Mechanistic approach to optimization of a kidney preservation solution

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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.