Abstract: FR-PO1091
In Vivo Single Cell Analysis of Senescence in Aging Kidney Disease and CKD
Session Information
- CKD Mechanisms: Progression, Fibrosis, and Beyond
November 03, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2303 CKD (Non-Dialysis): Mechanisms
Authors
- Aratani, Sae, Tokyo Daigaku Ikagaku Kenkyujo, Minato-ku, Tokyo, Japan
- Wang, Teh-Wei, Tokyo Daigaku Ikagaku Kenkyujo, Minato-ku, Tokyo, Japan
- Nakanishi, Makoto, Tokyo Daigaku Ikagaku Kenkyujo, Minato-ku, Tokyo, Japan
Background
It remains largely unknown how senescent cells affect kidney aging and chronic kidney diseases (CKD) in vivo. Previously, we generated a p16-CreERT2-tdTomato mouse model (p16-tdTomato mice hereafter) in which cells with high p16 expression, a prototypical senescent marker, were labeled with tdTomato through the administration of tamoxifen. In this study, we aimed to reveal the pathophysiological role of senescent cells in the kidney using p16-tdTomato mice.
Methods
We investigated three different kidney conditions; normal aging kidneys, adenine-induced kidney diseases, and rhabdomyolysis-induced kidney diseases model. Male 6-24-month-old p16-tdTomato mice and 2-3-month-old p16-tdTomato mice were used for the normal aging process and disease models, respectively. The adenine diet was administered for three weeks, while 50% glycerol was injected into each leg to induce rhabdomyolysis thrice weekly. All the mice were sacrificed after the five times of tamoxifen injections. For further analysis of single senescent cells, the kidneys were directly digested, and flow cytometry (FACS) separately sorted tdTomato-positive senescent or tdTomato-negative cells.
Results
In the normal aging process, 20-24-month-old mice showed an accumulation in tdTomato-positive senescent cells compared to young mice.
In both CKD models, the serum analysis demonstrated kidney dysfunction with increased BUN and Cr. Histopathological examinations showed tubular damage, interstitial fibrosis, and infiltration of immune cells, resembling CKD progression. Notably, the tdTomato-positive senescent cells increased following CKD; mainly, tdTomato-positive senescent PTECs were significantly detected. RNA-sequence analysis revealed that tdTomato-positive senescent PTECs collected from the CKD model by FACS showed high inflammatory signaling pathways, including hallmark TNFα signaling through NFkB, IL6 JAK-STAT, and TGFβ signaling. These inflammatory cascades are all related to senescence-associated secretory phenotype (SASP).
Conclusion
These data suggest that CKD progression might cause premature kidney aging. In particular, tdTomato-positive PTECs might contribute to an inflammatory environment in CKD by producing SASP. These tdTomato-positive PTECs might be a therapeutic target to prevent CKD progression and rejuvenate premature kidney aging.
Funding
- Government Support – Non-U.S.