SBIR Phase II: Dynamic Nanopores Enabling Non-toxic Cryopreservation to Advance Centralized Manufacturing & Transport of Regenerative Medicines

这个小企业创新研究第二阶段项目将继续开发尖端纳米孔技术，这将极大地加强干细胞、复杂组织和器官等再生药物的生物保存。这种技术有可能极大地改善按需细胞和组织的医疗保健的生物库和运输基础设施，改进大规模创伤护理和先进的个性化医疗程序。在再生医学中，几乎所有级别的再生医学都需要生物保存，包括原始材料的获取、分离、储存和将最终产品运送给患者。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，同时在超低温保存后保持高细胞活性和功能，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，在器官移植中，捐赠者和接受者之间的时间窗口(4-7小时)不足以正确匹配捐赠、筛查病原体或远距离运输。使美国能够在零下的温度下安全地储存器官，将显著提高国民医疗保健水平。美国面临着强大的商业和竞争力理由，需要投资于再生医学的方方面面，包括器官疗法。冷冻保存解决方案将间接地为医疗保健系统、患者和医疗保险公司节省大量成本，在美国，再生医学治疗每年节省的成本估计接近2500亿美元。目前生物保存技术的细胞毒性很大程度上与低效的冷冻保护剂和冷却期间跨细胞膜的水输送有关，从而导致冰层形成造成不可修复的细胞损伤。第一阶段建立了一种完全不同的冷冻保护剂(CPA)优化方法，开发了第一个以生物为灵感的纳米孔作为跨膜巨型高速公路，以促进在冷冻保存期间安全有效地在细胞内传递和移除低温保护剂，并显著提高解冻后细胞的产量、活力和功能。造孔剂有助于在低温下负载CPA，以弥补低温下生物蛋白通道的功能损失，减少极低浓度所需的CPA量，并扩大CPA的选择范围，所有这些都在生理温度下动态解离，以防止开孔毒性。进一步的研发工作将使分子设计多样化，优化铅分子，扩大生产，并对具有重大治疗意义的再生药物进行冷冻保存研究，如干细胞和复杂组织。这一奖项反映了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。

This Small Business Innovation Research Phase II project will continue development of cutting-edge nanopore technology that will greatly enhance biological preservation of regenerative medicines such as stem cells, complex tissues, and organs. Such a technology has the potential to greatly improve the biobanking and transport infrastructure of healthcare for on-demand cells and tissues, improved 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 while maintaining high cell viability and function after cryopreservation, 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 long distances. 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. Phase I established a fundamentally different approach to cryoprotective agent (CPA) 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 and dramatically increase post-thaw cell yield, viability, and function. The pore-formers assist CPA loading at reduced temperatures to make up the functional loss of biological protein channels at cold temperatures, reduce the amount of CPA required to remarkably low concentrations, and expand the selectable range of CPAs, all while dynamically dissociating at physiological temperature to prevent open-pore toxicity. Further R&D effort will diversify molecular design, optimize lead molecules, scale-up production, and perform cryopreservation studies on regenerative medicines of major therapeutic significance such as stem cells and complex tissues.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.