Genetically modified C3A cells with restored urea cycle for improved bioartificial liver

• Autorzy: Pluta Krzysztof Dariusz , Samluk Anna , Wencel Agnieszka , Zakrzewska Karolina Ewa , Gora Monika , Burzynska Beata , Ciezkowska Malgorzata , Motyl Joanna , Pijanowska Dorota Genowefa

Identyfikatory

DOI

10.1016/j.bbe.2019.12.006

• Języki publikacji: EN

Abstrakty

EN

The bioartificial liver, a hybrid device aimed at improving the survival of patients with fulminant liver failure, requires a cell source to replicate human liver function. However, liver support systems that utilize porcine or human hepatoma-derived cells felt short of expectations in clinical trials. Here we present engineered C3A cells, with a restored function of the urea cycle, which can be used in an efficacious bioartificial liver. The genetic modification was performed using a lentiviral-mediated gene transfer which led to effective integra- tion of the transgenes, coding for arginase I and ornithine transcarbamylase, into the target cell genomes. The engineered cells are more resistant to the oxidative/nitrosative stress induced by the presence of high concentrations of ammonia cations and produce more urea than their unmodified counterparts. Interestingly, the genetically modified cells secrete more albumin than control C3A cells and the synthesis of the protein is induced by increasing concentrations of ammonia. Although the physiological capabilities of the new cell line need to be further examined, at this stage of our study we may conclude that the genetically modified cells are able to convert ammonia to urea more effectively than regular C3A cells.

Słowa kluczowe

EN

bioartificial liver; genetically modified cell; lentiviral vector; urea cycle

PL

wątroba biosztuczna; komórka modyfikowana genetycznie; cykl mocznikowy

• Wydawca: Nałęcz Institute of Biocybernetics and Biomedical Engineering of the Polish Academy of Sciences ; Elsevier
• Czasopismo: Biocybernetics and Biomedical Engineering
• Rocznik: 2020
• Tom: Vol. 40, no. 1
• Strony: 378--387
• Opis fizyczny: Bibliogr. 35 poz., tab., wykr.

