International Registry of Werner Syndrome

项目摘要 沃纳综合征及相关疾病国际登记处(www.wernersyndrome.org) 作为一种资源来确定分离突变的核家系并对其进行基因分型 沃纳综合征(WS)和一系列其他节段性孕激素综合征。我们将建立和 超低温保存来自这些家系的生物材料，并将它们提供给世界各地的研究人员。 我们现在建议在全基因组范围内进行系统的基因突变研究 41例原因不明的孕激素病例，寻找治疗药物的证据。我们将聘请一名 SNP阵列和下一代测序的组合；这些已经成功地识别了新的突变 在少数情况下。这些发现继续支持基因组不稳定是一种主要的 生物衰老的机制。这些基因座突出了在DNA修复和复制中的主要作用：WRN(DNA 解旋酶/核酸外切酶)、POLD1(DNA聚合酶增量)和SPRTN(转位DNA募集 聚合酶)；核结构和染色质相互作用(LMNA)；P53抑制因子(MDM2)；调控 DNTP池(SAMHD1)；端粒维持(CTC1)。 我们还将调查为什么WS表型只有在青春期之后才表现出来。我们假设 可能存在代偿机制的激活，如其他RecQ解旋酶或DNA修复 早期发育过程中的路径。为了验证我们的假设，我们将产生人类多能干细胞系 使用人类多能干细胞(HPSCs)和CRISPR检测WRN病突变和不突变 进行转录组研究。分析将集中在这些问题上：其他RecQ解旋酶和DNA是如何 修复相关基因在WS hPSCs中的表达与对照hPSCs相比，这些表达如何变化 分化后，这些表达是否与细胞的表达相关 衰老基因。由这个项目产生的细胞系和所有可用的患者材料将 可供其他调查人员使用。

Project Summary The International Registry of Werner Syndrome & Related Disorders (www.wernersyndrome.org) serves as a resource to ascertain and genotype nuclear pedigrees segregating mutations responsible for Werner syndrome (WS) and a range of other segmental progeroid syndromes. We shall establish and cryopreserve biological materials from these pedigrees and provide them to investigators around the world. We now propose to conduct systematic genome-wide searches for the gene mutations responsible for 41 progeroid cases with unknown causes and to seek evidence for therapeutic agents. We will employ a combination of SNP arrays and next generation sequencing; these have successfully identified novel mutations in a small number of cases. Those findings continue to support the concept of genomic instability as a major mechanism of biological aging. These loci highlight major roles in DNA repair and replication: WRN (DNA helicase/exonuclease), POLD1 (DNA polymerase delta), and SPRTN (recruitment of translesional DNA polymerase); nuclear structure and chromatin interaction (LMNA); an inhibitor of p53 (MDM2); regulation of dNTP pools (SAMHD1); and telomere maintenance (CTC1). We will also investigate why WS phenotypes are manifested only after puberty. We hypothesize that there may be an activation of compensatory mechanisms such as other RecQ helicases or DNA repair pathways during early development. To test our hypothesis, we will generate human pluripotent stem cell lines with and without WRN disease mutations using human pluripotent stem cells (hPSCs) and CRISPR and conduct transcriptome studies. Analysis will focus on these questions: how other RecQ helicases and DNA repair related genes are expressed in WS hPSCs compared to control hPSCs, how these expressions change following differentiation; whether or not these expressions correlated with the expression of cellular senescence genes. Cell lines generated by this project and all available patient materials will be made available to other investigators.