P21-activated kinase 1 and 2 as therapeutic targets of the NF1 microenvironment

P21 激活激酶 1 和 2 作为 NF1 微环境的治疗靶点

基本信息

批准号：
8527331
负责人：
RACHELLE KOSOFF
金额：
$ 2.88万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-15 至 2014-09-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8527331
关键词：
Adhesions Adverse effects Biochemical Bone Marrow Bone Marrow Transplantation C-KIT Gene Cell Culture Techniques Cell Proliferation Cell Survival Cell physiology Cells Chemotactic Factors Clinic Collaborations Collagen Communication Data Deposition Dermal Disease Drug resistance Family Gene Mutation Genetic Growth Growth Factor Human Immigration In Vitro Infiltration Inflammatory Injection of therapeutic agent Intervention MAP Kinase Gene Malignant Childhood Neoplasm Malignant Neoplasms Measures Mediating Mediator of activation protein Modeling Mus Mutant Strains Mice NF1 gene Neurofibromatosis 1 Neurofibromatosis Type 1 Protein Operative Surgical Procedures Passive Cutaneous Anaphylaxis Pathway interactions Patients Peripheral Nerves Pharmaceutical Preparations Phenotype Phosphotransferases Play Protein Array Protein Isoforms Protein Tyrosine Kinase Proto-Oncogene Protein c-kit Research Role Safety Schwann Cells Signal Pathway Signal Transduction Skin Stromal Cells Transplantation Work angiogenesis cell growth cell motility cell type design drug efficacy in vivo inhibitor/antagonist mast cell migration mouse model mutant neoplastic cell neurofibroma p21-activated kinase 1 prevent public health relevance receptor research study response small molecule therapeutic target tumor tumor growth tumor initiation tumor microenvironment tumor progression tumorigenic

项目摘要

DESCRIPTION (provided by applicant): p21-activated kinase 1 and 2 as therapeutic targets of the NF1 microenvironment Neurofibromatosis Type I (NF1) is characterized by neurofibromas consisting of NF1-null schwann cells and a mixture of Nf1-heterozygous stromal cells. Cross talk between mutant stromal cells and schwann cells promotes angiogenesis, collagen deposition and mast cell infiltration. Therapeutic targeting of the c-kit signaling pathway in mas cells could potentially reduce neurofibromin formation. Nf1-heterozygous mast cells are hyperactive for c-kit receptor mediate migration, proliferation and chemoattraction to Schwann cells. By disrupting hyperactive c-kit signaling in mast cells and regulating the tumor microenvironment, we can disrupt tumor growth in NF1 and other tumors dependent on mast cells. Recent discoveries in our lab demonstrates that enhanced mast cell signaling downstream of Nf1 requires p21-activated kinase 1 (PAK1) for enhanced migration and proliferation in vitro and in vivo. Since mast cells promote tumor growth in NF1, and their deletion impairs tumor progression, PAK1 inhibition in mast cells is a desirable target for NF1 through small molecule inhibitors designed to inhibit Group A Pak family kinases (Pak1-3). Our preliminary studies show that another isoform, PAK2 is the dominant isoform in mast cells. This isoform has the opposite phenotype to PAK1 in mast cells in another mast cell receptor pathway. The role of Pak2 in c-kit/SCF/NF1 signaling axis has yet to be studied. The objective in our study is identify in vitro and in vivo if Pak2 functions opposite of Pak1 in c-kit/Nf1 signaling, and if so, identify the signaling differences. In vitro experiments on Pak2-deficient primary mast cells will measure mast cell migration, adhesion, proliferation and MAPK signaling to elucidate the differences between Pak1 and Pak2. With small molecule inhibitors being developed against Group A Paks, it is critical to identify differences that could result in reduced potency or potential side effects. In vivo studie using dermal mast cell migration to SCF and bone marrow transplantation in NF1 will elucidate the role that Pak2 plays in c-kit signaling in skin and in tumor microenvironment cells. By replacing Nf1 wildtype bone marrow with Nf1-heterozygous~Pak1-/- or Pak2-/- bone marrow in mice prone to neurofibromas, we will identify if they play opposing roles to support tumor growth. This research is necessary to understand potential side effects of inhibiting multiple Pak kinases simultaneously in NF1 patients wih drug inhibitors.

描述（由申请人提供）：p21激活激酶1和2作为NF1微环境的治疗靶标I型神经纤维瘤病（NF1）的特征是由NF1无效雪旺细胞和Nf1杂合基质细胞混合物组成的神经纤维瘤。突变基质细胞和雪旺细胞之间的相互作用促进血管生成、胶原蛋白沉积和肥大细胞浸润。针对mas细胞中的c-kit信号通路进行治疗可能会减少神经纤维蛋白的形成。 Nf1 杂合肥大细胞对 c-kit 受体过度活跃，介导迁移、增殖和对雪旺细胞的化学吸引。通过破坏肥大细胞中过度活跃的 c-kit 信号传导并调节肿瘤微环境，我们可以破坏 NF1 和其他依赖肥大细胞的肿瘤的生长。我们实验室的最新发现表明，Nf1 下游肥大细胞信号传导的增强需要 p21 激活激酶 1 (PAK1) 来增强体外和体内的迁移和增殖。由于肥大细胞促进 NF1 中的肿瘤生长，并且其缺失会损害肿瘤进展，因此通过设计用于抑制 A 组 Pak 家族激酶 (Pak1-3) 的小分子抑制剂，肥大细胞中的 PAK1 抑制是 NF1 的理想靶点。我们的初步研究表明，另一种异构体 PAK2 是肥大细胞中的主要异构体。该亚型与另一种肥大细胞受体途径中肥大细胞中的 PAK1 具有相反的表型。 Pak2在c-kit/SCF/NF1信号轴中的作用还有待研究。我们研究的目的是在体外和体内鉴定 Pak2 在 c-kit/Nf1 信号传导中是否与 Pak1 功能相反，如果是，则确定信号传导差异。 Pak2 缺陷型原代肥大细胞的体外实验将测量肥大细胞迁移、粘附、增殖和 MAPK 信号传导，以阐明 Pak1 和 Pak2 之间的差异。随着针对 A 组 Paks 的小分子抑制剂的开发，识别可能导致效力降低或潜在副作用的差异至关重要。使用真皮肥大细胞迁移至 SCF 和骨髓移植至 NF1 的体内研究将阐明 Pak2 在皮肤和肿瘤微环境细胞中的 c-kit 信号传导中所发挥的作用。通过在易患神经纤维瘤的小鼠中用 Nf1 杂合~Pak1-/- 或 Pak2-/- 骨髓替换 Nf1 野生型骨髓，我们将确定它们是否在支持肿瘤生长方面发挥相反的作用。这项研究对于了解使用药物抑制剂同时抑制多种 Pak 激酶对 NF1 患者的潜在副作用是必要的。