FAP-DMD: Elucidating the role of FAP cells in the process of muscle degeneration in patients with Duchenne muscular dystrophy

FAP-DMD：阐明FAP细胞在杜氏肌营养不良症患者肌肉退化过程中的作用

• 批准号: MR/W019086/1
• 负责人: Jordi Diaz-Manera
• 金额: $ 107.87万
• 依托单位: Newcastle University
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FW019086%2F1
• 关键词: FAP; DMD; Elucidating; role; cells

Duchenne muscular dystrophy (DMD) is a devastating disease produced by mutations in the dystrophin gene. Disease's onset with onset of weakness occurs before the age of 5 years and progresses leading to loose of ambulation during adolescence. Patients die during the third decade of life because of cardiac or respiratory complications. The only treatment approved so far is corticoids that slow down progression but without having a real impact in natural history. Despite huge efforts invested in developing new treatments, most of them have failed. At the moment, gene therapies based on the release of a healthy copy of the dystrophin gene to the muscle fibers are showing promising results, but it seems clear that they are not going to be able to stop disease's progression and that the process of muscle degeneration will remain active. The process of muscle degeneration in DMD has been studied for many years in animal models of the disease. Lack of dystrophin makes the fibers more susceptible to damage during normal muscle contraction. Damaged fibers are initially repaired by cells named satellite cells. However, repetitive damage triggers a series of consequences in the tissue including persistent infiltration by inflammatory cells that activates a type of cell known as fibro-adipogenic precursor cells (FAPs). FAPs contribute to muscle degeneration as they produce fat and fibrotic tissue which substitutes the damaged muscle fibers. During disease's progression, loss of fibers is associated to expansion of fibrotic and fatty tissue which impairs the capability of satellite cells to regenerate damaged muscle fibers. Although several molecules have been identified as key to regulate this process in mice, little is known in humans what is limiting the development of new treatments. We have developed a new protocol to isolate satellite cells and FAPs from muscle biopsies that were taken for diagnosis and are stored in biobanks. We have analyzed the cells obtained from DMD and healthy people and have identified different types of FAP cells. To summarize we have identified two main populations, one that is actively proliferating and retain stemness properties and another that is already committed to produce fibrotic tissue. In this proposal we will explore when these different populations appear in the progression of DMD patients using muscle biopsies of patients at different clinical stages and correlate their presence with tissue features to understand if these subpopulations are associated to changes such as fibrosis, inflammation or muscle fiber death. Then we will analyze the genes that are expressed in control and DMD muscle by fibers but also by other components, such as inflammatory cells, that could guide the changes in FAP subtype. Understanding the molecular pathways governing the changes in FAP subpopulations will provide potential new targets for therapies aimed to counteract the expansion of fibro-fatty tissue. In a third stage we will isolate the subpopulations of FAPs and study their properties including how they proliferate, move, or differentiate into fibrotic or fat producing cells and how they interact with satellite cells. These later studies provide information about how two cells interact in a dish, as it would happen when they are in the tissue. We will study this interaction both in standard 2D culture dishes, but also in 3D systems using printed molds that enable a more perdurable and structured generation of artificial muscles. These experiments will provide valuable information to understand if and how FAPs influence satellite cell function. Finally, we will test libraries of drugs able to counteract the function of key molecular pathways identified in the previous experiments to understand if they are able to modulate the function of FAPs.

Duchenne肌肉营养不良（DMD）是一种由肌营养不良蛋白基因突变产生的毁灭性疾病。疾病的发作随着弱点的发作发生，发生在5岁之前，导致青春期流动散落。由于心脏或呼吸并发症，患者在生命的第三个十年中死亡。到目前为止，唯一批准的治疗方法是降低降低进展但在自然史上没有真正影响的皮质类似。尽管为开发新疗法做出了巨大的努力，但其中大多数还是失败了。目前，基于释放肌营养不良蛋白基因对肌肉纤维的健康副本的基因疗法显示出令人鼓舞的结果，但很明显，它们无法阻止疾病的进展，并且肌肉退化的过程将保持活跃。 DMD中的肌肉变性过程已经在该疾病的动物模型中进行了多年研究。缺乏肌营养不良蛋白会使纤维在正常肌肉收缩期间更容易受到损伤。最初由名为卫星细胞的细胞修复受损的纤维。然而，重复损害会触发组织中的一系列后果，包括炎症细胞持续浸润，该细胞激活一种称为纤维辅助前体细胞（FAP）的细胞。 FAP会导致肌肉变性，因为它们产生脂肪和纤维化组织，替代受损的肌肉纤维。在疾病的进展过程中，纤维的损失与纤维化和脂肪组织的扩展有关，这会损害卫星细胞再生受损的肌肉纤维的能力。尽管几个分子被确定为调节小鼠过程的关键，但在人类中几乎没有什么限制新疗法的发展。我们开发了一种新方案，可以分离出用于诊断并存储在生物库中的肌肉活检中的卫星细胞和FAP。我们已经分析了从DMD和健康人员获得的细胞，并确定了不同类型的FAP细胞。总而言之，我们已经确定了两个主要种群，一个人群正在积极增殖和保留干性特性，而另一种已经承诺产生纤维化组织。在此提案中，我们将探讨这些不同的人群在DMD患者的进展中使用不同临床阶段的患者的肌肉活检，并将其存在与组织特征相关，以了解这些亚群是否与纤维化，炎症或肌肉纤维纤维死亡等变化有关。然后，我们将分析纤维在对照和DMD肌肉中表达的基因，还通过其他成分（例如炎症细胞）来指导FAP亚型的变化。了解管理FAP亚群变化的分子途径将为旨在抵消纤维脂肪组织扩张的疗法提供潜在的新靶标。在第三阶段，我们将隔离FAP的亚群，并研究其性质，包括它们如何增殖，移动或分化成纤维化或脂肪产生细胞，以及它们如何与卫星细胞相互作用。这些后来的研究提供了有关两个细胞在盘子中如何相互作用的信息，就像它们在组织中时会发生。我们将在标准的2D培养皿中研究这种相互作用，也将使用印刷模具在3D系统中研究这种相互作用，从而使人造肌肉具有更真实，结构化的生成。这些实验将提供有价值的信息，以了解FAP是否以及如何影响卫星细胞功能。最后，我们将测试能够抵消先前实验中确定的关键分子途径功能的药物文库，以了解它们是否能够调节FAP的功能。

