Kyle Willard1, Mary-Jessica Nancy Laguette1, Leonardo Alves de Souza Rios2, Caroline D'Alton1, Melissa Nel2, Sharon Prince2, Malcolm Collins1, Alison Victoria September3. 1. Division of Exercise Science and Sports Medicine, Department of Human Biology, University of Cape Town, South Africa; UCT Research Centre for Health through Physical Activity, Lifestyle and Sport, South Africa; International Federation of Sports Medicine (FIMS) Collaborative Centre of Sports Medicine, South Africa. 2. Division of Cell BiologyDepartment of Human Biology, University of Cape Town, South Africa. 3. Division of Exercise Science and Sports Medicine, Department of Human Biology, University of Cape Town, South Africa; UCT Research Centre for Health through Physical Activity, Lifestyle and Sport, South Africa; International Federation of Sports Medicine (FIMS) Collaborative Centre of Sports Medicine, South Africa. Electronic address: alison.september@uct.ac.za.
Abstract
OBJECTIVES: To investigate the functional effect of implicated variants within BGN and COL5A1 on gene expression of components of the extracellular matrix (ECM) in a TGF-β-stimulated risk model for musculoskeletal soft tissue injuries. DESIGN: Experimental research, laboratory study. METHODS: Skin biopsies were obtained from nine healthy participants with either a combined increased or reduced risk profile for COL5A1 rs12722 C>T and BGN rs1126499 C>T - rs1042103 G>A, and primary fibroblast cell lines were established. Total RNA was extracted at baseline (10% FBS), after serum starvation (1% FBS) and TGF-β1 treatment (1% FBS, 10ng/mL TGF-1β). Relative mRNA levels of BGN, COL5A1, DCN and VEGFA was quantified using Taqman® array pre-spotted plate assays (Applied Biosystems, Foster city, CA, USA). RESULTS: At baseline, the reduced risk group had 2.5, 1.9 and 2 fold increases (p<0.001) in relative BGN, COL5A1 and VEGFA mRNA levels respectively. In the serum starved experiments, except for a significant 1.5 fold (p=0.017) increase in relative DCN mRNA expression in the reduced risk group, similar observations were noted for the other three genes. After TGF-1β treatment, the reduced risk group had 1.3 (p=0.011) and 1.4 fold (p=0.001) increases in the relative COL5A1 and VEGFA mRNA levels, respectively. CONCLUSIONS: Altered mRNA levels associated with genetic risk profiles for musculoskeletal soft injury risk at baseline (BGN, COL5A1 and VEGFA), with serum starvation (DCN) and after TGF-β1 treatment (COL5A1 and VEGFA) provide additional functional evidence to support the association of implicated genetic loci with several musculoskeletal soft tissue injuries. Implication of altered gene expression profiles underpinning these genetic risk associated loci potentially highlight key therapeutic targets for management of these injuries.
OBJECTIVES: To investigate the functional effect of implicated variants within BGN and COL5A1 on gene expression of components of the extracellular matrix (ECM) in a TGF-β-stimulated risk model for musculoskeletal soft tissue injuries. DESIGN: Experimental research, laboratory study. METHODS: Skin biopsies were obtained from nine healthy participants with either a combined increased or reduced risk profile for COL5A1rs12722 C>T and BGNrs1126499 C>T - rs1042103 G>A, and primary fibroblast cell lines were established. Total RNA was extracted at baseline (10% FBS), after serum starvation (1% FBS) and TGF-β1 treatment (1% FBS, 10ng/mL TGF-1β). Relative mRNA levels of BGN, COL5A1, DCN and VEGFA was quantified using Taqman® array pre-spotted plate assays (Applied Biosystems, Foster city, CA, USA). RESULTS: At baseline, the reduced risk group had 2.5, 1.9 and 2 fold increases (p<0.001) in relative BGN, COL5A1 and VEGFA mRNA levels respectively. In the serum starved experiments, except for a significant 1.5 fold (p=0.017) increase in relative DCN mRNA expression in the reduced risk group, similar observations were noted for the other three genes. After TGF-1β treatment, the reduced risk group had 1.3 (p=0.011) and 1.4 fold (p=0.001) increases in the relative COL5A1 and VEGFA mRNA levels, respectively. CONCLUSIONS: Altered mRNA levels associated with genetic risk profiles for musculoskeletal soft injury risk at baseline (BGN, COL5A1 and VEGFA), with serum starvation (DCN) and after TGF-β1 treatment (COL5A1 and VEGFA) provide additional functional evidence to support the association of implicated genetic loci with several musculoskeletal soft tissue injuries. Implication of altered gene expression profiles underpinning these genetic risk associated loci potentially highlight key therapeutic targets for management of these injuries.
Keywords:
Alpha 1 chain of type V collagen; Biglycan; Decorin; Genetic susceptibility; Transfor; Vascular endothelial growth factor; ming growth factor
Authors: Daneil Feldmann; Christian D Bope; Jon Patricios; Emile R Chimusa; Malcolm Collins; Alison V September Journal: PLoS One Date: 2022-10-06 Impact factor: 3.752