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Genetic and Functional Studies of Human Ciliary Syndromes

人类睫状体综合征的遗传和功能研究

基本信息

批准号：
10436165
负责人：
Erica Ellen Davis
金额：
$ 44.12万
依托单位：
LURIE CHILDREN'S HOSPITAL OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-09-14 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10436165
关键词：
3&apos Untranslated Regions Ablation Affinity Alleles Arts Bardet-Biedl Syndrome Benign Binding Binding Sites Biochemical Biological Candidate Disease Gene Cell physiology Cells Cilia Clinical Code Complement Custom DNA Sequence Rearrangement Data Data Set Defect Development Diagnostic Disease Dissection Embryo Etiology Exhibits Family Functional disorder Genes Genetic Genetic Complementation Test Genetic Diseases Genetic Epistasis Genetic Variation Genotype Harvest Human Human Genetics In Vitro Incidence Kidney Knowledge Larva Length Libraries Link Measurement Measures Meckel-Gruber syndrome Mediating MicroRNAs Missense Mutation Modality Modeling Modification Molecular Mutate Mutation Organ Pathogenicity Pathway interactions Patients Phenotype Protein Isoforms Proteins RNA Splicing Refractory Regulation Reporter Reporting Retina Role Severities Severity of illness Signal Transduction Site Small Interfering RNA Specificity Syndrome Testing Therapeutic Tissue-Specific Splicing Tissues Transcript Variant WNT Signaling Pathway Work Zebrafish base causal variant ciliopathy cilium biogenesis cohort differential expression dosage exome sequencing experimental study gene discovery genetic architecture genome sequencing genome wide screen genome-wide improved in vivo kinetosome novel overexpression pleiotropism preference protein function rational design socioeconomics tool transcriptome transcriptomics whole genome

项目摘要

The ciliopathies are a group of >100 genetic disorders unified by overlapping structural and/or functional defects of the cilium and the basal body. The study of these disorders has highlighted fundamental developmental and homeostatic mechanisms, while offering the opportunity to develop computational and functional tools to discover new causal genes; to expand the phenotypic spectrum of known genes; and to design rational therapeutic paradigms. In the backdrop of striking progress during the past two decades, major challenges and opportunities remain. Specifically, despite a diagnostic rate that for some ciliopathies approaches 90%, the predictive power of the genotype at the primary recessive locus remains insufficient to inform clinical manifestation. This is in part because our understanding of the pathomechanisms of pleiotropy and variable expressivity remain poorly understood. Moreover, almost all ciliopathy discovery work to date has: (a) focused on coding mutations and genomic rearrangements; and (b) assumed that the pathogenicity of alleles is constant in all cellular contexts. In this Renewal, we aspire to improve upon these knowledge gaps. First, we will take advantage of a genome-wide siRNA screen on cells ablated for BBS4 that report on aberrant Wnt signaling. This experiment harvested 81 genes that, when suppressed, exacerbate ciliary cellular phenotypes and therefore represent de facto candidates for harboring either causal or epistatic mutations in patients with ciliary disease. We will test this hypothesis by sequencing our phenotypically diverse patient cohort and testing functionally resultant candidate genes and alleles using state-of-the-art in vivo tools. Critically, the majority of these 81 genes are not components of the ciliary apparatus, offering the opportunity to forge new biological links between ciliary dysfunction and other cellular processes. In parallel, we will pursue studies that will explore a new phenomenon of allele pathogenicity, in which bona fide pathogenic missense variants behave as deleterious to protein function in some splice isoforms but benign in others. Using a combination of in vivo complementation testing and in vitro biochemical studies, we will ask how common these phenomena are and whether subcellular localization or stability might represent informative biochemical drivers of these phenomena. Finally, through a genome-wide screen for miRNAs that regulate ciliogenesis, we discovered let-7b as a regulator of ciliary length. Subsequent transcriptomic analysis identified 42 ciliary genes with differential expression correlated to let-7b dosage, and which harbor predicted let-7b sites in their 3’ UTR. We will use this rich dataset to ask whether such miRNA binding sites in 3’ UTRs of known ciliopathy genes contribute to causality and modification in this group of disorders. Together, our studies will inform the genetic architecture of the ciliopathy disease entity by discovering sites that likely modulate the expressivity of disease; by exploring the context-dependent effect of pathogenic variation; and by interrogating the contribution to disease of a largely unexplored class of variants impacting regulatory mechanisms.

