Delineation factors governing production of multi-component cytokines in clinical grade cells

控制临床级细胞中多组分细胞因子产生的描绘因素

• 批准号: 2041107
• 负责人: Michael Diehl
• 金额: $ 40万
• 依托单位: William Marsh Rice University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2041107&HistoricalAwards=false
• 关键词: Delineation; factors; governing; production; multi

Mammalian cells can produce therapeutic molecules known as biologics. Biologics can be used to treat cancer, auto-immune disorders and infectious viral disease. This project will address critical gaps in the ability engineer mammalian cells. These gaps limit cell therapy applications where cells are delivered to specific disease sites in the body in order to facilitate local biologic production. New methods to engineer clinical-grade cells for these types of applications will be developed. Complex biologics that are composed of multiple protein complexes will be the engineered products of these cells. Undergraduates, especially those from underrepresented minorities, will be trained through participation in the research.The main goals of this project are to identify and develop biosynthetic routes to circumvent bottlenecks that limit the secretion levels of fully-bioactive macromolecules. Multi-fragment, pro-inflammatory cytokines and clinical-grade cell lines will be used as model biologic and cell platforms. Protein expression will be manipulated precisely to identify bottlenecks that affect cytokine synthesis and heterodimerization. Multiple-gene manipulation strategies will be used to modulate the expression of additional genes in order to probe downstream secretion bottlenecks. Processes of particularly interest are proteolytic protein processing, folding, and vesicular trafficking. These experiments will test the overarching hypothesis that maximized secretion of multi-fragment proteins requires precision multi-gene translational control combined with the upregulation of multiple, downstream effector proteins along the secretion pathway. Engineered cells will be encapsulated in biopolymers. Their secretion stability and robustness will be evaluated. The extent to which these changes are linked to endoplasmic reticulum driven unfolded protein responses (UPR) is of primary interest.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.

哺乳动物细胞可以产生被称为生物制剂的治疗分子。生物制剂可用于治疗癌症、自身免疫性疾病和传染性病毒性疾病。该项目将解决在能力工程哺乳动物细胞的关键差距。这些差距限制了细胞治疗应用，其中细胞被递送到体内的特定疾病部位，以促进局部生物生产。将开发用于这些类型应用的临床级细胞的新方法。由多种蛋白质复合物组成的复杂生物制剂将是这些细胞的工程产品。本科生，特别是那些来自代表性不足的少数民族，将通过参与研究进行培训。该项目的主要目标是确定和开发生物合成路线，以绕过限制完全生物活性大分子分泌水平的瓶颈。多片段促炎细胞因子和临床级细胞系将用作模型生物和细胞平台。蛋白质表达将被精确操纵，以确定影响细胞因子合成和异源二聚化的瓶颈。多基因操作策略将用于调节其他基因的表达，以探测下游分泌瓶颈。特别感兴趣的过程是蛋白水解蛋白质加工、折叠和囊泡运输。这些实验将测试总体假设，即多片段蛋白质的最大化分泌需要精确的多基因翻译控制与沿着分泌途径的多个下游效应蛋白沿着的上调相结合。工程细胞将被封装在生物聚合物中。将评价其分泌稳定性和耐用性。 这些变化在多大程度上与内质网驱动的未折叠蛋白反应（UPR）有关是主要的兴趣。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识价值和更广泛的影响审查标准进行评估来支持。

项目成果

Development of Serum-Free Media for Cryopreservation of Hydrogel Encapsulated Cell-Based Therapeutics

开发用于水凝胶封装的细胞疗法冷冻保存的无血清培养基

• DOI: 10.1007/s12195-022-00739-7
• 发表时间: 2022
• 期刊: Cellular and Molecular Bioengineering
• 影响因子: 2.8
• 作者: Cui, Yufei;Nash, Amanda M.;Castillo, Bertha;Sanchez Solis, Leonardo D.;Aghlara-Fotovat, Samira;Levitan, Maya;Kim, Boram;Diehl, Michael;Veiseh, Omid
• 通讯作者: Veiseh, Omid