The impact of TDP-43 on translation and the response to axonal damage in amyotrophic lateral sclerosis

TDP-43 对肌萎缩侧索硬化症轴突损伤反应的影响

• 批准号: MR/S006508/1
• 负责人: Pietro Fratta
• 金额: $ 245.45万
• 依托单位: University College London
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2019
• 资助国家: 英国
• 起止时间: 2019 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FS006508%2F1
• 关键词: impact; TDP; 43; translation; response

Amyotrophic lateral sclerosis (ALS), also known as motor neuron disease (MND), is a devastating neurodegenerative disorder that causes progressive loss of MNs, leading to impaired muscle function and paralysis. ALS is incurable and leads to death, usually caused by the inability to breathe, on average only 3 years after diagnosis, with a lifetime risk of about 1 in 400. Motor neurons (MNs) are the nerve cells that send signals from the spinal cord to our muscles. They are amongst the largest cells in the body, with their cell soma located in the spinal cord and its fiber (called 'axon') projecting outside to the muscles. All cell compartments, including axons, need a constant supply of newly synthesised proteins in order to function properly. On some occasions, for example when responding to an unpredicted event as cell damage, new and different proteins are absolutely needed to re-establish cell functionality and long term survival. This is particularly challenging for axons, as the RNA for these proteins, which is essential to make them, may need to be synthesized very far away in the nucleus. Multiple lines of evidence indicate that one of the crucial players in the disease mechanism of ALS is a protein called TDP-43, which is important for the specific transport of RNA to different locations in the axons and in the response of cells to stress and damage.In this Fellowship, I will test how TDP-43 impacts on the response of MNs to damage in the axons, and the relevance of this response pathway in ALS. To do so, I will combine novel mouse models of disease and patient cell lines with state-of-the-art biological tools that will allow the investigation of these very specific functions.In particular, we have developed novel strains of mice that carry specific mutations in the mouse TDP-43 gene and, as a result, develop crucial features of ALS. These mouse models of ALS will allow us, by looking over time at pre-symptomatic mice and at mice with overt ALS symptoms, to identify the molecular and cellular changes occurring during disease progression. Using an innovative and fully integrated approach, we will analyse the disease phenotype of these mice with new molecular biology and microscopic techniques to identify which types of RNAs and proteins are present in different MN regions, including the axon, and how these change in response to damage. I will therefore be able to investigate the cellular alterations triggered by TDP-43 in axonal damage response by studying: 1. MNs grown in special culture dishes where the axons are separated from the cell bodies; 2. The axons and their damage response in wild type and mutant mice; and 3. By validating our results in MNs derived from patient-induced stem cells and using post mortem brain samples donated to research by ALS patients. In summary, this project will contribute to understand how changes in TDP-43 impacts on MN survival. This long-awaited information is essential to develop effective therapeutics for motor neuron disorders.

