Characterization of Cell-Based Therapy for Congenital Heart Patients

该补助金是K08辅导职业发展奖提交支持博士的研究。为了推进他在先天性心脏病患者干细胞治疗方面的职业生涯，该资助包括两个重要领域的培训：（1）分子生物学技术（心脏干细胞的分离和表征，免疫组织化学）和（2）体内移植技术。这两个领域都是Kaushal博士以前的培训所没有的，但对于推进这一领域的研究至关重要。Kaushal博士的导师是西北大学的Doug Losordo博士，他在这一领域拥有丰富的研究经验。最近的证据已经确定了心脏本身内的细胞群作为细胞再生的潜在自体细胞来源，称为心脏干细胞（CSC）。到目前为止，这些CSC在发展为非缺血性心肌病的儿科患者中的临床应用尚不清楚。为了实现这一目标，我们将有三个具体目标，它们遵循一个渐进的顺序。目的1和2集中于来自心脏不同解剖部位的hCSC的生长、自我更新潜力和心肌分化特征的详细描述性特性的累积。此外，我们将确定患者的年龄或病理状态是否影响hCSC的先天特性。体内分析将涉及在多柔比星诱导的心肌病模型中检查定量和定性心肌再生，该模型复制了我们患有心力衰竭的儿童中存在的心肌病的许多显著特征。一旦我们确定了人心肌是否维持一般CSC或相反，更具心腔特异性的CSC（其可能受年龄和生理状态影响），我们将测试最佳鉴定的CSC以确定其在阿霉素诱导的心肌病模型中对于心肌细胞和冠状血管的再生是否上级、劣于或等于骨髓来源的细胞。我们坚信，阐明这些问题对于构建最强大的再生临床方案至关重要。相关性（见说明）：由于该提案中的所有实验都涉及人体组织，因此将产生非常强有力的转化结论，这将直接影响未来临床试验的发展。此外，这些实验对患有先天性心脏缺陷的患者具有直接的临床应用，因为这些患者的寿命更长，并且一些患者发展出可能受益于使用hCSC的基于细胞的治疗的心功能障碍。

This grant is a K08 Mentored Career Developmental Award submission supporting the research of Dr. Sunjay Kaushal. To advance his career in stem cell based therapy for congenital heart patients, this grant incorporates training in two vital fields: (1) molecular biology techniques (isolation and characterization of cardiac stem cells, immunohistiochemistry) and (2) in vivo transplantation techniques. Both of these fields is absent from Dr. Kaushal's previous training, yet are vital in advancing research in this field. Dr. Kaushal's mentor is Dr. Doug Losordo at Northwestern University who has an extensive research experience in this field. Recent evidence has identified a population of cells within the heart itself as a potential autologous cell source for cellular regeneration, termed cardiac stem cells (CSCs). As of yet, the clinical application of these CSCs for pediatric patients who develop non-ischemic cardiomyopathies is unclear. To accomplish this objective, we will have three Specific Aims which follow a progressive sequence. Aims 1 and 2 center on the accrual of detailed descriptive properties of the growth, self-renewal potential, and myocardial differentiation characteristics of hCSCs from different anatomic sites of the heart. In addition, we will determine whether age or the pathologic state of the patient affects the innate properties of the hCSCs. The in vivo analysis will involve examining quantitative and qualitative myocardial regeneration in a doxorubicin-induced cardiomyopathy model, which replicates many of the salient features of cardiomyopathy present in our children with heart failure. Once we have established whether the human myocardium maintains a generic CSC or, in contrast, a more cardiac chamber-specific CSC, which may be influenced by age and physiologic state, we will test the best identified CSC to determine whether it is superior, inferior, or equal to bone marrow-derived cells for the regeneration of cardiomyocytes and coronary vessels in the doxorubicin- induced cardiomyopathy model. We strongly believe that elucidating these issues is critical to constructing the most powerful regenerative clinical protocol. RELEVANCE (See instmctions): Since all of the experiments in this proposal involve human tissues, there will be very strong translational conclusions that will directly impact the development of future clinical trials. Furthermore, these experiments have direct clinical applications to patients who have congenital heart defects as these patients are living longer, and some develop cardiac dysfunction that may benefit from a cell-based therapy using hCSCs.