Anti-HIV NRTIs and the lysosomal toxicity

抗 HIV NRTI 和溶酶体毒性

• 批准号: 10273397
• 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
• 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/10273397
• 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/antagonist; 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

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.

项目摘要 核苷类逆转录酶抑制剂（NRTI）具有很强的抗HIV活性，但其治疗效果不理想， 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， 患者依从性差和药物疗效受损。我们的团队最近描述了一个不可或缺的 通过小鼠中ENT 3的缺失，溶酶体核苷转运蛋白ENT 3在溶酶体体内平衡中的作用。 有趣的是，ENT 3 KO小鼠表现出与NRTI ADR非常相似的临床表型。总体目标 本申请的第一个目的是评估ENT 3损失驱动的溶酶体毒性作为参与细胞毒性的推定机制。 NRTI的慢性不良后遗症。该提案的中心假设是，不干扰的NRTI 使用ENT 3支持的溶酶体稳态或包含溶酶体信号传导剂将最小化 发生NRTI ADR。目的1将评估避免NRTI毒性而不损害药物的策略 功效我们对此的工作假设是，ENT 3和NRTIs之间的相互作用的破坏， 包括溶酶体-自噬途径的药理学激动剂将减轻 NRTI ADR。初步研究表明，参与细胞表面NRTI转运蛋白 (e.g., ENT 1，CNT）而非溶酶体ENT 3对NRTI疗效的影响，AMPK和mTOR的失调 信号转导轴，以及使用免疫抑制剂对Ent 3-/-小鼠中多器官功能障碍的功能性拯救。 药理学AMPK激活剂AICAR;都支持这一目标。目标2将阐明 发生NRTI特异性ADR体征。我们的工作假设，这一目标是，NRTI，当目前在 临床相关的血液浓度，将抑制ENT 3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NRTIs将对 ENT 3在成人组织中发挥作用，引起不同的炎症、代谢和退行性变化， 干细胞改变，将解释临床观察到的NRTI ADR体征。初步研究表明， 证明了ENT 3转运许多产生ADR的NRTI，抑制溶酶体腺苷 在Ent 3-/-小鼠中通过NRTI的转运和腺苷的溶酶体再循环的扰动导致成年 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， 这两个因素合在一起支持了这一目标。该项目将利用新颖的生物化学和分子方法 ENT 3探针，新生成的ENT 3小鼠模型，代谢组学，组织工程，药效团 建模，合成和筛选程序和PKPD来实现目标。圆满完成 该项目的研究将为NRTI ADR的发生机制提供新的见解， 优化治疗的转化效益（如长期疗效、依从性、耐受性等）在 接受NRTI治疗的患者。