New humanised mouse models for dissecting the pathobiology of disease, using FUS-ALS as a paradigm

以 FUS-ALS 为范例，用于剖析疾病病理学的新型人源化小鼠模型

• 批准号: MR/L021056/1
• 负责人: Elizabeth Fisher
• 金额: $ 101.54万
• 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: 2014
• 资助国家: 英国
• 起止时间: 2014 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FL021056%2F1
• 关键词: New; humanised; mouse; models; dissecting

Humans and mice are descended from a common ancestor and so we share genes and have similar diseases caused by similar mutations in our equivalent genes (known as orthologs). Therefore to understand more about human disease we work with 'mouse models' that have the same mutations and manifest similar symptoms. Many neurodegenerative diseases run in families as 'dominant' diseases, so if your parent has the disorder there is a 50% chance you will inherit the mutant gene, and also succumb to the disease. These dominant disorders are modelled by mice called transgenics, which carry human mutant genes and so get the disease in a similar way to humans.Transgenic animals are made by incorporating the human mutant 'transgene' into the mouse DNA. Unfortunately when this happens the human mutant gene is usually incorporated in lots of copies, somewhere fairly random, not just a single copy as it would be normally in humans. One result is the mouse produces considerably more than the usual 'physiological' amount of mutant protein, and more often than not the mouse develops symptoms that simply come from having too much protein, rather than the actual effects of the mutation. This means the mouse is a far less accurate model than it could be.Also, sometimes the human and mouse biochemistry of the mutant protein can be different, so simply making the same change in the mouse gene does not produce an accurate disease model.We have developed a new type of mouse model in which we replace the mouse gene with the complete human gene, both the normal human gene and the mutated human gene in different mouse strains. Therefore we can look at what happens to the mouse when the human genetic material is expressed at normal levels, and this will give us new insight into disease processes. Our new mouse models may be a Refinement of existing models.As an example, we have chosen to work with a gene called FUS, which is mutated in an untreatable neurodegenerative disease called amyotrophic lateral sclerosis (ALS, also known as motor neuron disease). ALS is an incurable and relentless devastating neurodegenerative disorder which causes progressive loss of muscle function and paralysis. ALS leads to death, usually caused by the inability to breathe, on average only 3 years after diagnosis, with a lifetime risk of ~1 in 300 by 85 years old, in UK. ALS is generally a mid-life disease affecting people in their 40s and 50s although there is a wide age-range of symptom onset and aggressive FUS-caused ALS has been described in children as young as 11 years old. FUS is also occasionally mutated in other incurable neurological diseases.Now we are applying for a three year grant to:(1) extend our method to make a slightly different type of mouse called a conditional mutant, which allows us to turn the mutant protein on in different tissues at different times. This will help us work out what goes wrong in ALS and other disorders. And,(2) to fully characterise our new FUS mouse mutants, so we can learn more about the way that mutant FUS causes nerve cells to die. If we know this, we are step further along to therapies for ALS and other neurodegenerative diseases.Our technology can be used for any gene, not just FUS and for example, we are well along with using this method for SOD1, which is another 'ALS gene'. So we have developed a new technology that we believe will be widely used by other laboratories, and we want to characterise the mice we have already created to understand more of how neurodegeneration happens, so that we can better understand human disease and how to treat it.

