Mechanistic approach to optimization of a kidney preservation solution

优化肾脏保存溶液的机械方法

• 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
• 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

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不到10%的需求， 部分原因是目前技术可能存在的短保存期限制了器官的选择， 评估与分享。即使是潜在的功能，可移植的器官可能会被拒绝一个 一个接一个的移植中心，直到它的短期保存极限被超过。我们建议开发一种新的 基于我们的Unisol™解决方案的停滞鸡尾酒，针对肾脏储存进行了优化， 大陆器官匹配和交换，这反过来又可能改善成千上万患者的生活 患有终末期肾病。我们的技术改进了最可靠的低温疗法 在临床实践中，肾脏储存相对便宜，冰冷，并且由于不需要而易于空运 电池或电源。在本提案中，我们将评估Unisol™用于低温肾的优化 通过补充靶向特定缺血和再灌注损伤机制的化合物的储存 五个具体目标（SA）。在I期对照中，包括暴露于当前临床实践器官储存 将在两个SA中执行解决方案。在SA#1中，我们依次在体外测试了潜在的制剂补充剂 在几种人类肾细胞类型上，然后在体内大鼠肾移植模型中获得最佳结果。的in 体外研究将在我们位于南卡罗来纳州的实验室进行， 细胞，并将使用几种类型的原代人肾细胞证实结果。细胞活力测定 包括alamarBlue，活死染色，台盼蓝和MTT试验，以确定最佳的 补充制剂。在SA#2体内研究中，评价来自SA#1的最佳补充剂制剂， 在马里兰州的约翰霍普金斯使用原位大鼠肾移植模型在指导下进行 杰拉尔德·布兰达彻教授的成功证明24小时后动物/肾脏存活率为100%且>40% 储存36小时后和移植后28天将被视为进展可行性的证明 第二阶段，3个SA。在第II阶段，我们将扩大到离体猪肾（SA#3），然后是体内 移植（SA#4），然后用人肾进行离体测试（SA#5）。组织MALDI、蛋白质组学和 还将在SA 2-5中评估促炎细胞因子，以更好地了解结局 并潜在地建议配方修改。我们将从GMP生产过渡到cGMP Unisol™生产， 第二阶段。Larta，Inc.，我将协助组织测试技术有限责任公司进一步发展我们的第二阶段业务 在第一阶段的计划和过渡到第二阶段的市场。