Regulation of OATP1B1 and OATP1B3 by lysine acetylation and lysine deacetylase inhibitors

赖氨酸乙酰化和赖氨酸脱乙酰酶抑制剂对 OATP1B1 和 OATP1B3 的调节

基本信息

批准号：
10688231
负责人：
Wei Yue
金额：
$ 29万
依托单位：
UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-09-01 至 2026-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10688231
关键词：
Abbreviations Acetylation Adverse drug event Aging Antibiotics Antidiabetic Drugs Antineoplastic Agents Biochemical Blood Cardiac Glycosides Carrier Proteins Cell Line Cell model Clinical Clustered Regularly Interspaced Short Palindromic Repeats Data Deacetylase Deacetylation Disease Drug Interactions Drug Transport Epigenetic Process FDA approved Functional disorder Genetic Engineering Goals Hepatocyte Histone Acetylation Histone Deacetylase Inhibitor Histone Deacetylation Histones Human Immunosuppressive Agents Knock-out Knowledge Lipids Liver Liver diseases Lysine Mediating Membrane Messenger RNA Modeling Molecular Muscle Mutagenesis OATP Transporters Outcome Pharmaceutical Preparations Pharmacotherapy Physiological Post-Translational Protein Processing Post-Translational Regulation Protein Acetylation Proteins Proteomics Regulation Research Rhabdomyolysis Role Site Testing Toxic Environmental Substances Toxic effect Toxin Vorinostat Work clinical predictors clinically relevant drug disposition epigenetic regulation experimental study inhibitor inhibitor therapy loss of function new chemical entity novel novel strategies novel therapeutics rational design side effect stable cell line success tubacin uptake

项目摘要

Organic anion transporting polypeptides (OATP)1B1 and OATP1B3 (abbreviated as OATP1B1/3) are liver- specific drug transport proteins that mediate uptake, from blood into the liver, of a diverse array of endogenous compounds, environmental toxins, and many clinically important drugs (e.g., lipid-lowering statins and anticancer agents). OATP1B1/3 are important determinants of transport-mediated drug-drug interactions (DDIs) resulting in severe side effects, such as statin-induced rhabdomyolysis, a sometimes-fatal muscle toxicity. Dysfunction of OATP1B1/3 significantly contributes to altered drug disposition and adverse drug events. Our long-term goal is to delineate the molecular mechanisms underlying drug/toxin disposition through OATP1B1/3, and to predict and mitigate OATP-mediated drug-drug and drug-disease interactions. Although it is evident that factors (drugs, aging, disease) modulating OATP1B1/3 function could cause drug-drug or drug-disease interactions with OATP1B1/3 substrates, unfortunately, our ability to predict such interactions is hampered due the dearth of information on OATP1B1/3 regulation. In particular, modulating lysine acetylation, a major post-translational modification known to alter function of numerous target proteins, including histone for epigenetic regulation, has not been investigated for OATP1B1/3. The overall objective of this application is to determine the molecular mechanisms governing the regulation of OATP1B1/3 by lysine acetylation, and to evaluate the impact of lysine deacetylase (KDAC) inhibitors (KDACIs) on OATP1B1/3 expression and function. Our central hypothesis is that OATP1B1/3 are lysine-acetylated proteins, deacetylation of which involves KDAC6; OATP1B1/3 can be regulated at the epigenetic level by KDACIs targeting histone deacetylation and at the post-translational level by KDAC6 inhibition. Our novel preliminary data show that 1) OATP1B1/3 are lysine-acetylated; 2) mutagenesis mimicking hyper-acetylation of OATP1B1 and specific KDAC 6 inhibition significantly reduces OATP1B1/3 transport function; and 3) FDA-approved pan KDAC inhibitors (KDACIs) inhibiting histone deacetylation induce mRNA and protein levels of OATP1B1/3. Guided by preliminary results, two specific Aims are outlined. In Aim 1, we will elucidate the role of KDAC6 in regulation of OATP1B1/3 lysine acetylation and transporter function. In Aim 2, we will dissect the regulation of OATP1B1/3 by pan KDACI drugs and the interplay between epigenetic and posttranslational regulation of OATP1B1/3 via K-Ac. A combination of proteomics, biochemical, and genetic engineering approaches will be utilized in cell lines and in the physiologically relevant sandwich-cultured primary human hepatocytes. The outcomes of these experiments will elucidate novel posttranslational and epigenetic mechanism(s) involved in regulating OATP1B1/3. The knowledge gained from these studies will be invaluable toward the rational design of novel drugs and inhibitors to optimize drug therapy while avoiding unwanted drug interactions. This work will enhance our ability to predict altered OATP1B1/3 function by lysine-deacetylase modulators (e.g., drugs/candidates that are HDAC inhibitors/activators and liver disease states).

