SBIR Phase I: Developing Nanopores to Facilitate Delivery of Intracellular Cryoprotectants for Biopreservation at Low Temperature

SBIR第一阶段项目旨在开发一种尖端的纳米技术，该技术将大大增强再生药物（如干细胞、复杂组织和器官）的生物保存。这种技术有可能永远改变再生医疗保健。它将改变按需细胞和组织的生物库，改善大规模创伤护理和先进的个性化医疗程序。在再生医学中，几乎在源材料的获取、分离、储存和最终产品运输到患者的所有层面都需要生物保存。然而，至关重要的是，该领域缺乏安全有效地保存这些组织和药物的能力，严重限制了产品的保质期。没有任何地方比器官移植更明显地缺乏生物库技术，供体和受体之间的时间窗口（4 - 7小时）不足以正确匹配捐赠，筛查病原体或运输距离。更多的人将死于过早的器官衰竭而不是癌症。 使美国能够在零度以下的温度下安全地储存器官将大大提高国家医疗保健水平。 美国面临着强大的商业和竞争力理由，投资于再生医学的各个方面，包括器官治疗。冷冻保存解决方案将间接为医疗保健系统、患者和医疗保险公司，从再生医学治疗中节省的成本估计接近100万美元。目前生物保存技术的细胞毒性主要与冷却期间低效的冷冻保护剂和水输送穿过细胞膜相关，导致不可修复的细胞损伤。冰形成的伤害。该项目的目标是通过开发该领域的第一个生物启发纳米孔作为跨膜巨型高速公路，以促进冷冻保存期间冷冻保护剂的安全有效的细胞内递送和去除，来建立一种根本不同的冷冻保护剂优化方法。过去的研究已经证明了通过基于多个氢键、偶极-偶极和芳香族π-π堆叠相互作用的相互作用以及它们自插入脂质双层的形状持久性大环的强制堆叠来构建定义明确的纳米管组装体的可靠性。这些合理设计的有机纳米孔将在蛋白质通道在或低于3 ° C时发生故障时作为选择性跨膜通道。因此，细胞达到无冰冷冻保存温度的暴露时间将显著缩短。通过减少细胞内冰的形成，保存后的细胞产量和活力将大大提高。在复温时，这些有机纳米孔将有助于快速去除冷冻保护剂。在生理温度下，纳米孔将封闭，并从系统中冲洗出来，导致低毒性。将使用基于脂质体的葡萄糖转运、基于细胞的毒性和基于细胞的冷冻保存测定来检查纳米孔功能和效果。

This SBIR Phase I project aims to develop a cutting-edge nanotechnology that will greatly enhance biological preservation of regenerative medicines such as stem cells, complex tissues, and organs. Such a technology has the potential to changing regenerative healthcare forever. It would change biobanking for on-demand cells and tissues and improve mass trauma care and advanced personalized medical procedures. Biopreservation is required in regenerative medicine at nearly all levels in the acquisition of source material, isolation, storage and shipment of a final product to patient. Yet, critically, the field lacks the ability to safely and efficiently preserve these tissues and medicines severely limiting product shelf life. Nowhere is the absence of a biobanking technology more palpable than organ transplantation, where the time window between donor and recipient (4-7 hours) is not enough to properly match donations, screen for pathogens, or transport distances. More people will die from premature organ failure than cancer. Enabling the United States to safely bank organs at subzero temperature will significantly enhance national healthcare. The US faces strong commercial and competitiveness reasons to invest in all facets of regenerative medicine, including organ therapies. Cryopreservation solutions would indirectly enable significant savings to the healthcare system, the patient, and healthcare insurance companies with the cost savings from regenerative medicine treatments estimated to be nearly $250 billion per year in the U.S. The cytotoxicity of current biopreservation techniques is largely associated with inefficient cryoprotective agent and water delivery across the cell membrane during cooling leading to irreparable cell damage from ice formation. The goal of this project is to establish a fundamentally different approach to cryoprotective agent optimization by developing first-in-the-field bioinspired nanopores as transmembrane mega highways to facilitate safe and efficient intracellular delivery and removal of cryoprotective agents during cryopreservation. Past research has demonstrated the reliability of constructing well-defined nanotubular assemblies via the enforced stacking of shape-persistent macrocycles based on the interplay of multiple hydrogen-bonding, dipole-dipole, and aromatic pi-pi stacking interactions and their self-insertion into lipid bilayers. These rationally designed organic nanopores will serve as selective transmembrane channels when protein channels malfunction at or below 3 °C. As a result, the cell's exposure time to reach ice-free cryopreservation temperature will be significantly reduced. Post-preservation cell yield and viability will be greatly improved by reducing intracellular ice formation. Upon rewarming, these organic nanopores will facilitate rapid removal of the cryoprotective agents. At physiological temperature, the nanopores will seal off, and be washed out from the system resulting in low toxicity. Nanopore function and effect will be examined using liposome-based glucose transport, cell-based toxicity and cell-based cryopreservation assays.