Activation and Regulation Mechanisms of the RAF Kinase Family

RAF激酶家族的激活和调节机制

基本信息

批准号：
10798593
负责人：
Zhihong Wang
金额：
$ 2.76万
依托单位：
ROWAN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-10 至 2025-08-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10798593
关键词：
Address Affinity Allosteric Regulation BRAF gene Binding Binding Sites Biochemical Biological Assay Biological Process Biology CDK4 gene Cells Chemicals Client Clinical Co-Immunoprecipitations Complex Computing Methodologies Development Dimerization Disease Dissociation Drug resistance Elements Engineering Enzymes Event Family Goals Growth Factor Receptors Health Human In Vitro Intervention Kinetics Knowledge Length MAP Kinase Gene MAPK Signaling Pathway Pathway Malignant Neoplasms Mass Spectrum Analysis Measures Mediating Membrane Modeling Molecular Molecular Conformation Mutate Mutation Mutation Analysis N-terminal Nucleotides Oncogenic Outcome Output Pathologic Pathway interactions Peptides Pharmaceutical Preparations Phosphorylation Phosphotransferases Photoaffinity Labels Physiological Positioning Attribute Proliferating Protein Family Protein Isoforms Protein Kinase Protein-Serine-Threonine Kinases Proteins Proto-Oncogene Proteins B-raf RAS genes Ras/Raf Regulation Research Second Primary Cancers Serine Signal Transduction Sirolimus Specificity Structure System Tacrolimus Binding Proteins Therapeutic Variant Work X-Ray Crystallography biochemical tools biophysical tools cellular imaging chaperone machinery dimer inhibitor insight interdisciplinary approach live cell imaging member molecular dynamics multicatalytic endopeptidase complex mutant novel therapeutic intervention paralogous gene pharmacologic phosphoproteomics preference protein degradation prototype raf Kinases recruit success synergism targeted cancer therapy tumor tumorigenesis

项目摘要

BRAF, a member of the RAF family protein kinases, works in the MAPK signaling pathway, which controls broad cellular events like proliferation and differentiation. BRAF is the most frequently mutated kinase in human cancers. Furthermore, tumors lacking BRAF mutations are contingent on BRAF activity when there are mutations in upstream pathway members, such as RAS or growth factor receptors. Thus, BRAF represents an important target for cancer therapy. Despite three decades of intense research, only recently has a sufficient understanding of BRAF's mechanism creaked open the door to BRAF therapy. However, the current clinical BRAF inhibitors, vemurafenib and dabrafenib, are limited to the class I mutant BRAFV600E. Vemurafenib and dabrafenib, the two ATP-competitive inhibitors, can paradoxically activate non-BRAFV600E and wild-type BRAF, suggesting that BRAF has functions that are independent of its kinase activity. Therapeutic strategies for tumors with non- BRAFV600E mutations and wild-type BRAF are still lacking. Gaps in our knowledge contributed to the limitations of ATP-competitive BRAF drugs. The proposed work centers around three questions regarding RAF activation and regulation in health and disease states. Q1: how BRAF structural elements and ATP binding mediate the catalytic-independent functions of non-BRAFV600E and wild-type BRAF? Q2: what is the mechanism of action of the first BRAF allosteric inhibitor developed by our group and how can inhibitors targeting the catalytic and non- catalytic functions of BRAF be developed? Q3: What factors contribute to the non-overlapping functions of RAF isoforms, despite their structural similarity? The PI and her team will address these questions through multidisciplinary approaches, including X-ray crystallography, binding kinetics, phosphoproteomics, live cell imaging, chemical biology, and computational methods. Our findings will not only facilitate a better understanding of the complex biochemical mechanisms of the RAF kinase family, also provide a molecular basis for novel therapeutic approaches targeting BRAF-driven tumors.

RAF家族蛋白激酶的成员BRAF在MAPK信号通路中起作用，该途径控制宽阔细胞事件，例如增殖和分化。 BRAF是人类中最常见的激酶癌症。此外，缺乏BRAF突变的肿瘤是在有突变时的BRAF活动在上游途径成员中，例如RAS或生长因子受体。因此，BRAF代表一个重要的癌症治疗的靶标。尽管进行了三十年的深入研究，但直到最近才有足够的了解布拉夫（Braf）的机制吱吱作响，打开了BRAF治疗的大门。但是，当前的临床BRAF抑制剂， vemurafenib和dabrafenib仅限于I类突变Brafv600E。 Vemurafenib和Dabrafenib，两个 ATP兼容抑制剂可以矛盾地激活非BRAFV600E和野生型BRAF，这表明 BRAF具有独立于其激酶活性的功能。肿瘤的治疗策略 BRAFV600E突变和野生型BRAF仍然缺乏。我们知识的差距有助于局限 ATP竞争性BRAF药物。拟议的工作中心围绕有关RAF激活的三个问题和健康和疾病状态的调节。 Q1：BRAF结构元素和ATP结合如何介导非BRAFV600E和野生型BRAF的催化无关函数？ Q2：作用机制是什么我们小组开发的第一个BRAF变构抑制剂，以及如何针对催化和非抑制剂开发BRAF的催化功能？ Q3：哪些因素导致RAF的非重叠功能同工型，尽管结构相似？ PI和她的团队将通过多学科方法，包括X射线晶体学，结合动力学，磷蛋白质组学，活细胞成像，化学生物学和计算方法。我们的发现不仅会促进更好的理解 RAF激酶家族的复杂生化机理的复杂生化机制，也为新型提供了分子基础针对BRAF驱动肿瘤的治疗方法。