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

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两者组合的干扰时，海洋种群被囊动物Botryllus schlosseri细胞是如何进化的。主要的问题是：什么分子机制限制了细胞的永生？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，可用于生产具有保健作用的药物化合物、营养品、抗癌药物和其他生物活性化学物质。被囊贝在世界各地广泛分布，是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，增加了这种被囊生物适应症的效用。为处于所有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，重点是代表性不足的少数民族。本项目提出的主要假设是，特定的进化限制阻碍了海洋无脊椎动物的体细胞永生，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。该项目研究促进细胞增殖和对抗细胞衰老的条件和基因调控网络。schlos螺旋体血细胞和上皮单层的原代培养暴露于(1)环境因素（培养基补充和附着底物），(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遗传变异。目标是a)了解促进细胞增殖和体外抑制衰老的分子事件序列，b)为任何海洋无脊椎动物产生第一个细胞系。该项目利用系统生物学方法（转录组学、蛋白质组学）来表征基因调控网络如何控制相关的细胞表型（增殖、衰老）。使用网络建模和拓扑数据分析方法分析复杂数据集。此外，细胞从体内到体外过渡过程中mRNA：蛋白质协同调节的程度，可以深入了解暴露于环境应激和合成遗传背景下的海洋无脊椎动物细胞mRNA和蛋白质调节水平之间的非线性程度。该奖项反映了美国国家科学基金会的法定使命，并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，被认为值得支持。

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.