Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells
Atypical teratoid/rhabdoid tumor (ATRT) is a rare childhood malignancy that originates in the central nervous system. Over ninety-five percent of ATRT patients have biallelic inactivation of the tumor suppressor gene SMARCB1. ATRT has no standard treatment, and a major limiting factor in therapeutic...
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KeAi Communications Co., Ltd.
2024-01-01
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| Series: | Bioactive Materials |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2452199X23002487 |
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| author | Timothy Hua Yu Xue Drishty B. Sarker Sonia Kiran Yan Li Qing-Xiang Amy Sang |
| author_facet | Timothy Hua Yu Xue Drishty B. Sarker Sonia Kiran Yan Li Qing-Xiang Amy Sang |
| author_sort | Timothy Hua |
| collection | DOAJ |
| description | Atypical teratoid/rhabdoid tumor (ATRT) is a rare childhood malignancy that originates in the central nervous system. Over ninety-five percent of ATRT patients have biallelic inactivation of the tumor suppressor gene SMARCB1. ATRT has no standard treatment, and a major limiting factor in therapeutic development is the lack of reliable ATRT models. We employed CRISPR/Cas9 gene-editing technology to knock out SMARCB1 and TP53 genes in human episomal induced pluripotent stem cells (Epi-iPSCs), followed by brief neural induction, to generate an ATRT-like model. The dual knockout Epi-iPSCs retained their stemness with the capacity to differentiate into three germ layers. High expression of OCT4 and NANOG in neurally induced knockout spheroids was comparable to that in two ATRT cell lines. Beta-catenin protein expression was higher in SMARCB1-deficient cells and spheroids than in normal Epi-iPSC-derived spheroids. Nucleophosmin, Osteopontin, and Ki-67 proteins were also expressed by the SMARCB1-deficient spheroids. In summary, the tumor model resembled embryonal features of ATRT and expressed ATRT biomarkers at mRNA and protein levels. Ribociclib, PTC-209, and the combination of clofilium tosylate and pazopanib decreased the viability of the ATRT-like cells. This disease modeling scheme may enable the establishment of individualized tumor models with patient-specific mutations and facilitate high-throughput drug testing. |
| first_indexed | 2024-03-12T15:03:36Z |
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| institution | Directory Open Access Journal |
| issn | 2452-199X |
| language | English |
| last_indexed | 2024-03-12T15:03:36Z |
| publishDate | 2024-01-01 |
| publisher | KeAi Communications Co., Ltd. |
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| series | Bioactive Materials |
| spelling | doaj.art-981b6c5fed0d4886baa275cc660f7cf42023-08-13T04:54:32ZengKeAi Communications Co., Ltd.Bioactive Materials2452-199X2024-01-0131136150Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cellsTimothy Hua0Yu Xue1Drishty B. Sarker2Sonia Kiran3Yan Li4Qing-Xiang Amy Sang5Department of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, 32306-4390, USADepartment of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, 32306-4390, USADepartment of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, 32306-4390, USADepartment of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, 32306-4390, USADepartment of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, 32310-6046, USA; Institute of Molecular Biophysics, Florida State University, Tallahassee, FL, 32306-4380, USADepartment of Chemistry and Biochemistry, Florida State University, Tallahassee, FL, 32306-4390, USA; Institute of Molecular Biophysics, Florida State University, Tallahassee, FL, 32306-4380, USA; Corresponding author. Institute of Molecular Biophysics, Florida State University, Tallahassee, FL, 32306-4380, USA.Atypical teratoid/rhabdoid tumor (ATRT) is a rare childhood malignancy that originates in the central nervous system. Over ninety-five percent of ATRT patients have biallelic inactivation of the tumor suppressor gene SMARCB1. ATRT has no standard treatment, and a major limiting factor in therapeutic development is the lack of reliable ATRT models. We employed CRISPR/Cas9 gene-editing technology to knock out SMARCB1 and TP53 genes in human episomal induced pluripotent stem cells (Epi-iPSCs), followed by brief neural induction, to generate an ATRT-like model. The dual knockout Epi-iPSCs retained their stemness with the capacity to differentiate into three germ layers. High expression of OCT4 and NANOG in neurally induced knockout spheroids was comparable to that in two ATRT cell lines. Beta-catenin protein expression was higher in SMARCB1-deficient cells and spheroids than in normal Epi-iPSC-derived spheroids. Nucleophosmin, Osteopontin, and Ki-67 proteins were also expressed by the SMARCB1-deficient spheroids. In summary, the tumor model resembled embryonal features of ATRT and expressed ATRT biomarkers at mRNA and protein levels. Ribociclib, PTC-209, and the combination of clofilium tosylate and pazopanib decreased the viability of the ATRT-like cells. This disease modeling scheme may enable the establishment of individualized tumor models with patient-specific mutations and facilitate high-throughput drug testing.http://www.sciencedirect.com/science/article/pii/S2452199X23002487Atypical teratoid/rhabdoid tumorHuman induced pluripotent stem cellsCRISPR/Cas9 gene editingTumor suppressor genesSMARCB1Brain tumor modeling |
| spellingShingle | Timothy Hua Yu Xue Drishty B. Sarker Sonia Kiran Yan Li Qing-Xiang Amy Sang Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells Bioactive Materials Atypical teratoid/rhabdoid tumor Human induced pluripotent stem cells CRISPR/Cas9 gene editing Tumor suppressor genes SMARCB1 Brain tumor modeling |
| title | Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells |
| title_full | Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells |
| title_fullStr | Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells |
| title_full_unstemmed | Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells |
| title_short | Modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells |
| title_sort | modeling human brain rhabdoid tumor by inactivating tumor suppressor genes in induced pluripotent stem cells |
| topic | Atypical teratoid/rhabdoid tumor Human induced pluripotent stem cells CRISPR/Cas9 gene editing Tumor suppressor genes SMARCB1 Brain tumor modeling |
| url | http://www.sciencedirect.com/science/article/pii/S2452199X23002487 |
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