Validation and characterization of the identified variants associated with human longevity in mouse models

在小鼠模型中验证和表征与人类长寿相关的已识别变异

• 批准号: 10714393
• 负责人: JAN VIJG
• 金额: $ 56.38万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2028-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10714393
• 关键词: 5 year old; Acceleration; Adenosine; Affect; Age; Aging; Animals; Attenuated; Biological Models; CRISPR/Cas technology; Candidate Disease Gene; Cell Aging; Cells; Centenarian; Chronic Disease; Code; Collaborations; Complex; DNA Damage; DNA Repair Endonuclease; DNA Resequencing; DNA lesion; Data; Deacetylase; Defect; Diphosphates; Disease; Drug Targeting; ERCC1 gene; Epigenetic Process; Evaluation; FOXO3A gene; Frequencies; Funding; Gene Targeting; Genes; Genetic; Genetic study; Genome Stability; Genotoxic Stress; Goals; Grant; Health; Heterozygote; Human; I-kappa B Proteins; IGF1R gene; Intervention; Life; Link; Location; Longevity; MADH3 gene; Mammals; Measures; Medicine; Mitochondria; Modeling; Molecular; Molecular Target; Mouse Strains; Mus; Mutate; Mutation; NF-kappa B; Oxidative Stress; Pathology; Pathway interactions; Pattern; Pharmaceutical Preparations; Pharmacology Study; Phenotype; Point Mutation; Proteomics; Rapid screening; Reactive Oxygen Species; Reporter; Resources; Risk; Role; Sleep Deprivation; Syndrome; TNFRSF5 gene; Testing; Transgenes; Transgenic Model; Transgenic Organisms; Translations; Untranslated RNA; Validation; Variant; Virus Diseases; Wild Type Mouse; age related; aged; college; drug discovery; drug testing; efficacy testing; exome; follower of religion Jewish; frailty; fucoidan; gene product; genetic analysis; genetic approach; genetic variant; genome-wide; healthspan; healthy aging; improved; in vivo; inhibitor; innovation; macromolecule; mitochondrial dysfunction; mouse model; multiple chronic conditions; mutant; novel; novel therapeutic intervention; overexpression; oxidative damage; p65; pharmacologic; pre-clinical; rare variant; resilience; senescence; stem cell function; tool; transcriptomics

Abstract Although aging is extremely complex, the occurrence of genetic variants in humans that positively affect longevity and health offer an ideal starting point for the systematic identification of molecular targets for interventions that could extend human healthspan. Through genetic analysis of the Ashkenazi Jewish centenarian resource at the Albert Einstein College of Medicine, this U19 identified multiple rare, functional genetic variants significantly associated with healthy longevity, which represent potential targets for drug discovery. These include coding variants in IGF1R, SIRT6, ATM, USP35, UBE3C, and BLM, as well as functional non-coding variants in FOXO3A, SIRT6, SMAD3, and components of the IKK/NF-kB pathway, including p65, NFKB1/p50 and NFKB1a (IkBa). However, the mechanism by which these variant gene products extend healthspan and lifespan still need to be discovered and validated in model systems. During the previous funding period, we used the Ercc1-/∆ mouse model of accelerated aging and aged wild-type mice to demonstrate that reduction in ATM and NF-kB activity, achieved by ATM and p65/RelA haploinsufficiency, and overexpression of hSIRT6 reduced cellular senescence and extended healthspan of mice. In addition, treatment of Ercc1-/∆ mice (in collaboration with Project 4) with a novel IKK/NF-kB inhibitor (SR12343), a known ATM inhibitor (KU-55933), and a compound shown to stimulate the mono–adenosine 5′-diphosphate (ADP)–ribosyltransferase (mADPRT) activity of SIRT6 (fucoidan) reduce senescence and improve age-related phenotypes in mice. We also generated mice carrying a centenarian rare variant in IGF-1R (Igf1rhR407H), mice heterozygous for Smad3, Foxo3a, and Usp35, as well as mice carrying mutations in Sirt6 conferring loss of deacetylase or mADPRT activity, which are currently being analyzed. Our data suggests that many of these rare variants reduce DNA damage and/or cellular senescence. Here, we propose to expand these efforts to validate the role of additional rare variants identified by Projects 1 and 2 for their ability to extend healthspan using our murine model of a human progeroid syndrome to accelerate analyses. We also will continue to test compounds targeting the variant gene products and associated pathways provided by Project 4 in both progeroid and aged wild-type mice. Overall, the aim of Project 3 is to use genetic approaches to validate the pathways identified in Projects 1 and 2 as impacting healthspan in mice. We will determine the impact of haploinsufficiency, overexpression, or specific mutations on animal health, molecular endpoints associated with the hallmarks of aging, resilience, and geropathology in mice. We will also pharmacologically target these pathways in mice, in collaboration with Project 4, to determine the impact on the same endpoints. The successful completion of this project will validate the identified human variants as important for healthy aging and identify novel therapeutic strategies with the potential to extend healthspan.

