Anti-HIV NRTIs and the lysosomal toxicity

抗 HIV NRTI 和溶酶体毒性

基本信息

批准号：
10468143
负责人：
RAJGOPAL GOVINDARAJAN
金额：
$ 33.08万
依托单位：
OHIO STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10468143
关键词：
Address Adenosine Adherence Adult Agonist Anti-HIV Agents Autophagocytosis Biochemical Blood Bone Marrow Transplantation Cell surface Chronic Clinical Coupled Development Dose Drug Kinetics FDA approved FRAP1 gene Foundations Functional disorder Generations Genetically Engineered Mouse Goals HIV HIV Infections HIV therapy Hepatotoxicity Homeostasis Hyperpigmentation Impairment Inflammation Inflammatory Integrase Inhibitors Knock-out Knockout Mice Knowledge Lipodystrophy Lysosomes Maintenance Mass Spectrum Analysis Metabolic Mitochondria Modeling Modification Molecular Molecular Probes Mucous Membrane Mus Myelosuppression Nucleoside Transporter Nucleosides Organ Outcome Pancreatitis Pathogenesis Pathway interactions Patients Pericarditis Peripheral Nervous System Diseases Pharmaceutical Preparations Pharmacology Pre-Clinical Model Property Protease Inhibitor Public Health Quality of life Recycling Regimen Research Research Project Grants Reverse Transcriptase Inhibitors Role Screening procedure Sensorineural Hearing Loss Severities Signal Transduction Structure Symptoms Testing Therapeutic Tissue Differentiation Tissue Engineering Tissues Toxic effect Treatment Efficacy Vertebral column Work adult stem cell adverse drug reaction base chemical synthesis cheminformatics clinical phenotype clinically relevant compliance behavior drug development drug efficacy exhaustion experimental study inhibitor inhibitor therapy insight lipidomics metabolomics mouse model non-nucleoside reverse transcriptase inhibitors novel nucleoside analog pharmacokinetics and pharmacodynamics pharmacophore prevent regenerative screening small molecule libraries spatiotemporal stem cell function stem cell homeostasis stem cells tissue regeneration tissue repair treatment optimization

项目摘要

PROJECT SUMMARY The nucleoside reverse transcriptase inhibitors (NRTIs) have potent activities against HIV, but their therapeutic benefit in patients undergoing NRTI therapies is limited by significant adverse drug reactions (ADR), resulting in poor patient compliance and compromised drug efficacy. Our group has recently described an indispensable role of a lysosomal nucleoside transporter ENT3 in lysosomal homeostasis via deletion of ENT3 in mice. Intriguingly, ENT3 KO mice manifest clinical phenotypes closely resembling NRTI ADR. The overall objective of this application is to evaluate ENT3-loss driven lysosomal toxicity as a putative mechanism involved in the chronic adverse sequelae of NRTIs. The central hypothesis of this proposal is that NRTIs that do not interfere with ENT3-supported lysosomal homeostasis or the inclusion of lysosomal signaling agents will minimize the occurrence of NRTI ADR. Aim 1 will evaluate the strategies to avoid NRTI toxicity without compromising drug efficacy. Our working hypothesis for this aim is that disruption of interaction between ENT3 and NRTIs or the inclusion of pharmacological agonists of lysosomal-autophagy pathway will mitigate the onset and severity of NRTI ADR. The preliminary studies that demonstrate the involvement of the cell surface NRTI transporters (e.g., ENT1, CNT) and not the lysosomal ENT3 for NRTI efficacy, the misregulation of the AMPK and mTOR signaling axis in the Ent3-/- mice, and the functional rescue of multi-organ dysfunction in Ent3-/- mice using a pharmacological AMPK activator AICAR; all support this aim. Aim 2 will elucidate the mechanism(s) of occurrence of NRTI-specific ADR signs. Our working hypothesis for this aim is that NRTIs, when present at clinically relevant blood concentrations, will inhibit the ENT3-regulated adult stem cell functions resulting in disruption of tissue repair and regeneration. In addition, we hypothesize that NRTIs will differentially impact the ENT3 function in adult tissues to bring distinct inflammatory, metabolic and degenerative changes that coupled with stem cell alterations, will explain the clinically observed NRTI ADR signs. The preliminary studies that demonstrate the transport of many ADR-producing NRTIs by ENT3, the inhibition of lysosomal adenosine transport by NRTIs and the perturbation of lysosomal recycling of adenosine in Ent3-/- mice leading to adult stem cell exhaustion, tissue inflammation and degeneration, and breaches of mesodermal tissue integrity, which taken together supports this aim. The project will utilize biochemical and molecular approaches, novel ENT3 probes, newly generated ENT3 mouse models, metabolomics, tissue engineering, pharmacophore modeling, synthetic and screening procedures and PKPD to accomplish the goals. The successful completion of the project will provide new insights into the mechanisms of occurrence of NRTI ADR and may have translational benefit for optimizing treatments (such as long-term efficacy, adherence, tolerability, etc.) in patients undergoing NRTI therapies.

