title: Adenoviral overexpression of Lhx2 attenuates cell viability but does not preserve the stem cell like phenotype of hepatic stellate cells.
abstract: Hepatic stellate cells (HSC) are well known initiators of hepatic fibrosis. After liver cell damage, HSC transdifferentiate into proliferative myofibroblasts, representing the major source of extracellular matrix in the fibrotic organ. Recent studies also demonstrate a role of HSC as progenitor or stem cell like cells in liver regeneration. Lhx2 is described as stem cell maintaining factor in different organs and as an inhibitory transcription factor in HSC activation. Here we examined whether a continuous expression of Lhx2 in HSC could attenuate their activation and whether Lhx2 could serve as a potential target for antifibrotic gene therapy. Therefore, we evaluated an adenoviral mediated overexpression of Lhx2 in primary HSC and investigated mRNA expression patterns by qRT-PCR as well as the activation status by different in vitro assays. HSC revealed a marked increase in activation markers like smooth muscle actin alpha (αSMA) and collagen 1α independent from adenoviral transduction. Lhx2 overexpression resulted in attenuated cell viability as shown by a slightly hampered migratory and contractile phenotype of HSC. Expression of stem cell factors or signaling components was also unaffected by Lhx2. Summarizing these results, we found no antifibrotic or stem cell maintaining effect of Lhx2 overexpression in primary HSC.
authors.affiliation: Institute for Experimental Surgery, Rostock University Medical Center, Rostock, Germany.
authors.forename: Berit
authors.initials: B
authors.lastname: Genz
authors.affiliation: Dr. Margarete Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart, Germany.
authors.forename: Maria
authors.initials: M
authors.lastname: Thomas
authors.affiliation: Institute of Experimental Gene Therapy and Cancer Research, Rostock University Medical Center, Rostock, Germany.
authors.forename: Brigitte M
authors.initials: BM
authors.lastname: Pützer
authors.affiliation: Institute for Biostatistics and Informatics in Medicine and Ageing Research, Rostock University Medical Center, Rostock, Germany.
authors.forename: Marcin
authors.initials: M
authors.lastname: Siatkowski
authors.affiliation: Institute for Biostatistics and Informatics in Medicine and Ageing Research, Rostock University Medical Center, Rostock, Germany.
authors.forename: Georg
authors.initials: G
authors.lastname: Fuellen
authors.affiliation: Institute for Experimental Surgery, Rostock University Medical Center, Rostock, Germany.
authors.forename: Brigitte
authors.initials: B
authors.lastname: Vollmar
authors.affiliation: Institute for Experimental Surgery, Rostock University Medical Center, Rostock, Germany. Electronic address: kerstin.abshagen@uni-rostock.de.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Abshagen
mesh_terms: D000199:Actins / Q000235:genetics; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002470:Cell Survival / Q000235:genetics*; D054338:Cell Transdifferentiation / Q000235:genetics; D002478:Cells, Cultured; D024042:Collagen Type I / Q000235:genetics; D005260:Female; D057809:HEK293 Cells; D055166:Hepatic Stellate Cells / Q000378:metabolism*; D006801:Humans; D060850:LIM-Homeodomain Proteins / Q000235:genetics*; D008103:Liver Cirrhosis / Q000235:genetics; D008115:Liver Regeneration / Q000235:genetics; D051379:Mice; D008807:Mice, Inbred BALB C; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics*
chemical_list: D000199:Actins; D024042:Collagen Type I; D060850:LIM-Homeodomain Proteins; C489784:Lhx2 protein, mouse; D012333:RNA, Messenger; D014157:Transcription Factors; C544013:alpha-smooth muscle actin, mouse
keywords: Cell proliferation; Fluidigm qRT-PCR; Hepatic stem cells; Liver fibrosis