Engineering optimization and scaling enables high quality pancreatic islet cryopreservation for banking and transplant

工程优化和扩展可实现高质量胰岛冷冻保存以用于储存和移植

基本信息

批准号：
10680579
负责人：
JOHN C BISCHOF
金额：
$ 55.7万
依托单位：
UNIVERSITY OF MINNESOTA
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-22 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10680579
关键词：
Adopted Affect Biological Biology Cells Cellular Stress Cessation of life Chemicals Clinical Cryopreservation Cryoprotective Agents Crystallization Cyclic GMP Data Diabetes Mellitus Disease Dose Engineering Family suidae Freezing Glass Goals Health Health Care Costs Heating Human Ice Impairment In Vitro Individual Infusion procedures Injury Insulin Interdisciplinary Study Islets of Langerhans Islets of Langerhans Transplantation Knowledge Measures Methods Modeling Molecular Mus Organ Pathway interactions Personal Satisfaction Persons Phenotype Procedures Process Protocols documentation Recovery Reporting Resources Rewarming Risk Risk Reduction Source Survival Rate System Technology Testing Tissues Toxic effect Translations Transplantation Validation Whole Organism Xenograft Model advanced analytics analytical method cGMP production clinical translation cold temperature cost cryogenics human stem cells improved in vivo innovative technologies islet potency testing preservation prevent repaired response scale up stem cells success supply chain transplant centers

项目摘要

Project Summary Diabetes has a tremendous impact on the health and well-being of affected individuals, as well as a considerable overall societal burden. Pancreatic islet transplantation has the potential to cure diabetes, but one of the main problems limiting the success of this treatment is an inadequate supply of islets. Islets from a single donor are often insufficient to achieve insulin independence, and multiple infusions are often required, each with increasing risk. Two potential strategies exist to increase the number of islets available: (1) pool islets from multiple donors and perform single procedure, high-dose transplants; and (2) develop alternative sources such as stem-cell-derived islets. The availability of these limited resources becomes a supply chain problem, and for either approach, a method for islet preservation is essential. Our long-term objective is to develop a method for cryopreserving, or “banking,” islets prior to transplant. No previous strategy has achieved the high viability, function, and clinical scalability required for transplant in a single approach. To achieve long-term islet banking, we propose to use an alternative cryopreservation strategy, vitrification. That is, cryogenic storage in an ice-free glassy state. A significant challenge in the vitrification of biospecimens is that the cooling and heating rates needed for vitrifying and rewarming are tremendously high (>107 °C/min). These rates are reduced by adding cryoprotective agents (CPA) that inhibit ice formation, but these agents are themselves toxic to islets. Thus, the critical challenge in islet vitrification is achieving fast enough cooling and warming to avoid ice, while avoiding toxicity from the CPA, and doing so in a clinically scalable manner. Using engineering principles of heat and mass transfer, our multidisciplinary research team has developed an approach for vitrification and rewarming (VR) to solve this problem, termed “cryomesh VR,” for islets. Our central hypothesis is that the improved heat transfer achieved by cryomesh VR, combined with optimizations in CPA use, will enable ice-free vitrification and rewarming of islets while avoiding toxicity. Our preliminary data achieve cooling and warming rates far exceeding other methods, and we have shown CPA loading and unloading protocols with low toxicity in mouse, human, pig, and human stem-cell-derived (SC) islets. Indeed, in all cryopreserved islet models tested we have achieved viability, recovery, and function that meets or exceeds all previous reports and does so in a clinically scalable method. To further improve our approach and move towards clinical translation, we propose the following aims: Aim 1. Refine the optimal physical conditions for the cryomesh VR of mouse, human, and SC islets; Aim 2. Measure the viability, function, and in vivo potency of mouse, human, and SC islets following cryomesh VR; Aim 3. Define the molecular and cellular changes occurring in response to cryopreservation; and Aim 4. Scale-up cryomesh VR for clinical throughput and adapt the processes for cGMP production. If successful this approach could revolutionize how islets are isolated, allocated, and stored prior to transplant and increase utilization of deceased donor pancreases for the cure of diabetes.

