Peptide-mediated attachment of non-plateable hepatocytes to surfaces

描述（由申请人提供）：为了降低将新药推向市场的成本，必须在其开发的早期识别具有高失败概率的候选药物。制药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-药物相互作用。原代人肝细胞是评估药物代谢、肝毒性、药物-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，这导致研究人员开发了能够长期储存肝细胞的冷冻保存方法。尽管这项技术具有优势，但50%的肝细胞在冷冻保存后失去了附着在培养板上的能力。不像镀肝细胞可以存活数天到数周，悬浮的肝细胞只能存活数小时；因此，非可平板性肝细胞的使用仅限于短期代谢分析。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已经确定了肽序列特异性结合非平板冷冻保存的肝细胞。我们将利用这些肽将非板性肝细胞附着在胶原包被孔板上。我们预计这种附着将逆转与“分离诱导细胞死亡”相关的信号级联反应，最终使细胞有时间恢复与胶原底物的接触。

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.