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

基本信息

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
项目类别：
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

项目摘要

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（由申请人提供）：c-fes原癌基因编码细胞质蛋白酪氨酸激酶FES，其参与控制分化、存活、迁移和炎症的细胞信号级联反应。最近的全激酶组siRNA筛选将FES确定为必需的。 FES 也是多种血管生成因子的常见信号传导介质，这使得 FES 成为人类多发性骨髓瘤 (MM) 细胞系的潜在靶点。最近，我们发现了具有纳摩尔效力的一流小分子 FES 激酶抑制剂，利用这些抑制剂，我们确定了 FES 激酶活性在小鼠破骨细胞分化中的新作用。骨髓巨噬细胞，表明抑制这种 FES 依赖性途径可能对与 MM 相关的溶骨性骨丢失具有临床益处，这些观察结果表明，抑制 FES 激酶活性可能对 MM 具有三方面的益处。 MM：1) 直接抑制骨髓瘤细胞生长；2) 阻断 MM 驱动的破骨细胞分化；3) 抑制肿瘤血管生成，我们建议在 MM 的各个方面验证 FES 作为我们的抑制剂的靶标。具体目标如下：目标 1：检验 FES 蛋白酪氨酸激酶是多发性骨髓瘤中肿瘤内在药物靶点的假设，尽管 RNAi 介导的 FES 表达敲低会诱导细胞凋亡。在几种人骨髓瘤细胞系中，尚未探索 MM 中 FES 蛋白表达和激酶活性上调的频率，我们建议确定多种人 MM 细胞系（患者来源的 MM 细胞）的 FES 表达和活性谱。然后将测试MM细胞对我们的FES抑制剂在细胞增殖和存活方面的敏感性，验证FES作为抑制剂靶标将涉及通过工程化的异位表达来挽救抑制剂敏感性。抗抑制剂FES 离线，将活性 FES 引入对抑制剂不敏感、FES 阴性的 MM 细胞中可能会使它们对这些化合物敏感。目标 2：最近测试 FES 激酶活性是 MM 相关骨溶解和血管生成所必需的假设。我们意外地发现，多类 FES 激酶抑制剂可有效阻止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 抑制剂对原代人破骨细胞形成的抑制作用。巨噬细胞，并确定抑制剂是否也在骨吸收测定中抑制其溶骨活性。此外，我们将使用我们的 FES 抑制剂，利用人脐静脉内皮细胞探索 FES 激酶活性在内皮细胞增殖、迁移和毛细血管形成中的作用。最后，我们将使用 MM/HUVEC 共培养系统评估 FES 抑制对 MM 细胞粘附到血管内皮细胞的影响。