Integrated Manufacturing of Therapeutic Cardiac Cells

PI：Palecek，Sean P.提案：1743346心脏病是导致死亡的主要原因，是卫生保健系统的主要负担。 经历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后期医疗并发症的风险。 今天，没有恢复心脏机械功能的治疗方法，但从干细胞制造的心肌细胞移植代表了临床前研究和临床试验中有前途的方法。 干细胞衍生的心肌细胞目前的局限性是它们产生力并以比成人细胞更类似于胎儿细胞的方式传输电流。 我们建议探索的概念，即制造心肌细胞与干细胞衍生的心外膜细胞，这在心脏发育过程中提供了支持作用，将提高干细胞衍生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心肌与干细胞心外膜细胞之间的直接接触是否会改善心肌细胞功能，然后利用这些发现设计策略来制造大约10亿个心肌细胞，以取代心脏病发作期间失去的心脏细胞。该项目将能够从干细胞中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业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，与工业界合作进行制造业研究。该项目团队还将开发一个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，并完善一个针对研究生和行业员工的再生制造国际短期课程。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，但必须克服制造复杂活细胞的重大挑战。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（hPSC）中生产安全有效的治疗性心脏细胞。hPSC衍生的心肌细胞（CM）已经在临床前动物模型中证明了心室收缩功能的改善。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，导致昂贵的批次失败。此外，hPSC衍生的CM缺乏成熟成人CM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开发灵感的过程，该过程采用CM和心外膜细胞（EpiCs）的集成制造，这些细胞在心脏发育期间提供营养因子。该项目将测试EpiCs与CM的共分化将产生心肌细胞的假设，这些心肌细胞比单独分化的CM以更稳健、可扩展的方式表现出关键的成熟CQA。 研究计划根据3个目标组织：（1）比较CM和EpiC集成生产与单一培养生产的稳健性（作为差异化规模的函数）;（2）量化CM和EpiC集成生产对获得效价CQA的影响;和（3）开发CM和EpiC集成生产的可扩展混悬液工艺。该项目将更好地了解在从hPSC集成制造CM和EpiC期间与发育相关的内源性跨细胞通信。结果将建立新的原则，这种串扰如何影响批次间的稳定性的心脏细胞分化的制造工艺规模。此外，本研究将确定集成制造过程中EpiC-CM相互作用如何影响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.