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不到全球需求的10％， 部分是因为可能的短暂保存期可能具有当前的技术限制器官的选项 评估和共享。即使是潜在的功能性，可移植的器官也可能被一个 移植中心是另一个后方，直到其短暂的保存极限超过。我们提出了新的开发 基于我们的UNISOL™解决方案的Stasis Cocktail，优化用于肾脏的存储 大陆器官匹配和交换，反过来可能会改善数千名患者的生活 患有末期肾脏疾病。我们的技术改进了低温的最久经考验的方法 肾脏存储在临床实践中相对便宜，冷却且易于通过空气运送的肾脏存储 用于电池或电源。在此提案中，我们将评估UNISOL™对低温肾脏的优化 通过补充靶向缺血和再灌注损伤的特定机制的化合物来储存 在五个特定目标（SAS）中。在I阶段的控制中，包括暴露于当前临床实践器官存储 解决方案将在两个SAS中执行。在SA＃1中，我们在体外依次测试潜在的公式补充剂 在几种人类肾细胞类型上，然后在体内大鼠肾移植模型中取得最好的结果。 in 体外研究将在我们在南卡罗来纳州的实验室中采用原发性人类肾上皮进行 细胞和结果将使用几种类型的原代人肾细胞确认。细胞活力评估 将进行，包括Alamarblue，Live Dead Skins，Turpan Blue和MTT分析，以识别最好的 补充公式。在SA＃2体内研究中，以评估SA＃1的最佳补充配方 在马里兰州的约翰·霍普金斯（Johns Hopkins 杰拉尔德·布兰查教授。成功证明了24小时后100％动物/肾脏生存和> 40％ 36小时后，移植后28天将被视为证明进展可行性 使用3阶段的三个SAS。在第二阶段，我们将扩展到猪肾脏Ex Vivo（SA＃3），然后是体内 移植（SA＃4），然后用人肾脏进行体内测试（SA＃5）。组织MALDI，蛋白质组学和 促炎细胞因子也将在SAS 2-5中进行评估，以更好地了解结果 并有可能提出公式修改。我们将从GMP过渡到CGMP Unisol™生产 第二阶段。 Larta，Inc。将协助组织测试Technologies LLC进一步开发我们的II期业务 在第一阶段的计划，并在第二阶段过渡到市场。