Downstream of the “Androgen receptor-bottleneck”: the transcriptional and chromatin landscape of urogenital midline fusion at single cell resolution and its relevance to differences of sex development (DSD)

“雄激素受体瓶颈”的下游：单细胞分辨率下泌尿生殖中线融合的转录和染色质景观及其与性别发育差异（DSD）的相关性

• 批准号: 525378710
• 负责人: Professor Dr. Paul-Martin Holterhus
• 金额: --
• 依托单位: Institut für Humangenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/525378710?language=en
• 关键词: Downstream; Androgen; receptor; bottleneck; transcriptional

Differences of sex development (DSD) refer to conditions in which a person is born with reproductive or sexual anatomy that doesn’t fit the typical definitions of female or male. So far 75 disease genes have been associated with DSDs, but currently the majority of patients with DSD do not receive a molecular diagnosis even after Exome sequencing. This represents a huge challenge in clinical DSD-management due to an uncertain individual prognosis. We assume that missing knowledge about the genes and pathways involved in the androgen dependent genital midline closure program contributes significantly to this problem. The recent advances in the field of single-cell genomics have revolutionized how we study development and disease and could also help understanding the pathology of DSD. Here we hypothesize that joint profiling of single-cell transcriptome and chromatin accessibility in the external genital tubercle of a mouse model for DSD (TFM mouse) will reveal critical insights into the development of DSD and the regulatory architecture of the androgen pathway and help identify new disease genes and critical non-coding elements related to DSD. To investigate this hypothesis, we will apply the following three experimental approaches: Objective 1: To generate a joint single-cell transcriptome and chromatin accessibility atlas of external genital development and urethral tube formation in AR knock-out mice (XY TFM). Objective 2: To create an atlas of candidate genes and non-coding regulatory elements potentially associated with DSD Objective 3: To cross-validate candidate genes from the developed mouse reference atlas in DNA-samples established from cultured genital fibroblasts derived from the external genitalia of human DSD-patients by genome DNA sequencing enriched with patients with a pathological APOD assay indicating molecular disruption of cellular androgen signaling. Our study will generate new molecular insights into the development of DSD and identify new disease genes and non-coding enhancer elements that contribute to DSD. Mutations in these non-coding regulatory elements could in principle explain a substantial proportion of so far unresolved cases of DSD.

性发育差异 (DSD) 是指一个人出生时的生殖或性解剖结构不符合女性或男性的典型定义，迄今为止，已有 75 个疾病基因与 DSD 相关，但目前大多数是这种情况。由于个体预后不确定，DSD 患者即使在外显子组测序后也没有得到分子诊断，这对临床 DSD 管理来说是一个巨大的挑战。单细胞基因组学领域的最新进展彻底改变了我们研究发育和疾病的方式，也有助于理解 DSD 的病理学。在这里，我们对单细胞转录组和染色质可及性进行联合分析。 DSD 小鼠模型（TFM 小鼠）的外生殖器结节将揭示对 DSD 发展和雄激素途径调控架构的重要见解，并有助于识别与 DSD 相关的新疾病基因和关键非编码元件。根据这一假设，我们将应用以下三种实验方法： 目标 1：生成 AR 敲除小鼠（XY TFM）外生殖器发育和尿道管形成的联合单细胞转录组和染色质可及性图谱。创建可能与 DSD 相关的候选基因和非编码调控元件图谱 目标 3：在从培养的生殖器成纤维细胞建立的 DNA 样本中交叉验证已开发的小鼠参考图谱中的候选基因我们的研究将通过基因组 DNA 测序对人类 DSD 患者的外生殖器进行富集，并通过病理 APOD 检测表明细胞雄激素信号传导的分子破坏，并鉴定新的疾病基因和非编码增强子。这些非编码调控元件的突变原则上可以解释迄今为止尚未解决的 DSD 病例的很大一部分。