Single cell transcriptome sequencing of diabetic foot skin
Chronic wounds, including diabetic foot ulceration and venous ulcers, are a major problem that is associated with significant morbidity and mortality and financial cost. Recent studies by our unit and elsewhere have shown that chronic inflammation is one of the major factors that is associated with impaired wound healing2-4. Wound healing requires a well-coordinated action by numerous cell types, including neutrophils, monocyte/macrophages, fibroblasts, endothelial cells and keratinocytes. However, there is considerable lack of understanding of the molecular physiology of each of these cells that could lead to understanding of the pathophysiology of impaired wound healing. In this application, we propose to perform single-cell transcriptome sequencing from forearm and foot skin biopsies from healthy subjects and diabetic patients with foot ulcer. Our main will be to evaluate gene expression in various cell types and also explore possible differences in various body sites using highly robust drop-seq sequencing approach that allow evaluation of transcriptomic state of thousands of human cells in single experiment by measuring 4,000-7,000 genes. Finally, we will explore possible differences between diabetic and non-diabetic patients. Successful completion of this application will not only contribute to the development of a reference atlas of skin cells transcriptome state but can also play a major role in the development of new therapeutic approaches.