Genotyped and Single Cyst-derived Human ADPKD Cell Platforms for Industry and Academia

DESCRIPTION provided by applicant DiscoveryBioMed Inc DBM Inc or DBM specializes in normal and diseased human cell platform engineering DBM has assembled a consortium of experts whereby collectively we seek Fast Track SBIR funding to establish and offer a Product Line Menu of normal and ADPKD human cell platforms for the academic and industry marketplaces Existing and interested DBM clients will be presented in terms of which options in the Product Line Menu fit their research needs as highlighted in our Research Strategy and Commercialization Plan This highly anticipated Product Line Menu will include an array of a specialty cell culture components b primary normal and ADPKD human primary cultures c mixed immortal cultures and d clonal immortal cell lines that are well characterized genotyped and defined as to nephron segment of origin Immortal cell lines will be compared to the primary cultures from which they were derived to determine whether they are similar in bioassay performance in genotype and in nephron segment of origin DBM and collaborators have begun the collection and characterization of single cyst derived primary cultures and multicystic tissue derived primary cultures from human ADPKD huADPKD donor kidneys procured from commercial vendors in pilot Phase like studies These initial huADPKD primary cultures were combined with primary cultures from different donors from Dr Darren Wallaceandapos s PKD Research Biomaterials and Cellular Models Core at Kansas University Medical Center KUMC so that all samples could be genotyped by Dr Peter Harrisandapos Core at the Mayo PKD Center Each huADPKD primary culture has been profiled in D microtiter plate driven proliferation bioassays on tissue culture plastic as well as in D Matrigel
based cystogenesis bioassays Preliminary data are presented in support of the above pilot efforts Select primary cultures that display a compelling genotype and exhibit excellent utility i D and D bioassays will be immortalized by multiple genetic methods DBM has pledged to also immortalize KUMC primary cultures as necessary in collaboration Where possible nephron segment of origin will be identified in huADPKD cells Two hypotheses will be addressed within our applied science research Does primary huADPKD fibroblast support co culture conditioned medium both drive human ADPKD cystic epithelial cell primary cultures to be longer lived and or to attain specific phenotypes in D and D bioassays for PKD research and Do individual cysts within a given huADPKD donor kidney possess the same genotype or can they differ in genotype from cyst to cyst i e exploring the andapos one hitandapos vs andapos two htandapos hypothesis or the emergence of somatic mutations and the possible presence of haploinsufficiency DBM et al set milestones for our work to be supported by this possible Fast Track SBIR award Milestone will be the establishment of single cyst derived and multicystic tissue derived primary cultures from each huADPKD donor kidney tissue sample and with a final goal of different donors processed Milestone will be to genotype each individual cyst derived and multicystic tissue derived primary culture compare genotypes against the ADPKD genotype database and select candidate primary cultures for immortalization Milestone will be to immortalize select primary cultures using a well optimized Critical Path Milestone will be to characterize immortal clonal cell lines versus primary cultures wherefrom the cell lines originated to determine andapos usabilityandapos in different bioassays Milestone will be to offer custom packages of human normal and ADPKD kidney cell platforms from our Product Line Menu to the PKD academic and industry research communities in Years and of the award and beyond this term as a self sustaining business

PUBLIC HEALTH RELEVANCE DiscoveryBioMed Inc a small business concern is leading a collaboration with multiple academic centers cores and experts to establish useful human normal and autosomal dominant polycystic kidney disease ADPKD cell platforms for industry and academic marketplaces ADPKD is a debilitating inherited disease for which there are no specific FDA approved medicines to date ADPKD is the leading genetic cause of dialysis and renal transplantation worldwide affecting an estimated million diagnosed and undiagnosed individuals worldwide Diseased human ADPKD kidneys removed for dialysis or upon transplantation are a rich source of cystic kidney epithelial cells and fibroblasts that shoul be engineered into useful research tools and not discarded The cell cultures established by DBM and collaborators will be engineered into research platforms and will answer fundamental questions about ADPKD disease pathogenesis and genetics These platforms will also be used for therapeutics discovery specific to ADPKD a disease with unmet clinical need