纤毛病是一组超过 100 种遗传性疾病，由重叠的结构和/或功能统一起来。纤毛和基体的缺陷。对这些疾病的研究强调了基本的发育和稳态机制，同时提供发展计算和发现新因果基因的功能工具；扩大已知基因的表型谱；并到设计合理的治疗范式。在过去二十年取得显着进步的背景下，重大挑战和机遇依然存在。具体来说，尽管某些纤毛病的诊断率接近 90%，初级隐性基因座基因型的预测能力仍然不足以告知临床表现。这部分是因为我们对多效性病理机制的理解和变量表达性仍然知之甚少。此外，迄今为止几乎所有纤毛病发现工作都具有： (a) 重点关注编码突变和基因组重排； (b) 假设致病性等位基因在所有细胞环境中都是恒定的。在本次更新中，我们渴望弥补这些知识差距。首先，我们将利用对 BBS4 消融的细胞进行全基因组 siRNA 筛选，报告异常情况 Wnt 信号传导。该实验收获了 81 个基因，当这些基因被抑制时，会加剧睫状细胞表型，因此代表了携带因果突变或上位突变的事实上的候选者患有纤毛疾病的患者。我们将通过对表型多样化的患者进行测序来检验这一假设使用最先进的体内工具对候选基因和等位基因进行队列和测试功能。至关重要的是，这 81 个基因中的大多数都不是睫状体的组成部分，因此有机会在纤毛功能障碍和其他细胞过程之间建立新的生物学联系。与此同时，我们将追求研究将探索等位基因致病性的新现象，其中真正的致病性错义在某些剪接异构体中，变异体对蛋白质功能有害，但在其他剪接异构体中却是良性的。使用结合体内互补测试和体外生化研究，我们会问有多常见这些现象以及亚细胞定位或稳定性是否可能代表生化信息这些现象的驱动因素。最后，通过全基因组筛选调节纤毛发生的 miRNA，我们发现let-7b作为纤毛长度的调节剂。随后的转录组分析鉴定出 42 个纤毛基因差异表达与let-7b剂量相关，并且在其3'UTR中包含预测的let-7b位点。我们将使用这个丰富的数据集来询问已知纤毛病基因的 3' UTR 中是否存在此类 miRNA 结合位点有助于这组疾病的因果关系和改变。我们的研究将共同为遗传提供信息通过发现可能调节疾病表现的位点来了解纤毛病疾病实体的结构；通过探索致病变异的背景依赖性效应；并通过询问对影响调节机制的一类很大程度上未经探索的变异疾病。

项目成果

期刊论文数量（22）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（1）

CFAP45 deficiency causes situs abnormalities and asthenospermia by disrupting an axonemal adenine nucleotide homeostasis module.

DOI：
10.1038/s41467-020-19113-0
发表时间：
2020-11-02
期刊：
Nature communications
影响因子：
16.6
作者：
Dougherty GW;Mizuno K;Nöthe-Menchen T;Ikawa Y;Boldt K;Ta-Shma A;Aprea I;Minegishi K;Pang YP;Pennekamp P;Loges NT;Raidt J;Hjeij R;Wallmeier J;Mussaffi H;Perles Z;Elpeleg O;Rabert F;Shiratori H;Letteboer SJ;Horn N;Young S;Strünker T;Stumme F;Werner C;Olbrich H;Takaoka K;Ide T;Twan WK;Biebach L;Große-Onnebrink J;Klinkenbusch JA;Praveen K;Bracht DC;Höben IM;Junger K;Gützlaff J;Cindrić S;Aviram M;Kaiser T;Memari Y;Dzeja PP;Dworniczak B;Ueffing M;Roepman R;Bartscherer K;Katsanis N;Davis EE;Amirav I;Hamada H;Omran H
通讯作者：
Omran H

A human laterality disorder caused by a homozygous deleterious mutation in MMP21.

DOI：
10.1136/jmedgenet-2015-103336
发表时间：
2015-12
期刊：
Journal of medical genetics
影响因子：
4
作者：
Perles Z;Moon S;Ta-Shma A;Yaacov B;Francescatto L;Edvardson S;Rein AJ;Elpeleg O;Katsanis N
通讯作者：
Katsanis N

Thermosensory and mechanosensory perception in human genetic disease.

人类遗传疾病中的热感觉和机械感觉。