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: 2127517
• 负责人: Alison Gardell
• 金额: $ 49.99万
• 依托单位: University of Washington
• 依托单位国家: 美国
• 项目类别: Standard 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=2127517&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遗传操作或其组合的干扰时，海洋殖民被囊动物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具有作为“生物反应器”的巨大潜力，用于生产具有健康益处的药物化合物、营养品、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活动对沿海生态系统影响的共同哨兵物种。这项研究提供了分子工具，增加了这种被囊动物的生物指示作用。跨学科的培训，课程和专业发展的机会，提供给学生和研究人员在所有的职业生涯阶段，重点是代表性不足的minorities.The主要假设在这个项目中解决的是，特定的进化限制，防止体细胞永生化的海洋无脊椎动物和细胞永生化是由压力诱导的进化和/或遗传操作，消除这些限制。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。B的原代培养物。Schlosseri血细胞和上皮细胞单层暴露于（1）环境因素（培养基补充物和附着基质），（2）胁迫诱导的进化，（3）促增殖基因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中的可遗传变异。目标是a）理解在体外促进细胞增殖和抑制衰老的分子事件的顺序和B）产生任何海洋无脊椎动物的第一个细胞系。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）如何由基因调控网络控制。复杂数据集的分析使用网络建模和拓扑数据分析方法。此外，mRNA的程度：在细胞从体内到体外环境的转变过程中的蛋白质共调节提供了对非-海洋无脊椎动物细胞暴露于环境压力和合成遗传背景下的mRNA和蛋白质水平调节之间的线性关系。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进行评估，被认为值得支持审查标准。