Hepatitis C virus induced miR200c down modulates FAP-1, a negative regulator of Src signaling and promotes hepatic fibrosis.
Hepatitis C virus (HCV) induced liver disease is the leading indication for liver transplantation (LTx). Reinfection and accelerated development of fibrosis is a universal phenomenon following LTx. The molecular events that lead to fibrosis following HCV infection still remains poorly defined. In this study, we determined microRNA (miRNA) and mRNA expression profiles in livers from chronic HCV patients and normals using microarrays. Using Genego software and pathway finder we performed an interactive analysis to identify target genes that are modulated by miRNAs. 22 miRNAs were up regulated (>2 fold) and 35 miRNAs were down regulated (>2fold) compared to controls. Liver from HCV patients demonstrated increased expression of 306 genes (>3 fold) and reduced expression of 133 genes (>3 fold). Combinatorial analysis of the networks modulated by the miRNAs identified regulation of the phospholipase C pathway (miR200c, miR20b, and miR31through cellular proto-oncogene tyrosine-protein kinase Src (cSrc)), response to growth factors and hormones (miR141, miR107 and miR200c through peroxisome proliferator-activated receptor alpha and extracellular-signal-regulated kinases, and regulation of cellular proliferation (miR20b, miR10b, and miR141 through cyclin-dependent kinase inhibitor 1 or CDK-interacting protein 1 p21). Real time PCR (RT-PCR) validation of the miRNA in HCV infected livers demonstrated a 3.3 ±0.9 fold increase in miR200c. In vitro transfection of fibroblasts with miR200c resulted in a 2.2 fold reduction in expression of tyrosine-protein phosphatase non-receptor type 13 or FAS associated phosphatase 1 (FAP-1) and 2.3 fold increase in expression of cSrc. miR200c transfection resulted in significant increases in expression of collagen and fibroblast growth factor (2.8 and 3.4 fold, p<0.05). Therefore, we propose that HCV induced increased expression of miR200c can down modulate the expression of FAP1, a critical regulator of Src and MAP kinase pathway that play an important role in the production of fibrogenic growth factors and development of fibrosis.
Department of Surgery, Washington University School of Medicine, Saint Louis, Missouri, USA.
Sabarinathan
S
Ramachandran
Haseeb
H
Ilias Basha
Nayan J
NJ
Sarma
Yiing
Y
Lin
Jeffrey S
JS
Crippin
William C
WC
Chapman
Thalachallour
T
Mohanakumar
D000328:Adult; D000081247:CSK Tyrosine-Protein Kinase; D004789:Enzyme Activation; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation; D053263:Gene Regulatory Networks; D016174:Hepacivirus / Q000502:physiology*; D019698:Hepatitis C, Chronic / Q000150:complications / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D008103:Liver Cirrhosis / Q000209:etiology*; D008297:Male; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D008875:Middle Aged; D054595:Protein Tyrosine Phosphatase, Non-Receptor Type 13 / Q000235:genetics* / Q000378:metabolism; D000090063:Proto-Oncogene Mas; D034622:RNA Interference; D015398:Signal Transduction; D059467:Transcriptome; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000096:biosynthesis; D019061:src-Family Kinases / Q000378:metabolism*
C000715470:MAS1 protein, human; C533896:MIRN200 microRNA, human; D035683:MicroRNAs; D000090063:Proto-Oncogene Mas; D016212:Transforming Growth Factor beta; D000081247:CSK Tyrosine-Protein Kinase; D019061:src-Family Kinases; C000631346:CSK protein, human; C516742:PTPN13 protein, human; D054595:Protein Tyrosine Phosphatase, Non-Receptor Type 13