Department of Human Molecular Genetics and Biochemistry, Sackler Faculty of Medicine and Sagol School of Neurosciences, Tel Aviv University, Tel Aviv, Israel.
The Hadassah Human Embryonic Stem Cell Research Center, The Goldyne Savad Institute of Gene Therapy and Department of Gynecology, Jerusalem, Israel.
PLoS Biol. 2023 Jan 17;21(1):e3001924. doi: 10.1371/journal.pbio.3001924. eCollection 2023 Jan.
Tissue-specific transcription factors (TFs) control the transcriptome through an association with noncoding regulatory regions (cistromes). Identifying the combination of TFs that dictate specific cell fate, their specific cistromes and examining their involvement in complex human traits remain a major challenge. Here, we focus on the retinal pigmented epithelium (RPE), an essential lineage for retinal development and function and the primary tissue affected in age-related macular degeneration (AMD), a leading cause of blindness. By combining mechanistic findings in stem-cell-derived human RPE, in vivo functional studies in mice and global transcriptomic and proteomic analyses, we revealed that the key developmental TFs LHX2 and OTX2 function together in transcriptional module containing LDB1 and SWI/SNF (BAF) to regulate the RPE transcriptome. Importantly, the intersection between the identified LHX2-OTX2 cistrome with published expression quantitative trait loci, ATAC-seq data from human RPE, and AMD genome-wide association study (GWAS) data, followed by functional validation using a reporter assay, revealed a causal genetic variant that affects AMD risk by altering TRPM1 expression in the RPE through modulation of LHX2 transcriptional activity on its promoter. Taken together, the reported cistrome of LHX2 and OTX2, the identified downstream genes and interacting co-factors reveal the RPE transcription module and uncover a causal regulatory risk single-nucleotide polymorphism (SNP) in the multifactorial common blinding disease AMD.
组织特异性转录因子(TFs)通过与非编码调控区域(顺式作用元件)结合来控制转录组。确定决定特定细胞命运的 TF 组合、它们的特定顺式作用元件,并研究它们在复杂人类特征中的作用仍然是一个主要挑战。在这里,我们专注于视网膜色素上皮(RPE),它是视网膜发育和功能所必需的谱系,也是年龄相关性黄斑变性(AMD)的主要受累组织,AMD 是导致失明的主要原因之一。通过结合干细胞衍生的人类 RPE 中的机制研究、在小鼠中的体内功能研究以及全局转录组和蛋白质组学分析,我们揭示了关键的发育性 TF LHX2 和 OTX2 一起在包含 LDB1 和 SWI/SNF(BAF)的转录模块中发挥作用,以调节 RPE 的转录组。重要的是,通过使用报告基因检测进行功能验证,确定的 LHX2-OTX2 顺式作用元件与已发表的表达数量性状基因座、人类 RPE 的 ATAC-seq 数据以及 AMD 全基因组关联研究(GWAS)数据之间的交集,揭示了一个因果遗传变异,通过调节其启动子上的 LHX2 转录活性来改变 RPE 中的 TRPM1 表达,从而影响 AMD 风险。综上所述,所报道的 LHX2 和 OTX2 的顺式作用元件、确定的下游基因和相互作用的共同因子揭示了 RPE 的转录模块,并揭示了多因素常见致盲疾病 AMD 中的一个因果调节风险单核苷酸多态性(SNP)。