Genomic duplications in anophthalmia, microphthalmia and coloboma

项目摘要 小眼、无眼和眼缺损（MAC）是一组严重的眼部表型，其特征在于： 眼睛缩小或缺失，或涉及各种眼部结构的间隙（孔）。的成功率 确定MAC频谱的遗传诊断仍然不完整，使得新的发现 机制是重要的优先事项。拷贝数变异（CNV）是基因组DNA的缺失或重复。 可以遗传或作为显性从头变异发生的片段。有越来越多的发展清单 与CNV相关的表型，表明剂量不平衡是人类中的一个重要因素。 疾病在这一组中，由于许多因素，剂量增加变体的贡献仍然没有得到充分重视。 这些因素包括涉及多个基因，需要进行复杂的功能研究。在这一提议中， 我们将联合收割机结合我们在MAC和斑马鱼模型方面的专业知识，以揭示这些新的机制。 使人衰弱的表型。我们提供了两种不同剂量下剂量增加相关性的证据， 基因组区域，20 q11和3q 29，具有孤立和综合征MAC，确定了多个受累家族 在我们的队列中两个基因座都是如此。我们将利用我们广泛收集的DNA样本，从个人受影响， MAC进一步确定这些和其他候选区域对人类疾病的贡献，以及 发现新的致病剂量敏感位点。具体来说，我们将：1）识别驱动基因， 通过生成和研究剂量，参与与20 q11剂量变异相关的表型的途径 斑马鱼id 1、bcl 2l 1和tpx 2基因的增益/损失模型，被鉴定为该区域内的最佳候选基因; 2） 揭示与3q 29剂量变异相关的表型中涉及的驱动基因和途径， 探索位于受影响人体内的斑马鱼rubcn、dlg 1和/或bdh 1基因的剂量增加/损失模型 区域;以及3）通过额外的剂量敏感区域来揭示人类MAC谱中涉及的关键剂量敏感区域。 对先前鉴定的CNV作图以及探索一个大的队列（来自受影响的760个样本）， 个人及其亲属）的广泛测试，但无法解释的MAC家庭，然后解剖 斑马鱼中的新候选区域。该项目的成功完成将确定新的因素， 人眼发育，基因组区域/基因和受影响途径，并产生健壮的动物 这些复杂的人类表型的模型。

PROJECT SUMMARY Microphthalmia, anophthalmia and coloboma (MAC) is a group of severe ocular phenotypes characterized by a reduction in size or absence of the eye or a gap (hole) involving various ocular structures. The success rate for identifying a genetic diagnosis for MAC spectrum remains incomplete, making the discovery of novel mechanisms an important priority. Copy number variations (CNVs) are deletions or duplications of genomic segments that can be inherited or occur as dominant de novo variants. There is a growing list of developmental phenotypes associated with CNVs, indicating that dosage imbalance represents an important factor in human disease. Within this group, the contribution of dosage gain variants remains underappreciated due to many factors including the involvement of multiple genes necessitating complex functional studies. In this proposal, we will combine our expertise in MAC and the zebrafish model to reveal novel mechanisms of these debilitating human phenotypes. We present evidence for the association of dosage gain at two different genomic regions, 20q11 and 3q29, with isolated and syndromic MAC, with multiple affected families identified in our cohort for both loci. We will utilize our extensive collection of DNA samples from individuals affected with MAC to further define the contribution of these and other candidate regions to human disease, as well as to discover novel disease-causing dosage-sensitive loci. Specifically, we will: 1) identify the driver gene(s) and pathway(s) involved in phenotypes associated with 20q11 dosage variation by generating and studying dosage gain/loss models for the zebrafish id1, bcl2l1 and tpx2 genes, identified as top candidates within the region; 2) reveal driver gene(s) and pathway(s) involved in phenotypes associated with 3q29 dosage variation by exploring dosage gain/loss models for zebrafish rubcn, dlg1 and/or bdh1 genes, located in the affected human region; and 3) uncover critical dosage-sensitive regions involved in human MAC spectrum by additional mapping of previously identified CNVs as well as exploring a large cohort (760 samples from affected individuals and their relatives) of extensively tested but unexplained MAC families, followed by dissection of novel candidate regions in zebrafish. The successful completion of this project will identify novel factors in human ocular development, both genomic regions/genes and affected pathways, and generate robust animal models of these complex human phenotypes.