Single-cell RNA-seq reveals interferon-induced guanylate-binding proteins are linked with sarcopenia.
Single-cell RNA-seq reveals interferon-induced guanylate-binding proteins are linked with sarcopenia.
复制标题
DOI：
10.1002/jcsm.13091
复制
发表时间：
2022-12
期刊：
影响因子：
--
通讯作者：
中科院分区：
文献类型：
--
作者：
肌肉减少症定义为与年龄相关的肌肉质量和/或力量的进行性损失。虽然不同的因素可能导致这种疾病，但其潜在机制仍不清楚。我们以单细胞分辨率评估了肌肉减少症小鼠和对照小鼠骨骼肌中的转录异质性。建立了一种研究骨骼肌减少的小鼠模型。对从肌肉减少症小鼠和对照小鼠收集的胫骨前肌（TA）肌肉细胞进行单细胞RNA测序。进行了一系列生物信息学分析，以鉴定和比较不同条件下的不同细胞类型。使用免疫荧光染色和蛋白质印迹来验证来自单细胞实验的发现。进行管形成测定以进一步评估Gbp 2在血管生成期间对内皮细胞的影响。使用衰老加速小鼠品系（SAMP 6，n = 5）成功建立了小鼠肌肉减少症模型。通过给予地塞米松（20 mg/kg）和减少体力活动诱导肌肉减少症表型。招募具有相似活性但注射PBS的抗衰老小鼠品系（SAMR 1）和SAMP 6品系作为两个对照组。作为肌肉减少症的体征，肌肉减少症小鼠的体重、肌细胞计数和横截纤维面积均显著降低（P值分别= 0.004、0.03和0.035）。质量控制后，保留了13612个TA肌肉单细胞转录组用于分析。从分析的细胞中鉴定出14个细胞簇。其中，两种不同的内皮亚型分别在肌肉减少症组（42.2%细胞）和两个对照组（59.1%和47.9%细胞）中占主导地位。共检测到191个差异表达基因。肌肉减少症特异性内皮细胞亚型表现出干扰素家族基因和干扰素诱导型鸟苷酸结合蛋白（GBP）家族基因表达的显著增加。例如，减少肌肉的内皮细胞中的Igtp和Gbp 2分别是对照组的5.4和13.3倍。我们进一步验证了我们在肌肉减少症患者的肌肉标本中的发现，并观察到一部分患者（40例患者中的11例，27.5%）的内皮细胞中GBP 2水平升高，并且我们确定了显著更高的CD 31和GBP 2共定位（P值= 0.001128）。最后，我们在体外人脐静脉内皮细胞中过表达Gbp 2。具有升高的Gbp 2表达的内皮细胞显示受损的管形成。我们基于单细胞的结果表明，内皮细胞可能通过干扰素-GBP信号通路在肌肉减少症的发展中发挥关键作用，突出了减缓甚至逆转年龄相关性肌肉减少症的新治疗方向。
Sarcopenia is defined as an age‐related progressive loss of muscle mass and/or strength. Although different factors can contribute to this disease, the underlying mechanisms remain unclear. We assessed transcriptional heterogeneity in skeletal muscles from sarcopenic and control mice at single‐cell resolution. A mouse model was established to study sarcopenic skeletal muscles. Single‐cell RNA‐seq was performed on tibialis anterior (TA) muscle cells collected from sarcopenic and control mice. A series of bioinformatic analyses were carried out to identify and compare different cell types under different conditions. Immunofluorescence staining and western blotting were used to validate the findings from single‐cell experiments. Tube formation assays were conducted to further evaluate the effects of Gbp2 on endothelial cells during angiogenesis. A murine sarcopenia model was successfully established using a senescence‐accelerated mouse strain (SAMP6, n = 5). Sarcopenia phenotype was induced by administration of dexamethasone (20 mg/kg) and reduced physical activity. Senescence‐resistant mice strain (SAMR1) and SAMP6 strain with similar activity but injected with PBS were recruited as two control groups. As signs of sarcopenia, body weight, muscle cell counts and cross‐sectional fibre area were all significantly decreased in sarcopenic mice (P value = 0.004, 0.03 and 0.035, respectively). After quality control, 13 612 TA muscle single‐cell transcriptomes were retained for analysis. Fourteen cell clusters were identified from the profiled cells. Among them, two distinct endothelial subtypes were found to be dominant in the sarcopenia group (42.2% cells) and in the two control groups (59.1% and 47.9% cells), respectively. 191 differentially expressed genes were detected between the two endothelial subtypes. Sarcopenia‐specific endothelial cell subtype exhibited a dramatic increase in the interferon family genes and the interferon‐inducible guanylate‐binding protein (GBP) family gene expressions. For example, Igtp and Gbp2 in sarcopenic endothelial cells were 5.4 and 13.3 times higher than those in the control groups, respectively. We further validated our findings in muscle specimens of sarcopenia patients and observed that GBP2 levels were increased in endothelial cells of a subset of patients (11 of 40 patients, 27.5%), and we identified significantly higher CD31 and GBP2 co‐localization (P value = 0.001128). Finally, we overexpressed Gbp2 in human umbilical vein endothelial cells in vitro. The endothelial cells with elevated Gbp2 expression displayed compromised tube formation. Our single‐cell‐based results suggested that endothelial cells may play critical roles in sarcopenia development through interferon‐GBP signalling pathways, highlighting new therapeutic directions to slow down or even reverse age‐related sarcopenia.
登录
查看更多内容
影响因子：
4.6
作者：
Therakomen V;Petchlorlian A;Lakananurak N
通讯作者：
Lakananurak N
影响因子：
2.3
作者：
Chiu CS;Weber H;Adamski S;Rauch A;Gentile MA;Alves SE;Kath G;Flores O;Wilkinson HA
通讯作者：
Wilkinson HA
单细胞分析揭示成纤维细胞异质性以及成纤维细胞和壁细胞识别和区分的标准
影响因子：
16.6
作者：
Muhl, Lars;Genove, Guillem;Betsholtz, Christer
通讯作者：
Betsholtz, Christer
影响因子：
2.6
作者：
Chen, H.;Yao, X. F.;Shoumura, S.
通讯作者：
Shoumura, S.
影响因子：
5.5
作者：
Kargl, Christopher K.;Nie, Yaohui;Gavin, Timothy P.
通讯作者：
Gavin, Timothy P.