Investigating a role for eNAMPT-containing extracellular vesicles in mitigating age- and Alzheimer Disease-related cognitive decline

研究含有 eNAMPT 的细胞外囊泡在缓解年龄和阿尔茨海默病相关认知衰退中的作用

• 批准号: 10607376
• 负责人: Brian Vincent Lananna
• 金额: $ 7.18万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2026-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10607376
• 关键词: Abate; Acetylation; Adipocytes; Adipose tissue; Age; Age-associated memory impairment; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Amyloidosis; Anabolism; Behavior; Behavioral; Biological; Biology; Brain; C-terminal; Cell Culture Techniques; Cell Line; Cells; Ceramides; Characteristics; Cognitive; Cognitive deficits; Critical Pathways; Data; Deacetylase; Deposition; Deterioration; Engineering; Enzymes; Fellowship; Functional disorder; Future; Hippocampus; Hypothalamic structure; Impaired cognition; In Vitro; Injections; Knock-in Mouse; Knock-out; Label; Laboratories; Leadership; Limbic System; Location; Longevity; Mammals; Measures; Mediating; Membrane; Memory; Mentorship; Mus; NF-kappa B; Neurons; Neurosciences; Nicotinamide adenine dinucleotide; Pathologic; Pathology; Pathway interactions; Peripheral; Plasma; Play; Population; Preparation; Production; Proteins; Role; SIRT1 gene; Signal Pathway; Sirtuins; Stains; Symptoms; Synapses; Testing; Therapeutic; Tissues; Training; Universities; Viral; Washington; age related; aged; anti aging; behavior measurement; brain cell; career; cell type; cellular targeting; cognitive benefits; cognitive function; cognitive reappraisal; collaborative environment; density; effective therapy; effectiveness testing; empowerment; extracellular; extracellular vesicles; flexibility; healthspan; improved; in vivo; knock-down; morris water maze; mouse model; nicotinamide phosphoribosyltransferase; novel; overexpression; restoration; secondary endpoint; small hairpin RNA; success; synaptic function; tau Proteins; trafficking; transcriptome sequencing; ultra high resolution; uptake

PROJECT SUMMARY/ABSTRACT The steady erosion of cognitive function is a hallmark of aging and markedly exacerbated in Alzheimer Disease (AD). These deficits have no effective treatments. Depletion of nicotinamide adenine dinucleotide (NAD+) and reduction of NAD+-dependent sirtuin activity in aging and AD have been well documented. Restoring NAD+ or activating the sirtuin SIRT1 has induced abatement of aging symptoms including cognitive decline. However, current NAD+ replenishment strategies are non-specific and the success of SIRT1 activation as an anti-aging therapeutic may require tissue-specific activation and the concomitant restoration of NAD+. We have shown previously that nicotinamide phosphoribosyltransferase (NAMPT), the rate-limiting enzyme in the major mammalian NAD+ biosynthesis pathway, is contained in extracellular vesicles (EVs) and secreted to plasma. Treatment of aged mice with plasma-derived EVs containing extracellular NAMPT (eNAMPT) extends mouse lifespan and healthspan. Our preliminary data suggest that EV treatment can ameliorate cognitive dysfunction and rescue age-related decreases in hippocampal CA1 synapse counts in 24-month-old mice. EVs are also targeted to the brain and can increase NAD+ in cultured neurons. However, the tissue-specific EV targeting mechanism as well as the implications for EV treatment in the context of AD-related cognitive decline remain unknown. This study will test the hypothesis that EV-contained eNAMPT rescues age-related cognitive dysfunction via activating SIRT1 in key neuronal populations and that leveraging this pathway can lessen AD- associated cognitive decline. In Aim 1, cell culture-derived EVs will be utilized to test the necessity of eNAMPT in the targeting and uptake of EVs both in vitro and in vivo. In Aim 2, the possibility that the cognitive benefits of EV treatment occur through eNAMPT-mediated activation of SIRT1 will be evaluated. This determination will be accomplished via EV treatment with and without EV-contained eNAMPT and with and without hippocampal knockdown of Sirt1 by viral delivery of shRNA. Aim 3 will test the effectiveness of EVs in treating cognitive deficits and pathological progression in a mouse model of AD. Therefore, this study has the potential to elucidate a novel role for eNAMPT in facilitating both the cellular targeting of EVs and the efficacy of EVs in rescuing age- and AD-related cognitive dysfunction. This project may also establish eNAMPT-containing EVs as a viable biologic to treat cognitive deterioration in aging and AD. This fellowship proposal integrates the NAD+ biology and aging expertise of sponsor Dr. Shin Imai with comprehensive training in behavioral neuroscience, AD models, advanced synaptic analysis, EV engineering, and laboratory mentorship and leadership. This training will be further empowered by the intellectually stimulating and highly collaborative environment of Washington University in St. Louis. Thus, this fellowship will constitute ideal preparation for the applicant’s career as the future leader of an academic laboratory.

项目摘要/摘要 认知功能的稳定侵蚀是衰老的标志，在阿尔茨海默氏病中明显恶化 （广告）。这些定义没有有效的治疗方法。烟酰胺腺苷二核苷酸（NAD+）和 已经有充分的文献记录了衰老和AD中NAD+依赖性的Sirtuin活性。还原nad+或 激活Sirtuin Sirt1已导致减轻衰老症状，包括认知能力下降。然而， 当前的NAD+补充策略是非特异性的，SIRT1激活作为抗衰老的成功 治疗可能需要组织特异性的激活和NAD+的伴随恢复。我们已经显示了 以前是烟酰胺磷酸贝糖基转移酶（NAMPT），这是主要限制酶 哺乳动物NAD+生物合成途径包含在细胞外蔬菜（EV）中，并分泌为血浆。 用含有细胞外NAMPT（ENAMPT）的血浆来源的EV对老年小鼠进行治疗，扩展了小鼠 寿命和健康范围。我们的初步数据表明，EV治疗可以改善认知功能障碍 在24个月大的小鼠中，海马CA1突触计数中与年龄相关的救援年龄下降。电动汽车也是 针对大脑，可以增加培养的神经元中的NAD+。但是，组织特异性的EV靶向 在与广告相关的认知下降的背景下，机制以及对EV治疗的影响仍然存在 未知。这项研究将检验以下假设，即持续的eNAMPT拯救了与年龄有关的认知 通过在关键神经元种群中激活SIRT1的功能障碍，并且利用此途径可以减少AD- 相关的认知下降。在AIM 1中，将利用细胞培养的衍生电动汽车来测试必要的ENAMPT 在体外和体内的电动汽车的靶向和摄取中。在AIM 2中，认知益处的可能性 将评估通过ENAMPT介导的SIRT1激活进行EV处理。这种决心将 可以通过有或没有EV的ENAMPT以及没有海马的EV处理来完成 通过病毒递送shRNA敲除SIRT1。 AIM 3将测试电动汽车在治疗认知方面的有效性 AD小鼠模型中的缺陷和病理进展。因此，这项研究有可能 阐明了ENAMPT在促进电动汽车的细胞靶向和电动汽车效率方面的新作用 营救年龄和广告相关的认知功能障碍。该项目还可能建立含ENAMP的EVS 作为治疗衰老和AD认知确定的可行生物学。该奖学金提案集成了 NAD+赞助商Shin Imai博士的生物学和老龄化专业知识以及行为方面的全面培训 神经科学，广告模型，先进的突触分析，EV工程和实验室心态以及 领导。智能刺激和高度协作的培训将进一步授权 圣路易斯华盛顿大学的环境。那，该奖学金将构成理想的准备 申请人作为学术实验室的未来领导者的职业生涯。