Hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cells
Abstract Inflammasomes are multiprotein platforms responsible for the release of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. Mouse studies have identified inflammasome activation within dendritic cells (DC) as pivotal for driving tubulointerstitial fibrosis and inflammation, the hallma...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Nature Publishing Group
2022-08-01
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| Series: | Cell Death and Disease |
| Online Access: | https://doi.org/10.1038/s41419-022-05191-z |
| _version_ | 1811183267838164992 |
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| author | Kurt T. K. Giuliani Anca Grivei Purba Nag Xiangju Wang Melissa Rist Katrina Kildey Becker Law Monica S. Ng Ray Wilkinson Jacobus Ungerer Josephine M. Forbes Helen Healy Andrew J. Kassianos |
| author_facet | Kurt T. K. Giuliani Anca Grivei Purba Nag Xiangju Wang Melissa Rist Katrina Kildey Becker Law Monica S. Ng Ray Wilkinson Jacobus Ungerer Josephine M. Forbes Helen Healy Andrew J. Kassianos |
| author_sort | Kurt T. K. Giuliani |
| collection | DOAJ |
| description | Abstract Inflammasomes are multiprotein platforms responsible for the release of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. Mouse studies have identified inflammasome activation within dendritic cells (DC) as pivotal for driving tubulointerstitial fibrosis and inflammation, the hallmarks of chronic kidney disease (CKD). However, translation of this work to human CKD remains limited. Here, we examined the complex tubular cell death pathways mediating inflammasome activation in human kidney DC and, thus, CKD progression. Ex vivo patient-derived proximal tubular epithelial cells (PTEC) cultured under hypoxic (1% O2) conditions modelling the CKD microenvironment showed characteristics of ferroptotic cell death, including mitochondrial dysfunction, reductions in the lipid repair enzyme glutathione peroxidase 4 (GPX4) and increases in lipid peroxidation by-product 4-hydroxynonenal (4-HNE) compared with normoxic PTEC. The addition of ferroptosis inhibitor, ferrostatin-1, significantly reduced hypoxic PTEC death. Human CD1c+ DC activated in the presence of hypoxic PTEC displayed significantly increased production of inflammasome-dependent cytokines IL-1β and IL-18. Treatment of co-cultures with VX-765 (caspase-1/4 inhibitor) and MCC950 (NLRP3 inflammasome inhibitor) significantly attenuated IL-1β/IL-18 levels, supporting an NLRP3 inflammasome-dependent DC response. In line with these in vitro findings, in situ immunolabelling of human fibrotic kidney tissue revealed a significant accumulation of tubulointerstitial CD1c+ DC containing active inflammasome (ASC) specks adjacent to ferroptotic PTEC. These data establish ferroptosis as the primary pattern of PTEC necrosis under the hypoxic conditions of CKD. Moreover, this study identifies NLRP3 inflammasome signalling driven by complex tubulointerstitial PTEC-DC interactions as a key checkpoint for therapeutic targeting in human CKD. |
| first_indexed | 2024-04-11T09:45:12Z |
| format | Article |
| id | doaj.art-3f6627c645814f06a5a8676f3b366cb9 |
| institution | Directory Open Access Journal |
| issn | 2041-4889 |
| language | English |
| last_indexed | 2024-04-11T09:45:12Z |
| publishDate | 2022-08-01 |
| publisher | Nature Publishing Group |
| record_format | Article |
| series | Cell Death and Disease |
| spelling | doaj.art-3f6627c645814f06a5a8676f3b366cb92022-12-22T04:31:05ZengNature Publishing GroupCell Death and Disease2041-48892022-08-0113811310.1038/s41419-022-05191-zHypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cellsKurt T. K. Giuliani0Anca Grivei1Purba Nag2Xiangju Wang3Melissa Rist4Katrina Kildey5Becker Law6Monica S. Ng7Ray Wilkinson8Jacobus Ungerer9Josephine M. Forbes10Helen Healy11Andrew J. Kassianos12Conjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandFaculty of Medicine, University of QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandConjoint Internal Medicine Laboratory, Chemical Pathology, Pathology QueenslandAbstract Inflammasomes are multiprotein platforms responsible for the release of pro-inflammatory cytokines interleukin (IL)-1β and IL-18. Mouse studies have identified inflammasome activation within dendritic cells (DC) as pivotal for driving tubulointerstitial fibrosis and inflammation, the hallmarks of chronic kidney disease (CKD). However, translation of this work to human CKD remains limited. Here, we examined the complex tubular cell death pathways mediating inflammasome activation in human kidney DC and, thus, CKD progression. Ex vivo patient-derived proximal tubular epithelial cells (PTEC) cultured under hypoxic (1% O2) conditions modelling the CKD microenvironment showed characteristics of ferroptotic cell death, including mitochondrial dysfunction, reductions in the lipid repair enzyme glutathione peroxidase 4 (GPX4) and increases in lipid peroxidation by-product 4-hydroxynonenal (4-HNE) compared with normoxic PTEC. The addition of ferroptosis inhibitor, ferrostatin-1, significantly reduced hypoxic PTEC death. Human CD1c+ DC activated in the presence of hypoxic PTEC displayed significantly increased production of inflammasome-dependent cytokines IL-1β and IL-18. Treatment of co-cultures with VX-765 (caspase-1/4 inhibitor) and MCC950 (NLRP3 inflammasome inhibitor) significantly attenuated IL-1β/IL-18 levels, supporting an NLRP3 inflammasome-dependent DC response. In line with these in vitro findings, in situ immunolabelling of human fibrotic kidney tissue revealed a significant accumulation of tubulointerstitial CD1c+ DC containing active inflammasome (ASC) specks adjacent to ferroptotic PTEC. These data establish ferroptosis as the primary pattern of PTEC necrosis under the hypoxic conditions of CKD. Moreover, this study identifies NLRP3 inflammasome signalling driven by complex tubulointerstitial PTEC-DC interactions as a key checkpoint for therapeutic targeting in human CKD.https://doi.org/10.1038/s41419-022-05191-z |
| spellingShingle | Kurt T. K. Giuliani Anca Grivei Purba Nag Xiangju Wang Melissa Rist Katrina Kildey Becker Law Monica S. Ng Ray Wilkinson Jacobus Ungerer Josephine M. Forbes Helen Healy Andrew J. Kassianos Hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cells Cell Death and Disease |
| title | Hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cells |
| title_full | Hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cells |
| title_fullStr | Hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cells |
| title_full_unstemmed | Hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cells |
| title_short | Hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an NLRP3 inflammasome response in CD1c+ dendritic cells |
| title_sort | hypoxic human proximal tubular epithelial cells undergo ferroptosis and elicit an nlrp3 inflammasome response in cd1c dendritic cells |
| url | https://doi.org/10.1038/s41419-022-05191-z |
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