Tracking Human Cardiac Subtype Specification Using Molecular Beacons

描述（由申请人提供）：人体心脏组织的再生能力有限。心肌缺血或其他病理状态引起的心脏组织损伤可导致心力衰竭，心力衰竭是美国的主要死因。此外，由于先天性心脏缺陷、衰老和损害传导系统获得性疾病引起的起搏器组织中的细胞损失或功能障碍可引起严重心律失常。人类多能干细胞在心脏细胞治疗中具有巨大的前景。这些细胞可以分化为结型和工作型（心房和心室）心脏亚型，它们适用于不同的应用：富集的工作型细胞，而不污染心肌型细胞，用于修复受损的心室肌，富集的心肌型细胞用于开发生物起搏器。然而，现有的方法都不能产生特定亚型的同质细胞，并且用于分离和富集每种亚型的方法还没有得到很好的开发。由于缺乏可用于通量平台以区分和丰富心脏亚型的分析工具，心脏亚型规范的研究一直具有挑战性。这项R21提案旨在使用一种基于纳米技术的新工具来跟踪、富集和表征心脏亚型，而无需对细胞进行遗传修饰。这项技术将使我们能够产生高度同质的结节型或工作型细胞群，从而促进人类多能干细胞在心脏细胞治疗中的未来应用。它还可以帮助加速研究，以了解人类多能干细胞对心脏亚型特化的调节。

DESCRIPTION (provided by applicant): Human cardiac tissues have limited regenerative capacity. Damage in cardiac tissues from ischemia or other pathological conditions can result in heart failure, the leading cause of death in the United States. In addition, cell loss or dysfunctin in pacemaker tissues due to congenital heart defects, aging, and acquired diseases that damage the conduction system can cause severe arrhythmias. Human pluripotent stem cells hold great promise for cardiac cell therapy. These cells can be differentiated into nodal and working (atrial and ventricular) cardiac subtypes, which are suitable for different applications: enriched working-type cells without the contamination of nodal-type cells for repairing injured ventricular myocardium, and enriched nodal-type cells for developing a biological pacemaker. However, none of the existing methods can generate homogeneous cells for a specific subtype, and methods for the isolation and enrichment of each subtype have not been well developed. Research on cardiac subtype specification has been challenging due to the lack of an analysis tool that can be used in a throughput platform to distinguish and enrich cardiac subtypes. This R21 proposal aims to track, enrich and characterize cardiac subtypes using a novel nanotechnology based tool without genetically modifying the cells. This technology will allow us to generate highly homogeneous nodal- or working-type cell populations and therefore facilitate future applications of human pluripotent stem cells in cardiac cell therapy. It can also help accelerate studies to understand the regulation of cardiac subtype specification from human pluripotent stem cells.