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Article

The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells

by
Marcin Samiec
1,*,
Jerzy Wiater
2,*,
Kamil Wartalski
2,
Maria Skrzyszowska
1,
Monika Trzcińska
1,
Daniel Lipiński
3,
Jacek Jura
1,
Zdzisław Smorąg
1,
Ryszard Słomski
3,4 and
Małgorzata Duda
5
1
Department of Reproductive Biotechnology and Cryoconservation, National Research Institute of Animal Production, Krakowska 1 Street, 32-083 Balice, Poland
2
Department of Histology, Jagiellonian University Medical College, Kopernika 7 Street, 31-034 Kraków, Poland
3
Department of Biochemistry and Biotechnology, Poznań University of Life Sciences, Dojazd 11 Street, 60-647 Poznań, Poland
4
Institute of Human Genetics, Polish Academy of Sciences, Strzeszyńska 32 Street, 60-479 Poznań, Poland
5
Department of Endocrinology, Institute of Zoology and Biomedical Research, Faculty of Biology, Jagiellonian University in Krakow, Gronostajowa 9 Street, 30-387 Kraków, Poland
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2022, 23(18), 10296; https://doi.org/10.3390/ijms231810296
Submission received: 18 August 2022 / Revised: 1 September 2022 / Accepted: 4 September 2022 / Published: 7 September 2022
(This article belongs to the Special Issue Molecular Mechanism and Application of Somatic Cell Cloning in Mammals)
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Abstract

The present study sought to establish the mitotically stable adult cutaneous fibroblast cell (ACFC) lines stemming from hFUT2×hGLA×HLA-E triple-transgenic pigs followed by trichostatin A (TSA)-assisted epigenetically modulating the reprogrammability of the transgenes permanently incorporated into the host genome and subsequent comprehensive analysis of molecular signatures related to proteomically profiling the generated ACFC lines. The results of Western blot and immunofluorescence analyses have proved that the profiles of relative abundance (RA) noticed for both recombinant human α-galactosidase A (rhα-Gal A) and human leukocyte antigen-E (HLA-E) underwent significant upregulations in tri-transgenic (3×TG) ACFCs subjected to TSA-mediated epigenetic transformation as compared to not only their TSA-unexposed counterparts but also TSA-treated and untreated non-transgenic (nTG) cells. The RT-qPCR-based analysis of porcine tri-genetically engineered ACFCs revealed stable expression of mRNA fractions transcribed from hFUT2, hGLA and HLA-E transgenes as compared to a lack of such transcriptional activities in non-transgenic ACFC variants. Furthermore, although TSA-based epigenomic modulation has given rise to a remarkable increase in the expression levels of Galα1→3Gal (α-Gal) epitopes that have been determined by lectin blotting analysis, their semi-quantitative profiles have dwindled profoundly in both TSA-exposed and unexposed 3×TG ACFCs as compared to their nTG counterparts. In conclusion, thoroughly exploring proteomic signatures in such epigenetically modulated ex vivo models devised on hFUT2×hGLA×HLA-E triple-transgenic ACFCs that display augmented reprogrammability of translational activities of two mRNA transcripts coding for rhα-Gal A and HLA-E proteins might provide a completely novel and powerful research tool for the panel of further studies. The objective of these future studies should be to multiply the tri-transgenic pigs with the aid of somatic cell nuclear transfer (SCNT)-based cloning for the purposes of both xenografting the porcine cutaneous bioprostheses and dermoplasty-mediated surgical treatments in human patients.
Keywords: swine; trichostatin A; epigenetic transformation; ex vivo model; tri-genetically modified; ACFC line; HLA-E; rhα-Gal A; rhα1,2-FT; α-Gal antigenic determinant; porcine skin xenograft swine; trichostatin A; epigenetic transformation; ex vivo model; tri-genetically modified; ACFC line; HLA-E; rhα-Gal A; rhα1,2-FT; α-Gal antigenic determinant; porcine skin xenograft

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MDPI and ACS Style

Samiec, M.; Wiater, J.; Wartalski, K.; Skrzyszowska, M.; Trzcińska, M.; Lipiński, D.; Jura, J.; Smorąg, Z.; Słomski, R.; Duda, M. The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells. Int. J. Mol. Sci. 2022, 23, 10296. https://doi.org/10.3390/ijms231810296

AMA Style

Samiec M, Wiater J, Wartalski K, Skrzyszowska M, Trzcińska M, Lipiński D, Jura J, Smorąg Z, Słomski R, Duda M. The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells. International Journal of Molecular Sciences. 2022; 23(18):10296. https://doi.org/10.3390/ijms231810296

Chicago/Turabian Style

Samiec, Marcin, Jerzy Wiater, Kamil Wartalski, Maria Skrzyszowska, Monika Trzcińska, Daniel Lipiński, Jacek Jura, Zdzisław Smorąg, Ryszard Słomski, and Małgorzata Duda. 2022. "The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells" International Journal of Molecular Sciences 23, no. 18: 10296. https://doi.org/10.3390/ijms231810296

APA Style

Samiec, M., Wiater, J., Wartalski, K., Skrzyszowska, M., Trzcińska, M., Lipiński, D., Jura, J., Smorąg, Z., Słomski, R., & Duda, M. (2022). The Relative Abundances of Human Leukocyte Antigen-E, α-Galactosidase A and α-Gal Antigenic Determinants Are Biased by Trichostatin A-Dependent Epigenetic Transformation of Triple-Transgenic Pig-Derived Dermal Fibroblast Cells. International Journal of Molecular Sciences, 23(18), 10296. https://doi.org/10.3390/ijms231810296

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