FMSG: Bio: Advancing Extracellular Vesicle Biomanufacturing of CRISPR-Edited Human iPSC-derived MSCs with Next-Generation Purification

FMSG：生物：利用下一代纯化技术推进 CRISPR 编辑的人 iPSC 衍生的 MSC 的细胞外囊泡生物制造

• 批准号: 2229111
• 负责人: Zhen Ma
• 金额: $ 50万
• 依托单位: Syracuse University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-01-01 至 2024-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2229111&HistoricalAwards=false
• 关键词: FMSG; Bio; Advancing; Extracellular; Vesicle

Extracellular vesicles (EVs) produced by mesenchymal stem cells (MSCs) have been highlighted for their multifaceted therapeutic potentials. However, a great challenge of practical applications of EV therapy is the low production of EVs from therapeutic stem cells. To develop an advanced EV biomanufacturing process to meet the clinical need, this project aims to enable scale-up production of MSC-derived EVs by integrating human induced pluripotent stem cells (hiPSCs) for scalability in donor cell source, genome engineering for scalability in EV biogenesis, and advanced nano-membrane technology for scalability in EV purification. Furthermore, this project is designed to integrate research, education, industry, and diversity with an emphasis on strengthening research exposure to the students at multiple levels. It will be achieved by developing new collaborative classes across two campuses, providing research opportunities to graduate and undergraduate students, and leveraging existing programs at both institutes for outreach activities. The close industrial collaboration will also present industry internship opportunities to the graduate students.This project not only addresses the scalable issues in donor stem cell sources for therapeutic EV biomanufacturing, but also provides new insights to the field of EV biology and new solutions to the bottleneck of EV purification. Herein, this research brings together an interdisciplinary research team to (1) generate a stable hiPSC line with synthetic boosters of EV biogenesis and bioactivity, (2) establish a serum-free differentiation of perinatal tissue-like iMSCs as a potent EV factory, (3) optimize chemical priming conditions to enhance cell metabolic activities for boosting the EV production, and (4) introduce the nanopocket membrane technique for scalable EV purification with improved throughput and yield, comparing to the other standard techniques. Integration of hiPSC technology, genome engineering and membrane nanofabrication offers great potential to produce therapeutic EVs with high scalability, desired functions, and clinical relevance for future advancement of EV biomanufacturing.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

由间充质干细胞（MSC）产生的细胞外囊泡（EV）已因其多方面的治疗潜力而突出显示。然而，电动汽车疗法实际应用的巨大挑战是从治疗性干细胞中产生的电动汽车的产生较低。为了开发出一种满足临床需求的先进的EV生物制造过程，该项目旨在通过整合人类诱导的多能干细胞（HIPSC）来实现MSC衍生的电动汽车的扩展生产，以供供体细胞源的可伸缩性，基因组工程，以实现EV生物学的可伸缩性，以及高级纳米 - 膜技术的可伸缩能力，以伸缩EV含量。此外，该项目旨在整合研究，教育，行业和多样性，重点是加强对多个级别的学生的研究。这将是通过在两个校园中开发新的合作课程，为研究生和本科生提供研究机会，并利用两家学院的现有计划进行外展活动。紧密的工业合作还将向研究生展示行业实习机会。该项目不仅解决了供体干细胞源的可扩展问题的治疗性EV生物制造，而且还为EV生物学领域提供了新的见解，并为EV纯化的瓶颈提供了新的解决方案。在此，这项研究将一个跨学科研究团队汇集到（1）与eV生物发生和生物活性的合成助力器产生稳定的HIPSC线，（2）建立无血清组织的无血清分化，将类似于周围的IMSC作为有效的EV工厂，（3）为启动的化学启动条件，以增强EV的启动eV的作品（4），并启动EV型（4），并启用了NEB型（4），并启用了启动的NED（4），并构成了启动的NEB启用，并构成了启动的NEB启动，并构成了稳定的启动型号（4），并构成了启动的启动型号（4），并构成了启动的启动。与其他标准技术相比，用于可扩展的EV纯化，并改善吞吐量和产量。 Integration of hiPSC technology, genome engineering and membrane nanofabrication offers great potential to produce therapeutic EVs with high scalability, desired functions, and clinical relevance for future advancement of EV biomanufacturing.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.