Novel contribution to clubfoot pathogenesis: The possible role of extracellular matrix proteins

Abstract

Idiopathic pes equinovarus (clubfoot) is a congenital deformity of the feet and lower legs. Clubfoot belongs to a group of fibro‐proliferative disorders but its origin remains unknown. Our study aimed to achieve the first complex proteomic comparison of clubfoot contracted tissue of the foot (medial side; n = 16), with non‐contracted tissue (lateral side; n = 13). We used label‐free mass spectrometry quantification and immunohistochemistry. Seven proteins were observed to be significantly upregulated in the medial side (asporin, collagen type III, V, and VI, versican, tenascin‐C, and transforming growth factor beta induced protein) and four in the lateral side (collagen types XII and XIV, fibromodulin, and cartilage intermediate layer protein 2) of the clubfoot. Comparison of control samples from cadavers brought only two different protein concentrations (collagen types I and VI). We also revealed pathological calcification and intracellular positivity of transforming growth factor beta only in the contracted tissue of clubfoot. Most of the 11 differently expressed proteins are strongly related to the extracellular matrix architecture and we assume that they may play specific roles in the pathogenesis of this deformity. These proteins seem to be promising targets for future investigations and treatment of this disease. © 2019 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res

Integrated bioinformatics analysis of potential pathway biomarkers using abnormal proteins in clubfoot

“…Abnormal biological development, such as muscle, neurological, skeletal and vascular abnormalities, has been previously identified and confirmed (Basit & Khoshhal, 2018;Eckhardt et al, 2019;Herceg et al, 2006;Hester et al, 2009;Lovell & Morcuende, 2007;Ošt'ádal et al, 2015). The HOX and TBX families governed limb identity, and fibroblast growth factor participated in the formation of limb muscles (Ohuchi & Noji, 1999;Wang et al, 2008).…”

“…Select soft tissues in clubfoot are contracted, resulting in stiffness. Extracellular matrix proteins, such as asporin, collagen types III, V, and VI, versican, tenascin-C, and TGF-beta induced protein, are highly expressed in contracted tissues clubfoot (Eckhardt et al, 2019;Ošt'ádal et al, 2015). Additionally, the expression levels of growth factors TGF-beta and platelet-derived growth factor are high and a blockade of growth factors led to decreased collagen expression, proliferation, and chemotaxis (Li et al, 2001).…”

“…PPI analysis was performed using the MEx Interactome database. The parameters were referred Eckhardt et al (2019) and Ošt'ádal et al (2015) to the literature-curated comprehensive data from the Innate DB (Breuer et al, 2013). Gene-miRNA interactome analysis was carried out with comprehensive experimentally validated miRNA-gene interaction data collected from TarBase and miRTarBase (Chou et al, 2018;Vlachos et al, 2015).…”

“…Clubfoot is associated with neuromuscular lesions, heredity abnormalities, skeletal dysplasia, soft tissue contracture, vascular abnormalities, ECM abnormalities, and intrauterine growth retardation (Chesney, Barker & Maffulli, 2007;Eckhardt et al, 2019;Miedzybrodzka, 2003;Ošt'ádal et al, 2015;Poon, Li & Alman, 2009;Sodre et al, 1990;Wang et al, 2013a;Wang et al, 2013b). In addition, smoking and viral infections in pregnant women are also closely related to congenital clubfoot (Palma et al, 2013;Robertson Jr & Corbett, 1997).…”

Peer Review #2 of "Integrated bioinformatics analysis of potential pathway biomarkers using abnormal proteins in clubfoot (v0.1)"

“…Clubfoot is associated with neuromuscular lesions, heredity abnormalities, skeletal dysplasia, soft tissue contracture, vascular abnormalities, ECM abnormalities, and intrauterine growth retardation (Chesney et al, 2007;Eckhardt et al, 2019;Miedzybrodzka 2003;Ošt'ádal et al, 2015;Poon and Li, 2009;Sodre et al, 1990;. In addition, smoking and viral PeerJ reviewing PDF | (2019:08:40368:1:2:NEW 26 Nov 2019)…”