Friday, May 23, 2025
5/23/2025
Genotyped and Single Cyst-derived Human ADPKD Cell Platforms for Industry and Academia - 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 12 single cyst-derived primary cultures and 7 multicystic tissue-derived primary cultures from 2 human ADPKD (huADPKD) donor kidneys (procured from commercial vendors) in pilot Phase 1-like studies. These initial 19 huADPKD primary cultures were combined with primary cultures from 11 different donors from Dr. Darren Wallace’s PKD Research Biomaterials and Cellular Models Core at Kansas University Medical Center (KUMC) so that all 30 samples could be genotyped by Dr. Peter Harris’ Core at the Mayo PKD Center. Each huADPKD primary culture has been profiled in 2D microtiter plate-driven proliferation bioassays on tissue culture plastic as well as in 3D 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 in 2D and 3D 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: (1) 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 2D and 3D bioassays for PKD research? and (2) 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 ‘one hit’ vs. ‘two hit’ hypothesis or the emergence of somatic mutations and the possible presence of haploinsufficiency)? DBM et al. set 5 milestones for our work to be supported by this possible Fast Track SBIR award. Milestone 1 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 12-15 different donors processed. Milestone 2 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 3 will be to immortalize select primary cultures using a well-optimized Critical Path. Milestone 4 will be to characterize immortal clonal cell lines versus primary cultures wherefrom the cell lines originated to determine ‘usability’ in different bioassays. Milestone 5 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 2 and 3 of the award and beyond this term as a self-sustaining business.
HHS’ Tracking Accountability in Government Grants System (TAGGS) website provides access to detailed descriptions of grants, loans, aggregated direct payments and other types of financial assistance awarded by the HHS Operating Divisions (OPDIVs) and the Office of the Secretary Staff Divisions (STAFFDIVs).