FMSG: Enabling Technologies for Biomanufacturing EV-Based Therapeutics

FMSG：基于 EV 的生物制造治疗的使能技术

• 批准号: 2036809
• 负责人: Jamey Young
• 金额: $ 50万
• 依托单位: Vanderbilt University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-15 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2036809&HistoricalAwards=false
• 关键词: FMSG; Enabling; Technologies; Biomanufacturing; EV

Extracellular vesicles (EVs) are cell-made particles that provide a natural mechanism of information and material transfer between cells. There is growing interest in large-scale production of EVs that can be used as therapeutics due to their ability to communicate signals from producer cells and their potential use as carriers for delivery of drug molecules. EV therapeutics are being developed for treatment of a wide range of diseases including metabolic disorders, cancer, and neurodegeneration. Despite the excitement generated by several early-stage EV biotech companies, the technology to produce mass quantities of purified EVs with tunable properties is still in its infancy. The long-term goal of this project is to enable production of "designer EVs" that can be packaged with desired cargo molecules, decorated with tunable surface ligands, and secreted in high yield from specific producer cell types. The overall objective of the current project is to identify cellular processes that can be engineered to control the production, content, and in vivo trafficking of therapeutic EVs. Making this new class of drugs available to the public has potential to improve the health and quality of life of millions of patients in the US and around the world. The project will also provide the unique educational opportunity for trainees to engage in collaborative research with industry scientists. Other broader impacts will be accomplished through engaging undergraduate and high-school students in EV research, and through integration of the project with the Vanderbilt Program for EV Research and bioengineering courses led by the PIs. The investigators will focus their research program on loading and delivery of small RNAs, which are promising drug molecules that are not delivered efficiently to recipient cells using established nanoparticle-based carriers. The central hypothesis is that developing tissue-specific producer cells with tuned expression of EV-associated proteins and RNAs will enable researchers to maximize the product yield of specific EVs and the efficiency of RNA delivery from EVs to recipient cells. First, they will boost targeting of miRNAs to extracellular vesicles through modulating specific molecules at ER-contact sites. Second, they will optimize cargo delivery to target cells by engineering EVs for efficient endosomal escape and macrophage evasion properties. Third, they will develop a scalable platform for manufacturing tissue-specific EVs by differentiating induced pluripotent stem cells (iPSCs) to producer cells in 3D suspension cultures. The proposed research is innovative because it applies a multidisciplinary approach to address several critical barriers to commercial production of therapeutic EVs: cargo loading, cargo delivery, and scalable manufacturing. EVs secreted by MSCs are recognized as a viable alternative to overcome the potential risks from transplantation of primary MSCs. By developing the tools and strategies to customize and maximize EV production from iPSC-derived MSCs, the researchers will establish a flexible EV manufacturing platform that can expand to other relevant organ and tissue systems.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.

细胞外囊泡（EV）是细胞制成的颗粒，可提供自然的信息和材料转移的机理。由于能够从生产细胞中传达信号及其作为药物分子递送的携带者的潜在用途，因此人们对EV的大规模生产的兴趣日益增加。正在开发EV疗法，用于治疗多种疾病，包括代谢性疾病，癌症和神经退行性。尽管几家早期EV生物技术公司引起了人们的兴奋，但生产具有可调性能的纯净电动汽车的技术仍处于起步阶段。该项目的长期目标是实现可以用所需的货物分子包装的“设计师电动汽车”，并装饰有可调表面配体，并以高产量分泌于特定的生产者细胞类型。当前项目的总体目的是确定可以设计以控制治疗性电动汽车的生产，内容和体内运输的细胞过程。 使这类新的药物可供公众使用，有可能改善美国和世界各地数百万患者的健康和生活质量。该项目还将为学员提供与行业科学家进行合作研究的独特教育机会。通过使本科生和高中生参与EV研究，以及将项目与PIS领导的EV Research and Bioworgineering课程与Vanderbilt计划的整合，将实现其他更广泛的影响。 研究人员将把他们的研究计划集中在小型RNA的加载和递送上，这是有希望的药物分子，使用既定的纳米粒子载体没有有效地将其提供给受体细胞。中心假设是，开发具有EV相关蛋白质和RNA的调谐表达的组织特异性生产细胞将使研究人员能够最大程度地提高特定EV的产物产量以及RNA从EV到受体细胞的RNA递送效率。首先，它们将通过调节ER接触位点的特定分子来增强miRNA对细胞外囊泡的靶向。其次，他们将通过工程电动汽车来优化货物向目标细胞的运输，以实现有效的内体逃逸和巨噬细胞逃避特性。第三，他们将通过将诱导的多能干细胞（IPSC）与3D悬浮培养物中的生产细胞区分开来开发一个可扩展的平台，用于制造组织特异性EV。拟议的研究具有创新性，因为它采用了多学科方法来解决治疗性电动汽车商业生产的几个关键障碍：货物装载，货物交付和可扩展的制造业。 MSC分泌的EV被认为是克服原发性MSC移植的潜在风险的可行替代方法。通过开发从IPSC衍生的MSC自定义和最大化电动汽车生产的工具和策略，研究人员将建立一个灵活的EV制造平台，可以扩展到其他相关的器官和组织系统。该奖项反映了NSF的法定任务，并认为通过使用该基金会的知识分子和更广泛的影响来评估CRITERIA CRITERIA CRITERIA。

项目成果

Manufactured extracellular vesicles as human therapeutics: challenges, advances, and opportunities

• DOI: 10.1016/j.copbio.2022.102776
• 发表时间: 2022-08-27
• 期刊: CURRENT OPINION IN BIOTECHNOLOGY
• 影响因子: 7.7
• 作者: Estes, Scott;Konstantinov, Konstantin;Young, Jamey D.
• 通讯作者: Young, Jamey D.

Chemical and Biomolecular Strategies for STING Pathway Activation in Cancer Immunotherapy.

• DOI: 10.1021/acs.chemrev.1c00750
• 发表时间: 2022-03-23
• 期刊: Chemical reviews
• 影响因子: 62.1
• 作者: Garland KM;Sheehy TL;Wilson JT
• 通讯作者: Wilson JT

VAP-A and its binding partner CERT drive biogenesis of RNA-containing extracellular vesicles at ER membrane contact sites.

• DOI: 10.1016/j.devcel.2022.03.012
• 发表时间: 2022-04-25
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Barman, Bahnisikha;Sung, Bong Hwan;Krystofiak, Evan;Ping, Jie;Ramirez, Marisol;Millis, Bryan;Allen, Ryan;Prasad, Nripesh;Chetyrkin, Sergei;Calcutt, M. Wade;Vickers, Kasey;Patton, James G.;Liu, Qi;Weaver, Alissa M.
• 通讯作者: Weaver, Alissa M.