Department of Internal Medicine, Gangnam Severance Hospital, Yonsei University College of Medicine, Seoul, the Republic of Korea.
Severance Institute for Vascular and Metabolic Research, Yonsei University College of Medicine, Seoul, the Republic of Korea.
Stem Cell Rev Rep. 2022 Mar;18(3):980-992. doi: 10.1007/s12015-021-10141-x. Epub 2021 Mar 2.
Extracellular vesicles (EVs) shed from kidney mesenchymal stem cells (KMSCs) show protective effects against acute kidney injury and progressive kidney fibrosis via mRNA transfer. Previous studies report improvement of renal anemia following administration of genetically modified MSCs or peritoneal mesothelial cells that secrete erythropoietin (EPO). Here, we determined whether EPO-secreting KMSC-derived EVs (EPO-EVs) can improve renal anemia in mouse models of chronic kidney disease (CKD). The mouse CKD and renal anemia model was induced by electrocoagulation of the right renal cortex and sequential left nephrectomy. At six weeks post-nephrectomy, we observed significantly lower hemoglobin (10.4 ± 0.2 vs. 13.2 ± 0.2 g/dL) and significantly higher blood urea nitrogen and serum creatinine levels in CKD mice relative to controls (60.5 ± 0.5 and 0.37 ± 0.09 mg/dL vs. 19.9 ± 0.5 and 0.12 ± 0.02 mg/dL, respectively). Genetically engineered EPO-KMSCs secreted 71 IU/mL EPO/10 cells/24 h in vitro, and EPO-EVs isolated by differential ultracentrifugation expressed EPO mRNA and horizontally transferred EPO mRNA into target cells in vitro and in vivo. Furthermore, at two weeks post-injection of EPO-KMSCs or EPO-EVs into CKD mice with renal anemia, we observed significant increases in hemoglobin levels (11.7 ± 0.2 and 11.5 ± 0.2 vs. 10.1 ± 0.2 g/dL, respectively) and significantly lower serum creatinine levels at eight weeks in comparison to mice receiving vehicle control (0.30 ± 0.00 and 0.23 ± 0.03 vs. 0.43 ± 0.06 mg/dL, respectively). These results demonstrate that intraperitoneal administration of EPO-EVs significantly increased hemoglobin levels and renal function in CKD mice, suggesting the efficacy of these genetically engineered EVs as a promising novel strategy for the treatment of renal anemia.
细胞外囊泡（EVs）从肾脏间充质干细胞（KMSCs）中脱落，通过 mRNA 转移对急性肾损伤和进行性肾纤维化具有保护作用。先前的研究报告称，给予遗传修饰的间充质干细胞或分泌促红细胞生成素（EPO）的腹膜间皮细胞可改善肾性贫血。在这里，我们确定了 EPO 分泌的 KMSC 衍生的 EV（EPO-EV）是否可以改善慢性肾脏病（CKD）小鼠模型中的肾性贫血。通过对右肾皮质进行电凝和随后的左肾切除术诱导小鼠 CKD 和肾性贫血模型。在肾切除术后 6 周，与对照组相比，CKD 小鼠的血红蛋白（10.4±0.2 对 13.2±0.2 g/dL）明显降低，血尿素氮和血清肌酐水平明显升高（60.5±0.5 和 0.37±0.09 mg/dL 对 19.9±0.5 和 0.12±0.02 mg/dL，分别）。体外培养的基因工程 EPO-KMSC 分泌 71 IU/mL EPO/10 细胞/24 h，差速超速离心分离的 EPO-EV 表达 EPO mRNA，并在体外和体内将 EPO mRNA 水平转移到靶细胞。此外，在 EPO-KMSC 或 EPO-EV 注射到 CKD 合并肾性贫血的小鼠后两周，我们观察到血红蛋白水平明显升高（11.7±0.2 和 11.5±0.2 对 10.1±0.2 g/dL，分别），而在 8 周时接受载体对照的小鼠的血清肌酐水平明显降低（0.30±0.00 和 0.23±0.03 对 0.43±0.06 mg/dL，分别）。这些结果表明，腹腔内给予 EPO-EV 可显著提高 CKD 小鼠的血红蛋白水平和肾功能，表明这些基因工程 EV 作为治疗肾性贫血的一种有前途的新策略具有疗效。
