Abstract: FR-PO979
Chemokine Ligand 14 Is a Pro-Fibrotic Stimulus in CKD Progression Through Regulating the GATA2 Transcription Factor
Session Information
- CKD: Pathobiology - I
November 04, 2022 | Location: Exhibit Hall, Orange County Convention Center‚ West Building
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2203 CKD (Non-Dialysis): Mechanisms
Authors
- Lee, Sunhwa, Kangwon National University Hospital, Chuncheon, Kangwon, Korea (the Republic of)
- Kim, Yong Chul, Seoul National University Hospital, Jongno-gu, Seoul, Korea (the Republic of)
- Han, Seung Seok, Seoul National University Hospital, Jongno-gu, Korea (the Republic of)
- Kim, Dong Ki, Seoul National University Hospital, Jongno-gu, Korea (the Republic of)
- Kim, Yon Su, Seoul National University Hospital, Jongno-gu, Korea (the Republic of)
- Yang, Seung Hee, Seoul National University Hospital, Jongno-gu, Korea (the Republic of)
Background
Chemokine ligand 14 (CCL14), a ligand for CCR1, has been known as a M2 polarizing marker and chemotactic cytokine. By performing urine proteomic analysis of chronic kidney disease patients, we previously found out CCL14 expression tends to increase in patients with progressive CKD and GATA2 could be possible upstream transcription factor. To validate pathophysiologic role of CCL14 and its molecular pathway, we performed in vitro and in vivo evaluations.
Methods
To determine how CCL14 is involved in the fibrosis process, recombinant CCL14(9-74) were treated in primary cultured tubular epithelial cells (hTECs). Moreover, anti-CCL14 blocking antibody were treated in fibrosis induced hTECs using rTGFβ (2ng/ml) or Hypoxia (1% O2, 72hrs). Fibronectin, collagen IV, E-Cadherin, CCL14, CCR1 and GATA2 level was evaluated. And cell apoptosis and necrosis was also measured using an Annexin-PI assay.
For in vivo validation, we used C57BL/6 unilateral ureteral obstruction (UUO) model to re-inforce the CCL14/CCR1/GATA2 pathway in chronic kidney disease progression. We also further performed GATA2 immunohistochemistry staining in kidney specimens of CKD patients.
Results
Recombinant CCL14 itself increased the expression of fibronectin in hTECs. Adding anti-CCL14 with rTGFβ showed decreased expression level of collagen IV and increased cell junction marker, E-cadherin. Annexin-PI assay result showed decreased apoptotic cell count with anti-CCL14 stimulation.
In hypoxia-induced hTECs, anti-CCL14 antibody significantly suppressed the mRNA expression levels of fibronectin, αSMA, and periostin. In addition, decrease in the expression of CCR1 receptor and GATA2 transcription factor were also observed. In C57BL/6 UUO mouse model, on the 7th, and 14th day after operation, GATA2 mRNA expression were significantly increased. Similarly, in human kidney specimens, the expression level of GATA2 transcription factor increases as the chronic kidney disease stage progresses.
Conclusion
CCL14 could play a pro-fibrotic role in CKD progression via regulations of CCR1 receptor and GATA2 transcription factor. Since CCL14 acts not only as a biomarker of increased expression in CKD progression, but also as a direct pro-fibrotic stimulus, it can be considered as a target for the treatment of chronic kidney disease.
Funding
- Government Support – Non-U.S.