Platelet Derived Growth Factor-AA Correlates With Muscle Function Tests and Quantitative Muscle Magnetic Resonance in Dystrophinopathies

Alonso‐Jiménez, Alicia; Fernández‐Simón, Esther; Benito, Daniel Natera-de; Ortez, C.; García, Carmen; Montiel, Elena; Belmonte, Izaskun; Pedrosa, Irene; Segovia, Sonia; Piñol-Jurado, Patricia; Carrasco‐Rozas, Ana; Suárez‐Calvet, Xavier; Jiménez-Mallebrera, C.; Nascimento, A.; Llauger, Jaume; Nuñez‐Peralta, Claudia; Montesinos, Paula; Alonso‐Pérez, Jorge; Gallardo, Eduard; Illa, Isabel; Díaz‐Manera, Jordi

doi:10.3389/fneur.2021.659922

Cited by 5 publications

(4 citation statements)

References 61 publications

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“…To explore potential relationships between our differential gene expression and reported mechanisms of DMD pathology that can be mapped to individual genes, we analyzed the differential expression of previously reported human serum/blood DMD biomarkers (Hathout et al, 2014;Parolo et al, 2018;Spitali et al, 2018;Al-Khalili Szigyarto, 2020;Grounds et al, 2020;Alonso-Jiménez et al, 2021;Wagner et al, 2021;Lee-Gannon et al, 2022;Wu et al, 2022), and of genes modifying the phenotype of DMD in humans (Pegoraro et al, 2011;Flanigan et al, 2013;Bello et al, 2016;Hogarth et al, 2017;Li et al, 2018;Weiss et al, 2018;Spitali et al, 2020;Flanigan et al, 2023) or the mdx mouse (Deconinck et al, 1997;Wagner et al, 2002;Han et al, 2011;Morales et al, 2013;de Zélicourt et al, 2022), also known as genetic modifiers, across the VL and TA and identified the predominant cell type in which they were expressed.…”

Section: Genes That Are Previously Reported As Dmd Biomarkers and Gen...mentioning

confidence: 99%

Transcriptomic analysis of paired healthy human skeletal muscles to identify modulators of disease severity in DMD

Nieves‐Rodriguez¹,

Barthélémy²,

Woods³

et al. 2023

Front. Genet.

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Muscle damage and fibro-fatty replacement of skeletal muscles is a main pathologic feature of Duchenne muscular dystrophy (DMD) with more proximal muscles affected earlier and more distal affected later in the disease course, suggesting that different skeletal muscle groups possess distinctive characteristics that influence their susceptibility to disease. To explore transcriptomic factors driving differential gene expression and modulating DMD skeletal muscle severity, we characterized the transcriptome of vastus lateralis (VL), a more proximal and susceptible muscle, relative to tibialis anterior (TA), a more distal and protected muscle, in 15 healthy individuals using bulk RNA sequencing to identify gene expression differences that may mediate their relative susceptibility to damage with loss of dystrophin. Matching single nuclei RNA sequencing data was generated for 3 of the healthy individuals, to infer cell composition in the bulk RNA sequencing dataset and to improve mapping of differentially expressed genes to their cell source of expression. A total of 3,410 differentially expressed genes were identified and mapped to cell type using single nuclei RNA sequencing of muscle, including long non-coding RNAs and protein coding genes. There was an enrichment of genes involved in calcium release from the sarcoplasmic reticulum, particularly in the myofibers and these myofiber genes were higher in the VL. There was an enrichment of genes in “Collagen-Containing Extracellular Matrix” expressed by fibroblasts, endothelial, smooth muscle and pericytes, with most genes higher in the TA, as well as genes in “Regulation Of Apoptotic Process” expressed across all cell types. Previously reported genetic modifiers were also enriched within the differentially expressed genes. We also identify 6 genes with differential isoform usage between the VL and TA. Lastly, we integrate our findings with DMD RNA sequencing data from the TA, and identify “Collagen-Containing Extracellular Matrix” and “Negative Regulation Of Apoptotic Process” as differentially expressed between DMD compared to healthy. Collectively, these findings propose novel candidate mechanisms that may mediate differential muscle susceptibility in muscular dystrophies and provide new insight into potential therapeutic targets.

