Du Z, Bas-Cristóbal Menéndez A, Urban M, Hartley A, Ratsma D, Koedam M, van den Bosch T P P, Clahsen-van Groningen M, Gribnau J, Mulder J, Reinders M E J, Baan C C, van der Eerden B, Harbottle R P, Hoogduijn Martin J
Erasmus MC Transplant Institute, Department of Internal Medicine, University Medical Center, Wytemaweg 80, Rotterdam, 3015 CN, The Netherlands.
Department of Pediatrics, Sophia Children's Hospital, Erasmus University Medical Center, Rotterdam, The Netherlands.
Stem Cell Res Ther. 2025 Apr 12;16(1):174. doi: 10.1186/s13287-025-04282-w.
The kidney's endocrine function is essential for maintaining body homeostasis. Erythropoietin (EPO) is one of the key endocrine factors produced by the kidney, and kidney disease patients frequently experience anemia due to impaired EPO production. In the present study we explored the potential of human induced pluripotent stem cell (iPSC)-derived kidney organoids to restore EPO production.
EPO secretion by kidney organoids was examined under 1% and 20% oxygen levels. To increase the EPO secreting capacity of kidney organoids, iPSC were genetically engineered with a non-integrating scaffold/matrix attachment region (S/MAR) DNA vector containing the EPO gene and generated EPO-overexpressing (EPO+) kidney organoids. To assess the physiological effects of EPO + organoids, 2-8 organoids were implanted subcutaneously in immunodeficient mice.
Kidney organoids produced low amounts of EPO under 1% oxygen. EPO S/MAR DNA vectors persisted and continued to robustly express EPO during iPSC expansion and kidney organoid differentiation without interfering with cellular proliferation. EPO + iPSC demonstrated efficient differentiation into kidney organoids. One-month post-implantation, EPO + organoids displayed continuously elevated EPO mRNA levels and significantly increased endothelial cell numbers compared to control organoids. Hematocrit levels were notably elevated in mice implanted with EPO + organoids in an organoid number-dependent manner. EPO + organoids furthermore influenced bone homeostasis in their hosts, evidenced by a change in trabecular bone composition.
Kidney organoids modified by EPO S/MAR DNA vector allow stable long-term delivery of EPO. The observed physiological effects following the implantation of EPO + organoids underscore the potential of gene-edited kidney organoids for endocrine restoration therapy.
肾脏的内分泌功能对于维持机体稳态至关重要。促红细胞生成素（EPO）是肾脏产生的关键内分泌因子之一，肾病患者常因EPO生成受损而出现贫血。在本研究中，我们探讨了人诱导多能干细胞（iPSC）来源的肾类器官恢复EPO生成的潜力。
在1%和20%的氧水平下检测肾类器官的EPO分泌情况。为了提高肾类器官的EPO分泌能力，用含有EPO基因的非整合支架/基质附着区域（S/MAR）DNA载体对iPSC进行基因工程改造，生成过表达EPO（EPO+）的肾类器官。为了评估EPO+类器官的生理效应，将2 - 8个类器官皮下植入免疫缺陷小鼠体内。
肾类器官在1%氧气条件下产生少量EPO。EPO S/MAR DNA载体在iPSC扩增和肾类器官分化过程中持续存在并持续强劲表达EPO，而不干扰细胞增殖。EPO+iPSC表现出高效分化为肾类器官的能力。植入后一个月，与对照类器官相比，EPO+类器官的EPO mRNA水平持续升高，内皮细胞数量显著增加。植入EPO+类器官的小鼠的血细胞比容水平以类器官数量依赖的方式显著升高。EPO+类器官还影响其宿主的骨稳态，这在小梁骨组成的变化中得到证实。
经EPO S/MAR DNA载体修饰的肾类器官可实现EPO的稳定长期递送。植入EPO+类器官后观察到的生理效应强调了基因编辑肾类器官在内分泌恢复治疗中的潜力。
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