Twórcy

autor

Pluta Krzysztof Dariusz

• Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland

autor

Samluk Anna

• Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland

autor

Wencel Agnieszka

• Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland

autor

Zakrzewska Karolina Ewa

• Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland

autor

Gora Monika

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

autor

Burzynska Beata

• Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland

autor

Ciezkowska Malgorzata

• Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland

autor

Motyl Joanna

• Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland

autor

Pijanowska Dorota Genowefa

• Nalecz Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland

Bibliografia

• [1] Marcellin P, Kutala BK. Liver diseases: a major, neglected global public health problem requiring urgent actions and large-scale screening. Liver Int 2018;38:2–6.
• [2] Pimpin L, Cortez-Pinto H, Negro F, Corbould E, Lazarus JV, Webber L, et al. Burden of liver disease in Europe: epidemiology and analysis of risk factors to identify prevention policies. J Hepatol 2018;69:718–35.
• [3] Bernal W, Auzinger G, Dhawan A, Wendon J. Acute liver failure. Lancet 2010;376:190–201.
• [4] Zakrzewska KE, Samluk A, Wencel A, Dudek K, Pijanowska DG, Pluta KD. Liver tissue fragments obtained from males are the most promising source of human hepatocytes for cell-based therapies — flow cytometric analysis of albumin expression. PLoS One 2017;12e0182846.
• [5] Nibourg GA, Chamuleau RA, van Gulik TM, Hoekstra R. Proliferative human cell sources applied as biocomponent in bioartificial livers: a review. Expert Opin Biol Ther 2012;12:905–21.
• [6] Palakkan AA, Hay DC, Anil Kumar PR, Kumary TV, Ross JA. Liver tissue engineering and cell sources: issues and challenges. Liver Int 2013;33:666–76.
• [7] Rozga J, Williams F, Ro M-S, Neuzil DF, Giorgio TD, Backfisch G, et al. Development of a bioartificial liver: properties and function of a hollow-fiber module inoculated with liver cells. Hepatology 1993;17:258–65.
• [8] van Wenum M, Adam AAA, Hakvoort TBM, Hendriks EJ, Shevchenko V, van Gulik TM, et al. Selecting cells for bioartificial liver devices and the importance of a 3D culture environment: a functional comparison between the HepaRG and C3A cell lines. Int J Biol Sci 2016;12:964–78.
• [9] Nyberg SL, Remmel RP, Mann HJ, Peshwa MV, Hu WS, Cerra FB. Primary hepatocytes outperform Hep G2 cells as the source of biotransformation functions in a bioartificial liver. Ann Surg 1994;220:59–67.
• [10] Sussman NL, Chong MG, Koussayer T, He DE, Shang TA, Whisennand HH, et al. Reversal of fulminant hepatic failure using an extracorporeal liver assist device. Hepatology 1992;16:60–5.
• [11] Kelly JH, Koussayer T, He D, Chong MG, Shang TA, Whisennand HH, et al. Assessment of an extracorporeal liver assist device in anhepatic dogs. Artif Organs 1992;16:418–22.
• [12] Ellis AJ, Hughes RD, Wendon JA, Dunne J, Langley PG, Kelly JH, et al. Pilot-controlled trial of the extracorporeal liver assist device in acute liver failure. Hepatology 1996;24:1446–51.
• [13] Hughes RD, Nicolaou N, Langley PG, Ellis AJ, Wendon JA, Williams R. Plasma cytokine levels and coagulation and complement activation during use of the extracorporeal liver assist device in acute liver failure. Artif Organs 1998;22:854–8.
• [14] Khuu DN, Scheers I, Ehnert S, Jazouli N, Nyabi O, Buc- Calderon P, et al. In vitro differentiated adult human liver progenitor cells display mature hepatic metabolic functions: a potential tool for in vitro pharmacotoxicological testing. Cell Transplant 2011;20:287–302.
• [15] Stock P, Brückner S, Ebensing S, Hempel M, Dollinger MM, Christ B. The generation of hepatocytes from mesenchymal stem cells and engraftment into murine liver. Nat Protoc 2010;5:617–27.
• [16] Ren S, Irudayam JI, Contreras D, Sareen D, Talavera-Adame D, Svendsen CN, et al. Bioartificial liver device based on induced pluripotent stem cell-derived hepatocytes. J Stem Cell Res Ther 2015;5:263.
• [17] Mavri-Damelin D, Eaton S, Damelin LH, Rees M, Hodgson HJF, Selden C. Ornithine transcarbamylase and arginase I deficiency are responsible for diminished urea cycle function in the human hepatoblastoma cell line HepG2. Int J Biochem Cell Biol 2007;39:555–64.
• [18] Mavri-Damelin D, Damelin LH, Eaton S, Rees M, Selden C, Hodgson HJF. Cells for bioartificial liver devices: the human hepatoma-derived cell line C3A produces urea but does not detoxify ammonia. Biotechnol Bioeng 2008;99:644–51.
• [19] Pfaffl MW. Relative expression software tool (REST(C)) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 2002;30. 36e–36.
• [20] Bustin SA, Benes V, Garson JA, Hellemans J, Huggett J, Kubista M, et al. The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. Clin Chem 2009;55:611–22.
• [21] Millis JM, Cronin DC, Johnson R, Conjeevaram H, Conlin C, Trevino S, et al. Initial experience with the modified extracorporeal liver-assist device for patients with fulminant hepatic failure: system modifications and clinical impact. Transplantation 2002;74:1735–46.
• [22] Thompson J, Jones N, Al-Khafaji A, Malik S, Reich D, Munoz S, et al. Extracorporeal cellular therapy (ELAD) in severe alcoholic hepatitis: a multinational, prospective, controlled, randomized trial. Liver Transplant 2018;24:380–93.
• [23] Butterworth RF. Pathophysiology of hepatic encephalopathy: a new look at ammonia. Metab Brain Dis 2002;17:221–7.
• [24] Moedas MF, Adam AAA, Farelo MA, IJlst L, Chamuleau RAFM, Hoekstra R, et al. Advances in methods for characterization of hepatic urea cycle enzymatic activity in HepaRG cells using UPLC-MS/MS. Anal Biochem 2017;535:47–55.
• [25] Keshet R, Szlosarek P, Carracedo A, Erez A. Rewiring urea cycle metabolism in cancer to support anabolism. Nat Rev Cancer 2018;18:634–45.
• [26] Coward SM, Legallais C, David B, Thomas M, Foo Y, Mavri- Damelin D, et al. Alginate-encapsulated HepG2 cells in a fluidized bed bioreactor maintain function in human liver failure plasma. Artif Organs 2009;33:1117–26.
• [27] Tang N, Wang Y, Wang X, Zhou L, Zhang F, Li X, et al. Stable overexpression of arginase I and ornithine transcarbamylase in HepG2 cells improves its ammonia detoxification. J Cell Biochem 2012;113:518–27.
• [28] Zhang F-Y, Tang N-H, Wang X-Q, Li X-J, Chen Y-L. Simultaneous recovery of dual pathways for ammonia metabolism do not improve further detoxification of ammonia in HepG2 cells. Hepatobiliary Pancreat Dis Int 2013;12:525–32.
• [29] Wang X-Q, Tang N-H, Zhang F-Y, Li X-J, Chen Y-L. Therapeutic evaluation of a microbioartificial liver with recombinant HepG2 cells for rats with hepatic failure. Expert Opin Biol Ther 2013;13:1507–13.
• [30] Pluta K, Kacprzak MM. Use of HIV as a gene transfer vector. Acta Biochim Pol 2009;56:531–95.
• [31] Samluk A, Zakrzewska KE, Pluta KD. Generation of fluorescently labeled cell lines, C3A hepatoma cells, and human adult skin fibroblasts to study coculture models. Artif Organs 2013;37:E123–30.
• [32] Speir E. Cytomegalovirus gene regulation by reactive oxygen species. Agents in atherosclerosis. Ann N Y Acad Sci 2000;899:363–74.
• [33] Guzman-Lepe J, Cervantes-Alvarez E, Collin de l'Hortet A, Wang Y, Mars WM, Oda Y, et al. Liver-enriched transcription factor expression relates to chronic hepatic failure in humans. Hepatol Commun 2018;2:582–94.
• [34] The Journal of Gene Medicine. www.abedia.com/wiley/vectors.php. [Accessed 25 July 2019].
• [35] U.S. Food and Drug Administration. https://www.fda.gov/vaccines-blood-biologics/cellular-gene- therapy-products/kymriah-tisagenlecleucel. [Accessed 25 July 2019].

Uwagi

PL

Opracowanie rekordu ze środków MNiSW, umowa Nr 461252 w ramach programu "Społeczna odpowiedzialność nauki" - moduł: Popularyzacja nauki i promocja sportu (2020).