Department of Cell Biology, Naval Medical University (Second Military Medical University), Shanghai, China.
Department of Nephrology, Changhai Hospital, Naval Medical University (Second Military Medical University), Shanghai, China.
Theranostics. 2024 Feb 17;14(4):1764-1780. doi: 10.7150/thno.89174. eCollection 2024.
The present understanding of the cellular characteristics and communications in crystal nephropathy is limited. Here, molecular and cellular studies combined with single-cell RNA sequencing (scRNA-seq) were performed to investigate the changes in cell components and their interactions in glyoxylate-induced crystallized kidneys to provide promising treatments for crystal nephropathy. The transcriptomes of single cells from mouse kidneys treated with glyoxylate for 0, 1, 4, or 7 days were analyzed via 10× Genomics, and the single cells were clustered and characterized by the Seurat pipeline. The potential cellular interactions between specific cell types were explored by CellChat. Molecular and cellular findings related to macrophage-to-epithelium crosstalk were validated in sodium oxalate (NaOx)-induced renal tubular epithelial cell injury in vitro and in glyoxylate-induced crystal nephropathy in vivo. Our established scRNA atlas of glyoxylate-induced crystalline nephropathy contained 15 cell populations with more than 40000 single cells, including relatively stable tubular cells of different segments, proliferating and injured proximal tubular cells, T cells, B cells, and myeloid and mesenchymal cells. In this study, we found that Mrc1 macrophages, as a subtype of myeloid cells, increased in both the number and percentage within the myeloid population as crystal-induced injury progresses, and distinctly express IGF1, which induces the activation of a signal pathway to dominate a significant information flow towards injured and proliferating tubule cells. IGF1 promoted the repair of damaged tubular epithelial cells induced by NaOx in vitro, as well as the repair of damaged tubular epithelial cells and the recovery of disease outcomes in glyoxylate-induced nephrolithic mice in vivo. After constructing a cellular atlas of glyoxylate-induced crystal nephropathy, we found that IGF1 derived from Mrc1 macrophages attenuated crystal nephropathy through promoting renal tubule cell proliferation via the AKT/Rb signaling pathway. These findings could lead to the identification of potential therapeutic targets for the treatment of crystal nephropathy.
目前对结晶肾病中细胞特征和通讯的了解有限。在这里，进行了分子和细胞研究，并结合单细胞 RNA 测序 (scRNA-seq)，以研究在乙醛酸诱导结晶肾脏中细胞成分的变化及其相互作用，为结晶肾病提供有希望的治疗方法。用乙醛酸处理 0、1、4 或 7 天的小鼠肾脏的单细胞的转录组通过 10× Genomics 进行分析，并通过 Seurat 管道对单细胞进行聚类和特征分析。通过 CellChat 探索特定细胞类型之间的潜在细胞相互作用。分子和细胞发现与巨噬细胞到上皮细胞串扰有关，在体外草酸鈉 (NaOx) 诱导的肾小管上皮细胞损伤和体内乙醛酸诱导的结晶性肾病中得到验证。我们建立的乙醛酸诱导结晶性肾病的 scRNA 图谱包含 15 个细胞群，超过 40000 个单细胞，包括不同节段相对稳定的管状细胞、增殖和损伤的近端肾小管细胞、T 细胞、B 细胞以及髓细胞和间充质细胞。在这项研究中，我们发现 Mrc1 巨噬细胞作为髓细胞的一个亚型，在晶体诱导损伤进展过程中，在髓细胞群体中的数量和比例都增加，并且明显表达 IGF1，IGF1 诱导信号通路的激活，主导着流向受损和增殖的肾小管细胞的显著信息流。IGF1 促进了体外 NaOx 诱导的受损肾小管上皮细胞的修复，以及体内乙醛酸诱导的肾结石小鼠中受损肾小管上皮细胞的修复和疾病结局的恢复。在构建乙醛酸诱导结晶性肾病的细胞图谱后，我们发现 Mrc1 巨噬细胞衍生的 IGF1 通过 AKT/Rb 信号通路促进肾小管细胞增殖，从而减轻结晶性肾病。这些发现可能为治疗结晶性肾病确定潜在的治疗靶点。
