Integrated transcriptomic and proteomic analysis of the molecular cargo of extracellular vesicles derived from porcine adipose tissue-derived mesenchymal stem cells.
BACKGROUND
Mesenchymal stromal/stem cell (MSC) transplantation is a promising therapy for tissue regeneration. Extracellular vesicles (EVs) released by MSCs act as their paracrine effectors by delivering proteins and genetic material to recipient cells. To assess how their cargo mediates biological processes that drive their therapeutic effects, we integrated miRNA, mRNA, and protein expression data of EVs from porcine adipose tissue-derived MSCs.


METHODS
Simultaneous expression profiles of miRNAs, mRNAs, and proteins were obtained by high-throughput sequencing and LC-MS/MS proteomic analysis in porcine MSCs and their daughter EVs (n = 3 each). TargetScan and ComiR were used to predict miRNA target genes. Functional annotation analysis was performed using DAVID 6.7 database to rank primary gene ontology categories for the enriched mRNAs, miRNA target genes, and proteins. STRING was used to predict associations between mRNA and miRNA target genes.


RESULTS
Differential expression analysis revealed 4 miRNAs, 255 mRNAs, and 277 proteins enriched in EVs versus MSCs (fold change >2, p<0.05). EV-enriched miRNAs target transcription factors (TFs) and EV-enriched mRNAs encode TFs, but TF proteins are not enriched in EVs. Rather, EVs are enriched for proteins that support extracellular matrix remodeling, blood coagulation, inflammation, and angiogenesis.


CONCLUSIONS
Porcine MSC-derived EVs contain a genetic cargo of miRNAs and mRNAs that collectively control TF activity in EVs and recipient cells, as well as proteins capable of modulating cellular pathways linked to tissue repair. These properties provide the fundamental basis for considering therapeutic use of EVs in tissue regeneration.
Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, United States of America.
Alfonso
A
Eirin
Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, United States of America.
Xiang-Yang
XY
Zhu
Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, United States of America.
Amrutesh S
AS
Puranik
Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, United States of America.
John R
JR
Woollard
Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, United States of America.
Hui
H
Tang
Department of Health Sciences Research, Mayo Clinic, Rochester, Minnesota, United States of America.
Surendra
S
Dasari
Department of Cardiovascular Diseases, Mayo Clinic, Rochester, Minnesota, United States of America.
Amir
A
Lerman
Department of Orthopedic Surgery, Mayo Clinic, Rochester, Minnesota, United States of America.
Andre J
AJ
van Wijnen
Division of Nephrology and Hypertension, Mayo Clinic, Rochester, Minnesota, United States of America.|Department of Cardiovascular Diseases, Mayo Clinic, Rochester, Minnesota, United States of America.
Lilach O
http://orcid.org/0000-0002-3271-3887
LO
Lerman
D000273:Adipose Tissue / Q000737:chemistry* / Q000378:metabolism; D000818:Animals; D000067128:Extracellular Vesicles / Q000737:chemistry* / Q000378:metabolism; D020869:Gene Expression Profiling; D059630:Mesenchymal Stem Cells / Q000737:chemistry* / Q000378:metabolism; D035683:MicroRNAs / Q000032:analysis; D011506:Proteins / Q000032:analysis; D040901:Proteomics; D012333:RNA, Messenger / Q000032:analysis; D013552:Swine
D035683:MicroRNAs; D011506:Proteins; D012333:RNA, Messenger