Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma
Hepatocellular carcinoma (HCC) is an aggressive disease lacking effective treatment. Animal models of HCC are necessary for preclinical evaluation of the safety and efficacy of novel therapeutics. Large animal models of HCC allow testing image-guided locoregional therapies, which are widely used in...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-05-01
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| Series: | Frontiers in Oncology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fonc.2022.904031/full |
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| author | Lobna Elkhadragy Kimia Dasteh Goli William M. Totura Maximillian J. Carlino Maureen R. Regan Grace Guzman Lawrence B. Schook Lawrence B. Schook Lawrence B. Schook Ron C. Gaba Ron C. Gaba Kyle M. Schachtschneider Kyle M. Schachtschneider Kyle M. Schachtschneider |
| author_facet | Lobna Elkhadragy Kimia Dasteh Goli William M. Totura Maximillian J. Carlino Maureen R. Regan Grace Guzman Lawrence B. Schook Lawrence B. Schook Lawrence B. Schook Ron C. Gaba Ron C. Gaba Kyle M. Schachtschneider Kyle M. Schachtschneider Kyle M. Schachtschneider |
| author_sort | Lobna Elkhadragy |
| collection | DOAJ |
| description | Hepatocellular carcinoma (HCC) is an aggressive disease lacking effective treatment. Animal models of HCC are necessary for preclinical evaluation of the safety and efficacy of novel therapeutics. Large animal models of HCC allow testing image-guided locoregional therapies, which are widely used in the management of HCC. Models with precise tumor mutations mimicking human HCC provide valuable tools for testing precision medicine. AXIN1 and ARID1A are two of the most frequently mutated genes in human HCC. Here, we investigated the effects of knockout of AXIN1 and/or ARID1A on proliferation, migration, and chemotherapeutic susceptibility of porcine HCC cells and we developed subcutaneous tumors harboring these mutations in pigs. Gene knockout was achieved by CRISPR/Cas9 and was validated by Next Generation Sequencing. AXIN1 knockout increased the migration of porcine HCC cells but did not alter the cell proliferation. Knockout of ARID1A increased both the proliferation and migration of porcine HCC cells. Simultaneous knockout of AXIN1 and ARID1A increased the migration, but did not alter the proliferation of porcine HCC cells. The effect of gene knockout on the response of porcine HCC cells to two of the most commonly used systemic and locoregional HCC treatments was investigated; sorafenib and doxorubicin, respectively. Knockout of AXIN1 and/or ARID1A did not alter the susceptibility of porcine HCC cells to sorafenib or doxorubicin. Autologous injection of CRISPR edited HCC cells resulted in development of subcutaneous tumors in pigs, which harbored the anticipated edits in AXIN1 and/or ARID1A. This study elucidates the effects of CRISPR-mediated knockout of HCC-associated genes in porcine HCC cells, and lays the foundation for development and utilization of genetically-tailored porcine HCC models for in vivo testing of novel therapeutic approaches in a clinically-relevant large animal model. |
| first_indexed | 2024-12-12T16:09:11Z |
| format | Article |
| id | doaj.art-b843aa55c42d4b068273debefd588c9f |
| institution | Directory Open Access Journal |
| issn | 2234-943X |
| language | English |
| last_indexed | 2024-12-12T16:09:11Z |
| publishDate | 2022-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Oncology |
| spelling | doaj.art-b843aa55c42d4b068273debefd588c9f2022-12-22T00:19:14ZengFrontiers Media S.A.Frontiers in Oncology2234-943X2022-05-011210.3389/fonc.2022.904031904031Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular CarcinomaLobna Elkhadragy0Kimia Dasteh Goli1William M. Totura2Maximillian J. Carlino3Maureen R. Regan4Grace Guzman5Lawrence B. Schook6Lawrence B. Schook7Lawrence B. Schook8Ron C. Gaba9Ron C. Gaba10Kyle M. Schachtschneider11Kyle M. Schachtschneider12Kyle M. Schachtschneider13Department of Radiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Radiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Radiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Radiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Pathology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Radiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Animal Sciences, University of Illinois at Urbana-Champaign, Urbana, IL, United StatesNational Center for Supercomputing Applications, University of Illinois at Urbana-Champaign, Urbana, IL, United StatesDepartment of Radiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Pathology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Radiology, University of Illinois at Chicago, Chicago, IL, United StatesDepartment of Biochemistry and Molecular Genetics, University of Illinois at Chicago, Chicago, IL, United StatesNational Center for Supercomputing Applications, University of Illinois at Urbana-Champaign, Urbana, IL, United StatesHepatocellular carcinoma (HCC) is an aggressive disease lacking effective treatment. Animal models of HCC are necessary for preclinical evaluation of the safety and efficacy of novel therapeutics. Large animal models of HCC allow testing image-guided locoregional therapies, which are widely used in the management of HCC. Models with precise tumor mutations mimicking human HCC provide valuable tools for testing precision medicine. AXIN1 and ARID1A are two of the most frequently mutated genes in human HCC. Here, we investigated the effects of knockout of AXIN1 and/or ARID1A on proliferation, migration, and chemotherapeutic susceptibility of porcine HCC cells and we developed subcutaneous tumors harboring these mutations in pigs. Gene knockout was achieved by CRISPR/Cas9 and was validated by Next Generation Sequencing. AXIN1 knockout increased the migration of porcine HCC cells but did not alter the cell proliferation. Knockout of ARID1A increased both the proliferation and migration of porcine HCC cells. Simultaneous knockout of AXIN1 and ARID1A increased the migration, but did not alter the proliferation of porcine HCC cells. The effect of gene knockout on the response of porcine HCC cells to two of the most commonly used systemic and locoregional HCC treatments was investigated; sorafenib and doxorubicin, respectively. Knockout of AXIN1 and/or ARID1A did not alter the susceptibility of porcine HCC cells to sorafenib or doxorubicin. Autologous injection of CRISPR edited HCC cells resulted in development of subcutaneous tumors in pigs, which harbored the anticipated edits in AXIN1 and/or ARID1A. This study elucidates the effects of CRISPR-mediated knockout of HCC-associated genes in porcine HCC cells, and lays the foundation for development and utilization of genetically-tailored porcine HCC models for in vivo testing of novel therapeutic approaches in a clinically-relevant large animal model.https://www.frontiersin.org/articles/10.3389/fonc.2022.904031/fullliver cancerhepatocellular carcinomaporcine modelsARID1AAXIN1CRISPR |
| spellingShingle | Lobna Elkhadragy Kimia Dasteh Goli William M. Totura Maximillian J. Carlino Maureen R. Regan Grace Guzman Lawrence B. Schook Lawrence B. Schook Lawrence B. Schook Ron C. Gaba Ron C. Gaba Kyle M. Schachtschneider Kyle M. Schachtschneider Kyle M. Schachtschneider Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma Frontiers in Oncology liver cancer hepatocellular carcinoma porcine models ARID1A AXIN1 CRISPR |
| title | Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma |
| title_full | Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma |
| title_fullStr | Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma |
| title_full_unstemmed | Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma |
| title_short | Effect of CRISPR Knockout of AXIN1 or ARID1A on Proliferation and Migration of Porcine Hepatocellular Carcinoma |
| title_sort | effect of crispr knockout of axin1 or arid1a on proliferation and migration of porcine hepatocellular carcinoma |
| topic | liver cancer hepatocellular carcinoma porcine models ARID1A AXIN1 CRISPR |
| url | https://www.frontiersin.org/articles/10.3389/fonc.2022.904031/full |
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