Tracking the Molecular Scenarios for Tumorigenic Remodeling of Extracellular Matrix Based on Gene Expression Profiling in Equine Skin Neoplasia Models

Abstract

An important component of tissues is the extracellular matrix (ECM), which not only forms a tissue scaffold, but also provides the environment for numerous biochemical reactions. Its composition is strictly regulated, and any irregularities can result in the development of many diseases, including cancer. Sarcoid is the most common skin cancer in equids. Its formation results from the presence of the genetic material of the bovine papillomavirus (BPV). In addition, it is assumed that sarcoid-dependent oncogenic transformation arises from a disturbed wound healing process, which may be due to the incorrect functioning of the ECM. Moreover, sarcoid is characterized by a failure to metastasize. Therefore, in this study we decided to investigate the differences in the expression profiles of genes related not only to ECM remodeling, but also to the cell adhesion pathway, in order to estimate the influence of disturbances within the ECM on the sarcoid formation process. Furthermore, we conducted comparative research not only between equine sarcoid tissue bioptates and healthy skin-derived explants, but also between dermal fibroblast cell lines transfected and non-transfected with a construct encoding the E4 protein of the BP virus, in order to determine its effect on ECM disorders. The obtained results strongly support the hypothesis that ECM-related genes are correlated with sarcoid formation. The deregulated expression of selected genes was shown in both equine sarcoid tissue bioptates and adult cutaneous fibroblast cell (ACFC) lines neoplastically transformed by nucleofection with gene constructs encoding BPV1-E1^E4 protein. The identified genes (CD99, ITGB1, JAM3 and CADM1) were up- or down-regulated, which pinpointed the phenotypic differences from the backgrounds noticed for adequate expression pro-files in other cancerous or noncancerous tumors as reported in the available literature data. Unravelling the molecular pathways of ECM remodeling and cell adhesion in the in vivo and ex vivo models of epidermal/dermal sarcoid-related cancerogenesis might provide powerful tools for further investigations of genetic and epigenetic biomarkers for both silencing and re-initiating the processes of sarcoid-dependent neoplasia. Recognizing those biomarkers might insightfully explain the relatively high capacity of sarcoid-descended cancerous cell derivatives to epigenomically reprogram their nonmalignant neoplastic status in domestic horse cloned embryos produced by somatic cell nuclear transfer (SCNT).

Transcriptome and methylome sequencing reveals altered long non-coding RNA genes expression and their aberrant DNA methylation in equine sarcoids

“…We also found that predicted target genes in GO analysis were significantly associated with the extracellular matrix disassembly pathway. As we previously mentioned the changes in ECM composition are reported to play a critical role in the development of horse sarcoids (Martano et al 2016 ; Podstawski et al 2022 ). These results indicated that lncRNAs may play certain roles in the expression of ECM-related genes and play important roles in the pathogenesis of equine sarcoids.…”

“…For example, it was reported that the expression level of lncRNA H19 was significantly downregulated in the metastatic prostate cancer cell line and negatively correlated with the expression of the extracellular matrix protein TGFBI (Zhu et al 2014 ). Previous studies demonstrated that an imbalance of the ECM and an abnormal expression of its genes play a major role in sarcoid pathogenesis (Martano et al 2016 ; Podstawski et al 2022 ). It is therefore of interest, that our study showed upregulation of three lncRNAs — ENSECAG00000046740 , ENSECAG00000058013 , and ENSECAG00000048455 that may be connected with aberrant expression of their correlated genes, and thus indirectly affect the stability of the extracellular matrix during sarcoid formation and progression.…”