International Registry for Werner Syndrome

维尔纳综合症国际登记处

• 批准号: 10563164
• 负责人: JUNKO OSHIMA
• 金额: $ 36.43万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-02-04 至 2027-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10563164
• 关键词: Abbreviations; Acceleration; Adult; Affect; Aging; Autophagocytosis; BSCL2 gene; Basic Science; Biocompatible Materials; Biological; Biology of Aging; Blood specimen; Categories; Cell Culture Techniques; Cells; Chronic; Clinic; Clinical; Clinical Trials; Collaborations; Cryopreservation; Cultured Cells; DNA Damage; DNA Polymerase III; DNA-Directed DNA Polymerase; Development; Dideoxy Chain Termination DNA Sequencing; Disease; Enrollment; Epigenetic Process; Exhibits; FDA approved; Family; Family member; Fibroblasts; Follow-Up Studies; Genes; Genetic; Genetic Diseases; Genome Stability; Genomic Instability; Genotype; Goals; Human; Inflammation; International; Janus kinase; Japan; Japanese; Libraries; Lipids; Longevity; MADH4 gene; MDM2 gene; Maintenance; Malignant Neoplasms; Manuscripts; Mediating; Metformin; Microsatellite Instability; Mitochondria; Mutation; Non-Insulin-Dependent Diabetes Mellitus; Nuclear; Nuclear Structure; Nucleotide Excision Repair; Oxidative Stress; Parents; Pathogenesis; Pathogenicity; Pathway interactions; Patients; Pharmaceutical Preparations; Phenotype; Plasma; Play; Polymerase; Process; Production; Qualifying; Rare Diseases; Reagent; Registries; Regulation; Research; Resources; Role; STAT protein; Series; Siblings; Signal Transduction; Sirolimus; Small Interfering RNA; Specimen; Syndrome; TP53 gene; Testing; Therapeutic; Therapeutic Agents; Tissue Sample; Transforming Growth Factor beta; Translation Initiation; Translational Research; Variant; WRN gene; Washington; Werner Syndrome; age related; biological research; carcinogenesis; causal variant; clinical application; comparative genomic hybridization; comparison control; cytokine; disease phenotype; efficacy evaluation; established cell line; exome; exome sequencing; experimental study; gene discovery; genetic analysis; genetic pedigree; genetic variant; genome sciences; genome sequencing; genome-wide; genomic locus; helicase; high throughput screening; inhibitor; kinase inhibitor; mTOR Inhibitor; member; mutant; new therapeutic target; next generation sequencing; nicotinamide-beta-riboside; novel; participant enrollment; peripheral blood; preclinical study; recruit; screening; senescence; synergism; telomere; therapeutic target; transcriptome sequencing; whole genome

