Viable Banking of Human Tissues

人体组织的可行储存

基本信息

批准号：
10165714
负责人：
ALI EROGLU
金额：
$ 35.62万
依托单位：
AUGUSTA UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-09-15 至 2023-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10165714
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

项目摘要

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.

项目概要/摘要开发有效的人体组织冷冻保存技术对于解决关键问题至关重要通过解决全球范围内可移植人体组织的短缺和组织的保质期问题来满足生物医学需求阻碍大规模生产、储存、分配和安全/质量控制的工程组织结构。此外，开发可靠的组织保存方法将允许更好的组织匹配通过实现可行的储存，提高总体成功率并减轻免疫抑制的负担。患病组织，有效的保存方法也将促进生物标志物的发现和药物测试，从而实现针对癌症等毁灭性疾病的新有效疗法的个性化医疗。目前的保存策略不足以满足多细胞复杂组织的总体目标。拟议的研究旨在满足对有效、广泛适用的组织保存技术的迫切需求开发一种新的玻璃化方法，这是一种涉及液体凝固的冷冻保存策略使用高浓度冷冻保护剂（CPA）的玻璃状状态是一种有前途的方法。技术，但 CPA 具有固有的细胞毒性。事实上，CPA 的化学毒性被认为是主要的。复杂组织成功冷冻保存的障碍随着冷却的增加而降低 CPA 浓度。和升温速率可改善毒性，但足够快的降温和升温速率很难在组织并可能导致极端的热应力，从而导致组织破裂。目前的组织保存方法，我们建议使用一种方法来解决 CPA 的化学毒性问题我们的初步研究表明，有针对性的干预措施和生物物理建模的跨学科方法。首选 CPA 1,2-丙二醇 (PROH) 的主要毒性是通过线粒体 Ca2+ 发生的我们能够完全防止 PROH 对小鼠卵母细胞、人类肌瘤和小鼠的毒性。最近，我们还利用线粒体 Ca2+ 单向转运蛋白抑制剂建立了脑组织模型。优化了向内皮细胞添加高浓度 CPA 且细胞毒性最小的程序。这些令人鼓舞的初步数据，我们的中心假设是，跨学科方法结合通过数学建模和优化有针对性地抑制 CPA 毒性可以实现高 CPA 浓度，导致适用于不同组织的多功能组织玻璃化方法。提出的玻璃化方法是创新的，因为它规避了使用高 CPA 的主要障碍浓度（CPA 毒性），能够更好地抑制冰成核和失透；创新包括开发一种新型玻璃化介质，可以通过以下方式阻止冰晶生长合成聚合物并通过优化的抗氧化剂成分和使用来减轻自由基损伤基于生物物理学的新型数学优化策略，用于确定玻璃化毒性最小的方案。拟议的研究预计将为可行的人体组织储存铺平道路。

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Rapid quantification of multi-cryoprotectant toxicity using an automated liquid handling method.

DOI：
10.1016/j.cryobiol.2020.10.017
发表时间：
2021-03
期刊：
Cryobiology
影响因子：
2.7
作者：
Warner RM;Ampo E;Nelson D;Benson JD;Eroglu A;Higgins AZ
通讯作者：
Higgins AZ

Mathematical Modeling of Protectant Transport in Tissues.

DOI：
10.1007/978-1-0716-0783-1_5
发表时间：
2021
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
0
作者：
Warner RM;Higgins AZ
通讯作者：
Higgins AZ

Permeation of individual cryoprotectants and their different combinations into mouse liver tissue.

单个冷冻保护剂及其不同组合渗透到小鼠肝组织中。

DOI：
10.1016/j.cryobiol.2023.03.004
发表时间：
2023
期刊：
Cryobiology
影响因子：
2.7
作者：
Higgins,AdamZ;Lavarti,Rupa;Eroglu,Binnur;Ahmadkhani,Nima;Benson,JamesD;Eroglu,Ali
通讯作者：
Eroglu,Ali