项目概要核苷逆转录酶抑制剂 (NRTI) 具有有效的抗 HIV 活性，但其治疗效果较差接受 NRTI 治疗的患者的获益受到严重药物不良反应 (ADR) 的限制，导致患者依从性差，药物疗效受损。我们小组最近描述了一个不可或缺的溶酶体核苷转运蛋白 ENT3 通过缺失 ENT3 在小鼠溶酶体稳态中的作用。有趣的是，ENT3 KO 小鼠表现出与 NRTI ADR 非常相似的临床表型。总体目标该应用的目的是评估 ENT3 丢失驱动的溶酶体毒性作为参与 NRTI 的慢性不良后遗症。该提案的中心假设是 NRTI 不会干扰 ENT3支持的溶酶体稳态或包含溶酶体信号剂将最大限度地减少 NRTI ADR 的发生。目标 1 将评估在不影响药物的情况下避免 NRTI 毒性的策略功效。我们对此目标的工作假设是，ENT3 和 NRTI 之间的相互作用受到破坏，或者加入溶酶体自噬途径的药理学激动剂将减轻自噬的发生和严重程度 NRTI ADR。初步研究表明细胞表面 NRTI 转运蛋白的参与 (例如 ENT1、CNT) 而不是溶酶体 ENT3 的 NRTI 功效、AMPK 和 mTOR 的失调 Ent3-/- 小鼠中的信号轴，以及使用 Ent3-/- 小鼠多器官功能障碍的功能性挽救药理AMPK激活剂AICAR；所有人都支持这一目标。目标 2 将阐明以下机制：出现 NRTI 特异性 ADR 症状。我们对这一目标的工作假设是，当 NRTI 存在于临床相关的血液浓度，将抑制 ENT3 调节的成体干细胞功能，导致破坏组织修复和再生。此外，我们假设 NRTI 会对 ENT3 在成人组织中发挥作用，带来独特的炎症、代谢和退行性变化，随着干细胞的改变，将解释临床观察到的 NRTI ADR 迹象。初步研究表明证明 ENT3 可以转运许多产生 ADR 的 NRTI，并抑制溶酶体腺苷 NRTIs 的转运和 Ent3-/- 小鼠中腺苷溶酶体回收的扰动导致成年干细胞耗竭、组织炎症和变性以及中胚层组织完整性的破坏，综合起来支持这一目标。该项目将利用生物化学和分子方法，新颖 ENT3探针、新生成的ENT3小鼠模型、代谢组学、组织工程、药效团建模、合成和筛选程序以及 PKPD 来实现目标。顺利完成该项目将为 NRTI ADR 的发生机制提供新的见解，并可能对优化治疗的转化效益（例如长期疗效、依从性、耐受性等）接受 NRTI 治疗的患者。