Feasibility of Unfrozen Sub-zero Storage of Mammalian Tissues with Insect Anti-Fr

具有抗虫剂的哺乳动物组织的非冷冻零度以下储存的可行性

• 批准号: 7481796
• 负责人: Kelvin G.M. Brockbank
• 金额: $ 18.83万
• 依托单位: CELL AND TISSUE SYSTEMS, INC.
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-01 至 2009-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7481796
• 关键词: Animals; Antifreeze Proteins; Architecture; Biological Assay; Biological Preservation; Blood Vessels; Carotid Arteries; Cell Survival; Cells; Clinical; Condition; Control Groups; Cryopreservation; Development; Disruption; Dissection; Drug Formulations; Economics; Endothelium; Evaluation; Experimental Designs; Family suidae; Fishes; Freezing; Functional disorder; Goals; Harvest; Heart Transplantation; Histopathology; Hour; Ice; In Vitro; Injury; Insecta; Lead; Liver; Liver Failure; Logistics; Mental Depression; Metabolic; Metabolism; Methods; Modeling; Modification; Morphology; Organ; Organ Transplantation; Oryctolagus cuniculus; Outcome; Pancreas; Patients; Phase; Phase I Clinical Trials; Physiological; Procedures; Production; Relative (related person); Reporting; Research Design; Residual state; Risk; Screening procedure; Small Business Funding Mechanisms; Small Business Innovation Research Grant; Smooth Muscle; Solutions; Source; Structure; Structure of jugular vein; Surgical Replantation; Temperature; Testing; Time; Tissue Model; Tissue Viability; Tissues; Transplantation; Vascular Endothelium; Venous; base; cold temperature; day; design; liver transplantation; natural hypothermia; novel; pre-clinical; professor; research study; scale up; time interval

DESCRIPTION (provided by applicant): The long term goal is the development of better, longer hypothermic storage conditions for liver transplants than currently possible in clinical practice using novel insect antifreeze proteins. Hypothermic injury results in vascular dysfunction during liver storage, therefore vascular tissue models will be employed in this Phase I SBIR proposal. Blood vessel rings, derived from rabbit jugular veins and carotid arteries, will be employed because they will permit dissection of hypothermic storage effects upon the morphology and functional viability of vessels and because these studies can be performed with the small quantities of insect-derived antifreeze proteins that can be produced with our current manufacturing methods. Scale up of antifreeze protein production to achieve the quantities required for livers is not possible within the economic limitations of a Phase I SBIR proposal. The use of hypothermia as the principal means to suppress metared with controls without antifreeze proteins in order to increase the duration of hypothermic storage. The test temperature, -10¿C, was selected on the basis of the greater non-colligative freezing point depression produced by the insect antifreeze peptides that provide almost an order of magnitude greater activity compared with other previously described antifreeze proteins derived from other sources. The insect-derived antifreeze proteins will inhibit ice nucleation at -10¿C permitting mammalian tissue storage at much lower levels of endogenous metabolic activity. These storage conditions should lead to longer hypothermic storage periods for tissues and organs. In this Phase I SBIR proposal insect-derived antifreeze protein formulations, selected from research studies on thermal hysteresis by our consultant, Professor John Duman, and controls will be compared employing rabbit venous and arterial tissue models and a panel of assays for evaluation of the histopathology and physioloan livers in a subsequent Phase II SBIR proposal. PUBLICH HEALTH RELEVANCE: Livers destined for transplantation are currently stored using static cold storage in a chemically defined solution at 4¿C for periods of up to 6 hours. However, the application of static cold storage has proven to be insufficient to fulfill the organ demand for patients with irreversible liver failure, because marginal livers can not be preserved optimally, for long enough, by static cold storage to permit organ evaluation to occur. These limitations should be overcome by use of a lower, sub-zero hypothermic temperature (-10¿C) to further reduce metabolism in combination with insect-derived antifreeze proteins to inhibit ice nucleation resulting in increased availability of organs for transplant.

描述（由申请人提供）：长期目标是开发比目前临床实践中使用新型昆虫抗冻蛋白更好、更长的肝脏移植低温储存条件。低温损伤导致肝脏储存过程中血管功能障碍，因此，血管组织模型将用于本I期SBIR提案。将采用来自兔颈静脉和颈动脉的血管环，因为它们将允许解剖低温储存对血管形态和功能活力的影响，并且因为这些研究可以用我们目前的生产方法生产的少量昆虫来源的抗冻蛋白进行。在第一阶段SBIR提案的经济限制范围内，不可能扩大抗冻蛋白生产以达到肝脏所需的数量。使用低温作为主要手段来抑制与不含抗冻蛋白的对照的热交换，以增加低温储存的持续时间。选择试验温度-10 ℃是基于昆虫抗冻肽产生的更大的非依数性冰点降低，与先前描述的其他来源的抗冻蛋白相比，昆虫抗冻肽提供几乎一个数量级的更大活性。昆虫来源的抗冻蛋白将抑制-10 ° C下的冰成核，允许哺乳动物组织在低得多的内源代谢活性水平下储存。这些储存条件应该会导致组织和器官的低温储存期更长。在这第一阶段SBIR建议，昆虫衍生的抗冻蛋白制剂，从热滞后的研究，由我们的顾问，教授约翰杜曼，和控制的研究选择将比较采用兔静脉和动脉组织模型和一组试验的组织病理学和生理学的肝脏在随后的第二阶段SBIR建议的评价。公共卫生相关性：用于移植的肝脏目前使用静态冷藏在4 ° C的化学成分确定的溶液中储存长达6小时。然而，静态冷藏的应用已被证明不足以满足不可逆肝衰竭患者的器官需求，因为通过静态冷藏，边缘肝脏不能被最佳地保存足够长的时间以允许器官评估发生。这些限制应通过使用较低的零下低温（-10 ° C）来克服，以进一步减少代谢，并结合昆虫来源的抗冻蛋白来抑制冰核，从而增加器官移植的可用性。