Investigating neuronal RNA localisation and translational deficits as gain of function mechanisms in ALS.

研究神经元 RNA 定位和翻译缺陷作为 ALS 功能获得的机制。

• 批准号: MR/R005184/1
• 负责人: Elizabeth Fisher
• 金额: $ 71.03万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2017
• 资助国家: 英国
• 起止时间: 2017 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FR005184%2F1
• 关键词: Investigating; neuronal; RNA; localisation; translational

Motor neurons are the nerves that send signals from the spinal cord to our muscles. They are amongst the largest nerve cells in the body and are highly specialized, in that their cell body is located in the spinal cord and its fiber (called 'axon') projects outside to the muscles. Amyotrophic Lateral Sclerosis (ALS) is a relentless and incurable disease that can kill the motor neurons, resulting in progressive paralysis and death within 5 years of diagnosis. ALS has a lifetime risk in UK of 1 in 250 in the UK population.Every cell in our body contains DNA, RNA and proteins. DNA is in the nucleus of cells and carries the information on how to build the proteins. Proteins are the building block of our cells and they can also perform important tasks for the cell's functions. RNA is the intermediate and carries the instructions for making individual proteins from the DNA, to the various locations inside a cell where these proteins are made and needed for specific jobs. Relatively recently it has become clear that axons, the long projections from individual neurons, need proteins to be made throughout their length, including at the very end most distant from the cell body. We do not yet know why this is, but certainly the need to respond quickly to local injury of an axon at a distant site, is one good reason why proteins need to be made in a localized region of a neuron, along the axon. To have the capacity to be able to make proteins where they are needed, neurons have to transport RNA around the cell, especially along the axon. Motor neurons are some of the largest cells in the body and have tremendously long axons, in humans one single axon can extend from the spinal cord out to our feet, often for over 1 meter in length. So in these cells particularly, the movement of RNA, and the local production of proteins is extremely important.Unfortunately, currently, we do not know which RNAs are moved around motor neurons and nor do we know which proteins can be made as a result. Our lack of knowledge has been largely because of the shear technical difficulty of analyzing RNA and protein in the separate cell bodies and axons. But this knowledge is critical to understanding ALS, because it turns out that proteins that bind and transport RNA in motor neurons, including a protein called FUS, can be mutated in ALS. So to understand ALS, we need to know what happens to FUS and the RNA it binds and the proteins that it makes, in health and in ALS, in different regions of motor neurons.Here, we tackle the technical problems of looking at RNA and protein in different regions of motor neurons, including the very important axon, by combining new molecular biology and microscope techniques, with novel mouse model we have made, in an innovative approach to find out what RNA and proteins are present in different regions within motor neurons. We will look over time, at presymptomatic animals, and at animals with ALS, so that we can see the healthy state as mice age, and the progressive disease state. We can then validate our results in samples taken from human ALS patients. We badly need this information to understand the mechanisms and pathology of ALS and to develop effective therapeutics for motor neuron disorders.

运动神经元是将信号从脊髓发送到肌肉的神经。它们是身体中最大的神经细胞之一，并且是高度专业化的，因为它们的细胞体位于脊髓中，其纤维（称为“轴突”）伸出到肌肉之外。肌萎缩侧索硬化症（ALS）是一种无情的和不可治愈的疾病，可以杀死运动神经元，导致进行性瘫痪和死亡的5年内诊断。在英国，ALS的终生风险为1/250。我们体内的每个细胞都含有DNA，RNA和蛋白质。DNA位于细胞核中，携带如何构建蛋白质的信息。蛋白质是我们细胞的基石，它们也可以执行细胞功能的重要任务。RNA是中间体，它携带着从DNA制造单个蛋白质的指令，到达细胞内的各个位置，在那里制造这些蛋白质并进行特定的工作。相对而言，最近已经清楚的是，轴突，即来自单个神经元的长突起，需要在其整个长度上制造蛋白质，包括在离细胞体最远的末端。我们还不知道这是为什么，但可以肯定的是，对远处轴突局部损伤的快速反应的需要，是神经元局部区域（沿着轴突）制造蛋白质的一个很好的原因。为了能够在需要的地方制造蛋白质，神经元必须在细胞周围运输RNA，特别是沿着轴突。运动神经元是身体中最大的细胞之一，具有非常长的轴突，在人类中，一个轴突可以从脊髓延伸到我们的脚，通常超过1米长。因此，在这些细胞中，RNA的移动和蛋白质的局部产生是非常重要的。不幸的是，目前，我们不知道哪些RNA在运动神经元周围移动，也不知道哪些蛋白质可以被制造出来。我们缺乏知识，主要是因为在分离的细胞体和轴突中分析RNA和蛋白质的剪切技术困难。但这一知识对于理解ALS至关重要，因为事实证明，在运动神经元中结合和转运RNA的蛋白质，包括一种名为FUS的蛋白质，可以在ALS中发生突变。因此，为了了解ALS，我们需要知道FUS和它结合的RNA以及它在运动神经元的不同区域中产生的蛋白质在健康和ALS中发生了什么。在这里，我们解决了观察运动神经元不同区域中的RNA和蛋白质的技术问题，包括非常重要的轴突，通过结合新的分子生物学和显微镜技术，与我们制作的新型小鼠模型，用一种创新的方法来发现运动神经元内不同区域的RNA和蛋白质。我们将随着时间的推移，观察症状前动物和ALS动物，这样我们就可以看到随着小鼠年龄的增长，健康状态和疾病进展状态。然后，我们可以在从人类ALS患者身上采集的样本中验证我们的结果。我们迫切需要这些信息来了解ALS的机制和病理学，并为运动神经元疾病开发有效的治疗方法。

