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Comparative gene expression profiling of pig‐derived iPSC ‐like cells: Effects of induced pluripotency on expression of porcine endogenous retrovirus (PERV)
Author(s) -
Godehardt Antonia W.,
Petkov Stoyan,
Gulich Barbara,
Fischer Nicole,
Niemann Heiner,
Tönjes Ralf R.
Publication year - 2018
Publication title -
xenotransplantation
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.052
H-Index - 61
eISSN - 1399-3089
pISSN - 0908-665X
DOI - 10.1111/xen.12429
Subject(s) - endogenous retrovirus , biology , xenotransplantation , induced pluripotent stem cell , microbiology and biotechnology , retrovirus , reverse transcriptase , gene expression , gene expression profiling , cell culture , real time polymerase chain reaction , reprogramming , reverse transcription polymerase chain reaction , virology , gene , embryonic stem cell , genome , virus , rna , transplantation , genetics , medicine , surgery
Background Porcine induced pluripotent stem cells (pi PSC s) offer an alternative strategy in xenotransplantation ( XT x). As human endogenous retroviruses ( HERV ), particularly HERV ‐K, are highly expressed in natural human stem cells, we compared the expression of porcine endogenous retroviruses ( PERV ) and retrotransposon LINE ‐1 (L1) open reading frames 1 and 2 ( pORF 1 and pORF 2) in different pi PSC ‐like cell lines with their progenitors (porcine fetal fibroblasts, pFF ). Methods Cells reprogrammed via Sleeping Beauty‐transposed transcription factors were cultured and analyzed on a custom‐designed microarray representing the reference pig genome. Data were complemented by qRT ‐ PCR and reverse transcriptase ( RT ) assay. Results The expression profiles revealed that 8515 of 26 967 targets were differentially expressed. A total of 4443 targets showed log 2 expression ratio >1, and 4072 targets showed log 2 expression ratio less than −1 with 0.05 P ‐value threshold. Approximately ten percent of the targets showed highly significant expression ratios with log 2 ≥4 or ≤−4. Besides this general switch in cellular gene expression that was accompanied by an altered morphology, expression of both PERV and L1 pORF 1/ pORF 2 was significantly enhanced. pi PSC ‐like cells revealed a 10‐fold to 100‐fold higher transcription of the viral PERV ‐A and PERV ‐B envelope genes ( env ), viral protease/polymerase ( prt/pol ), and L1 elements. No functional retrovirus could be detected under these conditions. Conclusion Epigenetic reprogramming has functional impact on retrotransposons. Thus, the induction of pig‐derived pluripotent cells influences their PERV expression profile. Data emphasize the necessity to focus on animals, which show non‐functional endogenous viral background to ensure virological safety.