Human Genetic Variation and Disease

在这次更新中，我们将继续研究人类疾病的原因和疾病后果。 遗传变异我们已经从三代中产生了全基因组序列（WGS）数据， 犹他州CEPH家系纵向表征从头突变（DNM）的发生率和模式。我们 显示了家庭间突变累积率的显著差异，我们 表明10%的表观DNM实际上是导致嵌合现象的合子后事件。我们 还表明，年龄调整的生殖系DNM传播率较高， 寿命更短更年期更早这些结果意味着一个人的生殖系DNM率， 随着年龄的增长，与体细胞突变率有关。在下一个融资期内，我们将测试 这一假设直接使用血液来源的DNA样本收集在三个不同的时间点， 相同CEPH谱系成员在35年的时间内。据我们所知，这是第一个长期的 人类家系生殖细胞和体细胞突变率的纵向研究。我们还假设， DNA修复机制影响生殖细胞和体细胞突变率的变化。我们将测试 这一假设使用全基因组序列数据（由单独的资金支持）和RNA-seq数据 从新鲜采集的全血样本中。我们预测关键DNA修复的低表达 基因将导致更高的种系和体细胞突变率。 为了提高我们检测模式和疾病关联的能力，我们将（在单独的 资助）扩大CEPH谱系收藏，纳入第四代的700多名成员 这一代人，现在已经成年了。我们还将对第2代和第3代进行第三次重采样。我们将 在这些研究参与者中进行WGS和RNA-seq，创建一个独特的，公开可用的储存库 跨越四代的遗传信息。进行了180多项表型测量 对于20年前CEPH家系的第2代和第3代， 第四代。这些资源将使我们能够探索遗传变异的影响，包括突变 率和DNA修复，在几代人的表型的广泛分类。 此外，我们将继续努力开发新的和改进的基因组方法， 序列分析，包括检测移动的元件插入。我们将分析WGS， 从犹他州人群获得的大犹他州疾病谱系成员的RNA-seq数据 肌萎缩侧索硬化症和幼年特发性关节炎。这些项目 涉及WGS和RNA-seq数据分析以及体外和体内系统中的功能分析。

In this renewal, we will continue our studies of the causes and disease consequences of human genetic variation. We have generated whole-genome sequence (WGS) data from the three-generation Utah CEPH pedigrees to longitudinally characterize rates and patterns of de novo mutations (DNMs). We showed significant differences in the rates of accumulation of mutations among families, and we demonstrated that 10% of apparent DNMs are actually postzygotic events that result in mosaicism. We also showed that a higher rate of age-adjusted germline DNM transmission is significantly associated with a shorter lifespan and earlier menopause. These results imply that one’s germline DNM rate, which increases with age, is associated with the somatic mutation rate. In the next funding period, we will test this hypothesis directly using blood-derived DNA samples collected at three different time points in the same CEPH pedigree members over a 35-year time period. To our knowledge, this is the first long-term longitudinal study of germline and somatic mutation rates in human pedigrees. We also hypothesize that DNA repair mechanisms influence variation in both germline and somatic mutation rates. We will test this hypothesis using whole-genome sequence data (supported by separate funding) and RNA-seq data from freshly collected whole blood samples. We predict that lower expression of critical DNA repair genes will result in higher germline and somatic mutation rates. To increase our power to detect patterns and disease associations, we will (under separate funding) expand the CEPH pedigree collection to include more than 700 members of the fourth generation, who are now adults. We will also resample generations 2 and 3 for a third time. We will undertake WGS and RNA-seq in these study participants, creating a unique, publicly available repository of genetic information spanning four generations. More than 180 phenotypic measurements were made for generations 2 and 3 of the CEPH pedigrees 20 years ago, and the same measurements will be made for generation 4. These resources will allow us to explore the effects of genetic variation, including mutation rates and DNA repair, on a broad assortment of phenotypes across several generations. In addition, we will continue our work to develop new and improved methods for genome sequence analysis, including the detection of mobile element insertions. We will analyze WGS and RNA-seq data from members of large Utah disease pedigrees obtained from the Utah Population Database, focusing on amyotrophic lateral sclerosis and juvenile idiopathic arthritis. These projects involve analysis of WGS and RNA-seq data as well as functional analysis in in vitro and in vivo systems.