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，必须在其开发早期确定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，以发现不需要的有毒代谢物和有害的药物相互作用。原代人肝细胞是评估药物代谢、肝毒性、药物相互作用和CYP 450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，导致研究人员开发能够长期储存肝细胞的冷冻保存方法。尽管该技术具有优势，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。与可存活数天至数周的平板接种肝细胞不同，悬浮液中的肝细胞仅存活数小时;因此，不可平板接种的肝细胞仅限于用于短期代谢试验。最终，这减少了完全功能性肝细胞的可用供应，增加了肝细胞供应商和最终用户的成本。Affinergy已经鉴定了与非平板冻存肝细胞特异性结合的肽序列。我们将利用这些肽将不可接种的肝细胞附着到胶原包被的孔板上。我们预计这种附着将逆转与“炎症诱导的细胞死亡”相关的信号级联反应，最终使细胞有时间恢复与底层胶原基质的接触。

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.