Validation of the Fes Tyrosine Kinase as an Inhibitor Target in Multiple Myeloma

Fes 酪氨酸激酶作为多发性骨髓瘤抑制剂靶点的验证

• 批准号: 9017965
• 负责人: Thomas E. Smithgall
• 金额: $ 20.1万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-01 至 2018-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9017965
• 关键词: Adhesions; Angiogenesis Inhibitors; Angiogenic Factor; Apoptosis; Biological Assay; Biological Models; Blood capillaries; Bone Diseases; Bone Marrow; Bone Resorption; Cell Adhesion; Cell Line; Cell Proliferation; Cell Survival; Cell surface; Cells; Chemicals; Chromosomal translocation; Clinical; Coculture Techniques; Complement; Cytoplasmic Protein; Data; Drug Targeting; Ectopic Expression; Endothelial Cells; Engineering; FPS-FES Oncogene; Frequencies; Growth; Health; Hematopoietic Neoplasms; Human; In Vitro; Inflammation; Laboratories; Lesion; Macrophage Colony-Stimulating Factor; Malignant Neoplasms; Mediating; Mediator of activation protein; Multiple Myeloma; Mus; Osteoclasts; Osteolysis; Osteolytic; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Pharmacotherapy; Phenotype; Phosphotransferases; Protein Kinase; Protein Tyrosine Kinase; Proteins; Proto-Oncogenes; RNA Interference; RNA interference screen; Reporting; Resistance; Role; Signal Transduction; Signaling Protein; Small Interfering RNA; System; TNFSF11 gene; Testing; Therapeutic; Time; Tissue Microarray; Tumor Angiogenesis; Umbilical vein; Validation; Vascular Endothelial Cell; Vascular Endothelial Growth Factors; angiogenesis; base; bone; bone loss; capillary; cathepsin K; cell growth; drug candidate; inhibitor/antagonist; insight; kinase inhibitor; knock-down; macrophage; migration; mutant; nanomolar; neoplastic cell; novel therapeutics; osteoclastogenesis; protein expression; research study; small molecule; therapeutic target; tumor; tumorigenic

 DESCRIPTION (provided by applicant): The c-fes proto-oncogene encodes the cytoplasmic protein-tyrosine kinase FES, which participates in cellular signaling cascades that govern differentiation, survival, migration and inflammation. A recent kinome-wide siRNA screen identified FES as essential for the growth and survival of human multiple myeloma (MM) cell lines. FES is also a common signaling mediator for several angiogenic factors, making FES a potential target for anti-angiogenic therapy in MM and other cancers. Recently, we reported the discovery of first-in-class small-molecule FES kinase inhibitors with nanomolar potency. Using these inhibitors, we established a new role for FES kinase activity in osteoclast differentiation from mouse bone marrow macrophages, suggesting that inhibition of this FES-dependent pathway may be of clinical benefit in osteolytic bone loss associated with MM. Together, these observations suggest that inhibition of FES kinase activity may have a three-pronged benefit in MM: 1) direct inhibition of myeloma tumor cell growth; 2) block in MM-driven osteoclast differentiation; 3) suppression of tumor angiogenesis. Here we propose to validate FES as the target for our inhibitors in each of these aspects of MM with the following Specific Aims: Aim 1: Test the hypothesis that the FES protein-tyrosine kinase is a tumor-intrinsic drug target in multiple myeloma. Although RNAi-mediated knockdown of FES expression induces apoptosis in several human myeloma cell lines, the frequency with which FES protein expression and kinase activity are upregulated in MM has not been explored. We propose to determine FES expression and activity profiles for a diverse panel of human MM cell lines, patient- derived MM cells and myeloma tissue microarrays. MM cells will then be tested for sensitivity to our FES inhibitors in terms of cell proliferation and survival. Validation of FES as the inhibitor target wll involve rescue of inhibitor sensitivity by ectopic expression of engineered inhibitor-resistant FES mutants in sensitive cells. Conversely, introduction of active FES into inhibitor-insensitive, FES-negative MM cells may render them sensitive to these compounds. Aim 2: Test the hypothesis that FES kinase activity is required for MM-associated osteolysis and angiogenesis. Recently we made the unexpected discovery that multiple classes of FES kinase inhibitors potently block osteoclast differentiation from primary mouse bone marrow macrophages and cell lines. We propose to test our Fes inhibitors for suppression of osteoclast formation from primary human macrophages, and determine whether the inhibitors also suppress their osteolytic activity in bone resorption assays. In addition, we will use our FES inhibitors to explore the role of FES kinase activity in endothelial cell proliferation, migration and capillary formation using human umbilical vein endothelial cells (HUVECs). Finally, we will evaluate the consequences of FES inhibition on MM cell adhesion to vascular endothelial cells using a MM/HUVEC co-culture system.