项目成果

TDP-43 loss and ALS-risk SNPs drive mis-splicing and depletion of UNC13A.

• DOI: 10.1038/s41586-022-04436-3
• 发表时间: 2022-03
• 期刊: Nature
• 影响因子: 64.8
• 作者: Brown AL;Wilkins OG;Keuss MJ;Hill SE;Zanovello M;Lee WC;Bampton A;Lee FCY;Masino L;Qi YA;Bryce-Smith S;Gatt A;Hallegger M;Fagegaltier D;Phatnani H;NYGC ALS Consortium;Newcombe J;Gustavsson EK;Seddighi S;Reyes JF;Coon SL;Ramos D;Schiavo G;Fisher EMC;Raj T;Secrier M;Lashley T;Ule J;Buratti E;Humphrey J;Ward ME;Fratta P
• 通讯作者: Fratta P

TDP-43 mutations increase HNRNP A1-7B through gain of splicing function

TDP-43 突变通过获得剪接功能增加 HNRNP A1-7B

• DOI: 10.1093/brain/awy260
• 发表时间: 2018
• 期刊: Brain
• 影响因子: 14.5
• 作者: Sivakumar P

Humanized mutant FUS drives progressive motor neuron degeneration without aggregation in 'FUSDelta14' knockin mice.

• DOI: 10.1093/brain/awx248
• 发表时间: 2017-11-01
• 期刊: Brain : a journal of neurology
• 作者: Devoy A;Kalmar B;Stewart M;Park H;Burke B;Noy SJ;Redhead Y;Humphrey J;Lo K;Jaeger J;Mejia Maza A;Sivakumar P;Bertolin C;Soraru G;Plagnol V;Greensmith L;Acevedo Arozena A;Isaacs AM;Davies B;Fratta P;Fisher EMC
• 通讯作者: Fisher EMC

FUS ALS-causative mutations impact FUS autoregulation and the processing of RNA-binding proteins through intron retention

• DOI: 10.1101/567735
• 发表时间: 2019-03
• 期刊: bioRxiv
• 作者: J. Humphrey;N. Birsa;Carmelo Milioto;D. Robaldo;A. B. Eberle;Rahel Kräuchi;Matthew Bentham;A. Ule;Seth Jarvis;C. Bodo;M. G. Garone;A. Devoy;A. Rosa;I. Bozzoni;E. Fisher;M. Ruepp;O. Mühlemann;G. Schiavo;A. Isaacs;V. Plagnol;P. Fratta

Mice with endogenous TDP-43 mutations exhibit gain of splicing function and characteristics of amyotrophic lateral sclerosis.

• DOI: 10.15252/embj.201798684
• 发表时间: 2018-06-01
• 期刊: The EMBO journal
• 作者: Fratta P;Sivakumar P;Humphrey J;Lo K;Ricketts T;Oliveira H;Brito-Armas JM;Kalmar B;Ule A;Yu Y;Birsa N;Bodo C;Collins T;Conicella AE;Mejia Maza A;Marrero-Gagliardi A;Stewart M;Mianne J;Corrochano S;Emmett W;Codner G;Groves M;Fukumura R;Gondo Y;Lythgoe M;Pauws E;Peskett E;Stanier P;Teboul L;Hallegger M;Calvo A;Chiò A;Isaacs AM;Fawzi NL;Wang E;Housman DE;Baralle F;Greensmith L;Buratti E;Plagnol V;Fisher EM;Acevedo-Arozena A
• 通讯作者: Acevedo-Arozena A