Exosomal protein deficiencies: how abnormal RNA metabolism results in childhood-onset neurological diseases

外泌体蛋白缺乏：RNA 代谢异常如何导致儿童期发病的神经系统疾病

• 批准号: MR/N025431/2
• 负责人: Rita Horvath
• 金额: $ 29.74万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FN025431%2F2
• 关键词: Exosomal; protein; deficiencies; how; abnormal

All genes are copied into short-lived RNA molecules, which are then translated to protein, forming the building box of the cells in the body. The transcription of DNA, the processing of pre-mRNA into mature mRNA (splicing), the degradation of mRNA and the translation into proteins are all tightly regulated. Both too much and too little of a certain RNA species could be dangerous for a cell, but our understanding of the mechanisms to fine-tune the RNA amounts is still limited. The regulation of gene expression (RNA metabolism) is of utmost importance for normal cell function, nerve cells, however, seem less able to cope with errors. This is illustrated by an increasing number of severe inherited neurological diseases of infancy or childhood caused by a defect in the RNA production machinery; these disorders are characterized by abnormalities of the development and/or the structure of the brain and the central nervous system. The aim of this proposal is to study and characterize a novel form of RNA related neurological disorder due to dysfunction of a multi-protein complex called the human exosome. The role of the exosome consists of degradation and maturation of RNA. Mutations in the exosome subunit gene EXOSC3 were reported in patients with infantile-onset degeneration of the brainstem and cerebellum and muscle weakness due to spinal motor neuron dysfunction. We have recently identified mutations in EXOSC8, encoding another component of the human exosome, in infants with a similar severe neurological disease. In addition, these children also lose myelin in the brain and spinal cord, which normally coats and insulates nerves and serves to speed up signalling. We also identified a homozygous mutation in a novel gene RBM7 in a child with severe spinal motor neuron dysfunction. RBM7 is an interacting partner of the human exosome. The severe and isolated neurological symptoms in these children raise the possibility that the exosome is particularly important in neurons; however, there are still many open questions. The aim of this proposal is to further characterize the role of the exosome in nerve cells and will address the following aspects:1. Neuronal cells from patients: We will analyse RNA levels in patient derived skin cells and will transform them into neuronal cells with recently published methods. We will determine which RNA species are modified by changes in the exosome in the different cell types. 2. Genetically modified zebrafish: In parallel, we will create zebrafish models of exosomal protein deficiencies. By artificially removing single exosome components and by introducing mutations in zebrafish embryos we will explore its effect on the development and function of the brain and different types of neurons. In addition, we will also study the effect of modifying the exosome on myelin formation in zebrafish. Our results will identify the most important roles of the exosome in regulating gene expression and why is it more damaging for neurons. Combining the data in human cells and zebrafish will enable us to better define the important genes and interacting partners of the exosome. By modifying exosome components or some of the here identified interacting factors we may discover potential pathways to alter RNA degradation or processing in neurons, which can be further developed as a therapeutic approach in exosomal diseases or in other types of neurodegenerative diseases caused by abnormal RNA function.

所有基因都被复制成短命的 RNA 分子，然后翻译成蛋白质，形成体内细胞的构建盒。 DNA 的转录、前体 mRNA 加工成成熟 mRNA（剪接）、mRNA 降解以及翻译成蛋白质都受到严格调控。某种 RNA 种类过多或过少都可能对细胞造成危险，但我们对微调 RNA 量的机制的了解仍然有限。基因表达（RNA 代谢）的调节对于正常细胞功能至关重要，然而神经细胞似乎不太能够应对错误。由 RNA 生产机制缺陷引起的婴儿期或儿童期严重遗传性神经系统疾病数量的增加就说明了这一点；这些疾病的特征是大脑和中枢神经系统的发育和/或结构异常。该提案的目的是研究和表征一种新形式的 RNA 相关神经系统疾病，该疾病是由于称为人外泌体的多蛋白复合物功能障碍而引起的。外泌体的作用包括 RNA 的降解和成熟。据报道，患有婴儿期发病的脑干和小脑变性以及脊髓运动神经元功能障碍导致的肌肉无力的患者存在外泌体亚基基因 EXOSC3 的突变。我们最近在患有类似严重神经系统疾病的婴儿中发现了 EXOSC8 的突变，该突变编码人类外泌体的另一种成分。此外，这些儿童的大脑和脊髓也会失去髓磷脂，髓磷脂通常覆盖和隔离神经，并有助于加速信号传导。我们还在一名患有严重脊髓运动神经元功能障碍的儿童中发现了新基因 RBM7 的纯合突变。 RBM7 是人类外泌体的相互作用伙伴。这些儿童中严重且孤立的神经系统症状表明外泌体在神经元中特别重要的可能性；然而，仍有许多悬而未决的问题。该提案的目的是进一步表征外泌体在神经细胞中的作用，并将解决以下几个方面：1．来自患者的神经元细胞：我们将分析患者来源的皮肤细胞中的 RNA 水平，并使用最近发表的方法将其转化为神经元细胞。我们将确定哪些 RNA 种类会因不同细胞类型中外泌体的变化而被修饰。 2. 转基因斑马鱼：同时，我们将创建外泌体蛋白缺陷的斑马鱼模型。通过人工去除单个外泌体成分并在斑马鱼胚胎中引入突变，我们将探索其对大脑和不同类型神经元发育和功能的影响。此外，我们还将研究外泌体修饰对斑马鱼髓磷脂形成的影响。我们的结果将确定外泌体在调节基因表达中最重要的作用，以及为什么它对神经元的损害更大。结合人类细胞和斑马鱼的数据将使我们能够更好地定义外泌体的重要基因和相互作用的伙伴。通过修饰外泌体成分或一些此处确定的相互作用因子，我们可能会发现改变神经元中 RNA 降解或加工的潜在途径，这可以进一步开发为外泌体疾病或由异常 RNA 功能引起的其他类型神经退行性疾病的治疗方法。

项目成果

EURO-NMD registry: federated FAIR infrastructure, innovative technologies and concepts of a patient-centred registry for rare neuromuscular disorders.

