Integrated Manufacturing of Therapeutic Cardiac Cells

Pi：Palecek，Sean P.Proposal：心脏病是主要的死亡原因，是医疗保健系统的主要负担。心脏病发作或心力衰竭的患者表现出心脏机械功能的永久性丧失，降低了生活质量，并增加了未来医疗并发症的风险。今天还没有恢复心脏机械功能的治疗方法，但干细胞制造的心肌细胞移植在临床前研究和临床试验中是一种很有前途的方法。干细胞来源的心肌细胞的一个电流限制是它们产生的力量和传递电流的方式更类似于胎儿细胞，而不是成年细胞。我们建议探索这样的概念，即联合制造心肌细胞和干细胞来源的心外膜细胞将提高干细胞来源的心脏细胞的功能质量。该项目将探索这种与发育相关的化学因子交换或来自干细胞的心肌和心外膜细胞之间的直接接触是否会改善心肌细胞的功能，然后利用这些发现设计策略来制造大约10亿个必要的心肌细胞，以取代心脏病发作期间失去的心脏细胞。该项目将使干细胞能够制造足够数量的高质量心肌细胞，以促进临床前和临床研究，以恢复心脏病患者的心脏功能。该项目还解决了细胞治疗制造方面缺乏训练有素的劳动力的问题。项目活动将为本科生和研究生提供独特的培训机会，让他们与产业界合作进行制造研究。该项目团队还将开发一门多机构的细胞疗法制造课程，为K12学生创建与干细胞疗法相关的外展活动，并针对研究生和行业员工完善一门关于再生制造的国际短期课程。下一代疗法将涉及具有再生受损或患病人体组织能力的活细胞。美国有望成为细胞疗法的领导者，但在制造复杂的活细胞方面必须克服重大挑战。在这个项目中，我们将解决细胞治疗制造业面临的主要障碍，包括(1)缺乏强大的、可扩展的制造平台，(2)缺乏制造如何影响效力的关键质量属性(CQA)的知识，以及(3)缺乏训练有素的劳动力。该项目的重点是从人类多能干细胞(HPSCs)中制造安全有效的治疗性心脏细胞，该细胞具有健壮、可扩展和成本效益高的特点。HPSC来源的心肌细胞(CMS)在临床前动物模型中表现出改善的心室收缩功能。即将到来的临床试验的成功和有效的人类疗法的开发将需要以合理的成本生产高质量的细胞。然而，CM制造的规模扩大降低了差异化稳健性，导致代价高昂的批量失败。此外，hPSC来源的CMS缺乏成熟成人CMS的关键质量属性(CQA)。为了解决这些制造限制，研究小组将设计和评估一种受发育启发的工艺，该工艺采用CMS和心外膜细胞(EPICS)的集成制造，后者在心脏发育过程中提供营养因子。该项目将测试EPICS和CMS共分化将产生的心肌细胞比单独分化的CMS以更健壮、更可扩展的方式表现出关键成熟度CQA的假设。研究计划安排在三个目标下：(1)比较CMS和EPIC集成制造与单一培养制造作为差异化规模的函数的稳健性；(2)量化CMS和EPIC集成制造对获得有效性CQA的影响；以及(3)为CMS和EPIC集成制造开发可扩展的悬浮过程。该项目将更好地了解hPSC在CMS和EPIC的集成制造过程中与发育相关的内源性跨细胞通信。结果将建立新的原则，说明这种串扰如何随着制造过程的规模影响心脏细胞分化的批次到批次的稳健性。此外，这项研究将确定EPIC-CM在集成制造过程中的交互作用如何影响CM和EPIC产品的CQA。

PI: Palecek, Sean P.Proposal: 1743346Cardiac disease is a leading cause of death and represents a major burden on the health care system. Patients who experience a heart attack or heart failure exhibit a permanent loss in mechanical function in their hearts, reducing quality of life and increasing the risk of later medical complications. Today there is no treatment to restore mechanical function of the heart, but the transplantation of heart muscle cells manufactured from stem cells represents a promising approach in preclinical studies and clinical trials. A current limitation of stem cell-derived heart muscle cells is they generate forces and transmit electrical currents in a manner more similar to fetal cells than adult cells. We propose to explore the notion that manufacturing heart muscle cells in conjunction with stem cell-derived epicardial cells, which provide a supportive role during heart development, will enhance the functional quality of the stem cell-derived heart cells. This project will explore whether this developmentally relevant exchange of chemical factors or direct contact between the heart muscle and epicardial cells from stem cells will improve heart muscle cell function, then use these finding to design strategies to manufacture the approximately one billion heart muscle cells necessary to replace the heart cells lost during a heart attack. This project will enable the manufacturing of sufficient quantities of high quality heart muscle cells from stem cells to facilitate preclinical and clinical studies to restore heart function in persons who have experienced heart disease. This project also addresses the lack of a trained workforce in cell therapy manufacturing. Project activities will provide unique training opportunities to undergraduates and graduate students to perform manufacturing research in partnership with industry. The project team will also develop a multi-institutional course in cell therapy manufacturing, create outreach activities related to stem cell therapies for K12 students, and refine an international short course on regenerative manufacturing targeted to graduate students and industry employees.The next generation of therapeutics will involve living cells with the capacity to regenerate damaged or diseased human tissues. The US is poised to become a leader in cell therapies, but significant challenges in manufacturing complex living cells must be overcome. In this project we will address key roadblocks facing the cell therapy manufacturing industry including (1) lack of robust, scalable manufacturing platforms, (2) lack of knowledge of how manufacturing affects critical quality attributes (CQAs) of potency, and (3) the unavailability of a trained workforce. The project focuses on robust, scalable, and cost-effective manufacturing of safe and potent therapeutic cardiac cells from human pluripotent stem cells (hPSCs). hPSC-derived cardiomyocytes (CMs) have demonstrated improvement of ventricular contractile function in preclinical animal models. Success of impending clinical trials and development of effective human therapies will require production of high quality cells at a reasonable cost. However, scaleup of CM manufacturing reduces differentiation robustness, leading to expensive batch failures. Also, hPSC-derived CMs lack critical quality attributes (CQAs) of mature adult CMs. To address these manufacturing limitations the research team will design and evaluate a developmentally-inspired process that employs integrated manufacturing of CMs and epicardial cells (EpiCs), which provide trophic factors during heart development. This project will test the hypothesis that co-differentiation of EpiCs with CMs will generate cardiomyocytes that exhibit key maturity CQAs in a more robust, scalable manner than CMs differentiated alone. The Research Plan is organized under 3 objectives: (1) Compare robustness of integrated manufacturing of CMs and EpiCs with monoculture manufacturing as a function of differentiation scale; (2) Quantify effects of integrated manufacturing of CMs and EpiCs on acquisition of potency CQAs; and (3) Develop a scalable suspension process for integrated manufacturing of CMs and EpiCs. This project will provide a better understanding of developmentally-relevant endogenous, cross-cellular communication during integrated manufacturing of CMs and EpiCs from hPSCs. Outcomes will establish new principles for how this cross-talk affects batch-to-batch robustness of cardiac cell differentiation as the manufacturing process scales. Furthermore, this study will identify how EpiC-CM interactions during integrated manufacturing affect CQAs of both CM and EpiC products.