Viable Banking of Human Tissues

人体组织的可行储存

• 批准号: 9926261
• 负责人: ALI EROGLU
• 金额: $ 36.65万
• 依托单位: AUGUSTA UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-15 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9926261
• 关键词: Address; Antioxidants; Area; Biophysics; Calcium; Chemicals; Complex; Computer Simulation; Coupling; Cryopreservation; Cryopreserved Tissue; Cryoprotective Agents; Crystallization; Data; Derivation procedure; Development; Disease; Disease model; Electron Microscopy; Employment; Endothelial Cells; Engraftment; Fibroid Tumor; Free Radicals; Freezing; Generations; Genomics; Glass; Goals; Growth; Heart Valves; Histologic; Human; Ice; Immunosuppression; Intervention; Journals; Lead; Life; Liquid substance; Malignant Neoplasms; Mathematics; Measures; Medical; Methods; Mitochondria; Modeling; Mus; Oocytes; Organ; Outcome; Ovarian Tissue; Patients; Polymers; Procedures; Process; Production; Property; Propylene Glycols; Protocols documentation; Quality Control; Regenerative Medicine; Research; Safety; Skin Tissue; Techniques; Technology; Testing; Time; Tissue Engineering; Tissue Model; Tissue Preservation; Tissue Sample; Tissue Transplantation; Tissue Viability; Tissues; Toxic effect; Translational Research; Transplantation; Tumor Tissue; World Health Organization; base; biobank; biochemical model; biomarker discovery; biophysical model; brain tissue; cytotoxicity; design; disease mechanisms study; drug testing; effective therapy; ethylene glycol; human tissue; inhibitor/antagonist; innovation; interdisciplinary approach; mathematical model; myometrium; novel; novel strategies; novel therapeutic intervention; personalized medicine; preservation; prevent; success; thermal stress; transplantation medicine; treatment strategy

PROJECT SUMMARY/ABSTRACT Development of an effective cryopreservation technology for human tissues is necessary to address critical biomedical needs by solving a worldwide shortage of transplantable human tissues and shelf-life problems of engineered tissue constructs that hinder mass production, storage, distribution, and safety/quality control. Moreover, the development of a reliable tissue preservation method would allow better tissue matching toward increased overall success rates and reduced burden of immunosuppression. By enabling viable banking of diseased tissues, an effective preservation method would also advance biomarker discovery and drug testing, and thus personalized medicine toward new effective therapies of devastating diseases such as cancer. Current preservation strategies are inadequate for multicellular complex tissues. The overall goal of the proposed research is to meet the critical need for effective, widely applicable tissue preservation technology by developing a novel approach to vitrification, a cryopreservation strategy involving the solidification of liquid in a glass-like state using high concentrations of cryoprotective agents (CPAs). Vitrification is a promising technique, but CPAs have inherent cytotoxicity. In fact, chemical toxicity of CPAs is considered to be the main barrier to successful cryopreservation of complex tissues. Reduced CPA concentrations with increased cooling and warming rates ameliorate toxicity, but sufficiently fast cooling and warming rates are difficult to achieve in tissues and may lead to extreme thermal stresses, resulting in tissue cracking. To overcome the limitations of current tissue preservation approaches, we propose to address chemical toxicity of CPAs using an interdisciplinary approach of targeted interventions and biophysical modeling. Our preliminary studies revealed that the primary toxicity of 1,2-propanediol (PROH), a preferred CPA, occurs through mitochondrial Ca2+ overload. We were able to completely prevent PROH’s toxicity in mouse oocyte, human fibroid and mouse brain tissue models by using inhibitors of mitochondrial Ca2+ uniporter. Recently, we have also mathematically optimized a procedure to add high CPA concentrations to endothelial cells with minimal cytotoxicity. Based on these encouraging preliminary data, our central hypothesis is that an interdisciplinary approach combining targeted inhibition of CPA toxicity with mathematical modeling and optimization can enable employment of high CPA concentrations, leading to a versatile tissue vitrification method applicable to diverse tissues. The proposed vitrification approach is innovative, because it circumvents the main barrier to the use of high CPA concentrations (CPA toxicity), enabling better suppression of ice nucleation and devitrification; other innovations include the development of a novel vitrification medium that can block ice crystal growth through synthetic polymers and mitigate free radical damage by optimized composition of antioxidants and the use of a novel biophysics-based mathematical optimization strategy to identify minimally toxic protocols for vitrification. The proposed research is expected to pave the way for viable banking of human tissues.

项目总结/摘要 开发一种有效的人体组织冷冻保存技术是必要的，以解决关键的 通过解决可移植人体组织的全球短缺和 工程化组织构建体阻碍了大规模生产、储存、分配和安全/质量控制。 此外，可靠的组织保存方法的发展将允许更好的组织匹配， 提高了总体成功率并降低了免疫抑制的负担。通过使可行的银行业务， 疾病组织，一种有效的保存方法也将促进生物标志物的发现和药物测试， 从而使个体化医学朝着对诸如癌症的毁灭性疾病的新的有效疗法发展。 目前的保存策略不足以用于多细胞复杂组织。的总体目标 拟议的研究是为了满足有效的、广泛适用的组织保存技术的关键需求， 开发一种新的玻璃化方法，一种涉及液体固化的低温保存策略， 使用高浓度的冷冻保护剂（CPA）的玻璃样状态。玻璃化是一种很有前途的 技术，但CPA具有固有的细胞毒性。事实上，注册会计师的化学毒性被认为是主要的 成功冷冻保存复杂组织的障碍。随着冷却时间的增加，CPA浓度降低 和升温速率改善毒性，但足够快的冷却和升温速率是难以实现的， 组织，并可能导致极端的热应力，导致组织开裂。为了克服…的局限性 目前的组织保存方法，我们建议解决化学毒性的CPA使用 有针对性的干预和生物物理建模的跨学科方法。我们的初步研究显示 1，2-丙二醇（PROH）（优选的CPA）的主要毒性通过线粒体Ca 2+发生， 超载。我们能够完全预防PROH对小鼠卵母细胞、人子宫肌瘤和小鼠子宫内膜的毒性， 使用线粒体Ca 2+单向转运体抑制剂的脑组织模型。最近，我们还从数学上 优化了向内皮细胞添加高浓度CPA的程序，细胞毒性最小。基于 这些令人鼓舞的初步数据，我们的中心假设是，一个跨学科的方法结合 利用数学建模和优化的CPA毒性的靶向抑制可以使得能够使用高浓度的 CPA浓度，导致适用于不同组织的通用组织玻璃化方法。的 提出的玻璃化方法是创新的，因为它绕过了使用高CPA的主要障碍 浓度（CPA毒性），能够更好地抑制冰成核和失透;其他 创新包括开发一种新型的玻璃化介质， 合成聚合物和减轻自由基损伤的抗氧化剂的优化组合物和使用一种 新的基于生物制药学的数学优化策略，以确定玻璃化的最低毒性方案。 这项拟议中的研究有望为可行的人体组织库铺平道路。