TY - JOUR
T1 - Integrative analysis of miRNA and mRNA paired expression profiling of primary fibroblast derived from diabetic foot ulcers reveals multiple impaired cellular functions
AU - Liang, Liang
AU - Stone, Rivka C.
AU - Stojadinovic, Olivera
AU - Ramirez, Horacio
AU - Pastar, Irena
AU - Maione, Anna G.
AU - Smith, Avi
AU - Yanez, Vanessa
AU - Veves, Aristides
AU - Kirsner, Robert S.
AU - Garlick, Jonathan A.
AU - Tomic-Canic, Marjana
N1 - Funding Information:
We thank all members of the Tomic-Canic, Garlic and Veves laboratories for their help and support. We are very grateful to the Clinical Research Wound Healing Program (Carol Kittles, Aliette Espinosa, Dr. Alejandra Vivas, Dr. Katherine Barquerizo-Nole) for their assistance with patients and collection of specimens. We also thank Dr. Antonio Barrientos for generously sharing laboratory resources and equipment and we acknowledge the use of University of Miami Genomic Facility. Source of Funding: This work was funded by NIH grants DK098055 (JG), NR013881 (MT-C), University of Miami SAC Award SAC 2013-19 (MT-C), and the UMSDRC of the Department of Dermatology and Cutaneous Surgery at the University of Miami Miller School of Medicine. Conflict of Interest: The authors have no conflict of interest.
Publisher Copyright:
© 2016 by the Wound Healing Society
PY - 2016/11/1
Y1 - 2016/11/1
N2 - 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.
AB - 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.
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U2 - 10.1111/wrr.12470
DO - 10.1111/wrr.12470
M3 - Article
C2 - 27607190
AN - SCOPUS:84990220376
VL - 24
SP - 943
EP - 953
JO - Wound Repair and Regeneration
JF - Wound Repair and Regeneration
SN - 1067-1927
IS - 6
ER -