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Section: Genes That Are Previously Reported As Dmd Biomarkers and Gen...mentioning

confidence: 99%

Transcriptomic analysis of paired healthy human skeletal muscles to identify modulators of disease severity in DMD

Nieves‐Rodriguez¹,

Barthélémy²,

Woods³

et al. 2023

Front. Genet.

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“…Relative expression values were calculated by using the 2-CT method. Referring to the reference genes used in the previous study (De Pasquale et al, 2012;Hildyard et al, 2018;Alonso-Jimenez et al, 2021), we chose GAPDH, SDHA, HPRT1 and RPL13a as possible reference genes and determined the amount of their expressions in DMD and control groups by RT-qPCR. The web-based tool RefFinder 4 (Lu et al, 2018) was used to comprehensively evaluated the stability of candidate reference genes and GAPDH was shown as the most stable gene (Supplementary Figure 1), which was then chosen as an internal control.…”

Section: Real-time Quantitative Reverse Transcription Pcrmentioning

confidence: 99%

Identification of Auxiliary Biomarkers and Description of the Immune Microenvironmental Characteristics in Duchenne Muscular Dystrophy by Bioinformatical Analysis and Experiment

Han

Wang

et al. 2022

Front. Neurosci.

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BackgroundDuchenne muscular dystrophy (DMD) is a genetic muscle disorder characterized by progressive muscle wasting associated with persistent inflammation. In this study, we aimed to identify auxiliary biomarkers and further characterize the immune microenvironment in DMD.MethodsDifferentially expressed genes (DEGs) were identified between DMD and normal muscle tissues based on Gene Expression Omnibus (GEO) datasets. Bioinformatical analysis was used to screen and identify potential diagnostic signatures of DMD which were further validated by real-time quantitative reverse transcription PCR (RT-qPCR). We also performed single-sample gene-set enrichment analysis (ssGSEA) to characterize the proportion of tissue-infiltrating immune cells to determine the inflammatory state of DMD.ResultsIn total, 182 downregulated genes and 263 upregulated genes were identified in DMD. C3, SPP1, TMSB10, TYROBP were regarded as adjunct biomarkers and successfully validated by RT-qPCR. The infiltration of macrophages, CD4+, and CD8+ T cells was significantly higher (p < 0.05) in DMD compared with normal muscle tissues, while the infiltration of activated B cells, CD56dim natural killer cells, and type 17 T helper (Th17) cells was lower. In addition, the four biomarkers (C3, SPP1, TMSB10, TYROBP) were strongly associated with immune cells and immune-related pathways in DMD muscle tissues.ConclusionAnalyses demonstrated C3, SPP1, TMSB10, and TYROBP may serve as biomarkers and enhance our understanding of immune responses in DMD. The infiltration of immune cells into the muscle microenvironment might exert a critical impact on the development and occurrence of DMD.

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“…Treatment of murine models of DMD with tyrosine kinase inhibitors blocking the PDGFRα, such as imatinib, crenolanib, and nintedanib, decreased muscle fibrosis and improved muscle function 20–22 . Moreover, previous studies have shown that PDGF‐AA expression is higher in dystrophic compared with healthy muscles 23 and that PDGF‐AA serum levels are increased in DMD patients correlating with several muscle function tests 24 . However, the pathways activated by PDGF‐AA in FAPs in muscular dystrophies have not been described so far.…”

