Kidney Week

Abstract: TH-PO399

Selective Induction of Renal Interstitial Progenitor Cell Lineages from Human-Induced Pluripotent Stem Cells (hiPSCs) for Understanding of Mesangial and Erythropoietin (EPO)-Producing Cell Development and Kidney Regeneration

Session Information

• Development, Organoids, Injury, and Regeneration
  October 24, 2024 | Location: Exhibit Hall, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Development, Stem Cells, and Regenerative Medicine

• 600 Development, Stem Cells, and Regenerative Medicine

Authors

• Tsujimoto, Hiraku, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan

Hiraku Tsujimoto, MD, PhD

• Hoshina, Azusa, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan

Azusa Hoshina, MD, PhD

• Mae, Shin-ichi, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan

Shin-ichi Mae, PhD

• Araoka, Toshikazu, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan

Toshikazu Araoka, MD, PhD

• Graneli, Cecilia, AstraZeneca PLC, Cambridge, Cambridgeshire, United Kingdom

Cecilia Graneli, MS, PhD

• Woollard, Kevin, AstraZeneca PLC, Cambridge, Cambridgeshire, United Kingdom

Kevin Woollard, PhD

• Osafune, Kenji, Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan

Kenji Osafune, MD, PhD

Background

Recent regenerative studies using human pluripotent stem cells (hPSCs) have developed multiple kidney-lineage cells and organoids. To further form functional segments of the kidney, the interactions of epithelial and interstitial cells are required. By selectively generating one lineage, we can detail developmental signals in complex organ development. Renal interstitial progenitor cells (IPCs) are useful for elucidating the function and pathology of renal interstitium. Renin-producing mesangial lineage cells contribute to hypertension and EPO-producing cells are key in kidney fibrosis. We aim to develop a method to selectively induce renal IPCs and their derivatives from human induced pluripotent stem cells (hiPSCs) and reveal the developmental mechanisms of IPC-lineage cells.

Methods

We modified our previously reported induction method for nephron progenitor cells (NPCs) from hiPSCs by modulating developmental signals involved in the transition from primitive streak to renal IPCs, which were identified by single-cell RNA-sequencing (scRNA-seq) analysis of mouse embryos. We established OSR1-GFP/FOXD1-tdTomato double knock-in hiPSC lines.

Results

We developed a method to generate IPC-like cells (IPLCs) over 80% of induction efficiency. Our IPLCs combined with hiPSC-derived NPCs and nephric duct cells form nephrogenic niche- and mesangium-like structures in vitro. Furthermore, we successfully induced mesangial and EPO-producing cell lineages in vitro by screening differentiation-inducing factors. We confirmed that p38 MAPK and vascular endothelial growth factor (VEGF) signaling pathways are involved in the differentiation of mesangial-lineage cells. Re-analysis of public scRNA-seq data of human embryonic kidneys and an organ culture experiment using mouse embryonic kidneys supports p38 MAPK activation upon the differentiation of mesangial-lineage cells. Furthermore, using small molecules, we revealed that Sonic Hedgehog signaling is involved in the differentiation from IPLCs to EPO-producing cells in vitro.

Conclusion

Our IPC-lineage induction method contributes to kidney regeneration and developmental research.

Funding

• Commercial Support – Rege Nephro Co., Ltd., AstraZeneca plc