Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System
Karyomegalic interstitial nephritis (KIN) is a genetic kidney disease caused by mutations in the FANCD2/FANCI-Associated Nuclease 1 (<i>FAN1</i>) gene on 15q13.3, which results in karyomegaly and fibrosis of kidney cells through the incomplete repair of DNA damage. The aim of this study...
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MDPI AG
2023-09-01
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| Series: | Cells |
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| Online Access: | https://www.mdpi.com/2073-4409/12/18/2319 |
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| author | Sun Woo Lim Dohyun Na Hanbi Lee Xianying Fang Sheng Cui Yoo Jin Shin Kang In Lee Jae Young Lee Chul Woo Yang Byung Ha Chung |
| author_facet | Sun Woo Lim Dohyun Na Hanbi Lee Xianying Fang Sheng Cui Yoo Jin Shin Kang In Lee Jae Young Lee Chul Woo Yang Byung Ha Chung |
| author_sort | Sun Woo Lim |
| collection | DOAJ |
| description | Karyomegalic interstitial nephritis (KIN) is a genetic kidney disease caused by mutations in the FANCD2/FANCI-Associated Nuclease 1 (<i>FAN1</i>) gene on 15q13.3, which results in karyomegaly and fibrosis of kidney cells through the incomplete repair of DNA damage. The aim of this study was to explore the possibility of using a human induced pluripotent stem cell (hiPSC)-derived kidney organoid system for modeling <i>FAN1</i>-deficient kidney disease, also known as KIN. We generated kidney organoids using WTC-11 (wild-type) hiPSCs and <i>FAN1</i>-mutant hiPSCs which include KIN patient-derived hiPSCs and <i>FAN1</i>-edited hiPSCs (WTC-11 <sup>FAN1+/−</sup>), created using the CRISPR/Cas9 system in WTC-11-hiPSCs. Kidney organoids from each group were treated with 20 nM of mitomycin C (MMC) for 24 or 48 h, and the expression levels of Ki67 and H2A histone family member X (H2A.X) were analyzed to detect DNA damage and assess the viability of cells within the kidney organoids. Both WTC-11-hiPSCs and <i>FAN1</i>-mutant hiPSCs were successfully differentiated into kidney organoids without structural deformities. MMC treatment for 48 h significantly increased the expression of DNA damage markers, while cell viability in both <i>FAN1</i>-mutant kidney organoids was decreased. However, these findings were observed in WTC-11-kidney organoids. These results suggest that <i>FAN1</i>-mutant kidney organoids can recapitulate the phenotype of <i>FAN1</i>-deficient kidney disease. |
| first_indexed | 2024-03-10T22:56:22Z |
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| institution | Directory Open Access Journal |
| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T22:56:22Z |
| publishDate | 2023-09-01 |
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| record_format | Article |
| series | Cells |
| spelling | doaj.art-7f7b458a8c3343129a63470f190a78982023-11-19T10:00:21ZengMDPI AGCells2073-44092023-09-011218231910.3390/cells12182319Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid SystemSun Woo Lim0Dohyun Na1Hanbi Lee2Xianying Fang3Sheng Cui4Yoo Jin Shin5Kang In Lee6Jae Young Lee7Chul Woo Yang8Byung Ha Chung9Transplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaToolGen, Inc., Seoul 06591, Republic of KoreaToolGen, Inc., Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaTransplantation Research Center, College of Medicine, The Catholic University of Korea, Seoul 06591, Republic of KoreaKaryomegalic interstitial nephritis (KIN) is a genetic kidney disease caused by mutations in the FANCD2/FANCI-Associated Nuclease 1 (<i>FAN1</i>) gene on 15q13.3, which results in karyomegaly and fibrosis of kidney cells through the incomplete repair of DNA damage. The aim of this study was to explore the possibility of using a human induced pluripotent stem cell (hiPSC)-derived kidney organoid system for modeling <i>FAN1</i>-deficient kidney disease, also known as KIN. We generated kidney organoids using WTC-11 (wild-type) hiPSCs and <i>FAN1</i>-mutant hiPSCs which include KIN patient-derived hiPSCs and <i>FAN1</i>-edited hiPSCs (WTC-11 <sup>FAN1+/−</sup>), created using the CRISPR/Cas9 system in WTC-11-hiPSCs. Kidney organoids from each group were treated with 20 nM of mitomycin C (MMC) for 24 or 48 h, and the expression levels of Ki67 and H2A histone family member X (H2A.X) were analyzed to detect DNA damage and assess the viability of cells within the kidney organoids. Both WTC-11-hiPSCs and <i>FAN1</i>-mutant hiPSCs were successfully differentiated into kidney organoids without structural deformities. MMC treatment for 48 h significantly increased the expression of DNA damage markers, while cell viability in both <i>FAN1</i>-mutant kidney organoids was decreased. However, these findings were observed in WTC-11-kidney organoids. These results suggest that <i>FAN1</i>-mutant kidney organoids can recapitulate the phenotype of <i>FAN1</i>-deficient kidney disease.https://www.mdpi.com/2073-4409/12/18/2319<i>FAN1</i> geneinduced pluripotent stem cellskidney organoidkaryomegalic interstitial nephritis |
| spellingShingle | Sun Woo Lim Dohyun Na Hanbi Lee Xianying Fang Sheng Cui Yoo Jin Shin Kang In Lee Jae Young Lee Chul Woo Yang Byung Ha Chung Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System Cells <i>FAN1</i> gene induced pluripotent stem cells kidney organoid karyomegalic interstitial nephritis |
| title | Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System |
| title_full | Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System |
| title_fullStr | Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System |
| title_full_unstemmed | Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System |
| title_short | Modeling of <i>FAN1</i>-Deficient Kidney Disease Using a Human Induced Pluripotent Stem Cell-Derived Kidney Organoid System |
| title_sort | modeling of i fan1 i deficient kidney disease using a human induced pluripotent stem cell derived kidney organoid system |
| topic | <i>FAN1</i> gene induced pluripotent stem cells kidney organoid karyomegalic interstitial nephritis |
| url | https://www.mdpi.com/2073-4409/12/18/2319 |
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