RhoA/ROCK signalling activated by ARHGEF3 promotes muscle weakness via autophagy in dystrophic mdx mice.
RhoA/ROCK signalling activated by ARHGEF3 promotes muscle weakness via autophagy in dystrophic mdx mice.
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DOI：
10.1002/jcsm.13278
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发表时间：
2023-08
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中科院分区：
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Duchenne肌营养不良症(DMD)由肌营养不良蛋白缺乏引起，通过尚未完全破译的分子扰动导致进行性和致死性肌肉无力。新的证据表明，RhoA/Rho相关蛋白激酶(ROCK)信号与DMD的病理机制有关，但其在DMD肌肉功能中的直接作用及其相关机制尚不清楚。利用三维工程化的dystrophin缺陷型MDX骨骼肌和MDX小鼠，分别在体外和原位检测ROCK在DMD肌肉功能中的作用。通过建立Arhgef3基因敲除的MDX小鼠，研究了RhoA鸟嘌呤核苷酸交换因子(GEF)中的ARHGEF3在RhoA/ROCK信号转导和DMD病理中的作用。RhoA/ROCK信号在调节ARHGEF3功能中的作用是通过评估野生型或全环基金失活的ARHGEF3过表达与ROCK抑制剂处理的效果来确定的。为了获得更多的机制洞察，用氯喹评估了不同条件下的自噬通量和自噬的作用。用Y-27632抑制ROCK可以改善3D工程mdx肌肉(三个独立实验中的+25%，P<0.001)和小鼠(+25%，P<；<0.05)的肌力产生。与以前的研究不同的是，这种改善与肌肉分化或数量无关，而是与肌肉质量的提高有关。我们发现，ARHGEF3升高，并与MDX肌肉中RhoA/ROCK的激活有关，而在MDX小鼠中，耗尽ARHGEF3可以恢复肌肉质量(高达+36%，P<0.01)和形态，而不影响再生。相反，过度表达ARHGEF3进一步损害了MDX肌肉的质量(−为13%，与空载体对照相比，P<0.01)，这取决于全球环境基金的活动和岩石。值得注意的是，ARHGEF3/ROCK抑制通过挽救自噬发挥了作用，自噬在营养不良的肌肉中通常受到损害。我们的发现揭示了DMD肌肉无力的一个新的病理机制，涉及ARHGEF3-ROCK-自噬途径，以及靶向ARHGEF3在DMD中的治疗潜力。
Duchenne muscular dystrophy (DMD), caused by dystrophin deficiency, leads to progressive and fatal muscle weakness through yet‐to‐be‐fully deciphered molecular perturbations. Emerging evidence implicates RhoA/Rho‐associated protein kinase (ROCK) signalling in DMD pathology, yet its direct role in DMD muscle function, and related mechanisms, are unknown. Three‐dimensionally engineered dystrophin‐deficient mdx skeletal muscles and mdx mice were used to test the role of ROCK in DMD muscle function in vitro and in situ, respectively. The role of ARHGEF3, one of the RhoA guanine nucleotide exchange factors (GEFs), in RhoA/ROCK signalling and DMD pathology was examined by generating Arhgef3 knockout mdx mice. The role of RhoA/ROCK signalling in mediating the function of ARHGEF3 was determined by evaluating the effects of wild‐type or GEF‐inactive ARHGEF3 overexpression with ROCK inhibitor treatment. To gain more mechanistic insights, autophagy flux and the role of autophagy were assessed in various conditions with chloroquine. Inhibition of ROCK with Y‐27632 improved muscle force production in 3D‐engineered mdx muscles (+25% from three independent experiments, P < 0.05) and in mice (+25%, P < 0.001). Unlike suggested by previous studies, this improvement was independent of muscle differentiation or quantity and instead related to increased muscle quality. We found that ARHGEF3 was elevated and responsible for RhoA/ROCK activation in mdx muscles, and that depleting ARHGEF3 in mdx mice restored muscle quality (up to +36%, P < 0.01) and morphology without affecting regeneration. Conversely, overexpressing ARHGEF3 further compromised mdx muscle quality (−13% vs. empty vector control, P < 0.01) in GEF activity‐ and ROCK‐dependent manner. Notably, ARHGEF3/ROCK inhibition exerted the effects by rescuing autophagy which is commonly impaired in dystrophic muscles. Our findings uncover a new pathological mechanism of muscle weakness in DMD involving the ARHGEF3‐ROCK‐autophagy pathway and the therapeutic potential of targeting ARHGEF3 in DMD.
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影响因子：
9
作者：
通讯作者：
--
影响因子：
16.6
作者：
Milan, Giulia;Romanello, Vanina;Pescatore, Francesca;Armani, Andrea;Paik, Ji-Hye;Frasson, Laura;Seydel, Anke;Zhao, Jinghui;Abraham, Reimar;Goldberg, Alfred L.;Blaauw, Bert;DePinho, Ronald A.;Sandri, Marco
通讯作者：
Sandri, Marco
影响因子：
17.1
作者：
Rosenberg AS;Puig M;Nagaraju K;Hoffman EP;Villalta SA;Rao VA;Wakefield LM;Woodcock J
通讯作者：
Woodcock J
影响因子：
15.9
作者：
Alexander, Matthew S.;Carlos Casar, Juan;Kunkel, Louis M.
通讯作者：
Kunkel, Louis M.
影响因子：
2.7
作者：
Dellorusso, C;Crawford, RW;Brooks, SV
通讯作者：
Brooks, SV