Subzero Non-freezing preservation of whole mammalian organisms

摘要 实现人类的悬浮动画将对社会产生变革：在以下长期应用中 需要几个月的保存，它将使前往火星和其他地方的行星任务成为可能。在更直接的用途上 在某些情况下可用于延迟复苏。延长的假死时间通常是 在自然界中观察到的，包括几种在零度以下环境中冬眠的哺乳动物物种 温度低至-30摄氏度，而通常使用如此高的零下温度(这里定义为0？C至-30？C) 以自然界为手段建立整个生物体的长期保存，零度以下的非冷冻保存 在文献中没有对哺乳动物进行探索(见重要意义)，仍然是一个有利可图的研究领域。 我们的首要目标是通过使新陈代谢停滞和复苏来推动科学发现的边界 对于一个没有生死能力的生物体来说。本次NIGMS探索性研究的目的 对技术开发的资助是为了测试哺乳动物在零度以下的非冷冻(SZNF)储存的可行性 这是文学史上的第一次。这些研究还将在不存在的情况下产生初步数据，这反过来又会 在降低风险和机械目标后，可以继续使用更传统的筹资机制 确定为支持假设驱动的研究。这一目标是基于我们团队在 将零度以下的非冷冻保存扩展到人体肝脏，这为我们提供了关键的新技术，使控制 了解细胞内碳水化合物水平，了解内皮对缺血的敏感性和缓解策略，以及 机器辅助恢复/复活技术以减轻保存损伤。然而，从单一的 整个哺乳动物的器官是向前迈出的垂直一步，这是一个潜在的突破，但有相当大的 风险。因此，这个以技术设计为导向的建议书特别符合此探索性项目的计划要求 赠款机制。 作为这些研究的结果，我们希望开发能够保护新生儿和后遗症的新技术。 在零度以下的温度下出生的小鼠。为了做到这一点，我们的目标是开发几种新的方案和冷冻防腐剂，以避免 在整个生物体储存的情况下进行冷冻。这些结果预计将导致初步数据显示 在几天的零度以下存储后幸存下来，这将推动未来的研究，以及将支持公式制定的关键数据 提出新的假设，使之能够利用更传统的筹资机制。

ABSTRACT Realizing suspended animation for humans would be transformative for the society: in long term applications where months of preservation is needed, it would enable planetary missions to Mars and beyond. In the more immediate uses it could be enabling for delayed resuscitation purposes in certain cases. Extended suspended animation is commonly observed in nature including several mammalian species which hibernate at subzero environments with ambient temperature as low as -30 ºC. While such high subzero temperatures (defined here as 0 ºC to -30 ºC) is utilized commonly by nature as a means to establish whole organism preservation for extended durations, subzero non-freezing preservation has not been explored for mammalians in literature (see Significance) and remain a profitable area for research. Our overarching goal is to push the boundaries of scientific discovery by enabling metabolic stasis and reanimation for an organism without the natural capacity for suspended animation. The objective of this NIGMS exploratory research grant for technology development is to test the viability of sub-zero, non-freezing (SZNF) storage of a mammal for the first time in literature. The studies would also lead to preliminary data in a context where none exists, which in turn would enable continuation with a more traditional funding mechanism once the risks are reduced and mechanistic targets are identified to enable hypothesis driven research. This objective was formulated based on our team's prior success in scaling subzero nonfreezing preservation to human livers, which provided us with key new technologies enabling control of intracellular carbohydrate levels, understanding of endothelial sensitivity to ischemia and strategies for mitigation, and machine-assisted recovery/reanimation techniques to ameliorate preservation injury. However, the transition from a single organ to an entire mammalian organism presents a vertical step forward, a potential breakthrough but with considerable risk. As such, this technology design-directed proposal specifically matches the program requirements for this exploratory grant mechanism. As a result of these studies, we expect to develop novel technologies that enables preservation of neonatal and post- natal mice in subzero temperatures. To do so, we aim to develop several novel protocols and cryopreservatives to avoid freezing in the context of whole-organism storage. These results are expected to lead to preliminary data that shows survival after days of subzero storage, which will motivate future research, as well as key data that will enable formulation of new hypotheses that will enable the utilization of more traditional funding mechanisms.