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Biomaterials, Volume 283, April 2022, 121429 | https://doi.org/10.1016/j.biomaterials.2022.121429 Therapeutic correction of hemophilia A using 2D endothelial cells and multicellular 3D organoids derived from CRISPR/Cas9-engineered patient iPSCs

Jeong Sang Son^a,1, Chul-Yong Park^b,1, Gyunggyu Lee^a, Ji Young Park^a, Hyo Jin Kim^a, Gyeongmin Kim^a, Kyun Yoo Chi^a, Dong-Hun Woo^c, Choongseong Han^c, Sang Kyum Kim^d, Han-Jin Park^e, Dong-Wook Kim^b,*, Jong-Hoon Kim^a,*

^aLaboratory of Stem Cells and Tissue Regeneration, Department of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul, 02841, South Korea
^bDepartment of Physiology, Yonsei University College of Medicine, Seoul, 03722, South Korea ^cDepartment of Stem Cell Biology, NEXEL Co., Ltd., Seoul, 07802, South Korea ^dCollege of Pharmacy, Chungnam National University, Daejeon, 34134, South Korea ^eDepartment of Predictive Toxicology, Korea Institute of Toxicology, Daejeon, 34114, South Korea

¹These authors contributed equally: Jeong Sang Son, Chul-Yong Park.

^*Corresponding author.

Abstract

The bleeding disorder hemophilia A (HA) is caused by a single-gene (F8) defect and its clinical symptom can be substantially improved by a small increase in the plasma coagulation factor VIII (FVIII) level. In this study, we used F8-defective human induced pluripotent stem cells from an HA patient (F8d-HA hiPSCs) and F8-corrected (F8c) HA hiPSCs produced by CRISPR/Cas9 genome engineering of F8d–HA hiPSCs. We obtained a highly enriched population of CD157⁺ cells from CRISPR/Cas9-edited F8c-HA hiPSCs. These cells exhibited multiple cellular and functional phenotypes of endothelial cells (ECs) with significant levels of FVIII activity, which was not observed in F8d-HA hiPSC-ECs. After transplantation, the engineered F8c–HA hiPSC-ECs dramatically changed bleeding episodes in HA animals and restored plasma FVIII activity. Notably, grafting a high dose of ECs substantially reduced the bleeding time during multiple consecutive bleeding challenges in HA mice, demonstrating a robust hemostatic effect (90% survival). Furthermore, the engrafted ECs survived more than 3 months in HA mice and reversed bleeding phenotypes against lethal wounding challenges. We also produced F8c-HA hiPSC-derived 3D liver organoids by assembling three different cell types in microwell devices and confirmed its therapeutic effect in HA animals. Our data demonstrate that the combination of genome-engineering and iPSC technologies represents a novel modality that allows autologous cell-mediated gene therapy for treating HA.

논문정보

형식Research article
게재일2022년 04월 (BRIC 등록일 2022-03-30)
연구진국내 연구진
분야 생명과학 > 세포생물학

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