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.

纤毛病是由重叠结构和/或功能统一的> 100个遗传疾病的组纤毛和基本体的缺陷。这些疾病的研究突出了基本发展和稳态机制，同时提供了开发计算和发现新因果基因的功能工具；扩大已知基因的表型光谱；然后设计合理的治疗范例。在过去的二十年中，杰出进步的背景挑战和机遇仍然存在。具体而言，使命诊断率是某些纤毛病的诊断率接近90％，主要隐性基因座的基因型的预测能力仍然不足告知临床表现。这部分是因为我们对多效性的病理机理的理解并且可变表达性仍然很少理解。此外，迄今为止，几乎所有的纤毛病发现工作都有：（a）专注于编码突变和基因组重排；（b）假设等位基因在所有细胞环境中都是恒定的。在这种续约中，我们渴望改善这些知识差距。首先，我们将利用在烧开BBS4的细胞上的全基因组siRNA屏幕，以报告异常 wnt信号传导。该实验收集了81个基因，当抑制时，会加剧睫状细胞表型，因此代表了事实上的候选者，用于携带因果关系或鼻突变。患有睫状性疾病的患者。我们将通过对表型多样的患者进行测序来检验这一假设同类和测试在功能上由最先进的体内工具进行功能的候选基因和等位基因。至关重要的是，这81个基因中的大多数不是睫状设备的组成部分，为机会提供了机会在睫状功能障碍与其他细胞过程之间建立新的生物学联系。同时，我们将追求将探讨等位基因致病性新现象的研究，其中真正的致病错过在某些剪接同工型中，变体对蛋白质功能的表现是有害的，但在其他链接中却是良性的。使用体内完成测试和体外生化研究的结合，我们将询问如何普遍这些现象是以及亚细胞定位还是稳定性可能代表信息化的生化这些现象的驱动因素。最后，通过调节纤毛发生的miRNA的全基因组筛查，我们发现let-7b是睫状长度的调节剂。随后的转录组分析确定了42个睫状基因具有差异表达与Let-7b剂量相关，并且携带的3'UTR中预测了Let-7b位点。我们将使用这个丰富的数据集询问是否在3英寸UTR中的这种miRNA结合位点是否已知纤毛病基因在这组疾病中有助于偶然性和修改。一起，我们的研究将为通用通过发现可能调节疾病表现力的部位，纤毛病疾病实体的结构；通过探索致病性变异的上下文依赖性效应；并通过审问对影响调节机制的一系列意外类型的疾病。

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。