Elucidating the Understudied Kinase PNCK as a Prospective Drug Target in Renal Cell Carcinoma

项目总结 人类基因组是最成熟的可用药蛋白质家族之一。然而，在600多个Kinase中 目前只有一小部分是临床研究和化学探针开发的重点，而许多激酶 仍然没有得到充分的研究。激酶抑制剂在人类癌症的治疗中发挥着重要作用，例如 晚期肾细胞癌(RCC)是一种发病率不断增加且预后不良的恶性肿瘤。 虽然随着酪氨酸激酶抑制剂和免疫疗法的出现，肾细胞癌的临床结果正在改善，但大多数 患者最终会屈从于渐进性、抗药性疾病。我们的长期目标是找到晚期癌症的治疗方法 RCC通过识别和验证用于药物开发的新的RCC激酶靶点。 PNCK是一种未被充分研究的激酶。分析照亮可药物基因组(IDG)知识门户 Pharos等资料显示，PNCK是肾细胞癌中差异最大的过表达蛋白 并与特定疾病的存活率较差有关。在我们的初步研究中，PNCK过表达 导致肾癌细胞生长和增殖显著增加，而PNCK的抑制作用直接抑制肿瘤 通过抑制细胞生长，诱导肾细胞癌细胞周期停滞和凋亡，以及间接作用， 通过调节血管生成和DNA损伤反应通路的表达。这些发现突出了 肾癌中PNCK的临床和生物学意义该提案目标是生成并随后集成 肾细胞癌中PNCK的系统水平特征及靶向PNCK小分子抑制剂的激酶谱分析 通过两个具体目标为未来的化学探针和药物先导优化计划提供信息。 AIM 1将从PNCK的过度表达和敲除中产生转录和活性动态组签名 并通过多组学分析鉴定PNCK驱动的通路和信号网络。目标2将优化 然后使用计算模型和NanoBRET靶标来表征我们目前的PNCK抑制剂 接合后进行激酶分析，以评估选择性和非靶点。总而言之，系统级别 肾细胞癌中PNCK的特征和最佳化合物的靶向激酶图谱将进一步阐明 肾癌中PNCK的生物学和治疗潜力及后续化学探针和药物先导的信息 优化项目。 我们的团队结合了计算系统药理学/药物发现方面的互补专业知识 (Schürer)、转化型肾癌研究(Merchan)和多组学(包括化学蛋白质组学 MIB/MS)对癌症亲缘关系(Johnson)的功能表征，以成功完成拟议的 项目目标。

PROJECT SUMMARY The human kinome is one of the most established family of druggable proteins. Yet out of more than 600 kinases only a fraction is currently the focus of clinical studies and chemical probe development, and many kinases remain understudied. Kinase inhibitors play an important role in the treatment of human cancers, such as advanced renal cell carcinoma (RCC), a malignancy whose incidence is increasing and carries a poor prognosis. While RCC clinical outcomes are improving with tyrosine kinase inhibitors and immunotherapies, the majority of patients ultimately succumb to progressive, resistant disease. Our long-term goal is to find cures for advanced RCC by identifying and validating novel RCC kinase targets for drug development. PNCK is an understudied kinase. Analyzing the Illuminating the Druggable Genome (IDG) Knowledge Portal Pharos along with other resources revealed that PNCK is the most differentially overexpressed kinase in RCC patients and is associated with worse disease specific survival. In our preliminary studies PNCK overexpression led to significant increase in RCC cell growth and proliferation, while PNCK inhibition exerted direct antitumor effects, by inhibition of cell growth, induction of RCC cell cycle arrest and apoptosis, as well as indirect effects, by regulating expression of angiogenesis and DNA damage response pathways. These findings highlight the clinical and biological relevance of PNCK in RCC. The objective of this proposal is to generate and then integrate systems level signatures of PNCK in RCC with target-based kinase profiling of PNCK small molecule inhibitors to inform future chemical probe and drug lead optimization programs via two Specific Aims. Aim 1 will generate transcriptomics and active kinome signatures from PNCK overexpression and knockdown cell lines and identify PNCK-driven pathways and signaling networks by multi-omics analysis. Aim 2 will optimize and then characterize our current PNCK inhibitors using computational models and NanoBRET target engagement followed by kinase profiling to assess selectivity and off targets. Combined, the systems level signatures of PNCK in RCC and target-based kinase profiles of the best compounds will further elucidate the biology and therapeutic potential of PNCK in RCC and inform subsequent chemical probe and drug lead optimization projects. Our team combines complementary expertise in computational systems pharmacology / drug discovery (Schürer), translational kidney cancer research (Merchan), and multi-omics (including chemical proteomics MIB/MS) functional characterization of the cancer kinome (Johnson) to successfully complete the proposed project Aims.