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2016 ; 24
(6
): 943-953
Nephropedia Template TP
gab.com Text
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Integrative analysis of miRNA and mRNA paired expression profiling of primary
fibroblast derived from diabetic foot ulcers reveals multiple impaired cellular
functions
#MMPMID27607190
Liang L
; Stone RC
; Stojadinovic O
; Ramirez H
; Pastar I
; Maione AG
; Smith A
; Yanez V
; Veves A
; Kirsner RS
; Garlick JA
; Tomic-Canic M
Wound Repair Regen
2016[Nov]; 24
(6
): 943-953
PMID27607190
show ga
Diabetic foot ulcers (DFUs) are one of the major complications of diabetes. Its
molecular pathology remains poorly understood, impeding the development of
effective treatments. Although it has been established that multiple cell types,
including fibroblasts, keratinocytes, macrophages, and endothelial cells, all
contribute to inhibition of healing, less is known regarding contributions of
individual cell type. Thus, we generated primary fibroblasts from nonhealing DFUs
and evaluated their cellular and molecular properties in comparison to
nondiabetic foot fibroblasts (NFFs). Specifically, we analyzed both micro-RNA and
mRNA expression profiles of primary DFU fibroblasts. Paired genomic analyses
identified a total of 331 reciprocal miRNA-mRNA pairs including 21 miRNAs
(FC?>?2.0) along with 239 predicted target genes (FC?>?1.5) that are
significantly and differentially expressed. Of these, we focused on three miRNAs
(miR-21-5p, miR-34a-5p, miR-145-5p) that were induced in DFU fibroblasts as most
differentially regulated. The involvement of these microRNAs in wound healing was
investigated by testing the expression of their downstream targets as well as by
quantifying cellular behaviors in prospectively collected and generated cell
lines from 15 patients (seven DFUF and eight NFF samples). We found large number
of downstream targets of miR-21-5p, miR-34a-5p, miR-145-5p to be coordinately
regulated in mRNA profiles, which was confirmed by quantitative real-time PCR.
Pathway analysis on paired miRNA-mRNA profiles predicted inhibition of cell
movement and cell proliferation, as well as activation of cell differentiation
and senescence in DFU fibroblasts, which was confirmed by cellular assays. We
concluded that induction of miR-21-5p, miR-34a-5p, miR-145-5p in DFU dermal
fibroblasts plays an important role in impairing multiple cellular functions,
thus contributing to overall inhibition of healing in DFUs.