TNK 2、PTPN 11的抑制剂、PTPN 11/TNK 2活化的调节剂和下游调节剂的组合 使用细胞系和原代患者样品进行体外效应物。分子和生物化学研究将是 在克服治疗的样本上进行，以确定逃逸机制。这些研究将确定使用 联合治疗PTPN 11驱动的白血病，并提供关键的临床前数据， 这些组合的进一步发展。3)在动物模型中测试组合疗法。我们将测试 使用携带PTPN 11突变的小鼠模型以及原发性患者， 将具有PTPN 11突变的白血病样品异种移植到免疫缺陷小鼠中。这些研究将建立 联合靶向治疗的临床前基础，最终用于PTPN 11驱动的患者 白血病