دورية أكاديمية
أعرض في EDS

Standardized Preclinical In Vitro Blood-Brain Barrier Mouse Assay Validates Endocytosis-Dependent Antibody Transcytosis Using Transferrin-Receptor-Mediated Pathways.

التفاصيل البيبلوغرافية
العنوان: Standardized Preclinical In Vitro Blood-Brain Barrier Mouse Assay Validates Endocytosis-Dependent Antibody Transcytosis Using Transferrin-Receptor-Mediated Pathways.
المؤلفون: Morrison JI; Institutionen för Farmaci, Uppsala Universitet, Uppsala 752 37, Sweden., Petrovic A; Institutionen för Farmaci, Uppsala Universitet, Uppsala 752 37, Sweden., Metzendorf NG; Institutionen för Farmaci, Uppsala Universitet, Uppsala 752 37, Sweden., Rofo F; Institutionen för Farmaci, Uppsala Universitet, Uppsala 752 37, Sweden., Yilmaz CU; Institutionen för Farmaci, Uppsala Universitet, Uppsala 752 37, Sweden., Stenler S; Institutionen för Farmaci, Uppsala Universitet, Uppsala 752 37, Sweden., Laudon H; BioArctic AB, Stockholm 112 51, Sweden., Hultqvist G; Institutionen för Farmaci, Uppsala Universitet, Uppsala 752 37, Sweden.
المصدر: Molecular pharmaceutics [Mol Pharm] 2023 Mar 06; Vol. 20 (3), pp. 1564-1576. Date of Electronic Publication: 2023 Feb 21.
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: American Chemical Society Country of Publication: United States NLM ID: 101197791 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1543-8392 (Electronic) Linking ISSN: 15438384 NLM ISO Abbreviation: Mol Pharm Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Washington, DC : American Chemical Society, c2004-
مواضيع طبية MeSH: Blood-Brain Barrier*/metabolism , Percutaneous Coronary Intervention*, Mice ; Animals ; Brain/metabolism ; Receptors, Transferrin/metabolism ; Transcytosis ; Immunoglobulin G/metabolism ; Transferrins/metabolism
مستخلص: The presence of the blood-brain barrier (BBB) creates a nigh-on impenetrable obstacle for large macromolecular therapeutics that need to be delivered to the brain milieu to treat neurological disorders. To overcome this, one of the strategies used is to bypass the barrier with what is referred to as a "Trojan Horse" strategy, where therapeutics are designed to use endogenous receptor-mediated pathways to piggyback their way through the BBB. Even though in vivo methodologies are commonly used to test the efficacy of BBB-penetrating biologics, comparable in vitro BBB models are in high demand, as they benefit from being an isolated cellular system devoid of physiological factors that can on occasion mask the processes behind BBB transport via transcytosis. We have developed an in vitro BBB model (In-Cell BBB-Trans assay) based on the murine cEND cells that help delineate the ability of modified large bivalent IgG antibodies conjugated to the transferrin receptor binder scFv8D3 to cross an endothelial monolayer grown on porous cell culture inserts (PCIs). Following the administration of bivalent antibodies into the endothelial monolayer, a highly sensitive enzyme-linked immunosorbent assay (ELISA) is used to determine the concentration in the apical (blood) and basolateral (brain) chambers of the PCI system, allowing for the evaluation of apical recycling and basolateral transcytosis, respectively. Our results show that antibodies conjugated to scFv8D3 transcytose at considerably higher levels compared to unconjugated antibodies in the In-Cell BBB-Trans assay. Interestingly, we are able to show that these results mimic in vivo brain uptake studies using identical antibodies. In addition, we are able to transversely section PCI cultured cells, allowing for the identification of receptors and proteins that are likely involved in the transcytosis of the antibodies. Furthermore, studies using the In-Cell BBB-Trans assay revealed that transcytosis of the transferrin-receptor-targeting antibodies is dependent on endocytosis. In conclusion, we have designed a simple, reproducible In-Cell BBB-Trans assay based on murine cells that can be used to rapidly determine the BBB-penetrating capabilities of transferrin-receptor-targeting antibodies. We believe that the In-Cell BBB-Trans assay can be used as a powerful, preclinical screening platform for therapeutic neurological pathologies.
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فهرسة مساهمة: Keywords: 3R; TfR; blood−brain barrier; in vitro BBB; transcytosis; transferrin receptor
المشرفين على المادة: 0 (Receptors, Transferrin)
0 (Immunoglobulin G)
0 (Transferrins)
تواريخ الأحداث: Date Created: 20230222 Date Completed: 20230307 Latest Revision: 20230315
رمز التحديث: 20231215
مُعرف محوري في PubMed: PMC9997753
DOI: 10.1021/acs.molpharmaceut.2c00768
PMID: 36808999
قاعدة البيانات: MEDLINE
الوصف
تدمد:1543-8392
DOI:10.1021/acs.molpharmaceut.2c00768