有机阴离子转运多肽 (OATP)1B1 和 OATP1B3（缩写为 OATP1B1/3）是肝介导从血液到肝脏摄取多种内源性药物的特定药物转运蛋白化合物、环境毒素和许多临床重要药物（例如降脂他汀类药物和抗癌药物）代理）。 OATP1B1/3 是转运介导的药物相互作用 (DDI) 的重要决定因素严重的副作用，例如他汀类药物引起的横纹肌溶解症，这是一种有时致命的肌肉毒性。功能障碍 OATP1B1/3 显着影响药物处置的改变和药物不良事件。我们的长期目标是描述通过 OATP1B1/3 进行药物/毒素处置的分子机制，并预测和减轻 OATP 介导的药物与药物以及药物与疾病的相互作用。尽管很明显因素（药物、衰老、疾病）调节 OATP1B1/3 功能可能会导致药物与药物或药物与疾病之间的相互作用不幸的是，由于缺乏 OATP1B1/3 底物，我们预测这种相互作用的能力受到了阻碍。有关 OATP1B1/3 调节的信息。特别是调节赖氨酸乙酰化，这是一种主要的翻译后活性已知修饰可以改变许多靶蛋白的功能，包括用于表观遗传调控的组蛋白，尚未对 OATP1B1/3 进行研究。本应用的总体目标是确定分子赖氨酸乙酰化调节 OATP1B1/3 的机制，并评估赖氨酸的影响脱乙酰酶 (KDAC) 抑制剂 (KDACIs) 对 OATP1B1/3 表达和功能的影响。我们的中心假设是 OATP1B1/3 是赖氨酸乙酰化蛋白，其脱乙酰化涉及 KDAC6； OATP1B1/3 可以在表观遗传水平上通过针对组蛋白脱乙酰化的 KDACIs 进行调节，并在翻译后水平上通过 KDAC6 抑制。我们的新初步数据表明 1) OATP1B1/3 是赖氨酸乙酰化的； 2）诱变模拟 OATP1B1 的过度乙酰化和特异性 KDAC 6 抑制显着降低 OATP1B1/3 运输功能； 3) FDA 批准的泛 KDAC 抑制剂 (KDACIs) 抑制组蛋白脱乙酰化诱导 OATP1B1/3 的 mRNA 和蛋白质水平。在初步结果的指导下，概述了两个具体目标。瞄准 1，我们将阐明KDAC6在调节OATP1B1/3赖氨酸乙酰化和转运蛋白功能中的作用。在目标2，我们将剖析泛KDACI药物对OATP1B1/3的调节以及表观遗传之间的相互作用以及通过 K-Ac 对 OATP1B1/3 进行翻译后调节。蛋白质组学、生物化学和遗传学的结合工程方法将用于细胞系和生理相关的三明治培养原代细胞人类肝细胞。这些实验的结果将阐明新的翻译后和表观遗传参与调节 OATP1B1/3 的机制。从这些研究中获得的知识将是无价的合理设计新药和抑制剂以优化药物治疗，同时避免不需要的药物互动。这项工作将增强我们预测赖氨酸脱乙酰酶改变的 OATP1B1/3 功能的能力调节剂（例如，HDAC 抑制剂/激活剂和肝脏疾病状态的药物/候选药物）。