Robriquet Florence, Lardenois Aurélie, Babarit Candice, Larcher Thibaut, Dubreil Laurence, Leroux Isabelle, Zuber Céline, Ledevin Mireille, Deschamps Jack-Yves, Fromes Yves, Cherel Yan, Guevel Laetitia, Rouger Karl
INRA, UMR703 PAnTher, Nantes, France; LUNAM Université, Oniris, École nationale vétérinaire, agro-alimentaire et de l'alimentation Nantes-Atlantique, Nantes, France; Université de Nantes, Nantes, France.
INRA, UMR703 PAnTher, Nantes, France; LUNAM Université, Oniris, École nationale vétérinaire, agro-alimentaire et de l'alimentation Nantes-Atlantique, Nantes, France.
PLoS One. 2015 May 8;10(5):e0123336. doi: 10.1371/journal.pone.0123336. eCollection 2015.
Several adult stem cell populations exhibit myogenic regenerative potential, thus representing attractive candidates for therapeutic approaches of neuromuscular diseases such as Duchenne Muscular Dystrophy (DMD). We have recently shown that systemic delivery of MuStem cells, skeletal muscle-resident stem cells isolated in healthy dog, generates the remodelling of muscle tissue and gives rise to striking clinical benefits in Golden Retriever Muscular Dystrophy (GRMD) dog. This global effect, which is observed in the clinically relevant DMD animal model, leads us to question here the molecular pathways that are impacted by MuStem cell transplantation. To address this issue, we compare the global gene expression profile between healthy, GRMD and MuStem cell treated GRMD dog muscle, four months after allogenic MuStem cell transplantation.
In the dystrophic context of the GRMD dog, disease-related deregulation is observed in the case of 282 genes related to various processes such as inflammatory response, regeneration, calcium ion binding, extracellular matrix organization, metabolism and apoptosis regulation. Importantly, we reveal the impact of MuStem cell transplantation on several molecular and cellular pathways based on a selection of 31 genes displaying signals specifically modulated by the treatment. Concomitant with a diffuse dystrophin expression, a histological remodelling and a stabilization of GRMD dog clinical status, we show that cell delivery is associated with an up-regulation of genes reflecting a sustained enhancement of muscle regeneration. We also identify a decreased mRNA expression of a set of genes having metabolic functions associated with lipid homeostasis and energy. Interestingly, ubiquitin-mediated protein degradation is highly enhanced in GRMD dog muscle after systemic delivery of MuStem cells.
Overall, our results provide the first high-throughput characterization of GRMD dog muscle and throw new light on the complex molecular/cellular effects associated with muscle repair and the clinical efficacy of MuStem cell-based therapy.
多个成体干细胞群体表现出肌源性再生潜力，因此是治疗诸如杜氏肌营养不良症（DMD）等神经肌肉疾病的有吸引力的候选细胞。我们最近发现，系统性递送在健康犬中分离得到的骨骼肌驻留干细胞——Mu干细胞，可使肌肉组织重塑，并在金毛猎犬肌营养不良症（GRMD）犬中产生显著的临床益处。在具有临床相关性的DMD动物模型中观察到的这种整体效应，促使我们在此质疑受Mu干细胞移植影响的分子途径。为解决这个问题，我们比较了同种异体Mu干细胞移植四个月后，健康、GRMD以及经Mu干细胞治疗的GRMD犬肌肉之间的全基因组表达谱。
在GRMD犬的营养不良背景下，观察到与多种过程相关的282个基因出现疾病相关的失调，这些过程包括炎症反应、再生、钙离子结合、细胞外基质组织、代谢和凋亡调节。重要的是，基于31个显示出受治疗特异性调节信号的基因选择，我们揭示了Mu干细胞移植对多种分子和细胞途径的影响。伴随着弥漫性肌营养不良蛋白表达、组织学重塑以及GRMD犬临床状态稳定，我们表明细胞递送与反映肌肉再生持续增强的基因上调相关。我们还鉴定出一组具有与脂质稳态和能量相关代谢功能的基因的mRNA表达降低。有趣的是，在系统性递送Mu干细胞后，GRMD犬肌肉中泛素介导的蛋白质降解显著增强。
总体而言，我们的结果首次对GRMD犬肌肉进行了高通量表征，并为与肌肉修复相关的复杂分子/细胞效应以及基于Mu干细胞的治疗的临床疗效提供了新的见解。