 描述（由适用提供）：C-FES原始癌基因编码细胞质蛋白 - 酪氨酸激酶FES，它参与了控制分化，存活，迁移和炎症的细胞信号级联。近期整个Kinome siRNA筛查对人类多发性骨髓瘤（MM）细胞系的生长和存活至关重要。 FES也是几种血管生成因子的常见信号介体，使FES成为MM和其他癌症中抗血管生成治疗的潜在靶标。最近，我们报告了具有纳摩尔效力的第一类小分子FES激酶抑制剂。使用这些抑制剂，我们确定了FES激酶活性在与小鼠骨髓巨噬细胞分化中的新作用，这表明抑制这种FES依赖性途径在与MM相关的骨化骨损失中可能具有临床益处。总之，这些观察结果表明，对FES激酶活性的抑制作用可能在MM中具有三pon依的益处：1）直接抑制骨髓瘤肿瘤细胞生长； 2）在MM驱动的破骨细胞分化中块； 3）抑制肿瘤血管生成。 Here we propose to validate FES as the target for our inhibitors in each of these aspects of MM with the following Specific Aims: Aim 1: Test the hypothesis that the FES protein-tyrosine kinase is a tumor-intrinsic Although RNAi-mediated knockdown of FES expression induces apoptosis in several human myeloma cell lines, the frequency with which FES protein expression and kinase activity are updated in MM has not been探索。我们建议确定人类MM细胞系，患者衍生的MM细胞和骨髓瘤组织微阵列的潜水员面板的FES表达和活性谱。然后，将MM细胞在细胞增殖和存活方面对我们的FES抑制剂进行敏感性。将FES作为抑制剂靶标的验证涉及通过耐药FES的生态表达来挽救抑制剂敏感性 敏感细胞中的突变体。相反，将活性FES引入对抑制剂不敏感的FES阴性MM细胞可能会使它们对这些化合物敏感。 AIM 2：检验假设的假设，即MM相关的骨溶解和血管生成需要FES激酶活性。最近，我们提出了一个意外的发现，即多种FES激酶抑制剂可能会阻断与原代小鼠骨髓巨噬细胞和细胞系的破骨细胞分化。我们建议测试我们的FES抑制剂以抑制原代人巨噬细胞的破骨细胞形成，并确定抑制剂是否还抑制了其在骨骼分辨率测定中的骨化活性。此外，我们将使用我们的FES抑制剂探索FES激酶活性在内皮细胞增殖，迁移和毛细血管形成中使用人脐静脉内皮细胞（HUVECS）的作用。最后，我们将使用MM/HUVEC共培养系统评估FES抑制对MM细胞粘附对血管内皮细胞的后果。

项目成果

Dual inhibition of Fes and Flt3 tyrosine kinases potently inhibits Flt3-ITD+ AML cell growth.

• DOI: 10.1371/journal.pone.0181178
• 发表时间: 2017
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Weir MC;Hellwig S;Tan L;Liu Y;Gray NS;Smithgall TE
• 通讯作者: Smithgall TE