项目概要糖尿病对受影响个体的健康和福祉产生巨大影响，并且胰岛移植有可能治愈糖尿病，但其带来的整体社会负担相当大。限制这种治疗成功的主要问题之一是胰岛供应不足。捐献者往往不足以实现胰岛素独立，并且常常需要多次输注，每次输注存在两种潜在的策略来增加可用胰岛的数量：（1）池胰岛多个捐赠者并进行单一程序、高剂量移植；(2) 开发替代来源，例如作为干细胞衍生的胰岛，这些有限资源的可用性成为一个供应链问题。无论采用哪种方法，胰岛保存的方法都是至关重要的，我们的长期目标是开发一种方法。移植前冷冻保存或“储存”胰岛，之前没有任何策略能够实现高存活率。通过单一方法进行移植所需的功能和临床可扩展性。为了实现长期胰岛银行，我们建议使用另一种冷冻保存策略，即玻璃化。也就是说，在无冰玻璃态下的低温储存是生物样本玻璃化的重大挑战。玻璃化和复温所需的冷却和加热速率非常高（>107°C/min）。通过添加抑制冰形成的冷冻保护剂 (CPA) 可以降低这些速率，但这些试剂因此，胰岛玻璃化的关键挑战是实现足够快的冷却和冷冻。加热以避免结冰，同时避免 CPA 的毒性，并以临床可扩展的方式进行。利用传热传质工程原理，我们的多学科研究团队开发了一种用于解决胰岛问题的玻璃化和复温 (VR) 方法，称为“cryomesh VR”。中心假设是 Cromesh VR 结合优化实现了改进的传热在 CPA 中使用，将实现胰岛的无冰玻璃化和复温，同时避免毒性。数据实现的降温和升温速率远远超过其他方法，并且我们已经显示了 CPA 加载和事实上，在小鼠、人类、猪和人类干细胞衍生（SC）胰岛中具有低毒性的卸载方案。我们测试的所有冷冻胰岛模型都达到或超过了活力、恢复和功能所有以前的报告，并以临床可扩展的方法进行，以进一步改进我们的方法和行动。临床转化，我们提出以下目标：目标 1. 细化最佳身体条件，以实现小鼠、人类和 SC 胰岛的冷冻网 VR 目标 2. 测量胰岛的活力、功能和体内效力冷冻网 VR 后的小鼠、人类和 SC 胰岛目标 3. 定义发生的分子和细胞变化；响应冷冻保存；目标 4. 扩大冷冻网 VR 的临床通量并调整如果成功的话，这种方法可能会彻底改变胰岛的分离、分配和使用方式。并在移植前储存，并提高已故供体胰腺的利用率，以治疗糖尿病。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Pancreatic islet cryopreservation by vitrification achieves high viability, function, recovery and clinical scalability for transplantation.

玻璃化胰岛冷冻保存可实现移植的高活力、功能、恢复和临床可扩展性。

DOI：
发表时间：
2022-04
期刊：
影响因子：
82.9
作者：
Zhan, Li;Rao, Joseph Sushil;Sethia, Nikhil;Slama, Michael Q;Han, Zonghu;Tobolt, Diane;Etheridge, Michael;Peterson, Quinn P;Dutcher, Cari S;Bischof, John C;Finger, Erik B
通讯作者：
Finger, Erik B

Vitrification and Rewarming of Magnetic Nanoparticle-Loaded Rat Hearts.

装载磁性纳米粒子的大鼠心脏的玻璃化和复温。

DOI：
发表时间：
2022-03
期刊：
影响因子：
6.8
作者：
Gao, Zhe;Namsrai, Baterdene;Han, Zonghu;Joshi, Purva;Rao, Joseph Sushil;Ravikumar, Vasanth;Sharma, Anirudh;Ring, Hattie L;Idiyatullin, Djaudat;Magnuson, Elliott C;Iaizzo, Paul A;Tolkacheva, Elena G;Garwood, Michael;Rabin, Yoed;Etheridge, Mic
通讯作者：
Etheridge, Mic

Vitrification and nanowarming enable long-term organ cryopreservation and life-sustaining kidney transplantation in a rat model.

玻璃化和纳米加温可以在大鼠模型中实现长期器官冷冻保存和维持生命的肾移植。

DOI：
发表时间：
2023-06-09
期刊：
影响因子：
16.6
作者：
Han, Zonghu;Rao, Joseph Sushil;Gangwar, Lakshya;Namsrai, Bat;Pasek;Etheridge, Michael L;Wolf, Susan M;Pruett, Timothy L;Bischof, John C;Finger, Erik B
通讯作者：
Finger, Erik B

Cryopreservation of Whole Rat Livers by Vitrification and Nanowarming.

通过玻璃化和纳米加温冷冻保存整个大鼠肝脏。

DOI：
发表时间：
2023-03
期刊：
影响因子：
3.8
作者：
Sharma, Anirudh;Lee, Charles Y;Namsrai, Bat;Han, Zonghu;Tobolt, Diane;Rao, Joseph Sushil;Gao, Zhe;Etheridge, Michael L;Garwood, Michael;Clemens, Mark G;Bischof, John C;Finger, Erik B
通讯作者：
Finger, Erik B