Focal Adhesion Kinase - Tumor Biology and Therapeutics

项目摘要 粘着斑激酶（FAK）是一种多功能的非受体酪氨酸激酶和支架蛋白， 在许多实体瘤包括黑色素瘤中过表达，而在正常组织中表达最低。FAK 由于其在肿瘤进展的多个方面的贡献， 包括粘附、侵袭、增殖、存活、转移、血管生成和免疫细胞抑制。 然而，FAK抑制剂的开发主要集中在抑制FAK的激酶， 抑制关键的脚手架相互作用。特别是，在发现和发现的过程中所做的努力有限， 对FAK的粘着斑靶向（FAT）支架结构域（结构域）抑制剂的生物学评价 所需的FAK本地化局灶性粘连。在这段支持期间，我们确定了第一个 发现了基于钉合肽的FAK抑制剂（UA-1907），其与FAT结构域结合并共结晶， 并竞争性抑制FAK-桩蛋白结合。我们已经鉴定了肉豆蔻酰化衍生物（UA-2012）， 改善的细胞效能、有利的药物样性质和体内功效;并开发了二价肽 （UA-2023）具有低纳摩尔的与FAT的结合。然而，这些新型FAT之间的机械差异 结构域肽和传统的FAK激酶抑制剂对粘着斑复合物和FAK扰动的影响 相互作用组尚未阐明。此外，我们有初步的数据表明，脂肪抑制， 在NRAS突变黑素瘤细胞中提供选择性抗癌作用。黑色素瘤是最致命的皮肤病 目前还没有针对NRAS突变型黑色素瘤的有效靶向疗法，这代表了 约30%的患者。我们假设FAK FAT结构域抑制剂对细胞凋亡具有不同的生物学效应。 与FAK激酶抑制剂相比，癌细胞中的粘着斑复合物; NRAS信号通路的改变对FAK FAT支架具有分子依赖性， 存活，从而促进选择性抗癌功效。在具体目标1中，我们将确定 FAK FAT支架抑制剂和FAK激酶抑制剂对粘着斑调节的差异 复合物和FAK相互作用组。在具体目标2中，我们将定义 NRAS和BRAF激活突变的黑色素瘤细胞对FAK FAT抑制的敏感性/抗性 途径。在具体目标3中，我们将在小鼠模型中评估新型FAK FAT结构域抑制剂的体内功效 NRAS和BRAF驱动的癌症。总的来说，在这个项目中，我们将利用FAT钉合肽来验证， 与FAK相比，FAT结构域靶向对粘着斑复合物提供了额外的生物学功效。 激酶抑制剂，且NRAS突变型黑素瘤对FAKFAT抑制具有独特敏感性。 .

PROJECT ABSTRACT Focal Adhesion Kinase (FAK) is a multifunctional non-receptor tyrosine kinase and scaffolding protein that is overexpressed in numerous solid tumors including melanoma, while minimally expressed in normal tissue. FAK has been widely investigated as a cancer drug target due its contribution in multiple aspects of tumor progression, including adhesion, invasion, proliferation, survival, metastasis, angiogenesis, and immune cell suppression. However, development of FAK inhibitors has largely focused on inhibition of the kinase enzyme of FAK opposed to inhibition of key scaffolding interactions. Particularly, limited efforts have been made at the discovery and biological evaluation of inhibitors of the focal adhesion targeting (FAT) scaffolding domain of FAK, the domain required for FAK localization to focal adhesions. During this period of support, we have identified the first discovered stapled peptide-based FAK inhibitor (UA-1907) that binds to and co-crystallizes with the FAT domain, and competitively inhibits FAK-paxillin binding. We have identified a myristoylated derivative (UA-2012) with improved cellular potency, favorable drug-like properties, and in vivo efficacy; and developed a bivalent peptide (UA-2023) with low nanomolar binding to FAT. However, the mechanistic differences between these novel FAT domain peptides and traditional FAK-kinase inhibitors on perturbation of focal adhesion complexes and the FAK interactome have yet to be elucidated. Furthermore, we have preliminary data showing that FAT inhibition provides selective anti-cancer effects in NRAS mutant melanoma cells. Melanoma is the deadliest form of skin cancer and there are no current effective targeted therapies against NRAS mutant melanoma, which represents ~30% of all patients. We hypothesize that FAK FAT domain inhibitors have distinct biological effects on the focal adhesion complex in cancer cells compared to FAK kinase inhibitors; and cancer cells with alterations in NRAS signaling pathways have a molecular dependence on FAK FAT scaffolding for survival, thus promoting selective anti-cancer efficacy. In specific aim 1, we will identify the unique differences between FAK FAT scaffold inhibitors and FAK kinase inhibitors on modulation of the focal adhesion complex and the FAK interactome. In specific aim 2, we will define the molecular mechanisms of sensitivity/resistance to FAK FAT inhibition in melanoma cells with activating mutations in NRAS and BRAF pathways. In specific aim 3, we will evaluate in vivo efficacy of novel FAK FAT domain inhibitors in mouse models of NRAS and BRAF driven cancer. Overall, in this project we will utilize FAT stapled peptides to validate that FAT domain targeting provides additional biological efficacy on the focal adhesion complex compared to FAK- kinase inhibitors and that NRAS mutant melanoma has a unique sensitivity to FAK FAT inhibition. .