SRP404116 PRJNA893389 Identification of tRNA-derived small RNAs using small RNA sequencing and tRF-Gly-CCC-039 inhibits diabetics wound healing Delayed skin healing in diabetic wounds are a major clinical problem. The tRNA-derived small RNAs (tsRNAs) were reported to associate with diabetes. However, the role of tsRNAs in diabetic wound healing is unclear. Our study was designed to explore the tsRNAs expression profile, mine key potential tsRNAs and its mechanism in diabetic wound. Skin tissue of diabetic foot ulcers and healthy control were subjected to small RNA sequencing. The role of candidate tsRNA was explored by loss- and gain-function experiments in HUVECs. A total of 55 differentially expressed tsRNAs were identified, including 12 up-regulated and 43 down-regulated in diabetes group compared with control group. These tsRNAs were mainly concentrated in intercellular interactions and neural function regulation in GO terms, and enriched in MAPK, insulin, FoxO, calcium, Ras, ErbB, Wnt, T cell receptor, cGMP-PKG signaling pathway. The tRF-Gly-CCC-039 expression was up-regulated both in vivo and in vitro during diabetic model. High glucose injured endothelial function in HUVECs, and tRF-Gly-CCC-039 mimics further harmed HUVECs function, characterized by suppression of proliferation, migration, tube formation, and the expression of Coll1a1, Coll4a2, and MMP9. Conversely, tRF-Gly-CCC-039 inhibitor could attenuate high glucose-induced endothelial injury to HUVECs. We investigated the tsRNAs expression profile in diabetic foot ulcers and defined an impairment role of tRF-Gly-CCC-039 in endothelial function in HUVECs. This study may provide a new insight into accelerating diabetic skin wound healing.