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

用于工业界和学术界的基因分型和单囊肿衍生的人类 ADPKD 细胞平台

基本信息

批准号：
9139596
负责人：
Erik Mills Schwiebert
金额：
$ 36.62万
依托单位：
DISCOVERYBIOMED, INC.
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-04-01 至 2017-05-04
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9139596
关键词：
Academia Academic Medical Centers Address Affect Applied Research Autosomal Dominant Polycystic Kidney Award Baltimore Basic Science Biological Assay Biomaterials Research Businesses Cell Culture Techniques Cell Line Cell model Cells Client Clinical Coculture Techniques Collaborations Collection Communities Complement Conditioned Culture Media Critical Pathways Custom Cyst Cystic kidney Data Databases Diagnosis Dialysis procedure Differentiation Antigens Disease Engineering Epithelial Cells Exhibits FDA approved Fibroblasts Funding Genetic Genotype Goals Heterogeneity Human Individual Industry Inherited Kansas Kidney Kidney Transplantation Life Medicine Methods Mutation Nephrons Normal Cell Pathogenesis Performance Phase Phenotype Population Principal Investigator Process Research Research Personnel Sampling Small Business Innovation Research Grant Somatic Mutation Source Therapeutic Uses Tissue Sample Tissues Translational Research Transplantation Vendor Work base commercialization immunocytochemistry interest kidney cell kidney epithelial cell matrigel medical specialties meetings public health relevance tissue culture tool usability

项目摘要

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 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 i 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 ht' 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.

描述（由申请人提供）：DiscoveryBioMed, Inc.（DBM, Inc. 或 DBM）专门从事正常和患病人类细胞平台工程，DBM 已组建了一个专家联盟；因此，我们共同寻求快速通道 SBIR 资金来建立和实施。为学术和行业市场现有的和感兴趣的 DBM 客户提供正常和 ADPKD 人类细胞平台的产品线菜单，并将根据产品线菜单中的哪些选项适合他们的研究进行展示。正如我们的研究战略和商业化计划中所强调的，这一备受期待的产品线菜单将包括一系列：(a) 特种细胞培养成分，(b) 原代正常和 ADPKD 人类原代培养物，(c) 混合永生培养物和(d) 将经过充分表征、基因分型并定义为肾单位起源片段的克隆永生细胞系与它们来源的原代培养物进行比较，以确定它们在生物测定性能方面是否相似。基因型，并且在DBM 和合作者已开始在试验阶段 1 中收集和表征来自 2 个人类 ADPKD (huADPKD) 供体肾脏（从商业供应商采购）的 12 个单囊肿来源的原代培养物和 7 个多囊组织来源的原代培养物。这些最初的 19 个 huADPKD 原代培养物与来自 Darren Wallace 博士的 PKD 研究生物材料和细胞模型的 11 个不同捐赠者的原代培养物相结合。堪萨斯大学医学中心 (KUMC) 的核心，以便 Mayo PKD 中心 Peter Harris 博士的核心可以对所有 30 个样本进行基因分型，每个 huADPKD 原代培养物均已在组织培养塑料上通过 2D 微量滴定板驱动的增殖生物测定进行了分析。以及 3D 基质胶基于胞囊发生生物测定的初步数据支持了上述试点工作，显示出令人信服的基因型并表现出卓越实用性的 2D 和 3D 生物测定将通过多种遗传方法永生化。在必要的情况下，我们的应用科学研究将在 huADPKD 细胞中鉴定肾单位起源片段：（1）原发性 huADPKD 是否存在。成纤维细胞支持（共培养、条件培养基，两者）使人 ADPKD 囊性上皮细胞原代培养物寿命更长和/或在 PKD 研究的 2D 和 3D 生物测定中获得特定表型，以及 (2) 单个囊肿是否存在于单个囊肿中？考虑到 huADPKD 供体肾脏具有相同的基因型，或者囊肿与囊肿之间的基因型是否不同（即，探索“一次打击”与“两次 ht”假说或出现体细胞突变和可能存在的单倍体不足）？ DBM 等人为我们的工作设定了 5 个里程碑，以获得这一可能的快速通道 SBIR 奖项的支持，其中里程碑 1 将是建立来自单囊肿和多囊组织的原代培养物。每个 huADPKD 供体肾组织样本，最终目标是处理 12-15 个不同的供体，里程碑 2 将对每个囊肿来源和多囊组织来源的原代培养物进行基因分型，进行比较。里程碑 3 将是使用优化的关键路径使选定的原代培养物永生化，并根据 ADPKD 基因型数据库选择候选原代培养物。里程碑 4 将表征永生克隆细胞系与细胞系来源的原代培养物。确定不同生物测定中的“可用性”将是在第 2 年和第 2 年向 PKD 学术和行业研究社区提供人类正常和 ADPKD 肾细胞平台的定制包。 3 奖项及此期限后作为自给自足的企业。