Synthetic Antifreeze Glycoproteins for Cellular Cryopreservation

非技术摘要：生活在极地地区的动物已经适应了冰冻的条件。他们的策略之一是生产专门的抗冻蛋白，阻止他们的血液和细胞中形成冰。这些蛋白质可以防止冰晶造成的损害和死亡。当出于医疗和研究目的而冷冻组织和细胞时，会添加化学物质来防止冰的形成和造成损害。然而，这些化学物质可能会对医学上重要的细胞有害，例如干细胞，因此需要更好的策略。这项研究项目的目标是创造出功能类似于动物体内的抗冻蛋白的聚合物材料。这些材料将接受测试，看看它们是否可以阻止冰晶的形成，以及细胞是否可以在这些材料的存在下被冷冻存活。我们还将检查是否有任何毒性影响或基因变化。作为这一研究项目的一部分，本科生和研究生将接受培训，所有调查人员都将参与社区外展工作。这将包括对女性和代表性不足的群体的支持和培训，以鼓励她们在STEM中追求职业生涯。技术摘要：寒冷气候中的极端微生物产生专门的抗冻糖蛋白(AFGP)，以保护他们的组织免受冻害。随着研究和再生医学的应用，这些蛋白质在预防细胞和组织的冷冻损伤方面显示出巨大的潜力。然而，AFGP在结构上是不同的，只能从北极鱼的血液中分离出少量的AFGP。该项目的总体目标是产生能够捕捉天然AFGP的化学和生物物理性质的变革性材料，并探索其结合冰和冷冻保存的活动。这将通过开发一种可扩展和可调的sAFGP结构变体的合成路线并研究其冰结合、冰成形和膜稳定性能来实现。我们还将检查生物活性，包括冷冻保存后的细胞活性和功能，细胞定位，任何遗传变化，以及代谢停滞的诱导。此外，我们的目标是将EDI原则纳入研究实验室、课堂和社区外展的生物材料和糖科学教育和指导中。该奖项反映了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。

Non-technical Abstract:Animals living in polar regions have adapted to survive freezing conditions. One of their strategies is to produce specialized antifreeze proteins that stop ice from forming in their blood and cells. These proteins prevent damage and death caused by ice crystals. When tissues and cells are frozen for medical and research purposes, chemicals are added to prevent ice from forming and causing damage. However, these chemicals can be harmful to cells that are important for medical purposes, such as stem cells, and there is a need for better strategies. The goal of this research project is to create polymer materials that work like the antifreeze proteins found in animals. The materials will be tested to see if they can stop ice crystals from forming and whether cells can survive being frozen in the presence of these materials. We will also examine if there are any toxic effects or genetic changes. As part of this research project, undergraduate and graduate students will be trained and all investigators will participate in community outreach efforts. This will include support and training efforts to females and underrepresented groups to encourage them to pursue careers in STEM.Technical Abstract:Extremophile organisms in cold climates produce specialized antifreeze glycoproteins (AFGPs) to defend their tissues against freezing damage. These proteins have shown great potential for prevention of cryopreservation damage of cells and tissues with research and regenerative medicine applications. However, AFGPs are heterogeneous in structure and can only be isolated in small quantities from the blood of polar fish. The overall goals of this project are to generate transformative materials that capture the chemical and biophysical properties of natural AFGPs and probe their ice binding and cryopreservation activity. This will be achieved by developing a scalable and tunable synthetic route to sAFGP structural variants and studying their ice binding, ice shaping, and membrane stabilizing properties. We will also examine bioactivity, including cellular viability and function after cryopreservation, cellular localization, any genetic changes, and induction of metabolic stasis. Additionally, we aim to incorporate EDI principles into biomaterials and glycoscience education and mentoring in the research lab, the classroom, and in community outreach.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.