Single-Cell RNA Sequencing Identifies Response of Renal Lymphatic Endothelial Cells to Acute Kidney Injury

Abstract

Significance Statement The renal lymphatic vasculature and the lymphatic endothelial cells that make up this network play important immunomodulatory roles during inflammation. How lymphatics respond to AKI may affect AKI outcomes. The authors used single-cell RNA sequencing to characterize mouse renal lymphatic endothelial cells in quiescent and cisplatin-injured kidneys. lymphatic endothelial cell gene expression changes were confirmed in ischemia–reperfusion injury and in cultured lymphatic endothelial cells, validating renal lymphatic endothelial cells single-cell RNA sequencing data. This study is the first to describe renal lymphatic endothelial cell heterogeneity and uncovers molecular pathways demonstrating lymphatic endothelial cells regulate the local immune response to AKI. These findings provide insights into previously unidentified molecular pathways for lymphatic endothelial cells and roles that may serve as potential therapeutic targets in limiting the progression of AKI. Background The inflammatory response to AKI likely dictates future kidney health. Lymphatic vessels are responsible for maintaining tissue homeostasis through transport and immunomodulatory roles. Owing to the relative sparsity of lymphatic endothelial cells in the kidney, past sequencing efforts have not characterized these cells and their response to AKI. Methods Here, we characterized murine renal lymphatic endothelial cell subpopulations by single-cell RNA sequencing and investigated their changes in cisplatin AKI 72 hours postinjury. Data were processed using the Seurat package. We validated our findings by quantitative PCR in lymphatic endothelial cells isolated from both cisplatin-injured and ischemia–reperfusion injury, by immunofluorescence, and confirmation in in vitro human lymphatic endothelial cells. Results We have identified renal lymphatic endothelial cells and their lymphatic vascular roles that have yet to be characterized in previous studies. We report unique gene changes mapped across control and cisplatin-injured conditions. After AKI, renal lymphatic endothelial cells alter genes involved in endothelial cell apoptosis and vasculogenic processes as well as immunoregulatory signaling and metabolism. Differences between injury models were also identified with renal lymphatic endothelial cells further demonstrating changed gene expression between cisplatin and ischemia–reperfusion injury models, indicating the renal lymphatic endothelial cell response is both specific to where they lie in the lymphatic vasculature and the kidney injury type. Conclusions In this study, we uncover lymphatic vessel structural features of captured populations and injury-induced genetic changes. We further determine that lymphatic endothelial cell gene expression is altered between injury models. How lymphatic endothelial cells respond to AKI may therefore be key in regulating future kidney disease progression.

Bibliometric and visual analysis of immunisation associated with acute kidney injury from 2003 to 2023

“…The occurrence and development of renal ischemia-reperfusion injury (IRI) might mediate inflammatory response through inflammation-related signaling pathways, and blocking inflammatory response plays a very important role in improving the pathological status of AKI ( Li et al, 2022 ; Creed et al, 2024 ). Relevant studies ( Vande Walle and Lamkanfi, 2024 ; Xu K. et al, 2024 ) have shown that pyroptosis is a key mechanism of renal IRI.…”

Kidney Endothelial Cell Biology in Health and Disease

“…2 In this issue of JASN, Creed et al provide additional insights into the implications of lymphatic biology for kidney diseases. 3 By exploring the altered gene expression changes in response to cisplatin-induced AKI in male mice, the investigators identified the lymphatic endothelial cells (LECs) molecular signature as being most consistent with vascular endothelial growth factor receptor signaling, T-cell differentiation, and regulation of inflammatory response, supporting the notion that LECs respond to AKI by undergoing lymphangiogenesis and upregulating their interactions with immune cells. In addition, by comparing the molecular responses of LECs to distinct injury stimuli (cisplatin versus ischemia-reperfusion injury), the investigators revealed that the response of LECs is likely influenced by the specific mode and context of injury, thereby substantiating the existing evidence of conflicting outcomes in the literature across different models.…”