Tracking Human Cardiac Subtype Specification Using Molecular Beacons

描述（由申请人提供）：人类心脏组织具有有限的再生能力。心脏组织因缺血或其他病理状况造成的损伤可导致心力衰竭，这是美国人死亡的主要原因。此外，由于先天性心脏缺陷、衰老和获得性疾病导致的起搏器组织的细胞损失或功能障碍会损害传导系统，从而导致严重的心律失常。人类多能干细胞在心脏细胞治疗中具有很大的前景。这些细胞可分化为结型和工作型（心房和心室）心脏亚型，适用于不同的应用：未污染结型细胞的富集工作型细胞用于修复损伤的心室心肌，富集结型细胞用于发展生物起搏器。然而，现有的方法都不能为特定亚型产生均匀的细胞，并且每种亚型的分离和富集方法还没有很好的发展。由于缺乏一种可用于通量平台的分析工具来区分和丰富心脏亚型，因此心脏亚型规范的研究一直具有挑战性。这项R21提案旨在使用一种新的基于纳米技术的工具来跟踪、丰富和表征心脏亚型，而不需要对细胞进行基因修饰。这项技术将使我们能够产生高度均匀的节点型或工作型细胞群，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了解人类多能干细胞对心脏亚型规范的调节。

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.