Cellular Engineering for Metabolic Stasis

代谢停滞的细胞工程

• 批准号: 6796449
• 负责人: Mehmet Toner
• 金额: $ 48.08万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2008-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6796449
• 关键词: 3T3 cells; adenosine triphosphate; apoptosis; bioengineering /biomedical engineering; biotechnology; enzyme activity; fluorescence recovery after photobleaching; gene expression; laboratory rat; liver cells; metabolism; peptidylprolyl isomerase; serine threonine protein kinase; technology /technique development; temperature; thermodynamics; tissue /organ preservation; tissue engineering

DESCRIPTION (provided by applicant): With the advancements being made in tissue engineering, cell transplantation, stem cell biology, and gene therapy, the clinical demand for effective long-term storage methods for cells and tissues will continue to increase. We propose to develop novel methods to biostabilization of mammalian cells for long-term preservation in a desiccated state at ambient temperature. In nature, many animals and organisms down regulate their metabolism and may enter into a state of stasis by either desiccation through removal of water from their cells (i.e., anhydrobiosis) or by a developmentally programmed arrest under full hydration (i.e., diapause). The ability to enter diapause prior to desiccation is crucial for the survivorship of many organisms that undergo natural states of dormancy. Furthermore, a common theme is that desiccation-tolerant animals accumulate large amounts of disaccharides, especially trehalose and sucrose. These sugars provide protective effects by forming stable sugar glasses at high water contents, and by stabilizing biological membranes and proteins through direct interaction with polar residues. We, therefore, hypothesize that metabolic preconditioning of mammalian cells to induce diapause-like state followed by controlled drying, storage, and rehydration conditions (i.e., physicochemical, biochemical, and metabolic) can be used to achieve desiccation tolerance in mammalian cells and tissues. To this end, our 3 distinct, but interactive, specific aims are: 1) To develop optimal physicochemical conditions to stabilize desiccated cells. 2) To metabolically precondition mammalian cells to improve survivorship during storage. 3) To develop metabolic and biophysical strategies to accelerate recovery of desiccated cells. This project is one of the first attempts to apply engineering and quantitative concepts to achieving anhydrobiotic state in mammalian cells. It provides a systems view of the metabolic and cellular changes a cell encounters before, during, and after desiccation. This project is inspired by nature and it uses engineering concepts and approaches to translate nature's solution to long-term storage or "suspended animation" for mammalian systems. The proposed studies will significantly impact on human health by providing a solution to the problem of providing long-term storage of blood cells, stem cells, tissue engineered products, and cell-based biosensors for use in regenerative medicine, tissue engineering, and bioterrorism. In the short term, it will help increase the treatment modalities available to liver failure by providing stable, long-term stabilized cells for bioartificial liver assist devices. The longer-range outcome of the proposed research is to translate the information gained from these studies into whole organ preservation.

描述（申请人提供）：随着组织工程、细胞移植、干细胞生物学和基因治疗的进步，临床对细胞和组织的有效长期储存方法的需求将不断增加。我们建议开发新的方法来生物稳定哺乳动物细胞，以便在环境温度下干燥状态下长期保存。在自然界中，许多动物和生物体下调其新陈代谢，并可能通过从细胞中去除水分而干燥（即脱水）或在完全水合的情况下发育程序停滞（即滞育）而进入停滞状态。在干燥之前进入滞育的能力对于许多经历自然休眠状态的生物体的生存至关重要。此外，一个共同的主题是耐干燥的动物会积累大量的二糖，尤其是海藻糖和蔗糖。这些糖通过在高含水量下形成稳定的糖玻璃以及通过与极性残基的直接相互作用来稳定生物膜和蛋白质来提供保护作用。因此，我们假设对哺乳动物细胞进行代谢预处理以诱导滞育样状态，然后控制干燥、储存和补水条件（即物理化学、生物化学和代谢）可用于实现哺乳动物细胞和组织的干燥耐受性。为此，我们的 3 个不同但相互作用的具体目标是：1) 开发最佳的物理化学条件来稳定干燥的细胞。 2) 对哺乳动物细胞进行代谢预处理，以提高储存期间的存活率。 3）制定代谢和生物物理策略以加速干燥细胞的恢复。该项目是应用工程和定量概念在哺乳动物细胞中实现脱水状态的首次尝试之一。它提供了细胞在干燥之前、期间和之后所经历的代谢和细胞变化的系统视图。该项目受到大自然的启发，它使用工程概念和方法将大自然的解决方案转化为哺乳动物系统的长期存储或“假死”。拟议的研究将为用于再生医学、组织工程和生物恐怖主义的血细胞、干细胞、组织工程产品和基于细胞的生物传感器的长期储存问题提供解决方案，从而对人类健康产生重大影响。短期内，它将通过为生物人工肝辅助装置提供稳定、长期稳定的细胞，有助于增加肝衰竭的治疗方式。拟议研究的长期成果是将这些研究中获得的信息转化为整个器官的保存。