项目成果

Analysis of muscle magnetic resonance imaging of a large cohort of patient with VCP-mediated disease reveals characteristic features useful for diagnosis.

• DOI: 10.1007/s00415-023-11862-4
• 发表时间: 2023-12
• 期刊: JOURNAL OF NEUROLOGY
• 影响因子: 6
• 作者: Esteller, Diana;Schiava, Marianela;Villar-Quiles, Rocio-Nur;Dibowski, Boris;Venturelli, Nadia;Laforet, Pascal;Alonso-Perez, Jorge;Olive, Montse;Dominguez-Gonzalez, Cristina;Paradas, Carmen;Velez, Beatriz;Kostera-Pruszczyk, Anna;Kierdaszuk, Biruta;Rodolico, Carmelo;Claeys, Kristl;Pal, Endre;Malfatti, Edoardo;Souvannanorath, Sarah;Alonso-Jimenez, Alicia;de Ridder, Willem;De Smet, Eline;Papadimas, George;Papadopoulos, Constantinos;Xirou, Sofia;Luo, Sushan;Muelas, Nuria;Vilchez, Juan J.;Ramos-Fransi, Alba;Monforte, Mauro;Tasca, Giorgio;Udd, Bjarne;Palmio, Johanna;Sri, Srtuhi;Krause, Sabine;Schoeser, Benedikt;Fernandez-Torron, Roberto;Lopez de Munain, Adolfo;Pegoraro, Elena;Farrugia, Maria Elena;Vorgerd, Mathias;Manousakis, Georgious;Chanson, Jean Baptiste;Nadaj-Pakleza, Aleksandra;Cetin, Hakan;Badrising, Umesh;Warman-Chardon, Jodi;Bevilacqua, Jorge;Earle, Nicholas;Campero, Mario;Diaz, Jorge;Ikenaga, Chiseko;Lloyd, Thomas E.;Nishino, Ichizo;Nishimori, Yukako;Saito, Yoshihiko;Oya, Yasushi;Takahashi, Yoshiaki;Nishikawa, Atsuko;Sasaki, Ryo;Marini-Bettolo, Chiara;Guglieri, Michela;Straub, Volker;Stojkovic, Tanya;Carlier, Robert Y.;Diaz-Manera, Jordi
• 通讯作者: Diaz-Manera, Jordi

Nintedanib Reduces Muscle Fibrosis and Improves Muscle Function of the Alpha-Sarcoglycan-Deficient Mice.

• DOI: 10.3390/biomedicines10102629
• 发表时间: 2022-10-19
• 期刊: BIOMEDICINES
• 影响因子: 4.7
• 作者: Alonso-Perez, Jorge;Carrasco-Rozas, Ana;Borrell-Pages, Maria;Fernandez-Simon, Esther;Pinol-Jurado, Patricia;Badimon, Lina;Wollin, Lutz;Lleixa, Cinta;Gallardo, Eduard;Olive, Montse;Diaz-Manera, Jordi;Suarez-Calvet, Xavier
• 通讯作者: Suarez-Calvet, Xavier

RhoA/ROCK2 signalling is enhanced by PDGF-AA in fibro-adipogenic progenitor cells: implications for Duchenne muscular dystrophy.

• DOI: 10.1002/jcsm.12923
• 发表时间: 2022-04
• 期刊: Journal of cachexia, sarcopenia and muscle
• 作者: Fernández-Simón E;Suárez-Calvet X;Carrasco-Rozas A;Piñol-Jurado P;López-Fernández S;Pons G;Bech Serra JJ;de la Torre C;de Luna N;Gallardo E;Díaz-Manera J
• 通讯作者: Díaz-Manera J

Muscle magnetic resonance imaging of a large cohort of distal hereditary motor neuropathies reveals characteristic features useful for diagnosis

• DOI: 10.1016/j.nmd.2023.08.010
• 发表时间: 2023-11-02
• 期刊: NEUROMUSCULAR DISORDERS
• 影响因子: 2.8
• 作者: Esteller，Diana;Morrow，Jasper;Diaz-Manera，Jordi
• 通讯作者: Diaz-Manera，Jordi