Peptide-mediated attachment of non-plateable hepatocytes to surfaces

肽介导的不可电镀肝细胞与表面的附着

基本信息

批准号：
8780418
负责人：
Bruce Lamb
金额：
$ 28.05万
依托单位：
AFFINERGY, LLC
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2015-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8780418
关键词：
Adherence Albumins Binding Binding Sites Biological Assay CYP1A2 gene CYP3A4 gene Cell Communication Cell Culture Techniques Cell Death Cell physiology Cell surface Cells Chemicals Collagen Collagen Type I Complex Cryopreservation Deposition Development Drug Industry Drug Interactions Enzyme Induction Enzymes Excretory function Exhibits Extracellular Matrix Failure Health Hepatocyte Hepatotoxicity Hour Human In Vitro Incubated Integrins Lead Libraries Liver Longevity Marketing Measures Mediating Membrane Metabolic Metabolism Methods Modeling Morphology Peptides Pharmaceutical Preparations Pharmacologic Substance Phase Probability Process Production Proteins Protocols documentation Research Personnel Signal Transduction Sterilization Surface Suspension substance Suspensions Techniques Testing Time absorption cost culture plates density drug candidate drug metabolism enzyme activity monolayer protein aminoacid sequence public health relevance success tissue culture

项目摘要

DESCRIPTION (provided by applicant): In order to reduce the cost of bringing a new drug to market, drug candidates with a high probability for failure must be identified early in their development. The pharmaceutical industry employs in vitro liver models to screen new drugs for undesirable toxic metabolites and harmful drug-drug interactions. Primary human hepatocytes are the preferred model for assessing drug metabolism, hepatoxicity, drug-drug interactions, and CYP450 enzyme induction. However, the availability of fresh human hepatocytes is unpredictable, leading researchers to develop cryopreservation methods that enable long-term hepatocyte storage. Despite the advantages of this technique, 50% of hepatocytes lose their ability to attach to culture plates following cryopreservation. Unlike plated hepatocytes that remain viable for days to weeks, hepatocytes in suspension are only viable for a few hours; thus, non-plateable hepatocytes are limited in use to only short-term metabolic assays. Ultimately, this diminishes the available supply of fully functional hepatocytes, increasing costs for both hepatocyte suppliers and end users. Affinergy has identified peptide sequences that bind specifically to non- plateable cryopreserved hepatocytes. We will utilize these peptides to attach non-plateable hepatocytes to collagen-coated well plates. We expect that this attachment will reverse signaling cascades associated with "detachment-induced cell death", ultimately giving the cells time to restore contact with the underlying collagen substrates.

描述（由申请人提供）：为了降低新药上市的成本，必须在开发早期就确定失败可能性较高的候选药物。制药行业采用体外肝脏模型来筛选新药中不良的有毒代谢物和有害的药物间相互作用。原代人肝细胞是评估药物代谢、肝毒性、药物相互作用和 CYP450 酶诱导的首选模型。然而，新鲜人肝细胞的可用性是不可预测的，因此研究人员开发了能够长期储存肝细胞的冷冻保存方法。尽管该技术具有诸多优点，但 50% 的肝细胞在冷冻保存后失去了附着于培养板的能力。与平板肝细胞可存活数天至数周不同，悬浮肝细胞仅可存活数小时；因此，不可接种的肝细胞仅限于短期代谢测定。最终，这减少了全功能肝细胞的可用供应，增加了肝细胞供应商和最终用户的成本。 Affinergy 已鉴定出与不可电镀的冷冻保存肝细胞特异性结合的肽序列。我们将利用这些肽将不可电镀的肝细胞附着到胶原蛋白包被的孔板上。我们预计这种附着将逆转与“脱离诱导的细胞死亡”相关的信号级联，最终使细胞有时间恢复与下面的胶原蛋白基质的接触。