Understanding Ice Recrystallization Inhibitors - Design, Synthesis and Mechanism(s) of Action

了解冰重结晶抑制剂 - 设计、合成和作用机制

• 批准号: RGPIN-2017-06298
• 负责人: Ben, Robert
• 金额: $ 4.37万
• 依托单位: University of Ottawa
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2018
• 资助国家: 加拿大
• 起止时间: 2018-01-01 至 2019-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=645112
• 关键词: Understanding; Ice; Recrystallization; Inhibitors; Design

Cell-based therapeutics emerged as a critical aspect of modern healthcare in regenerative medicine and transfusion. Unfortunately, current cryopreservation protocols remain suboptimal and limit the effectiveness of such therapies. Loss of cell viability during cryopreservation has been shown to dramatically reduce potency of progenitor cells such as those used in hematopoietic stem cell (HSC) transplantation and red blood cell (RBC) transfusion resulting in poor engraftment or a need for additional transfusions. Recent studies have indicated that the most significant cause of decreased post-thaw cell viability and loss of function is cellular damage from ice recrystallization during freezing and thawing. A detailed understanding of this process is fundamental to the natural sciences since practical strategies to inhibit it are lacking. This multidisciplinary proposal will address this issue using in vitro approaches relying on advanced spectroscopic (confocal Raman) and modern cellular/biophyscial techniques (nanoparticle tracking, Ektacytometric analysis, cryomicroscopy etc.). Specific objectives are; 1) study and correlate the ability of our novel ice recrystallization inhibitors (IRIs) to their physical parameters (hydrophobicity, self-assembly and hydration properties) and 2) study the in vitro cellular effects of these molecules on cellular membranes, microparticle (MP) formation, nucleation of ice and the recrystallization of intracellular ice. The knowledge gained from understanding how these inhibitors function will significantly advance our understanding of ice recrystallization (both in the presence and absence of cells). This knowledge will be directly relevant to other processes such as biomineralization, and calcification and lead to the generation of novel methods to control these physiologically important processes.***Significance/Impact:***Our integrated approach of using in vitro strategies relying on advanced spectroscopic (confocal Raman) and modern cellular/biophyscial techniques (nanoparticle tracking, Ektacytometric analysis, cryomicroscopy etc.) to study small molecule IRIs will expedite an understanding of how these inhibitors function. In addition, the design of new and improved cryoprotectants is timely and will address many of the storage problems associated with technologies in the areas of regenerative medicine, cellular therapy and tissue engineering.

基于细胞的治疗方法成为再生医学和输血现代医疗保健的一个重要方面。不幸的是，目前的冷冻保存方案仍然是次优的，并限制了这种疗法的有效性。已经显示在冷冻保存期间细胞活力的丧失显著降低祖细胞的效力，所述祖细胞例如用于造血干细胞（HSC）移植和红细胞（RBC）输注的祖细胞，导致移植不良或需要额外的输注。最近的研究表明，解冻后细胞活力下降和功能丧失的最重要原因是冻融过程中冰重结晶造成的细胞损伤。对这一过程的详细了解是自然科学的基础，因为缺乏抑制它的实用策略。这个多学科的建议将解决这个问题，使用体外方法依赖于先进的光谱（共焦拉曼）和现代细胞/生物荧光技术（纳米粒子跟踪，Ektacytometric分析，低温显微镜等）。 具体目标是; 1）研究我们的新型冰重结晶抑制剂（IRI）的能力并将其与它们的物理参数（疏水性、自组装和水合性质）相关联; 2）研究这些分子对细胞膜、微粒（MP）形成、冰的成核和细胞内冰的重结晶的体外细胞效应。 从了解这些抑制剂如何发挥作用中获得的知识将大大促进我们对冰重结晶的理解（无论是在细胞存在还是不存在的情况下）。 这些知识将直接关系到其他过程，如生物矿化和钙化，并导致产生新的方法来控制这些重要的生理过程。重要性/影响：* 我们使用体外策略的综合方法依赖于先进的光谱（共焦拉曼）和现代细胞/生物荧光技术（纳米颗粒跟踪，Ektacytometric分析，低温显微镜等）。研究小分子IRI将加速对这些抑制剂如何起作用的理解。此外，新的和改进的冷冻保护剂的设计是及时的，并将解决与再生医学，细胞治疗和组织工程领域的技术相关的许多储存问题。