Elucidating the protective effects of the KL-VS variant using isogenic hiPSCs

使用同基因 hiPSC 阐明 KL-VS 变体的保护作用

• 批准号: 10354135
• 负责人: DAVID A BRAFMAN
• 金额: $ 43.18万
• 依托单位: ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-15 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10354135
• 关键词: 3-Dimensional; AD transgenic mice; Address; Aging; Alzheimer disease prevention; Alzheimer' s Disease; Alzheimer' s disease patient; Alzheimer' s disease risk; Amyloid; Amyloid beta-Protein; Animal Model; Attenuated; Biochemical; Biomedical Engineering; Calcium; Cell Line; Cells; Coculture Techniques; Cognitive; Data; Development; Disease; Disease model; Ensure; Environment; Exhibits; Future; Generations; Genes; Genetic; Genetic Polymorphism; Genetic Transcription; Gold; Haplotypes; Heterozygote; Human; Human Engineering; Impaired cognition; Individual; Inflammatory; Inflammatory Response; Investigation; Karyotype; Knock-out; Learning; Link; Mediating; Methods; Modeling; Molecular; Mus; Mutation; Nature; Neuraxis; Neurodegenerative Disorders; Nitric Oxide; Onset of illness; Oxidative Stress; Pathologic; Pathology; Phenotype; Probability; Process; Production; Regulation; Reporter; Reporting; Research; Risk; Risk Factors; Series; Signal Pathway; Signaling Protein; Single Nucleotide Polymorphism; Stimulus; Synapses; System; Testing; Therapeutic Intervention; Variant; Veins; Work; age related; attenuation; base; base editing; biological adaptation to stress; design; gene regulatory network; genetic variant; genome editing; genome wide association study; in vivo; induced pluripotent stem cell; loss of function; molecular targeted therapies; mouse model; neuroprotection; neurotoxic; non-demented; overexpression; pluripotency; protective effect; relating to nervous system; stem cell model; stem cells; tau Proteins; tau phosphorylation; therapeutic target; transcriptomics; uptake; β-amyloid burden

PROJECT SUMMARY ABSTRACT Genome-wide association studies (GWAS) studies have identified several risk factors associated with altered probability of sporadic AD (SAD) onset. However, the majority of these studies have identified genetic variants that increase risk of AD and, consequently, downstream therapeutic interventions have focused on mitigating these effects. On the other hand, the study of variants that decrease risk of AD might provide an alternative to identifying promising therapeutic target for AD prevention. In this regard, polymorphisms in the Klotho (KL) gene have been associated with a reduced risk of AD onset and age-related progression. KL was first identified as suppressing numerous age-related processes as KL deficient mice exhibited accelerated aging. With regards to AD, KL overexpression in AD transgenic mouse models significantly ameliorates AD-related pathological and cognitive phenotypes. Interestingly, it has been shown that two single nucleotide polymorphisms in the KL gene, rs9536314 (F352V) and rs9527025 (C370S) segregate together to form a haplotype KL-VS. Specifically, KL-VS heterozygosity has not only been associated with greater cortical volumes and elevated synaptic connectivity but also attenuation of AD-related phenotypes. As such, understanding the mechanism by which the KL-VS variant mitigates AD onset and age-related progression will have a significant impact on therapeutic interventions. In this proposal, we will use our collective expertise in stem cell bioengineering, neurodegenerative disease modeling, and highly efficient genome editing to elucidate the specific mechanisms by which KL-VS heterozygosity mitigates AD risk. In the first aim, we will use our recently developed gene editing methods to introduce the KL-VS variant into isogenic hiPSCs from healthy non-demented control (NDC) and AD patients. In the second aim, we will employ these isogenic hiPSC lines in a 3-D co-culture model to (i) determine if KL-VS heterozygosity mitigates the presence of AD-related phenotypes and (ii) exerts global transcriptional changes in signaling pathways and gene regulatory networks that would enhance neuroprotection against AD. In addition, through the use of KL knockout hiPSCs we will be able to determine if these KL-VS heterozygosity induces its effects occur through gain- or loss-of-function mechanisms. Overall, a better understanding of potential genetic factors, such as KL-VS heterozygosity, that enhance neuroprotection against AD will provide new avenues for therapeutic interventions.

项目总结摘要 全基因组关联研究(GWAS)研究已经确定了几个与动脉粥样硬化相关的危险因素 散发性阿尔茨海默病(SAD)发病的概率。然而，这些研究中的大多数已经确定了遗传变异 这增加了AD的风险，因此，下游治疗干预措施的重点是减轻 这些影响。另一方面，对降低阿尔茨海默病风险的变种的研究可能会提供一种替代 为AD预防寻找有前景的治疗靶点。在这方面，Klotho(KL)基因的多态 与AD发病和年龄相关进展的风险降低有关。吉隆坡最初被确认为 抑制许多与年龄相关的过程，因为KL缺乏的小鼠表现出加速衰老。关于…… AD转基因小鼠模型中AD、KL的过表达显著改善AD相关的病理和病理改变 认知表型。有趣的是，已发现KL基因中的两个单核苷酸多态性， Rs9536314(F352V)和rs9527025(C370S)分离形成单倍型KL-VS。具体来说，KL-VS 杂合性不仅与更大的皮质体积和突触连接性有关 而且AD相关表型的衰减也是如此。因此，了解KL-VS的机制 变种可缓解AD的发病和年龄相关的进展将对治疗产生重大影响 干预措施。在这项提案中，我们将利用我们在干细胞生物工程、神经退行性疾病方面的集体专业知识 疾病建模和高效基因组编辑以阐明KL-VS 杂合性可降低AD风险。在第一个目标中，我们将使用我们最近开发的基因编辑方法来 将KL-VS变异体导入健康非痴呆对照(NDC)和AD患者的等基因HiPSCs。在……里面 第二个目标，我们将在三维共培养模型中使用这些等基因的HiPSC株来(I)确定KL-VS 杂合性减轻AD相关表型的存在和(Ii)在 信号通路和基因调控网络将增强对AD的神经保护。此外, 通过使用KL基因敲除的HiPSC，我们将能够确定这些KL-VS杂合性是否诱导其 效应通过增益性或失效性机制发生。总体而言，更好地理解潜在的基因 KL-vs杂合性等增强对AD神经保护的因素将为以下方面提供新的途径 治疗性干预。