Genomic and epigenomic mechanisms of pediatric ocular disorders

医学遗传学和眼科基因组实验室将研究计划的目标和目标推进如下： 1.利用CRISPR/Cas9基因编辑以及发育、分子和细胞生物学研究，在斑马鱼、小鼠和体外建立了眼部症状与编码基因变异的新关联，以阐明疾病机制。我们目前正在动物和细胞模型中模拟几种新的人类疾病基因。一个例子是CSDE1，它是一个包含RNA结合程序的冷休克结构域。我们已经证明，在人类中，该基因的单倍性不足会导致一种新的神经发育障碍，具有自闭症样特征、身材矮小、相对小头畸形，在某些情况下还会出现虹膜缺陷症。这与人类相邻的基因缺失是一致的，这表明CSDE1是1p13.2微缺失综合征的关键基因。这份手稿目前正在印刷中(《科学进展》，2019)。我们小组还合作定义了多种新的疾病-基因关联，包括SMPD4、BMPR1A和MYRF(见参考文献)。 2.使用功能基因组学技术确定非编码基因组和眼睛组织中的疾病关联。使用RNAseq、ATACseq和Hi-C，以及单细胞测序技术，我们正在绘制来自诱导多能细胞系的人眼细胞的活性基因组图，以根据人类测序数据进行不同的优先排序。这些转录学工作促进了眼整合，这是一个用于从眼组织中获取人类测序数据的公共数据库，并使用机器学习生成了预测网络(眼整合.nei.nih.gov；见参考文献)。

The Medical Genetics and Ophthalmic Genomics Laboratory has advanced the goals and objectives of the research program as follows: 1. Modeling novel associations of ocular syndromes with coding gene variation in zebrafish, mouse, and in vitro using CRISPR/Cas9 gene editing and developmental, molecular, and cellular biology investigations to elucidate disease mechanisms. We are currently modeling several new human disease genes in animal and cellular models. One example is CSDE1, a cold-shock domain containing RNA-binding program. We have shown that haploinsufficiency of this gene in humans causes a novel neurodevelopemntal disorder with autistic like features, short stature, relative microcephaly, and in some cases iris coloboma. This is consistent with contiguous gene deletions in humans, framing CSDE1 as the critical gene for 1p13.2 microdeletion syndrome. This manuscript is currently in press (Science Advances, 2019). Our group has also acted in collaboration to define multiple additional new disease-gene associations, including SMPD4, BMPR1A, and MYRF (see bibliography). 2. Defining disease-associations in the noncoding genome and in eye tissues using functional genomics techniques. Using RNAseq, ATACseq, and Hi-C, along with single-cell sequencing technologies, we are mapping the active genome in human ocular cells derived from induced pluripotent cell lines for variant prioritization from human sequencing data. These transcriptomic efforts contributed to eyeIntegration, a public database for human sequencing data from ocular tissues and generation of predictive networks using machine learning (eyeintegration.nei.nih.gov; see bibliography).