Mechanism of transplanted neonatal cardiac progenitor cells to repair ischemic myocardium

利用我们独特的人类新生儿心脏组织资源，并通过我们之前的R01提供资金，我们 已有研究表明，新生CPC在修复受损心脏方面具有较好的疗效。 与任何其他类型的细胞相比，这是由于部分由热休克因子1控制的更强大的分泌体 (HSF1)。NCPC临床成功的关键将是确定他们的心肌恢复机制。我们 最近报告了aCDC和aCPC之间的正面比较，证明aCPC 在基于细胞和体内再生的检测中，ACDCs的表现优于ACDCs。以非侵入性的方式监控 体内移植的aCPC或aCDC，我们纯化并询问了祖细胞特异性外切体(EXOS) 从受者的血浆中提取全部EXOS。通过使用我们之前发布的计算模型，该模型采用 主成分分析(PCA)和偏最小二乘回归分析(PLSR)的优点 在CPC衍生的循环EXOS中发现了潜在的有影响力的miRNA签名，这些签名推动了 受损心肌的修复机制至少涉及两个重要过程：抗纤维化和 血管生成增加。出于这些原因，我们认为移植的祖细胞的miRNA图谱- 从受体血浆中分离的EXOS更准确地预测了干细胞的临床结果 治疗比培养的祖细胞或其EXOS的RNA图谱更重要。然而，确定的直接作用是 移植的CPC体内的miRNAs还没有被确定，血浆miRNAs是如何变化的 在细胞移植后的术后阶段，以及最终该方法的有效性和 大型临床前动物模型的计算建模。因此，我们假设等离子体EXOS 反映了亲代移植的祖细胞恢复后触发的特定分子通路 损伤的心肌。AIM1将验证以下预测的血管生成和抗纤维化MIRS CPC的计算模型对于恢复缺血心肌是必不可少的。目标2将决定 手术后循环EXOS MIR成分如何变化的计算模型。Aim3将 在临床前的大型动物中使用移植的神经干细胞来扩展我们的计算模型的预测能力 猪心肌梗死模型。成功完成将证明使用来自新生儿的祖细胞 组织具有最高的再生能力，这可能是临床成功的关键。此外，我们还将 为基于细胞的治疗确定一种更定量的方法的新范例，以揭示一种非侵入性 窗口进入移植细胞的条件状态。总而言之，这些发现将证明 循环祖细胞特异性外切体作为液体活检提供非侵入性窗口的可能性 进入移植的新生儿CPC的条件状态。这些数据暗示了监控的可能性 用于其他基于同种异体细胞治疗的细胞特异性外切体。

Using our unique resource of human neonatal cardiac tissue and funded through our previous R01, we have consistently demonstrated that neonatal CPCs (nCPCs) have superior efficacy in repairing the injured heart compared to any other cell type due to a more potent secretome controlled partly by the heat shock factor 1 (HSF1). Critical to the nCPC’s clinical success will be determining their mechanism of myocardial recovery. We have recently reported a head to head comparison between aCDCs and aCPCs that demonstrated that aCPCs outperformed the aCDCs in cell-based and in vivo regenerative assays. To noninvasively monitor the activity of the transplanted aCPCs or aCDCs in vivo, we purified and interrogated progenitor-specific exosomes (EXOs) from the recipient total plasma EXOs. By using our previously published computational modeling which takes advantage of principal component analysis (PCA) and partial least squares regression analysis (PLSR), we identified potentially impactful miRNA signatures within aCPCs–derived circulating EXOs that drives mechanisms of repair in the injured myocardium involving at least two important processes: antifibrosis and increased angiogenesis. For these reasons, we believe that miRNA profiling of transplanted progenitor cell– derived EXOs isolated from recipient plasma more accurately predicts the clinical outcomes seen with stem cell therapy than the RNA profiles of cultured progenitor cells or their EXOs. However, the direct role of the identified miRNAs within transplanted CPCs have not yet been determined in vivo, how the plasma miRNAs change during the post-operative period after cell transplantation, and finally the validity of this methodology and computational modeling in a large preclinical animal model. Thus, we hypothesize that the plasma EXOs reflects specific molecular pathways triggered by the parent transplanted progenitor cells that recovers the injured myocardium. Aim1 will validate whether angiogenesis and antifibrosis mIRs predicted by computational modeling for CPCs are essential for recovering the ischemic myocardium. Aim 2 will determine how the circulating EXOs mIR composition changes post-operatively by computational modeling. Aim3 will expand the predictive capacity of our computational model using transplanted nCPCs in a large animal preclinical porcine MI model. Successful completion will demonstrate that the use of progenitor cells derived from neonatal tissue has the highest regenerative abilities which maybe critical for the clinical success. In addition, we will determine a new paradigm for a more quantitative methodology for cell based therapies to reveal a noninvasive window into the conditional state of the transplanted cells. Collectively, these findings will demonstrate the potential of circulating progenitor cell–specific exosomes as a liquid biopsy that provides a noninvasive window into the conditional state of the transplanted neonatal CPCs. These data implicate the surveillance potential of cell-specific exosomes for other allogeneic cell based therapies.