Collaborative Research: NSF-BSF: Somatic cell adaptation towards immortalization in a marine tunicate

合作研究：NSF-BSF：海洋被囊动物体细胞对永生的适应

• 批准号: 2127516
• 负责人: Dietmar Kültz
• 金额: $ 98.06万
• 依托单位: University of California-Davis
• 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: 2021-08-01 至 2025-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2127516&HistoricalAwards=false
• 关键词: Collaborative; Research; NSF; BSF; Somatic

The research investigates how cells from the marine colonial tunicate Botryllus schlosseri evolve when critical molecular networks that control cell growth and programmed cell death are disturbed by either stress or genetic manipulation or a combination thereof. The main question addressed is: What molecular mechanisms constrain cell immortalization? Immortal cell lines of marine invertebrates have great potential as “bioreactors” for producing pharmaceutical compounds with health benefits, nutraceuticals, anticancer drugs, and other biologically active chemicals. The tunicate B. schlosseri is widely distributed throughout the world and used as a common sentinel species for assessing marine pollution and other anthropogenic impacts on coastal ecosystems. The research contributes molecular tools that increases the utility of this tunicate for bioindication. Opportunities for cross-disciplinary training, curricular, and professional development are provided to students and researchers at all career stages with emphasis on underrepresented minorities.The main hypothesis addressed in this project is that specific evolutionary constraints prevent somatic cell immortalization in marine invertebrates and that cell immortalization is facilitated by stress-induced evolution and/or genetic manipulation that removes these constraints. The project investigates the conditions and gene regulatory networks that promote cell proliferation and counteract cellular senescence. Primary cultures of B. schlosseri hemocytes and epithelial monolayers are exposed to (1) environmental factors (media supplements and attachment substrates), (2) stress-induced evolution, (3) synthetic manipulation of pro- and anti-proliferative genes, and (4) cell fusion technology to increase heritable variation in proteome networks and cellular phenotypes that natural selection can act on. The goal is to a) understand the sequence of molecular events that promotes cell proliferation and inhibits senescence in vitro and b) to generate the first cell line for any marine invertebrate. This project utilizes systems biology approaches (transcriptomics, proteomics) to characterize how relevant cellular phenotypes (proliferation, senescence) are controlled by gene regulatory networks. Complex datasets are analyzed using network modeling and topological data analysis approaches. Moreover, the extent of mRNA: protein coregulation during transition of cells from in vivo to in vitro contexts provides insight into the degree of non-linearity between mRNA and protein levels of regulation in marine invertebrate cells exposed to environmental stress and synthetic genetic backgrounds.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.

该研究调查了当控制细胞生长和程序性细胞死亡的关键分子网络受到压力或基因操作或其组合的干扰时，来自海洋殖民地被囊动物 Botryllus schlosseri 的细胞如何进化。解决的主要问题是：什么分子机制限制细胞永生化？海洋无脊椎动物的永生细胞系作为“生物反应器”具有巨大的潜力，可用于生产具有健康益处的药物化合物、营养保健品、抗癌药物和其他生物活性化学品。被囊动物 B. schlosseri 广泛分布于世界各地，并被用作评估海洋污染和其他人为对沿海生态系统影响的常见哨兵物种。该研究提供了分子工具，提高了这种被囊动物的生物指示效用。为各个职业阶段的学生和研究人员提供跨学科培训、课程和专业发展的机会，重点关注代表性不足的少数群体。该项目提出的主要假设是，特定的进化限制阻止了海洋无脊椎动物的体细胞永生化，而压力诱导的进化和/或消除这些限制的基因操作促进了细胞永生化。该项目研究促进细胞增殖和抵抗细胞衰老的条件和基因调控网络。史氏芽孢杆菌血细胞和上皮单层的原代培养物暴露于（1）环境因素（培养基补充剂和附着底物），（2）应激诱导的进化，（3）促增殖和抗增殖基因的合成操作，以及（4）细胞融合技术，以增加自然选择可以作用的蛋白质组网络和细胞表型的可遗传变异。目标是a）了解体外促进细胞增殖和抑制衰老的分子事件序列，b）为任何海洋无脊椎动物生成第一个细胞系。该项目利用系统生物学方法（转录组学、蛋白质组学）来表征相关细胞表型（增殖、衰老）如何受基因调控网络控制。使用网络建模和拓扑数据分析方法来分析复杂的数据集。此外，细胞从体内环境过渡到体外环境期间的 mRNA：蛋白质共调节程度可以深入了解暴露于环境压力和合成遗传背景的海洋无脊椎动物细胞中 mRNA 和蛋白质调节水平之间的非线性程度。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力价值和更广泛的影响审查进行评估，被认为值得支持 标准。