Identification of new therapeutic targets for ADPKD

ADPKD 新治疗靶点的确定

• 批准号: 10462701
• 负责人: EDWARD Y SKOLNIK
• 金额: $ 50.82万
• 依托单位: NEW YORK UNIVERSITY SCHOOL OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10462701
• 关键词: Affect; Affinity; Animal Model; Autosomal Dominant Polycystic Kidney; Binding; Biological Assay; Clinical Trials; Collaborations; Combined Modality Therapy; Couples; Cyclic GMP; Cyst; Data; Doxycycline; Drug Targeting; End stage renal failure; Epithelial cyst; Extracellular Matrix; Fibrosis; Focal Adhesion Kinase 1; Funding; Genes; Genetic; Genetic Transcription; Goals; Grant; Growth; Human; Kidney; Kidney Diseases; Lipids; Mass Spectrum Analysis; Mediating; Medicine; Messenger RNA; Mus; Mutation; PTK2 gene; Pathogenesis; Patients; Persons; Pharmaceutical Preparations; Pharmacology; Phosphotransferases; Play; Proprotein Convertase 1; Proprotein Convertase 2; Proteins; Renal function; Role; Signal Pathway; Signal Transduction; Techniques; Technology; Testing; Therapeutic; Time; Toxic effect; Translating; Universities; base; discoidin domain receptor 1; drug candidate; genetic approach; human disease; in vivo; inhibitor; insight; interest; interstitial; kinase inhibitor; mouse model; new therapeutic target; novel; novel strategies; polycystic kidney disease 1 protein; preservation; tandem mass spectrometry; therapeutic target; tolvaptan

Autosomal-dominant polycystic kidney disease (ADPKD) affects more than 12 million people worldwide and is a common cause of end-stage kidney disease (ESKD). ADPKD is caused by mutations in one of two genes, PKD1 or PKD2, which encodes polycystin 1 (PC) and PC2 respectively. Loss of PC1 or PC2 results in activation of numerous kinases and downstream signaling pathways, which is central to the pathogenesis of cyst growth in ADPKD. In addition, pharmacologically inhibiting a number of different kinases up-regulated in PKD kidneys has been shown to slow cyst growth in animal models of PKD making kinase inhibitors among the most promising class of drug candidates to treat patients with ADPKD. However, while the human kinome consists of more than 500 kinases, only a fraction of these kinases have been tested to determine if they play a role in ADPKD pathogenesis. As a result, there are likely many kinases that are active in ADPKD kidneys that play prominent roles in cyst growth that are yet-to-be discovered and may be good therapeutic targets. In collaboration with James Duncan at Temple University, we have now adapted a novel approach to broadly screen PKD kidneys in an unbiased manner for kinases that are more active in lysates from PKD kidneys compared with lysates from wild type kidneys. In this assay, active kinases are affinity captured by passing lysates over multiplex inhibitor beads (MIB) containing a cocktail of kinase inhibitors. Bound kinases are then identified by LC separation followed by quantitative tandem mass spectrometry (LC- MS/MS). Using this approach, we have now identified a number of both known and unknown kinases specifically up-regulated and down-regulated in PKD kidneys. The major goal of this proposal is to assess the role of several of the kinases identified thus far whose activity is increased in PKD kidneys and determine whether inhibiting or activating any of the kinases identified slows cyst growth, inhibits interstitial fibrosis, and preserves renal function in animal models of ADPKD. We will then take both biased and unbiased approaches to identify the signaling pathways regulated by these kinases that are critical to pathogenesis with the goal of developing a more complete picture of the relevant signaling hubs and networks that are aberrantly activated in PKD kidneys. In addition, we will use this technology to broadly screen PKD kidneys from a variety of different “early” and “late” mouse models of ADPKD at different stages in cyst formation and in kidneys from human patients with ADPKD to identify in an unbiased manner additional kinases that are activated and inhibited in PKD kidneys to determine the similarities and differences between mouse models, which kinases may be most relevant to human disease, and whether distinct sets of kinases are activated early post cyst induction and function as early “drivers” of cyst growth. Ultimately, we hope this new information will identify new safe drug targets and rational approaches to combination therapies to slow cyst growth that can then be translated into clinical trials.

常染色体主导的多囊肾脏疾病（ADPKD）影响了全球超过1200万人 终末期肾脏疾病（ESKD）的常见原因。 ADPKD是由两个基因之一突变引起的， PKD1或PKD2，分别编码Polycystin 1（PC）和PC2。 PC1或PC2的丢失导致 激活众多激酶和下游信号通路，这对于 ADPKD中的囊肿生长。此外，药理学抑制了许多上调的不同激酶 PKD Kidsneys已显示在PKD动物模型中降低囊肿的生长，从而使激酶抑制剂在 治疗ADPKD患者的最有希望的候选药物类别。但是，虽然人类的活力 由500多个激酶组成，仅测试了这些激酶的一小部分，以确定它们是否玩 在ADPKD发病机理中的作用。结果，可能有许多活跃在ADPKD儿童中的激酶 在尚未发现的囊肿生长中扮演着重要角色，可能是良好的治疗靶标。 与Temple University的James Duncan合作，我们现在已经改编了一种新颖的方法 广泛的筛选PKD儿童尼斯，以一种无偏的方式，用于激酶，这些激酶在裂解物中更为活跃 与野生型儿童的裂解物相比，PKD Kidneys。在此测定中，活动激酶是亲和力 通过将裂解物传递到含有激酶抑制剂鸡尾酒的多重抑制剂珠（MIB）上而捕获。 然后通过LC分离鉴定结合激酶，然后进行定量串联质谱法（LC- MS/MS）。使用这种方法，我们现在已经确定了许多已知和未知激酶 特别是在PKD Kidneys中上调和下调。 该提案的主要目的是评估迄今为止鉴定出的几种激酶的作用 在PKD肾脏中增加，并确定抑制或激活任何鉴定出的速度 囊肿生长，抑制间质纤维化并保留ADPKD动物模型中的肾功能。然后我们会 采用有偏见和公正的方法来识别这些激酶调节的信号传导途径 对发病机理至关重要，目的是开发相关信号中心的更完整的图片 以及在PKD Kidneys中异常激活的网络。此外，我们将使用这项技术广泛 屏幕PKD儿童来自各种不同阶段的各种不同的“早期”和“晚”鼠标模型 囊肿形成和来自ADPKD患者的肾脏，以公正的方式识别 激活和抑制PKD Kidneys的激酶，以确定相似性和差异 小鼠模型，激酶可能与人类疾病最相关，以及不同的激酶。 被激活后囊肿后的早期诱导和功能作为囊肿生长的早期“驱动因素”。最终，我们希望这个 新信息将确定新的安全药物靶标和合理方法来减缓疗法 然后可以将囊肿生长转化为临床试验。