<< Back to the abstract archive
Epigenetic Mapping of Wound Edge from Chronic Wound Patients using Next Generation Sequencing
Saba Tabasum, Kanhaiya Singh, Sashwati Roy, Durba Pal, Subhadip Ghatak, Dolly Khona, Savita Khanna, Chandan K Sen
University
2019-02-15
Presenter: Saba Tabasum
Affidavit:
I certify that the material proposed for presentation in this abstract has not been published in any scientific journal or previously presented at a major meeting.
Director Name: CKS
Author Category: Other Specialty Resident
Presentation Category: Basic Science Research
Abstract Category: General Reconstruction
The loud biochemical microenvironment of the chronic wound sharply departs from that of the skin under homeostatic conditions and is likely to induce epigenetic changes thus influencing wound healing outcomes. Unbiased whole-genome DNA methylation (methylome) was studied in normal skin (NS) and in wound-edge of chronic wound (CW) patients. DNA isolated from CW or NS was enriched for the study of methylated DNA. Single-end 50bp sequencing was performed using Illumina HiSeq 2500. Methylation status of proximal promoter (1 Kb) was calculated using MethylCap-Seq data analysis. Whole-genome RNA sequencing and microRNA nanoString analysis over CW and NS samples was performed to qualify the methylation data. Genes involved in epithelial to mesenchymal transition (EMT) were hypermethylated in CW compared to NS. Bisulfite sequencing was used to validate hypermethylation of predicted upstream regulators. microRNA promoters were differentially methylated in CW compared to NS (71 hyper methylated; 20 hypomethylated). A sum total of 1281 genes were found to be differentially expressed in CW compared to NS (n=3, p<0.05). Comparison between hypermethylated and downregulated expressed genes identified common candidate genes of EMT pathway like ADAM17, TWIST1 and SMURF1 obtained through two independent global screening approaches. Topical application of DNA demethylation agent 5'-azacytidine successfully rescued ischemic wounds of mice by increasing the expression of ADAM17. Lentiviral overexpression of ADAM17 alone rescued ischemic wounds by enabling the P53-ADAM17 cascade. Epigenetic activity is high at the chronic wound site. Such activity has significant impact on gene expression at the wound-edge and is therefore likely to influence healing outcomes.
