Manufacturing USA: Biomanufacturing of Cell Microparticles from Stem Cells As Novel Thrombotic Therapeutic Agents

细胞和基因疗法可用于治疗多种疾病。治疗的基础是使用全细胞或病毒将基因或蛋白质传递给患者。膜囊又称胞外囊泡(EVS)，是一种很有前途的新型给药系统。它们可以针对特定类型的细胞运送货物，并可以携带蛋白质、脂肪和核糖核酸。该项目将开发基于骨髓中产生血小板的特殊细胞的电动汽车技术。这项技术的发展可以通过提供稳定和丰富的血小板来源，为解决输血医学中长期存在的问题带来新的战略。来自代表性不足群体的高中生和大学生将参与研究经验，以激发他们对工程和生物制造职业的兴趣。该项目的目标是发展基础科学和工程学，以制备功能性胞外囊泡(EVS)，从而实现新的细胞和基因治疗。EVS是一种亚微米膜泡，携带来自其母细胞的RNA、蛋白质和脂类。细胞使用EV与其他细胞交流，通过其内容物传递信号，并通过基于表面蛋白的受体识别机制靶向相当有限的细胞类型。现在人们普遍认为，细胞外小泡是实现安全和有效的细胞和基因治疗的极佳候选者。该项目将专注于巨核细胞微粒(MPS；一种细胞外小泡)，它可以靶向造血干细胞和祖细胞(HSPC)并向其运送货物。这些MPS最重要的特征是它们专门针对HSPC，并对它们进行编程，以产生更多的巨核细胞，即血小板的前体。该项目将开发这些MP的表征方法和分析方法，以便它们可以用于开发产生大量MP的可靠协议。最后，该项目还将使用一个简单的小鼠模型来评估这些MPS作为血栓剂的生物学效果。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

Cell and gene therapies can be used to treat a large array of diseases. Therapy is based on using whole cells or viruses to deliver genes or proteins to the patient. Membrane sacs, known as extracellular vesicles (EVs), are a promising new delivery system. They can target specific types of cells for delivery of their cargo, and can carry proteins, fats and ribonucleic acids. This project will develop EV technology based on specialized cells in the bone marrow that generate platelets. Development of this technology could lead to new strategies for solving long-standing problems in Transfusion Medicine by providing a stable and abundant source of platelets. High school and college students from underrepresented groups will engage in research experiences to stimulate their interest in engineering and biomanufacturing careers. The goal of this project is to develop foundational science and engineering for preparing functional extracellular vesicles (EVs) to enable novel cell and gene therapies. EVs are submicron membrane vesicles that carry RNAs, proteins and lipids from their parent cells. Cells use EVs to communicate with other cells, delivering signals through their content, and targeting a rather narrow range of cell types through receptor-recognition mechanisms based on surface proteins. It is now widely accepted that extracellular vesicles are excellent candidates for enabling safe and potent cell and gene therapies. This project will focus on megakaryocytic microparticles (MPs; a type of extracellular vesicles), which target and deliver cargo to hematopoietic stem & progenitor cells (HSPCs). The most important feature of these MPs is that they specifically target HSPCs and program them to generate more megakaryocytes, the precursors of platelets. This project will develop characterization methods and assays for these MPs so that they can be used to develop robust protocols for generating large MP quantities. Finally, the project will also assess the biological efficacy of these MPs as thrombotic agents using a simple murine model.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.