Mechanistic approach to optimization of a kidney preservation solution

优化肾脏保存溶液的机械方法

基本信息

批准号：
10545982
负责人：
Kelvin G.M. Brockbank
金额：
$ 38.55万
依托单位：
TISSUE TESTING TECHNOLOGIES, LLC
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-08-19 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10545982
关键词：
Air AlamarBlue Animal Model Animals Antioxidants Apoptosis Inhibitor Biological Assay Blood Vessels Businesses Capital Cardiac Cardiac Death Cell Survival Clinical Clinical Data Cryopreservation Cyclic GMP Data Development Devices Dialysis procedure End stage renal failure Epithelial Cells European Evaluation Exposure to Family suidae Formulation Future Gold Grant Heart Heart Transplantation Histopathology Hour Human Hydrogen Sulfide Ice In Vitro Investigation Kidney Kidney Transplantation Laboratories Letters Longevity Manuscripts Maryland Methodology Methods Modification Mus Organ Organ Procurements Organ Transplantation Outcome Patients Perfusion Persons Phase Postoperative Period Power Sources Preparation Production Proteomics Quality of life Rattus Regulation Renal function Reperfusion Injury Research Safety Ships Small Business Innovation Research Grant South Carolina Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization Stains Supplementation Technology Testing Time Tissues Transplantation Trypan Blue Universities University of Wisconsin-lactobionate solution Waiting Lists Work World Health Organization base blood perfusion cell type clinical practice cytokine efficacy evaluation efficacy trial experience glucose analog improved in vivo inflammatory marker kidney cell kidney epithelial cell kidney preservation meetings porcine model post-transplant preclinical evaluation preservation professor prospective protocol development renal epithelium scale up screening stress tolerance time use transplant centers transplant model

项目摘要

The World Health Organization estimates that organ transplants are meeting less than 10% of global demand, in part because the short preservation periods possible with current technology limits options for organ assessment and sharing. Even potentially functional, transplantable organs may be turned down by one transplant center after another until its short preservation limit is exceeded. We propose development of a new stasis cocktail, based upon our Unisol™ solution, optimized for storage of kidneys that would enable cross continent organ matching and exchange that in turn would potentially improve the lives of thousands of patients with end stage renal disease. Our technology improves upon the most tried and true method for hypothermic kidney storage in clinical practice that is relatively inexpensive, ice cooled, and easy to ship by air due to no need for batteries or power supply. In this proposal we will assess optimization of Unisol™ for hypothermic kidney storage by supplementation with compounds targeting specific mechanisms of ischemia and reperfusion injury in five specific aims (SAs). In Phase I controls consisting of exposure to current clinical practice organ storage solutions will be performed in both SAs. In SA#1 we sequentially test potential formulation supplements in vitro on several human kidney cell types and then the best outcomes in an in vivo rat kidney transplant model. The in vitro studies will be performed in our laboratories in South Carolina employing primary human renal epithelial cells and the results will be confirmed using several types of primary human kidney cells. Cell viability assays will be employed, including alamarBlue, live dead stains, Trypan blue, and the MTT assay, to identify the best supplement formulations. In SA#2 in vivo studies to evaluate the best supplement formulations from SA#1 will be performed at Johns Hopkins in Maryland using an orthotopic rat kidney transplant model under the direction of Professor Gerald Brandacher. Successful demonstration of 100% animal/ kidney survival after 24h and >40% after 36h of storage and 28 days post-transplant will be considered demonstration of feasibility for progression to Phase II with 3 SAs. In Phase II we will scale up to porcine kidneys ex vivo (SA#3), followed by in vivo transplantation (SA#4) and then ex vivo testing with human kidneys (SA#5). Tissue MALDI, proteomics and proinflammatory cytokines will be also be assessed in SAs 2-5 to provide a better understanding of outcomes and potentially suggest formulation modifications. We will transition from GMP to cGMP Unisol™ production for Phase II. Larta, Inc., will assist Tissue Testing Technologies LLC in further development of our Phase II business plans during Phase I and transitioning to the market place in Phase II.

世界卫生组织估计，器官移植只能满足全球需求的不到 10%，部分原因是当前技术可能保存时间较短，限制了器官的选择即使是具有潜在功能的可移植器官也可能被拒绝。一个又一个移植中心，直到超过其短期保存限制，我们建议开发一个新的。 stasiscocktail，基于我们的 Unisol™ 解决方案，针对肾脏储存进行了优化，可实现交叉大陆器官匹配和交换可能会改善数千名患者的生活我们的技术改进了最可靠的低温治疗方法。临床实践中的肾脏储存相对便宜，冰冷，并且由于不需要而易于空运在本提案中，我们将评估 Unisol™ 对于低温肾脏的优化。通过补充针对缺血和再灌注损伤特定机制的化合物进行储存在五个具体目标（SA）中，第一阶段的控制包括暴露于当前的临床实践器官储存。解决方案将在两个 SA 中进行。在 SA#1 中，我们依次在体外测试潜在的配方补充剂。对几种人类肾细胞类型进行研究，然后在体内大鼠肾移植模型中获得最佳结果。体外研究将在我们位于南卡罗来纳州的实验室中使用原代人肾上皮进行细胞，结果将通过几种类型的原代人肾细胞细胞活力测定来证实。将采用包括 alamarBlue、活死染色剂、台盼蓝和 MTT 测定在内的方法来确定最佳在 SA#2 体内研究中，评估 SA#1 的最佳补充剂配方。在马里兰州约翰霍普金斯大学使用原位大鼠肾移植模型在指导下进行 Gerald Brandacher 教授成功证明 24 小时后动物/肾脏存活率达到 100%，并且 >40% 储存 36 小时和移植后 28 天后将被视为进展可行性的证明进入具有 3 个 SA 的第二阶段在第二阶段，我们将扩大到离体猪肾 (SA#3)，然后是体内。移植 (SA#4)，然后用人体肾脏进行离体测试 (SA#5)、蛋白质组学和促炎细胞因子也将在 SA 2-5 中进行评估，以更好地了解结果并可能建议配方修改，我们将从 GMP 过渡到 cGMP Unisol™ 生产。第二阶段，Larta, Inc. 将协助 Tissue Test Technologies LLC 进一步发展我们的第二阶段业务。第一阶段的计划和第二阶段过渡到市场。