人类和老鼠是共同祖先的后代，因此我们共享基因，并有类似的疾病，这些疾病是由我们同等基因(称为同源基因)的相似突变引起的。因此，为了更多地了解人类疾病，我们研究了具有相同突变和相似症状的“小鼠模型”。许多神经退行性疾病在家族中表现为“显性”疾病，所以如果你的父母患有这种疾病，你有50%的可能性会遗传突变基因，同时也会死于这种疾病。这些显性疾病是由被称为转基因的小鼠建立模型的，这种小鼠携带人类突变基因，因此患上这种疾病的方式与人类相似。转基因动物是通过将人类突变的“转基因”整合到小鼠DNA中而产生的。不幸的是，当这种情况发生时，人类突变基因通常会被合并到许多拷贝中，在相当随机的地方，而不是像通常在人类中那样只有一个拷贝。其中一个结果是，小鼠产生的突变蛋白远远超过通常的“生理”量，而且通常情况下，小鼠出现症状只是因为蛋白质过多，而不是突变的实际影响。这意味着小鼠的模型远不是一个准确的模型。此外，有时人类和小鼠突变蛋白的生物化学可能不同，因此简单地对小鼠基因进行相同的改变并不能产生准确的疾病模型。我们开发了一种新型的小鼠模型，在不同的小鼠品系中，我们用完整的人类基因替换小鼠基因，包括正常的人类基因和突变的人类基因。因此，我们可以观察当人类遗传物质在正常水平表达时，小鼠会发生什么，这将使我们对疾病过程有新的了解。我们的新小鼠模型可能是现有模型的改进。例如，我们选择了一种名为FUS的基因，这种基因在一种名为肌萎缩侧索硬化症(ALS，也称为运动神经元病)的不可治愈的神经退行性疾病中突变。肌萎缩侧索硬化症是一种无法治愈和无情的破坏性神经退行性疾病，会导致肌肉功能的进行性丧失和瘫痪。在英国，肌萎缩侧索硬化症会导致死亡，通常是由于呼吸困难引起的，平均在确诊后3年内，到85岁时，一生中有1/300的风险。肌萎缩侧索硬化症通常是一种中年疾病，影响40多岁和50多岁的人，尽管有广泛的年龄范围的症状出现，侵袭性肌萎缩侧索硬化症被描述为年仅11岁的儿童。在其他无法治愈的神经疾病中，FUS偶尔也会发生突变。现在，我们正在申请一项为期三年的拨款：(1)扩展我们的方法，制造一种略有不同的类型的小鼠，称为条件性突变，这允许我们在不同的组织中在不同的时间启动突变蛋白。这将帮助我们找出肌萎缩侧索硬化症和其他疾病的问题所在。以及，(2)全面描述我们新的FUS小鼠突变体，这样我们就可以更多地了解突变FUS导致神经细胞死亡的方式。如果我们知道了这一点，我们就可以更进一步地治疗肌萎缩侧索硬化症和其他神经退行性疾病。我们的技术可以用于任何基因，而不仅仅是FUS，例如，我们可以很好地将这种方法用于SOD1，这是另一种“肌萎缩侧索硬化基因”。因此，我们开发了一项新技术，我们相信这项技术将被其他实验室广泛使用，我们想要描述我们已经创造的小鼠的特征，以更多地了解神经退化是如何发生的，这样我们就可以更好地了解人类疾病以及如何治疗它。

项目成果

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

Screening a UK amyotrophic lateral sclerosis cohort provides evidence of multiple origins of the C9orf72 expansion.

• DOI: 10.1016/j.neurobiolaging.2014.07.037
• 发表时间: 2015-01
• 期刊: Neurobiology of aging
• 影响因子: 4.2
• 作者: Fratta P;Polke JM;Newcombe J;Mizielinska S;Lashley T;Poulter M;Beck J;Preza E;Devoy A;Sidle K;Howard R;Malaspina A;Orrell RW;Clarke J;Lu CH;Mok K;Collins T;Shoaii M;Nanji T;Wray S;Adamson G;Pittman A;Renton AE;Traynor BJ;Sweeney MG;Revesz T;Houlden H;Mead S;Isaacs AM;Fisher EM
• 通讯作者: Fisher EM

Generation and analysis of innovative genomically humanized knockin SOD1, TARDBP (TDP-43), and FUS mouse models.

• DOI: 10.1016/j.isci.2021.103463
• 发表时间: 2021-12-17
• 期刊: iScience
• 影响因子: 5.8
• 作者: Devoy A;Price G;De Giorgio F;Bunton-Stasyshyn R;Thompson D;Gasco S;Allan A;Codner GF;Nair RR;Tibbit C;McLeod R;Ali Z;Noda J;Marrero-Gagliardi A;Brito-Armas JM;Williams C;Öztürk MM;Simon M;O'Neill E;Bryce-Smith S;Harrison J;Atkins G;Corrochano S;Stewart M;Gilthorpe JD;Teboul L;Acevedo-Arozena A;Fisher EMC;Cunningham TJ
• 通讯作者: Cunningham TJ

Profilin1 E117G is a moderate risk factor for amyotrophic lateral sclerosis.

• DOI: 10.1136/jnnp-2013-306761
• 发表时间: 2014-05
• 期刊: Journal of neurology, neurosurgery, and psychiatry
• 作者: Fratta P;Charnock J;Collins T;Devoy A;Howard R;Malaspina A;Orrell R;Sidle K;Clarke J;Shoai M;Lu CH;Hardy J;Plagnol V;Fisher EM
• 通讯作者: Fisher EM

Mutation in the FUS nuclear localisation signal domain causes neurodevelopmental and systemic metabolic alterations.

• DOI: 10.1242/dmm.050200
• 发表时间: 2023-10-01
• 期刊: Disease models & mechanisms
• 影响因子: 4.3