Synthetic Antifreeze Glycoproteins for Cellular Cryopreservation

摘要：生活在极地地区的动物已经适应了在寒冷的环境中生存。它们的策略之一是产生专门的抗冻蛋白，阻止血液和细胞中的冰形成。这些蛋白质可以防止冰晶造成的伤害和死亡。当组织和细胞因医学和研究目的而冷冻时，会添加化学物质以防止冰的形成和造成损害。然而，这些化学物质可能对干细胞等对医学有重要意义的细胞有害，因此需要更好的策略。这个研究项目的目标是创造出像动物体内的防冻蛋白一样工作的高分子材料。将对这些材料进行测试，看看它们是否能阻止冰晶的形成，以及细胞是否能在这些材料存在的情况下存活。我们还将检查是否有任何毒性作用或基因变化。作为该研究项目的一部分，本科生和研究生将接受培训，所有研究人员将参与社区外展工作。这将包括为女性和代表性不足的群体提供支持和培训，以鼓励她们从事STEM领域的职业。技术摘要：寒冷气候下的极端生物产生专门的抗冻糖蛋白（AFGPs）来保护其组织免受冻害。这些蛋白在预防细胞和组织的低温保存损伤的研究和再生医学应用中显示出巨大的潜力。然而，afgp在结构上是异质的，只能从极地鱼类的血液中少量分离出来。该项目的总体目标是生成能够捕获天然afgp的化学和生物物理特性的变革性材料，并探测其冰结合和低温保存活性。这将通过开发可扩展和可调的sAFGP结构变体合成路线，并研究其冰结合、冰成型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力和功能，细胞定位，任何遗传变化和诱导代谢停滞。此外，我们的目标是将EDI原则纳入生物材料和糖科学的教育和指导，在研究实验室，课堂和社区外展。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。

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.