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

多囊卵巢综合征（PCOS）是一种高度遗传的复杂生殖和代谢疾病，影响全球高达15%的育龄妇女。PCOS的病因仍然未知，因此诊断标准，例如NIH和Rotter-dam，是基于专家意见，而不是疾病机制的知识。我们最近对欧洲（EA）血统病例的全基因组关联研究（GWAS）进行的荟萃分析发现， 不同诊断标准所定义的PCOS的发病率基本相似。这一发现表明， 不能识别生物学上不同的疾病亚型。相比之下，使用无监督的层次聚类分析 在EA PCOS中，我们确定了两种PCOS亚型：一种是“生殖”组，其特征是较高的促黄体生成素（LH）和性激素结合球蛋白（SHBG）水平，相对较低的BMI和胰岛素水平; “代谢”组的特点是较高的BMI以及葡萄糖和胰岛素水平与相对较低的SHBG LH水平。我们在另一个EA PCOS队列中复制了这些亚型。我们执行了GWAS， 亚型，并发现6个新的基因座在全基因组意义，5个基因座与生殖亚型 和一个与代谢亚型相关的基因座。此外，这些位点的效应量基本上是 大于现有标准下与PCOS诊断相关的GWAS位点。我们有令人兴奋的初步数据表明，这些亚型存在于非洲裔美国人（AA），西班牙裔（HA）和东方人（HA）的PCOS病例中。 亚洲（韩国，KA）血统。我们的总体假设是，PCOS有表型亚型， 独特的基因结构我们将：（1）检验在额外的EA中存在PCOS亚型的假设 表型多样性PCOS的队列，并评估这些亚型的遗传结构。我们将执行 在额外的EA癌症PCOS病例对照队列中进行生殖和代谢数量性状的无监督分层聚类分析。我们将正式评估遗传结构的差异，并对GWAS数据进行精细映射，以选择Aim 3功能研究的变体。(2)检验亚型是 存在于非洲，西班牙裔和东亚血统的PCOS中，并评估这些遗传结构 亚型聚类分析，GWAS与亚型，遗传结构的评估和精细定位将是 在AA、HA和KA PCOS病例对照队列中按照目标1进行。将进行跨种族荟萃分析，以利用血统差异进行基因发现。(3)测试高优先级变体 与PCOS亚型相关的基因在与疾病发病机制相关的组织中起作用。我们将确定 来自目标1和2的非编码遗传变异体以高通量产生遗传关联信号， 我们在人卵泡膜、颗粒层和前脂肪细胞系中开发了报告基因测定。受影响的基因将 使用基于CRISPR/Cas9的测定进行研究。这项研究将产生持续和持久的影响， 通过定义PCOS的生物学相关亚型和确定关键途径中的因果变异， 在不同人群中与PCOS发病机制相关。

PCOS (polycystic ovary syndrome) 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.