Tracking Human Cardiac Subtype Specification Using Molecular Beacons

描述(申请人提供)：人体心脏组织再生能力有限。缺血或其他病理条件对心脏组织的损害可能会导致心力衰竭，这在美国是主要的死亡原因。此外，由于先天性心脏缺陷、衰老和后天性疾病损害传导系统，起搏器组织中的细胞丢失或功能障碍可能会导致严重的心律失常。人类多能干细胞为心脏细胞治疗带来了巨大的希望。这些细胞可分为交界型和工作(房室)型，适用于不同的用途：用于修复受损心肌的富集型工作型细胞和用于研制生物起搏器的富集型结型细胞。然而，现有的方法都不能为特定的亚型产生均一细胞，而且每个亚型的分离和浓缩方法也没有很好地开发出来。由于缺乏可用于吞吐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改。这项技术将使我们能够产生高度同质的结节或工作型细胞群，从而促进未来人类多能干细胞在心脏细胞治疗中的应用。它还有助于加速研究，以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.