Section: Introductionmentioning

confidence: 99%

“… 20 , 22 Moreover, previous studies have shown that PDGF‐AA expression is higher in dystrophic compared with healthy muscles 23 and that PDGF‐AA serum levels are increased in DMD patients correlating with several muscle function tests. 24 However, the pathways activated by PDGF‐AA in FAPs in muscular dystrophies have not been described so far. This knowledge could help us to identify new targets for therapies aiming to reduce fibrosis in DMD and slow down the on‐going degenerative process.…”

Section: Introductionmentioning

confidence: 99%

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

Fernández‐Simón

Suárez‐Calvet

Carrasco‐Rozas

et al. 2022

J cachexia sarcopenia muscle

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BackgroundThe lack of dystrophin expression in Duchenne muscular dystrophy (DMD) induces muscle fibre and replacement by fibro-adipose tissue. Although the role of some growth factors in the process of fibrogenesis has been studied, pathways activated by PDGF-AA have not been described so far. Our aim was to study the molecular role of PDGF-AA in the fibrotic process of DMD. Methods Skeletal muscle fibro-adipogenic progenitor cells (FAPs) from three DMD treated with PDGF-AA at 50 ng/mL were analysed by quantitative mass spectrometry-based proteomics. Western-blot, immunofluorescence, and G-LISA were used to confirm the mass spectrometry results. We evaluated the effects of PDGF-AA on the activation of RhoA pathway using two inhibitors, C3-exoenzyme and fasudil. Cell proliferation and migration were determined by BrdU and migration assay. Actin reorganization and collagen synthesis were measured by phalloidin staining and Sircol assay, respectively. In an in vivo proof of concept study, we treated dba/2J-mdx mice with fasudil for 6 weeks. Muscle strength was assessed with the grip strength. Immunofluorescence and flow cytometry analyses were used to study fibrotic and inflammatory markers in muscle tissue. Results Mass spectrometry revealed that RhoA pathway proteins were up-regulated in treated compared with nontreated DMD FAPs (n = 3, mean age = 8 ± 1.15 years old). Validation of proteomic data showed that Arhgef2 expression was significantly increased in DMD muscles compared with healthy controls by a 7.7-fold increase (n = 2, mean age = 8 ± 1.14 years old). In vitro studies showed that RhoA/ROCK2 pathway was significantly activated by PDGF-AA (n = 3, 1.88-fold increase, P < 0.01) and both C3-exoenzyme and fasudil blocked that activation (n = 3, P < 0.05 and P < 0.001, respectively). The activation of RhoA pathway by PDGF-AA promoted a significant increase in proliferation and migration of FAPs (n = 3, P < 0.001), while C3-exoenzyme and fasudil inhibited FAPs proliferation at 72 h and migration at 48 and 72 h (n = 3, P < 0.001). In vivo studies showed that fasudil improved muscle function (n = 5 non-treated dba/2J-mdx and n = 6 treated dba/2J-mdx, 1.76-fold increase, P < 0.013), and histological studies demonstrated a 23% reduction of collagen-I expression area (n = 5 non-treated dba/2J-mdx and n = 6 treated dba/2Jmdx, P < 0.01). Conclusions Our results suggest that PDGF-AA promotes the activation of RhoA pathway in FAPs from DMD patients. This pathway could be involved in FAPs activation promoting its proliferation, migration, and actin reorganization, which represents the beginning of the fibrotic process. The inhibition of RhoA pathway could be considered as a potential therapeutic target for muscle fibrosis in patients with muscular dystrophies.

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Platelet Derived Growth Factor-AA Correlates With Muscle Function Tests and Quantitative Muscle Magnetic Resonance in Dystrophinopathies

Cited by 5 publications

References 61 publications

Transcriptomic analysis of paired healthy human skeletal muscles to identify modulators of disease severity in DMD

Transcriptomic analysis of paired healthy human skeletal muscles to identify modulators of disease severity in DMD

Identification of Auxiliary Biomarkers and Description of the Immune Microenvironmental Characteristics in Duchenne Muscular Dystrophy by Bioinformatical Analysis and Experiment

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

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