Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted Form
Polycystic kidney disease (PKD) is one of the most common inherited diseases and is characterized by the development of fluid-filled cysts along multiple segments of the nephron. Autosomal dominant polycystic kidney disease (ADPKD) is the most common form of PKD, which is caused by mutations in eith...
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MDPI AG
2022-01-01
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| Online Access: | https://www.mdpi.com/2073-4409/11/3/483 |
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| author | Hsin-Yin Chuang Wen-Yih Jeng Ellian Wang Si-Tse Jiang Chen-Ming Hsu Hsiu Mei Hsieh-Li Yuan-Yow Chiou |
| author_facet | Hsin-Yin Chuang Wen-Yih Jeng Ellian Wang Si-Tse Jiang Chen-Ming Hsu Hsiu Mei Hsieh-Li Yuan-Yow Chiou |
| author_sort | Hsin-Yin Chuang |
| collection | DOAJ |
| description | Polycystic kidney disease (PKD) is one of the most common inherited diseases and is characterized by the development of fluid-filled cysts along multiple segments of the nephron. Autosomal dominant polycystic kidney disease (ADPKD) is the most common form of PKD, which is caused by mutations in either <i>PKD1</i> or <i>PKD2</i> genes that encode polycystin-1 (PC1) and polycystin-2 (PC2), respectively. As ADPKD progresses, cysts enlarge and disrupt normal kidney architecture, eventually leading to kidney failure. Our previous study showed that overexpression of exogenous kidney-specific neutrophil gelatinase-associated lipocalin (NGAL) reduced cyst progression and prolonged the lifespan of ADPKD mice (<i>Pkd1<sup>L3/L3</sup></i>, 2L3 for short). In this study, we attempted to explore the underlying mechanism of reduced cyst progression in the presence of NGAL using immortalized 2L3 cells. The results of MTT and BrdU incorporation assays showed that recombinant mouse NGAL (mNGAL) protein significantly decreased the viability and proliferation of 2L3 cells. Flow cytometry and western blot analyses showed that mNGAL inhibited activation of the ERK and AKT pathways and induced apoptosis and autophagy in 2L3 cells. In addition, a 3D cell culture platform was established to identify cyst progression in 2L3 cells and showed that mNGAL significantly inhibited cyst enlargement in 2L3 cells. Overexpression of secreted mNGAL (pN + LS) and nonsecreted mNGAL (pN − LS) repressed cell proliferation and cyst enlargement in 2L3 cells and had effects on markers involved in proliferation, apoptosis, and autophagy. However, secreted mNGAL had a more pronounced and consistent effect than that of nonsecreted form. These results reveal that secreted mNGAL has stronger ability to inhibit cyst enlargement of ADPKD cells than that of nonsecreted form. These findings could help to identify strategies for the future clinical treatment of ADPKD. |
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| issn | 2073-4409 |
| language | English |
| last_indexed | 2024-03-10T00:03:24Z |
| publishDate | 2022-01-01 |
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| spelling | doaj.art-8c9c2aa9a6544bf6ae2edc7217f2c6e92023-11-23T16:12:43ZengMDPI AGCells2073-44092022-01-0111348310.3390/cells11030483Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted FormHsin-Yin Chuang0Wen-Yih Jeng1Ellian Wang2Si-Tse Jiang3Chen-Ming Hsu4Hsiu Mei Hsieh-Li5Yuan-Yow Chiou6Department of Life Science, National Taiwan Normal University, Taipei 11677, TaiwanUniversity Center for Bioscience and Biotechnology, National Cheng Kung University, Tainan 70101, TaiwanDivision of Pediatric Nephrology, Department of Pediatrics, National Cheng Kung University Hospital, Tainan 70403, TaiwanInstitute of Clinical Medicine, Medical College, National Cheng Kung University, Tainan 70101, TaiwanDepartment of Life Science, National Taiwan Normal University, Taipei 11677, TaiwanDepartment of Life Science, National Taiwan Normal University, Taipei 11677, TaiwanDivision of Pediatric Nephrology, Department of Pediatrics, National Cheng Kung University Hospital, Tainan 70403, TaiwanPolycystic kidney disease (PKD) is one of the most common inherited diseases and is characterized by the development of fluid-filled cysts along multiple segments of the nephron. Autosomal dominant polycystic kidney disease (ADPKD) is the most common form of PKD, which is caused by mutations in either <i>PKD1</i> or <i>PKD2</i> genes that encode polycystin-1 (PC1) and polycystin-2 (PC2), respectively. As ADPKD progresses, cysts enlarge and disrupt normal kidney architecture, eventually leading to kidney failure. Our previous study showed that overexpression of exogenous kidney-specific neutrophil gelatinase-associated lipocalin (NGAL) reduced cyst progression and prolonged the lifespan of ADPKD mice (<i>Pkd1<sup>L3/L3</sup></i>, 2L3 for short). In this study, we attempted to explore the underlying mechanism of reduced cyst progression in the presence of NGAL using immortalized 2L3 cells. The results of MTT and BrdU incorporation assays showed that recombinant mouse NGAL (mNGAL) protein significantly decreased the viability and proliferation of 2L3 cells. Flow cytometry and western blot analyses showed that mNGAL inhibited activation of the ERK and AKT pathways and induced apoptosis and autophagy in 2L3 cells. In addition, a 3D cell culture platform was established to identify cyst progression in 2L3 cells and showed that mNGAL significantly inhibited cyst enlargement in 2L3 cells. Overexpression of secreted mNGAL (pN + LS) and nonsecreted mNGAL (pN − LS) repressed cell proliferation and cyst enlargement in 2L3 cells and had effects on markers involved in proliferation, apoptosis, and autophagy. However, secreted mNGAL had a more pronounced and consistent effect than that of nonsecreted form. These results reveal that secreted mNGAL has stronger ability to inhibit cyst enlargement of ADPKD cells than that of nonsecreted form. These findings could help to identify strategies for the future clinical treatment of ADPKD.https://www.mdpi.com/2073-4409/11/3/483ADPKDneutrophil gelatinase-associated lipocalin3D culture |
| spellingShingle | Hsin-Yin Chuang Wen-Yih Jeng Ellian Wang Si-Tse Jiang Chen-Ming Hsu Hsiu Mei Hsieh-Li Yuan-Yow Chiou Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted Form Cells ADPKD neutrophil gelatinase-associated lipocalin 3D culture |
| title | Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted Form |
| title_full | Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted Form |
| title_fullStr | Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted Form |
| title_full_unstemmed | Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted Form |
| title_short | Secreted Neutrophil Gelatinase-Associated Lipocalin Shows Stronger Ability to Inhibit Cyst Enlargement of ADPKD Cells Compared with Nonsecreted Form |
| title_sort | secreted neutrophil gelatinase associated lipocalin shows stronger ability to inhibit cyst enlargement of adpkd cells compared with nonsecreted form |
| topic | ADPKD neutrophil gelatinase-associated lipocalin 3D culture |
| url | https://www.mdpi.com/2073-4409/11/3/483 |
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