摘要 虽然衰老是极其复杂的，但人类基因变异的发生对长寿有积极影响， 和健康提供了一个理想的起点，系统地确定分子目标的干预措施， 可以延长人类的健康寿命。通过对德系犹太百岁老人的遗传分析， 阿尔伯特·爱因斯坦医学院，这U19确定了多种罕见的，功能性遗传变异显着 与健康长寿相关，这代表了药物发现的潜在目标。其中包括编码 IGF 1 R、SIRT 6、ATM、USP 35、UBE 3C和BLM中的变体，以及FOXO 3A中的功能性非编码变体， SIRT 6、SMAD 3和IKK/NF-kB通路的组分，包括p65、NFKB 1/p50和NFKB 1a（IkBa）。 然而，这些变异基因产物延长健康寿命和寿命的机制仍然需要进一步研究。 在模型系统中发现和验证。在上一个资助期间，我们使用了Ercc 1-/Mouse 加速老化和老化的野生型小鼠模型，以证明ATM和NF-kB活性的降低， 通过ATM和p65/RelA单倍不足实现，hSIRT 6过表达减少细胞衰老 和延长小鼠的健康寿命。此外，用抗肿瘤药物治疗Ercc 1-/Ercc 2小鼠（与项目4合作）， 新的IKK/NF-kB抑制剂（SR 12343），一种已知的ATM抑制剂（KU-55933），和一种显示刺激 SIRT 6（岩藻依聚糖）单腺苷5′-二磷酸（ADP）-核糖基转移酶（mADPRT）活性降低 衰老和改善小鼠中与年龄相关的表型。我们还培养了携带百岁老人罕见基因的老鼠， IGF-1 R变异体（Igf 1 rhR 407 H）、Smad 3、Foxo 3a和Usp 35杂合小鼠以及携带 Sirt 6突变导致脱乙酰酶或mADPRT活性丧失，目前正在分析。我们 数据表明，许多这些罕见的变异减少DNA损伤和/或细胞衰老。这里我们 我建议扩大这些努力，以验证项目1和2确定的其他罕见变异的作用， 他们的能力，以延长健康寿命使用我们的小鼠模型的人类早衰综合征，以加速分析。 我们还将继续测试针对所提供的变异基因产物和相关途径的化合物 通过项目4在早老性和老年野生型小鼠中进行。总体而言，项目3的目的是使用遗传方法 验证项目1和2中确定的影响小鼠健康寿命的途径。康贝特人将以 单倍不足、过表达或特定突变对动物健康的影响，分子终点 与小鼠的衰老、恢复力和老年病理学特征相关。我们也将 与项目4合作，在小鼠中靶向这些途径，以确定对相同终点的影响。 该项目的成功完成将验证已确定的人类变异对健康老龄化的重要性 并确定具有延长健康寿命潜力的新治疗策略。