肌萎缩性侧索硬化症（ALS），也被称为运动神经元疾病（MND），是一种毁灭性的神经退行性疾病，导致MNs逐渐丧失，导致肌肉功能受损和瘫痪。ALS是无法治愈的，通常会导致死亡，通常是由于无法呼吸，平均仅在诊断后3年就会死亡，终生风险约为400分之一。运动神经元（MNs）是从脊髓向肌肉发送信号的神经细胞。它们是人体中最大的细胞之一，它们的细胞体位于脊髓中，其纤维（称为“轴突”）向外突出到肌肉中。所有的细胞区室，包括轴突，都需要新合成蛋白质的持续供应才能正常工作。在某些情况下，例如，当对细胞损伤等不可预测的事件作出反应时，绝对需要新的和不同的蛋白质来重建细胞功能和长期生存。这对轴突来说尤其具有挑战性，因为制造这些蛋白质所必需的RNA可能需要在离细胞核很远的地方合成。多种证据表明，一种名为TDP-43的蛋白质在ALS的发病机制中起着至关重要的作用，它对RNA向轴突不同位置的特异性运输以及细胞对应激和损伤的反应起着重要作用。在本研究中，我将测试TDP-43如何影响MNs对轴突损伤的反应，以及这种反应途径在ALS中的相关性。为此，我将把新的疾病小鼠模型和患者细胞系与最先进的生物工具结合起来，这将允许研究这些非常具体的功能。特别是，我们已经开发出携带小鼠TDP-43基因特定突变的新型小鼠菌株，从而发展出ALS的关键特征。这些肌萎缩性侧索硬化症的小鼠模型将允许我们，通过观察出现症状前的小鼠和有明显肌萎缩性侧索硬化症症状的小鼠，来识别疾病进展过程中发生的分子和细胞变化。使用一种创新的和完全集成的方法，我们将用新的分子生物学和显微技术分析这些小鼠的疾病表型，以确定哪些类型的rna和蛋白质存在于不同的MN区域，包括轴突，以及这些如何响应损伤而变化。因此，我将能够通过以下研究来研究TDP-43在轴突损伤反应中引发的细胞改变：1。在轴突与细胞体分离的特殊培养皿中培养的MNs；2. 野生型和突变型小鼠轴突及其损伤反应和3。通过验证从患者诱导的干细胞中提取的MNs的结果，并使用ALS患者捐献的死后大脑样本进行研究。总之，该项目将有助于了解TDP-43的变化如何影响MN的生存。这一期待已久的信息对于开发运动神经元疾病的有效治疗方法至关重要。

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

FUS-ALS mutants alter FMRP phase separation equilibrium and impair protein translation.

• DOI: 10.1126/sciadv.abf8660
• 发表时间: 2021-07
• 期刊: Science advances
• 影响因子: 13.6
• 作者: Birsa N;Ule AM;Garone MG;Tsang B;Mattedi F;Chong PA;Humphrey J;Jarvis S;Pisiren M;Wilkins OG;Nosella ML;Devoy A;Bodo C;de la Fuente RF;Fisher EMC;Rosa A;Viero G;Forman-Kay JD;Schiavo G;Fratta P
• 通讯作者: Fratta P

HnRNP K mislocalisation is a novel protein pathology of frontotemporal lobar degeneration and ageing and leads to cryptic splicing.

• DOI: 10.1007/s00401-021-02340-0
• 发表时间: 2021-10
• 期刊: Acta neuropathologica
• 影响因子: 12.7
• 作者: Bampton A;Gatt A;Humphrey J;Cappelli S;Bhattacharya D;Foti S;Brown AL;Asi Y;Low YH;Foiani M;Raj T;Buratti E;Fratta P;Lashley T
• 通讯作者: Lashley T

The role of hnRNPs in frontotemporal dementia and amyotrophic lateral sclerosis.

• DOI: 10.1007/s00401-020-02203-0
• 发表时间: 2020-11
• 期刊: Acta neuropathologica
• 影响因子: 12.7
• 作者: Bampton A;Gittings LM;Fratta P;Lashley T;Gatt A
• 通讯作者: Gatt A

Corrigendum: Neuregulin 1 type III improves peripheral nerve myelination in a mouse model of congenital hypomyelinating neuropathy.

勘误表：Neuregulin 1 III 型可改善先天性低髓鞘神经病小鼠模型的周围神经髓鞘形成。

• DOI: 10.1093/hmg/ddz037
• 发表时间: 2019
• 期刊: Human molecular genetics
• 影响因子: 3.5
• 作者: Belin,Sophie;Ornaghi,Francesca;Shackleford,Ghjuvan'Ghjacumu;Wang,Jie;Scapin,Cristina;Lopez-Anido,Camila;Silvestri,Nicholas;Robertson,Neil;Williamson,Courtney;Ishii,Akihiro;Taveggia,Carla;Svaren,John;Bansal,Rashmi;Schwab,MarkusH
• 通讯作者: Schwab,MarkusH