Project Summary The International Registry of Werner Syndrome recruits cases of Werner syndrome (WS) and a range of other segmental progeroid syndromes from around the world with the goal of elucidating underlying mechanisms of accelerated aging. Detailed clinical information, the results of genetic analyses, and biological specimens are made available to a wide range of qualified geroscientists. In this application, we propose to extend our previous studies to include systematic genome-wide searches for the genetic variants responsible for the 78 progeroid cases that we have so far been unable to genetically characterize. An additional important extension of our research agenda is to initiate translational research that can lead to potential therapeutic agents. We will employ a combination of next generation sequencings, array CGH, and Sanger sequencing, followed by confirmatory Western analysis and quantitative PCR. These approaches have successfully identified novel pathogenic variants of WRN (a DNA helicase), LMNA (a component of nuclear structure), POLD1 (DNA polymerase delta), SPRTN (recruiter of DNA polymerase), ERCC4 (nucleotide excision repair), CTC1 (telomere replication), MDM2 (an inhibitor of P53) and SAMHD1 (regulation of dNTP pools). These loci highlight major roles for genome instability, now widely accepted as one of the hallmarks of aging. We also made progress in the identification of disease mutations that suggest other mechanisms of accelerated aging, such as BSCL2 (lipid droplet formation) and SMAD4 (intracellular signaling of a component of SASP, TGFβ). As indicated above, we will pursue translational research with the potential for the development of ameliorative therapies for our progeroid patients. Based on our previous studies, our collaborator, Dr. Yokote Koutaro at the Japanese Werner Consortium, is evaluating the efficacy of an NAD intermediate and an mTOR inhibitor, metformin, in WS patients. We shall begin independent studies of the effects of suppressors of chronic inflammation, namely Janus kinase (JAK) inhibitors, in cultures from WS patients and controls. This effort was motivated by our findings that SMAD4 mutant fibroblasts exhibited increased accumulation of DNA damage and that WRN mutant fibroblasts showed elevated SMAD4 expressions and dramatically higher levels of SASP compared to control cells, suggesting that a synergy of persistent DNA damage and inflammation may be one of the common key mechanisms leading to the accelerated aging. Concordant experiments will explore the effects of these novel therapeutic targets with high throughput screening of cell cultures from patients with other progeroid syndromes. An initial small scale experiment involving siRNA screening has revealed that siRNAs and drugs that alter the intranuclear dNTP concentration are able to modulate the cellular disease phenotypes of POLD1 mutants. Larger scale siRNA screening will be employed to identify additional novel relevant target pathways as well as previously unknown functional interactions of progeroid genetic loci.

项目摘要 Werner综合征国际登记处招募了Werner综合征（WS）病例， 来自世界各地的其他节段性早老样综合征，目的是阐明潜在的 加速老化的机制。详细的临床信息，遗传分析结果，以及生物学 样本提供给各种合格的老年科学家。 在本申请中，我们建议扩展我们以前的研究，包括系统的全基因组 寻找导致78例早衰症的遗传变异，我们迄今为止还无法 基因特征我们的研究议程的另一个重要扩展是启动翻译 研究可能导致潜在的治疗药物。 我们将采用下一代测序、阵列CGH和桑格测序的组合， 随后进行确证性Western分析和定量PCR。这些方法成功地 鉴定了WRN（DNA解旋酶）、LMNA（核结构的组分） POLD 1（DNA聚合酶δ），SPRTN（DNA聚合酶募集），ERCC 4（核苷酸切除修复）， CTC 1（端粒复制）、MDM2（P53的抑制剂）和SAMHD 1（dNTP库的调节）。这些基因座 强调基因组不稳定性的主要作用，现在被广泛接受为衰老的标志之一。我们也 在识别疾病突变方面取得了进展，这些突变表明了加速衰老的其他机制， 如BSCL 2（脂滴形成）和SMAD 4（SASP成分TGFβ的细胞内信号传导）。 如上所述，我们将继续进行具有发展潜力的转化研究， 为我们的早衰症患者提供改善治疗。根据我们之前的研究，我们的合作者，横手博士 日本沃纳财团的Koutaro正在评估NAD中间体和mTOR的功效， 抑制剂二甲双胍治疗WS患者。我们将开始独立的研究， 慢性炎症，即Janus激酶（JAK）抑制剂，在来自WS患者和对照的培养物中。这 我们发现SMAD4突变的成纤维细胞表现出增加的DNA积累， WRN突变成纤维细胞显示SMAD 4表达升高， SASP与对照细胞相比，表明持续的DNA损伤和炎症的协同作用， 可能是导致加速老化的共同关键机制之一。协调一致的实验将 探索这些新的治疗靶点的作用，高通量筛选细胞培养物， 患有其他早衰综合征的患者。一个涉及siRNA筛选的初始小规模实验， 揭示了siRNA和改变核内dNTP浓度的药物能够调节细胞内dNTP浓度。 POLD1突变体的疾病表型。将采用更大规模的siRNA筛选来鉴定另外的siRNA。 新的相关靶向通路以及先前未知的早衰症遗传基因座的功能相互作用。