Illuminating the biological functions of CDKL1 Kinase

阐明 CDKL1 激酶的生物学功能

• 批准号: 10217716
• 负责人: Navjot Pabla
• 金额: $ 15.6万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-04-15 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217716
• 关键词: Ablation; Acute Renal Failure with Renal Papillary Necrosis; Antibodies; Atlases; Biochemical; Biological; Biological Assay; Biological Process; Biology; Brain; Cell Culture Techniques; Cell Death; Cell physiology; Cells; Cellular Stress; Clinical; Communication; Cyclic AMP-Dependent Protein Kinases; Cyclin-Dependent Kinases; Dependence; Development; Disease; Enzymes; Epithelial; Epithelial Cells; Etiology; Family; Family member; Functional disorder; Future; G-Protein-Coupled Receptors; Gene Deletion; Gene Family; Genes; Genetic Transcription; Genome; Goals; Health; Human Genome; Hypoxia; Inflammatory; Injury; Injury to Kidney; Kidney; Knockout Mice; Lead; Link; Malignant - descriptor; Mediating; Modeling; Molecular; Molecular Target; Morbidity - disease rate; Mus; Nature; Nuclear; Oncogenic; Pathogenesis; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Pharmacology; Phosphorylation; Phosphotransferases; Physiological; Property; Protein Kinase; Protein-Serine-Threonine Kinases; Proteins; RNA Interference; RNA interference screen; Renal function; Research; Risk; Role; SOX11 gene; Safety; Severities; Signal Transduction; Stress; Syndrome; Testing; Therapeutic; Tissues; Tubular formation; base; cancer therapy; chemical genetics; clinical translation; genetic approach; human disease; in vivo; innovation; interest; kinase inhibitor; knock-down; member; mortality; multidisciplinary; nephrotoxicity; novel; protein activation; renal epithelium; screening; small molecule; therapeutic target; tool

ABSTRACT: Protein kinases control most cellular processes through signal transduction linked phosphorylation networks. Importantly, dysregulated protein kinase signaling is both the cause and consequence of numerous human diseases. Due to the crucial role of kinases in disease pathogenesis, as well as suitable pharmacological properties and clinical safety profile of kinase inhibitors, protein kinases have emerged as attractive therapeutic targets, especially for anti-cancer therapy. However, the underlying biology of the majority of protein kinases, a key gene family within the druggable genome, remains unclear. In addition, the role of protein kinases in the pathogenesis of non-oncological diseases, especially acute kidney injury (AKI), remains underexplored. AKI is a common disorder associated with toxic, inflammatory, and hypoxic insults to tubular epithelial cells. Using a kinome-wide RNAi screen, we recently identified Cdkl5 kinase as a critical regulator of epithelial cell death associated with acute kidney injury (AKI). While Cdkl5 kinase has been previously studied for its role in brain development, very little is known about the other four members of the cyclin-dependent kinase-like (Cdkl) family. Our preliminary results show that Cdkl1 kinase is activated under stress conditions associated with AKI and RNAi mediated Cdkl1 knockdown protects epithelial cells from nephrotoxic injury. Through chemical genetics approach, we have also identified Sox11 as a novel Cdkl1 substrate. We hypothesize that Cdkl1 is a stress- responsive kinase that phosphorylates Sox11 resulting in suppression of a protective transcriptional regulator, contributing to epithelial dysfunction and cell death. In the current proposal, we aim to test our hypothesis by examining the consequences of Cdkl1 gene ablation on the severity of AKI (Aim 1) and utilize biochemical and cell biological approaches to probe the functional consequences of Cdkl1 mediated Sox11 phosphorylation on stress-induced renal epithelial cell death (Aim 2). These studies will lead to the development of critical experimental tools that are expected to accelerate future studies into the biological functions of Cdkl1 kinase. Finally, these studies are expected to reveal the role of Cdkl1 kinase in kidney injury and illuminate the physiological role of an important member of the enigmatic Cdkl family of kinases.

摘要： 蛋白激酶通过信号转导连接的磷酸化网络控制大多数细胞过程。 重要的是，失调的蛋白激酶信号传导是许多人类疾病的原因和后果。 疾病由于激酶在疾病发病机制中的关键作用，以及合适的药理学作用， 由于激酶抑制剂的性质和临床安全性，蛋白激酶已成为有吸引力的治疗药物， 靶点，尤其是用于抗癌治疗。然而，大多数蛋白激酶的基础生物学， 关键基因家族在可药用基因组中，仍然不清楚。此外，蛋白激酶在 非肿瘤疾病，特别是急性肾损伤（阿基）的发病机制仍然未被探索。阿基是 一种与肾小管上皮细胞的毒性、炎症和缺氧损伤相关的常见疾病。使用 通过全激酶组RNAi筛选，我们最近鉴定了Cdkl 5激酶作为上皮细胞死亡的关键调节因子 急性肾损伤（阿基）。虽然Cdkl 5激酶先前已经研究了其在脑中的作用， 在发育过程中，对细胞周期蛋白依赖性激酶样（Cdkl）家族的其他四个成员知之甚少。 我们的初步结果表明，Cdkl 1激酶在与阿基相关的应激条件下被激活， RNAi介导的Cdkl 1敲低保护上皮细胞免受肾毒性损伤。通过化学遗传学 方法，我们还确定了Sox 11作为一种新的Cdkl 1底物。我们假设Cdkl 1是一种应激- 磷酸化Sox 11导致抑制保护性转录调节因子的应答性激酶， 导致上皮功能障碍和细胞死亡。在目前的提议中，我们的目标是通过以下方式来检验我们的假设： 检查Cdkl 1基因切除对阿基严重程度的影响（目的1），并利用生物化学和 细胞生物学方法来探测Cdkl 1介导的Sox 11磷酸化对 应激诱导的肾上皮细胞死亡（Aim 2）。这些研究将导致关键的发展 这些实验工具有望加速未来对Cdkl 1激酶生物学功能的研究。 最后，这些研究有望揭示Cdkl 1激酶在肾损伤中的作用，并阐明Cdkl 1激酶在肾损伤中的作用。 这是激酶的神秘Cdkl家族的重要成员的生理作用。