HIGH THROUGHPUT GENOTYPING AND DNA SEQUENCING FOR STUDYING THE GENETIC CONTRIBUTIONS TO HUMAN HEALTH AND DISEASE: Whole Genome Sequencing of Bilateral Cleft Lip and Palate families from Africa

口面裂（OFC）影响全世界每700名出生婴儿中的一名，是人类最常见的出生缺陷之一。对OFC的遗传和分子病因学的研究是预防的必要前提，并将对与OFC相关的经济、教育、医疗、心理和文化问题产生巨大影响，从而带来重大的公共卫生效益。我们通过7项CL/P全基因组关联研究（GWAS）、3项CL/P荟萃分析、3项仅腭裂（CPO）全基因组关联研究和2项全基因组测序（WGS）研究，在人类OFC遗传学方面取得了合理的进展，确定了60多个风险基因座和几种新生变异。提出的具体目标将使我们能够识别和测试增加CL/P风险的常见和罕见遗传变异，并识别与人类中积累了最大遗传变异的人群中特定OFC表型相关的遗传变异。使用全基因组测序来鉴定罕见的功能变体有望大大扩展已经进行的OFC的少数外显子组研究的结果。我们假设双侧完全性唇腭裂（BCLP），临床上最严重的OFC形式，与较轻的形式相比，与更高的突变负荷相关，专注于BCLP将有助于发现新的风险变体。这种方法是稳健的，因为我们使用临床同质队列，以尽量减少遗传异质性，并增加发现BCLP遗传因素的可能性。这项申请将由Butali博士作为主要研究者领导。他将与约翰霍普金斯大学的陶布博士和NHGRI的Adeyemo博士合作。总的来说，本提案中概述的实验设计和方法、PI和co-Is以及环境提供了改善口腔和颅面健康的催化剂。

Orofacial clefts (OFCs) affects one out of every 700 infants born worldwide and are some of the most common birth defects in humans. Investigations into the genetic and molecular etiology of OFC is an essential pre- requisite for prevention and will have a huge impact on financial, educational, medical, psychological, and cultural problems associated with OFCs—leading to significant public health benefits. We have made reasonable progress in human OFC genetics through seven Genome Wide Association Studies (GWAS) for CL/P, three meta-analyses for CL/P, three GWAS for cleft palate only (CPO), and two Whole Genome Sequencing (WGS) study identifying over 60 risk loci and several de novo variants. The proposed specific aims will allow us to identify and test both common and rare genetic variants that elevate risk for CL/P, and to identify genetic variants associated with specific OFC phenotypes in the population that has accumulated the greatest genetic variation in the human race. The use of whole-genome sequencing to identify rare functional variants is expected to substantially expand the findings from the few exome studies of OFC that have been conducted. We hypothesize that bilateral complete cleft lip and palate(BCLP), the most clinically severe form of OFC, is associated with a higher mutation load than less severe forms and focusing on BCLP will facilitate the discovery of novel risk variants. This approach is robust in that we are using a clinically homogeneous cohort in order to minimize genetic heterogeneity and increases the likelihood of discovering genetic factors for BCLP. This application will be led by Dr Butali as Principal Investigator. He will collaborate with Dr. Taub at John Hopkins University and Dr. Adeyemo at the NHGRI. Collectively, the experimental design and approach, the PI and co-Is, and the environment outlined in this proposal provide the catalyst to improve oral and craniofacial health.