Overline: Kidney DiseaseOne Sentence Summary:iPSC-derived cells secrete functional erythropoietin in a physiological manner and ameliorate renal anemia in a mouse model. Cell therapy for renal anemia using iPSCsErythropoietin dysregulation is a hallmark of renal anemia. Although recombinant erythropoietin treatment is beneficial and safe, more physiological therapies are required. Hitomi et al. developed a differentiation protocol for erythropoietin-producing cells from human and mouse iPSCs/ESCs.These cells produced and secreted functional erythropoietin protein in a hypoxia-dependent manner. Transplantation of these cells into a mouse model improved renal anemia. From the perspective of basic research, erythropoietin cells may be a useful tool for investigating the molecular mechanisms of erythropoietin production and secretion. From the perspective of clinical research, these results may provide a physiological therapeutic agent for renal anemia. AbstractThe production of erythropoietin (EPO), a principal hormone for the hematopoietic system, by the kidneys is reduced in patients with chronic kidney disease (CKD), eventually resulting in severe anemia. Although recombinant human EPO treatment improves anemia in patients with CKD, returning to full red blood cell production without fluctuations does not always occur. In this study, we established a method to generate EPO-producing cells from human induced pluripotent stem cells (hiPSCs) by modifying previously reported hepatic differentiation protocols. These cells showed increased EPO expression and secretion in response to low oxygen conditions, prolyl hydroxylase domain-containing enzymes inhibitors and insulin-like growth factor-1. The EPO protein secreted from hiPSC-derived EPO-producing (hiPSC-EPO) cells induced the erythropoietic differentiation of human umbilical cord blood progenitor cells in vitro. Furthermore, transplantation of hiPSC-EPO cells into mice with CKD induced by adenine treatment improved renal anemia. Thus, hiPSC-EPO cells may be a useful tool for clarifying the mechanisms of EPO production and may be useful as a therapeutic strategy for treating renal anemia.

Cell therapies using human induced pluripotent stem cell (hiPSC)-derived nephron progenitor cells (NPCs) are expected to ameliorate acute kidney injury (AKI). However, using hiPSC-derived NPCs clinically is a challenge because hiPSCs themselves are tumorigenic. LIN28A, ESRG, CNMD and SFRP2 transcripts have been used as a marker of residual hiPSCs for a variety of cell types undergoing clinical trials. In this study, by reanalyzing public databases, we found a baseline expression of LIN28A, ESRG, CNMD and SFRP2 in hiPSC-derived NPCs and several other cell types, suggesting LIN28A, ESRG, CNMD and SFRP2 are not always reliable markers for iPSC detection. As an alternative, we discovered a lncRNA marker gene, MIR302CHG, among many known and unknown iPSC markers, as highly differentially expressed between hiPSCs and NPCs, by RNA sequencing and quantitative RT-PCR (qRT-PCR) analyses. Using MIR302CHG as an hiPSC marker, we constructed two assay methods, a combination of magnetic bead-based enrichment and qRT-PCR and digital droplet PCR alone, to detect a small number of residual hiPSCs in NPC populations. The use of these in vitro assays could contribute to patient safety in treatments using hiPSC-derived cells.

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Erythropoietin (EPO) is a crucial hormone for erythropoiesis and produced by adult kidneys. Insufficient EPO production in chronic kidney disease (CKD) can cause renal anemia. Although hypoxia-inducible factors (HIFs) are known as a main regulator, the mechanisms of EPO production have not been fully elucidated. In this study, we aimed to examine the roles of retinoic acid (RA) in EPO production using EPO-producing cells derived from human induced pluripotent stem cells (hiPSC-EPO cells) that we previously established. RA augmented EPO production by hiPSC-EPO cells under hypoxia or by treatment with prolyl hydroxylase domain-containing protein (PHD) inhibitors that upregulate HIF signals. Combination treatment with RA and a PHD inhibitor improved renal anemia in vitamin A-depleted CKD model mice. Our findings using hiPSC-EPO cells and CKD model mice may contribute to clarifying the EPO production mechanism and developing efficient therapies for renal anemia.

Background and aimsive Kind of Operation Number of Cases (M/F) Spine Surgeries 10 (5/5) Artificial Knee Joint Replacement 6 (3/3) Total Hip Arthroplasty 4 (0/4) Amputation of Toe Necrosis 1 (1/0) Bone Fracture Surgeries 3 (2/1) foramen magnum decompression 1 (0/1)