Elucidating the Genetic Architecture of Metabolic and Reproductive PCOS Subtypes in Diverse Populations

多囊卵巢综合征是一种高度可遗传的复杂生殖和代谢疾病，影响多达15%的生殖- 世界各地的老年妇女。多囊卵巢综合征的病因尚不清楚，因此诊断标准，如NIH和Rotter- 是基于专家的意见，而不是疾病机制的知识。我们最近的荟萃分析 对欧洲人(EA)祖先进行的全基因组关联研究(GWAS)发现，遗传结构 不同诊断标准所定义的多囊卵巢综合征的诊断结果大体相似。这一发现表明，这些标准 不要识别生物学上截然不同的疾病亚型。相比之下，使用无监督的层次聚类分析 在EA PCOS中，我们发现了两种PCOS亚型：一种以高黄体化水平为特征的“生殖”型； 体重指数和胰岛素水平相对较低的黄体生成素和性激素结合球蛋白水平； “代谢”组的特点是BMI较高，血糖和胰岛素水平较低，SHBG相对较低 和促黄体生成素水平。我们在另一个EA PCOS队列中复制了这些亚型。我们使用 发现了6个具有全基因组意义的新基因座，其中5个与生殖亚型相关 和一个与代谢亚型相关的基因座。此外，这些基因座的效应大小基本上是 大于现有标准中与PCOS诊断相关的GWA位点数。我们有激动人心的预演- 初步数据表明，这些亚型存在于非裔美国人(AA)、西班牙裔(HA)和东方人的多囊卵巢综合征病例中 亚洲人(韩国人，Ka人)血统。我们的主要假设是，存在PCOS的表型亚型 独特的遗传结构。我们将：(1)检验在附加EA中存在PCOS亚型的假设 对表型不同的多囊卵巢综合征的队列，并评估这些亚型的遗传结构。我们将表演 生殖代谢数量性状的非监督系统聚类分析 CESTRESS多囊卵巢综合征病例对照队列。我们将正式评估遗传架构的差异，并进行精细- 为AIM 3功能研究选择变异体绘制Gwas数据图。(2)验证子类型为 存在于非洲、西班牙裔和东亚血统的多囊卵巢综合征中，并评估这些血统的遗传结构 子类型。将进行聚类分析、带亚型的遗传多样性分析、遗传结构评估和精细作图 按照目标1在AA、HA和KA多囊卵巢综合征病例对照队列中执行。跨文化的荟萃分析将继续- 以利用祖先的差异来发现基因。(3)检验高优先级变量的假设 与PCOS相关的亚型在与疾病发病机制相关的组织中具有功能。我们将确定 来自AIMS 1和2的非编码遗传变体导致遗传关联信号与高通量 我们在人卵泡膜、颗粒和前脂肪细胞系中建立了报告基因分析。受影响的基因将 使用基于CRISPR/CAS9的检测方法进行调查。这项研究将对 通过定义与生物相关的多囊卵巢综合征亚型和确定关键途径中的因果变异来研究这一领域 与不同人群中多囊卵巢综合征的发病机制有关。

PCOS is a highly heritable, complex reproductive and metabolic disorder affecting up to 15% of reproductive- age women worldwide. The etiology of PCOS remains unknown so the diagnostic criteria, e.g. NIH and Rotter- dam, are based on expert opinion rather than on knowledge of disease mechanisms. Our recent meta-analysis of genomewide association studies (GWAS) of European (EA) ancestry cases found that the genetic architecture of PCOS defined by the different diagnostic criteria was generally similar. This finding suggests that these criteria do not identify biologically distinct disease subtypes. In contrast, using unsupervised hierarchical cluster analysis in EA PCOS, we identified two PCOS subtypes: a “reproductive” group characterized by higher luteinizing hor- mone (LH) and sex hormone binding globulin (SHBG) levels with relatively low BMI and insulin levels; and a “metabolic” group characterized by higher BMI as well as glucose and insulin levels with relatively low SHBG and LH levels. We replicated these subtypes in an additional EA PCOS cohort. We performed GWAS with the subtypes and found six novel loci at genomewide significance, five loci associated with the reproductive subtype and one locus associated with the metabolic subtype. Further, the effect sizes for these loci were substantially greater than those GWAS loci associated with PCOS diagnosis by the existing criteria. We have exciting pre- liminary data that these subtypes are present in PCOS cases of African American (AA), Hispanic (HA) and East Asian (Korean, KA) ancestry. Our overarching hypothesis is that there are phenotypic subtypes of PCOS with distinct genetic architecture. We will: (1) Test the hypothesis that there are subtypes of PCOS in additional EA cohorts of phenotypically diverse PCOS and assess the genetic architecture of these subtypes. We will perform unsupervised hierarchical cluster analysis of reproductive and metabolic quantitative traits in additional EA an- cestry PCOS case-control cohorts. We will formally assess differences in genetic architecture and conduct fine- mapping of GWAS data to select variants for Aim 3 functional studies. (2) Test the hypothesis that subtypes are present in PCOS of African, Hispanic and East Asian ancestry and assess the genetic architecture of these subtypes. Cluster analysis, GWAS with subtypes, assessment of genetic architecture and fine-mapping will be performed as in Aim 1 in AA, HA and KA PCOS case-control cohorts. Transethnic meta-analysis will be con- ducted to leverage differences in ancestry for gene discovery. (3) Test the hypothesis that high priority variants associated with PCOS subtypes are functional in tissues relevant to disease pathogenesis. We will identify the noncoding genetic variants from Aims 1 and 2 causing the genetic association signals with a high-throughput reporter assay we developed in human theca, granulosa and preadipocyte cell lines. The genes impacted will be investigated using CRISPR/Cas9-based assays. This research will have a sustained and lasting impact on the field by defining biologically relevant subtypes of PCOS and identifying causal variants in key pathways related to PCOS pathogenesis in diverse populations.