Strategic Center of Cell & Bio Therapy, Seoul National University Hospital, Seoul, South Korea.
Department of Molecular Medicine and Biopharmaceutical Sciences, Graduate School of Convergence Science and Technology, and College of Medicine or College of Pharmacy, Seoul National University, Seoul, South Korea.
Stem Cells Transl Med. 2022 Mar 31;11(3):332-342. doi: 10.1093/stcltm/szab015.
Discovering cell-surface markers based on a comprehensive understanding of development is utilized to isolate a particular cell type with high purity for therapeutic purposes. Given that latrophilin-2 (Lphn2) substantially contributes to cardiac differentiation, we examined whether Lphn2 regulates functional significance in heart development and repair. We performed whole-mount immunostaining followed by clearing technique of embryo, RNA sequencing related to Lphn2-knockout (KO) embryo, and in vivo functional analyses of Lphn2+ cells using echocardiography. After immunostaining the cleared embryo sample, Lphn2 was exclusively observed in cardiac cells expressing α-sarcomeric actinin at embryonic days E9.5 and E10.5. Homozygous Lphn2-KO mice were embryonically lethal and showed underdevelopment of the ventricular myocardium. However, Lphn2 was not required to develop vessels, including endothelial cells and smooth muscle cells. For the purpose of cardiac regeneration, we transplanted pluripotent stem cell (PSC)-derived Lphn2+ cells into the infarcted heart. PSC-derived Lphn2+ cells differentiated into cardiomyocytes and regenerated the myocardium when transplanted into the infarcted heart, unlike Lphn2- cells. Transplanted Lphn2+ cells improved left-ventricle systolic function and reduced infarct size. We demonstrated that Lphn2 exhibits potential as a cardiomyogenic marker to facilitate targeted stem cell therapy for heart repair in clinical practice.
基于对发育的全面理解来发现细胞表面标志物，用于为治疗目的分离具有高纯度的特定细胞类型。鉴于拉普罗林-2（Lphn2）对心脏分化有重要贡献，我们研究了 Lphn2 是否调节心脏发育和修复中的功能意义。我们进行了胚胎全胚胎免疫染色，随后进行胚胎清除技术、与 Lphn2 敲除（KO）胚胎相关的 RNA 测序，以及使用超声心动图对 Lphn2+细胞进行体内功能分析。在对清除的胚胎样本进行免疫染色后，Lphn2 仅在表达α-横纹肌肌动蛋白的胚胎第 9.5 天和第 10.5 天的心脏细胞中观察到。杂合 Lphn2-KO 小鼠在胚胎期致死，并表现出心室心肌发育不良。然而，Lphn2 对于血管的发育不是必需的，包括内皮细胞和平滑肌细胞。为了进行心脏再生，我们将多能干细胞（PSC）衍生的 Lphn2+细胞移植到梗死的心脏中。与 Lphn2-细胞不同，PSC 衍生的 Lphn2+细胞在移植到梗死心脏中时分化为心肌细胞并再生心肌。移植的 Lphn2+细胞改善了左心室收缩功能并减少了梗死面积。我们证明 Lphn2 具有作为心肌生成标志物的潜力，有助于在临床实践中针对心脏修复的靶向干细胞治疗。