EURO-NMD 注册中心：联合 FAIR 基础设施、创新技术和以患者为中心的罕见神经肌肉疾病注册中心概念。

• DOI: 10.1186/s13023-024-03059-3
• 发表时间: 2024
• 期刊: Orphanet journal of rare diseases
• 影响因子: 3.7
• 作者: Atalaia A

100,000 Genomes Pilot on Rare-Disease Diagnosis in Health Care - Preliminary Report.

• DOI: 10.1056/nejmoa2035790
• 发表时间: 2021-11-11
• 期刊: The New England journal of medicine
• 作者: 100,000 Genomes Project Pilot Investigators;Smedley D;Smith KR;Martin A;Thomas EA;McDonagh EM;Cipriani V;Ellingford JM;Arno G;Tucci A;Vandrovcova J;Chan G;Williams HJ;Ratnaike T;Wei W;Stirrups K;Ibanez K;Moutsianas L;Wielscher M;Need A;Barnes MR;Vestito L;Buchanan J;Wordsworth S;Ashford S;Rehmström K;Li E;Fuller G;Twiss P;Spasic-Boskovic O;Halsall S;Floto RA;Poole K;Wagner A;Mehta SG;Gurnell M;Burrows N;James R;Penkett C;Dewhurst E;Gräf S;Mapeta R;Kasanicki M;Haworth A;Savage H;Babcock M;Reese MG;Bale M;Baple E;Boustred C;Brittain H;de Burca A;Bleda M;Devereau A;Halai D;Haraldsdottir E;Hyder Z;Kasperaviciute D;Patch C;Polychronopoulos D;Matchan A;Sultana R;Ryten M;Tavares ALT;Tregidgo C;Turnbull C;Welland M;Wood S;Snow C;Williams E;Leigh S;Foulger RE;Daugherty LC;Niblock O;Leong IUS;Wright CF;Davies J;Crichton C;Welch J;Woods K;Abulhoul L;Aurora P;Bockenhauer D;Broomfield A;Cleary MA;Lam T;Dattani M;Footitt E;Ganesan V;Grunewald S;Compeyrot-Lacassagne S;Muntoni F;Pilkington C;Quinlivan R;Thapar N;Wallis C;Wedderburn LR;Worth A;Bueser T;Compton C;Deshpande C;Fassihi H;Haque E;Izatt L;Josifova D;Mohammed S;Robert L;Rose S;Ruddy D;Sarkany R;Say G;Shaw AC;Wolejko A;Habib B;Burns G;Hunter S;Grocock RJ;Humphray SJ;Robinson PN;Haendel M;Simpson MA;Banka S;Clayton-Smith J;Douzgou S;Hall G;Thomas HB;O'Keefe RT;Michaelides M;Moore AT;Malka S;Pontikos N;Browning AC;Straub V;Gorman GS;Horvath R;Quinton R;Schaefer AM;Yu-Wai-Man P;Turnbull DM;McFarland R;Taylor RW;O'Connor E;Yip J;Newland K;Morris HR;Polke J;Wood NW;Campbell C;Camps C;Gibson K;Koelling N;Lester T;Németh AH;Palles C;Patel S;Roy NBA;Sen A;Taylor J;Cacheiro P;Jacobsen JO;Seaby EG;Davison V;Chitty L;Douglas A;Naresh K;McMullan D;Ellard S;Temple IK;Mumford AD;Wilson G;Beales P;Bitner-Glindzicz M;Black G;Bradley JR;Brennan P;Burn J;Chinnery PF;Elliott P;Flinter F;Houlden H;Irving M;Newman W;Rahman S;Sayer JA;Taylor JC;Webster AR;Wilkie AOM;Ouwehand WH;Raymond FL;Chisholm J;Hill S;Bentley D;Scott RH;Fowler T;Rendon A;Caulfield M
• 通讯作者: Caulfield M

A guide to writing systematic reviews of rare disease treatments to generate FAIR-compliant datasets: building a Treatabolome

撰写罕见疾病治疗系统评价以生成符合 FAIR 标准的数据集的指南：构建 Treatabolome

• DOI: 10.17863/cam.56498
• 发表时间: 2020
• 作者: Atalaia A

Mitochondrial and nuclear disease panel (Mito-aND-Panel): Combined sequencing of mitochondrial and nuclear DNA by a cost-effective and sensitive NGS-based method.

• DOI: 10.1002/mgg3.500
• 发表时间: 2018-11
• 期刊: Molecular genetics & genomic medicine
• 影响因子: 2
• 作者: Abicht A;Scharf F;Kleinle S;Schön U;Holinski-Feder E;Horvath R;Benet-Pagès A;Diebold I
• 通讯作者: Diebold I

Multifocal demyelinating motor neuropathy and hamartoma syndrome associated with a de novo PTEN mutation.

• DOI: 10.1212/wnl.0000000000005566
• 发表时间: 2018-05-22
• 期刊: Neurology
• 影响因子: 9.9
• 作者: Bansagi B;Phan V;Baker MR;O'Sullivan J;Jennings MJ;Whittaker RG;Müller JS;Duff J;Griffin H;Miller JAL;Gorman GS;Lochmüller H;Chinnery PF;Roos A;Swan LE;Horvath R
• 通讯作者: Horvath R