Department of Nuclear Medicine and PET-CT Center, The Second Hospital, Zhejiang University School of Medicine, Zhejiang, 310009, Hangzhou, China.
Key Laboratory of Medical Molecular Imaging of Zhejiang Province, Zhejiang, 310009, Hangzhou, China.
Eur J Nucl Med Mol Imaging. 2021 Mar;48(3):708-720. doi: 10.1007/s00259-020-05087-0. Epub 2020 Nov 20.
To investigate the post-transplantation behaviour and therapeutic efficacy of human urinary-induced pluripotent stem cell-derived cardiomyocytes (hUiCMs) in infarcted heart.
We used clustered regularly interspaced short palindromic repeats/CRISPR-associated nuclease 9 (CRISPR/Cas9) technology to integrate a triple-fusion (TF) reporter gene into the AAVS1 locus in human urine-derived hiPSCs (hUiPSCs) to generate TF-hUiPSCs that stably expressed monomeric red fluorescent protein for fluorescence imaging, firefly luciferase for bioluminescence imaging (BLI) and herpes simplex virus thymidine kinase for positron emission tomography (PET) imaging.
Transplanted cardiomyocytes derived from TF-hUiPSCs (TF-hUiCMs) engrafted and proliferated in the infarcted heart as monitored by both BLI and PET imaging and significantly improved cardiac function. Under ischaemic conditions, TF-hUiCMs enhanced cardiomyocyte (CM) glucose metabolism and promoted angiogenic activity.
This study established a CRISPR/Cas9-mediated multimodality reporter gene imaging system that can determine the dynamics and function of TF-hUiCMs in myocardial infarction, which is helpful for investigating the application of stem cell therapy.
研究人尿源性诱导多能干细胞衍生的心肌细胞（hUiCMs）在梗死心脏中的移植后行为和治疗效果。
我们使用成簇规律间隔短回文重复序列/CRISPR 相关核酸酶 9（CRISPR/Cas9）技术将一个三重融合（TF）报告基因整合到人尿源性 hiPSC（hUiPSC）的 AAVS1 基因座中，生成稳定表达单体红色荧光蛋白的 TF-hUiPSC，用于荧光成像、萤火虫荧光素酶用于生物发光成像（BLI）和单纯疱疹病毒胸苷激酶用于正电子发射断层扫描（PET）成像。
移植的来源于 TF-hUiPSC（TF-hUiCMs）的心肌细胞在梗死心脏中植入和增殖，通过 BLI 和 PET 成像监测，并显著改善了心脏功能。在缺血条件下，TF-hUiCMs 增强了心肌细胞（CM）的葡萄糖代谢并促进了血管生成活性。
本研究建立了一种 CRISPR/Cas9 介导的多模态报告基因成像系统，可确定 TF-hUiCMs 在心肌梗死中的动态和功能，有助于研究干细胞治疗的应用。
