Canhão Pedro G M, Snoeys Jan, Geerinckx Suzy, van Heerden Marjolein, Van den Bergh An, Holm Camden, Markus Jan, Ayehunie Seyoum, Monshouwer Mario, Evers Raymond, Augustijns Patrick, Kourula Stephanie
Preclinical Sciences & Translational Safety, Johnson & Johnson, Turnhoutseweg 30, 2340, Beerse, Belgium; Drug Delivery and Disposition, KU Leuven, Gasthuisberg O&N II, Herestraat 49 - box 921, 3000 Leuven, Belgium.
Preclinical Sciences & Translational Safety, Johnson & Johnson, Turnhoutseweg 30, 2340, Beerse, Belgium.
Eur J Pharm Sci. 2025 Jun 1;209:107025. doi: 10.1016/j.ejps.2025.107025. Epub 2025 Jan 24.
The purpose of this study was to evaluate EpiColon, a novel human organotypic 3D colon microtissue prototype, developed to assess colonic drug disposition, with a particular focus on permeability ranking, and compare its performance to Caco-2 monolayers. EpiColon was characterized for barrier function using transepithelial electrical resistance (TEER), morphology via histology and immunohistochemistry, and functionality through drug transport studies measuring apparent permeability (P). Cutoff thresholds for the permeability of FITC-dextran 4 kDa (FD4), FITC-dextran 10 kDa (FD10S), and [C]mannitol were established to monitor microtissue integrity. Permeability of EpiColon for 20 benchmark drugs was compared with Caco-2 data, and the activity of pivotal efflux transporters, including multidrug resistance protein 1/P-glycoprotein (MDR1/P-gp), along with multidrug resistance protein 2 (MRP2) and breast cancer resistance protein (BCRP), was evaluated using selective substrates. EpiColon exhibited a physiological barrier function (272.0 ± 53.05 Ω x cm) and effectively discriminated between high (e.g., budesonide and [H]metoprolol) and low permeable compounds (e.g., [H]atenolol and [C]mannitol). The model demonstrated functional activity for key efflux transporters, with efflux ratios of 2.32 for [H]digoxin (MDR1/P-gp) and 3.34 for sulfasalazine (MRP2 and BCRP). Notably, EpiColon showed an enhanced dynamic range in the low permeability range, differentiating P between FD4 and FD10S, in contrast to Caco-2 monolayers. Significant positive correlations were observed between human fraction absorbed (f) and logarithmically transformed P [AP-BL] values for both EpiColon (r = 0.68) and Caco-2 (r = 0.68). Furthermore, EpiColon recapitulates some essential phenotypic and cellular features of the human colon, including the expression of critical marker genes (Pan-Cytokeratin: epithelial/colonocytes, Vimentin: mesenchymal/fibroblast, and Alcian Blue: goblet cell/mucus). In conclusion, EpiColon is a promising platform that offers a valuable complement to conventional Caco-2 monolayers for studying colonic drug disposition. However, the presence of flat and some cuboidal cells, along with low throughput, must be addressed to improve its applicability in both academic research and pharmaceutical industry.
本研究的目的是评估EpiColon，一种新型的人体器官型3D结肠微组织原型，其开发用于评估结肠药物处置，特别关注通透性排序，并将其性能与Caco-2单层细胞进行比较。使用跨上皮电阻（TEER）对EpiColon的屏障功能进行表征，通过组织学和免疫组织化学研究其形态，并通过测量表观通透性（P）的药物转运研究评估其功能。建立了4 kDa异硫氰酸荧光素 - 葡聚糖（FD4）、10 kDa异硫氰酸荧光素 - 葡聚糖（FD10S）和[C]甘露醇通透性的截止阈值，以监测微组织的完整性。将EpiColon对20种基准药物的通透性与Caco-2的数据进行比较，并使用选择性底物评估关键外排转运蛋白的活性，包括多药耐药蛋白1/ P - 糖蛋白（MDR1 / P - gp）以及多药耐药蛋白2（MRP2）和乳腺癌耐药蛋白（BCRP）。EpiColon表现出生理屏障功能（272.0±53.05Ωx cm），并有效区分高通透性化合物（如布地奈德和[H]美托洛尔）和低通透性化合物（如[H]阿替洛尔和[C]甘露醇）。该模型展示了关键外排转运蛋白的功能活性，[H]地高辛（MDR1 / P - gp）的外排率为2.32，柳氮磺胺吡啶（MRP2和BCRP）的外排率为3.34。值得注意的是，与Caco-2单层细胞相比，EpiColon在低通透性范围内显示出增强的动态范围，能够区分FD4和FD10S之间的P值。在EpiColon（r = 0.68）和Caco-2（r = 0.68）中，均观察到人体吸收分数（f）与对数转换后的P [AP - BL]值之间存在显著正相关。此外，EpiColon概括了人类结肠的一些基本表型和细胞特征，包括关键标记基因的表达（泛细胞角蛋白：上皮/结肠细胞，波形蛋白：间充质/成纤维细胞，阿尔辛蓝：杯状细胞/黏液）。总之，EpiColon是一个有前景的平台，为研究结肠药物处置提供了对传统Caco-2单层细胞有价值的补充。然而，必须解决扁平细胞和一些立方体细胞的存在以及低通量问题，以提高其在学术研究和制药行业中的适用性。
