Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo delivery
Although a considerable number of disease-related biomolecular interactions occur in the cytosol, therapeutic and diagnostic application of target-specific binding proteins is largely confined to surface-exposed or extracellular targets. Therefore, protein-cargo delivery approaches, including cell-p...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-03-01
|
| Series: | Molecular Therapy: Oncology |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2950329924001735 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832595259840790528 |
|---|---|
| author | Carolin Sophie Dombrowsky Felix Klaus Geyer Diana Zakharchuk Harald Kolmar |
| author_facet | Carolin Sophie Dombrowsky Felix Klaus Geyer Diana Zakharchuk Harald Kolmar |
| author_sort | Carolin Sophie Dombrowsky |
| collection | DOAJ |
| description | Although a considerable number of disease-related biomolecular interactions occur in the cytosol, therapeutic and diagnostic application of target-specific binding proteins is largely confined to surface-exposed or extracellular targets. Therefore, protein-cargo delivery approaches, including cell-penetrating peptides and cytosol-penetrating antibodies, are being explored to overcome this limitation. In this context, we have developed a modular approach for cytosolic penetration of tumor cells based on bispecific antibodies containing a masked cytosol-penetrating Fab on one arm and a tumor-targeting scFv linked via an endosomal cleavable linker on the other arm. The relevance of the antigen-specific binding, internalization, and cytosolic cargo delivery was demonstrated in several in vitro assays using different cell lines with anti-B7-H3 scFv, the well-characterized trastuzumab (HER2), and inotuzumab (CD22) as examples. In addition, presence of the masking moiety to prevent non-specific surface binding, as well as the activation of cytosol-penetrating capabilities in the tumor microenvironment upon release by tumor-specific proteases was confirmed using the catalytic domain of Pseudomonas exotoxin as model cargo for cytosol delivery. Tumor microenvironment-dependent as well as tumor-associated antigen-specific cytosol-penetrating antibodies of the type developed here have the potential to serve as a modular platform to deliver macromolecular cargoes for addressing intracellular targets in tumor cells. |
| format | Article |
| id | doaj-art-7721cb9ee1734da7883f9d0532b4ea2d |
| institution | Kabale University |
| issn | 2950-3299 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Molecular Therapy: Oncology |
| spelling | doaj-art-7721cb9ee1734da7883f9d0532b4ea2d2025-01-19T06:27:06ZengElsevierMolecular Therapy: Oncology2950-32992025-03-01331200931Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo deliveryCarolin Sophie Dombrowsky0Felix Klaus Geyer1Diana Zakharchuk2Harald Kolmar3Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Peter-Grünberg-Strasse 4, 64287 Darmstadt, GermanyInstitute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Peter-Grünberg-Strasse 4, 64287 Darmstadt, GermanyInstitute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Peter-Grünberg-Strasse 4, 64287 Darmstadt, GermanyInstitute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Peter-Grünberg-Strasse 4, 64287 Darmstadt, Germany; Centre for Synthetic Biology, Technical University of Darmstadt, 64287 Darmstadt, Germany; Corresponding author: Harald Kolmar, Institute for Organic Chemistry and Biochemistry, Technical University of Darmstadt, Peter-Grünberg-Strasse 4, 64287 Darmstadt, Germany.Although a considerable number of disease-related biomolecular interactions occur in the cytosol, therapeutic and diagnostic application of target-specific binding proteins is largely confined to surface-exposed or extracellular targets. Therefore, protein-cargo delivery approaches, including cell-penetrating peptides and cytosol-penetrating antibodies, are being explored to overcome this limitation. In this context, we have developed a modular approach for cytosolic penetration of tumor cells based on bispecific antibodies containing a masked cytosol-penetrating Fab on one arm and a tumor-targeting scFv linked via an endosomal cleavable linker on the other arm. The relevance of the antigen-specific binding, internalization, and cytosolic cargo delivery was demonstrated in several in vitro assays using different cell lines with anti-B7-H3 scFv, the well-characterized trastuzumab (HER2), and inotuzumab (CD22) as examples. In addition, presence of the masking moiety to prevent non-specific surface binding, as well as the activation of cytosol-penetrating capabilities in the tumor microenvironment upon release by tumor-specific proteases was confirmed using the catalytic domain of Pseudomonas exotoxin as model cargo for cytosol delivery. Tumor microenvironment-dependent as well as tumor-associated antigen-specific cytosol-penetrating antibodies of the type developed here have the potential to serve as a modular platform to deliver macromolecular cargoes for addressing intracellular targets in tumor cells.http://www.sciencedirect.com/science/article/pii/S2950329924001735MT: Regular Issuecytosol-penetrating antibodycytosolic deliveryendosomal escapemasked cytosol-penetrating antibodytumor-associated antigen specificity |
| spellingShingle | Carolin Sophie Dombrowsky Felix Klaus Geyer Diana Zakharchuk Harald Kolmar Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo delivery Molecular Therapy: Oncology MT: Regular Issue cytosol-penetrating antibody cytosolic delivery endosomal escape masked cytosol-penetrating antibody tumor-associated antigen specificity |
| title | Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo delivery |
| title_full | Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo delivery |
| title_fullStr | Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo delivery |
| title_full_unstemmed | Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo delivery |
| title_short | Tumor-specific cytosol-penetrating antibodies for antigen- and TME-dependent intracellular cargo delivery |
| title_sort | tumor specific cytosol penetrating antibodies for antigen and tme dependent intracellular cargo delivery |
| topic | MT: Regular Issue cytosol-penetrating antibody cytosolic delivery endosomal escape masked cytosol-penetrating antibody tumor-associated antigen specificity |
| url | http://www.sciencedirect.com/science/article/pii/S2950329924001735 |
| work_keys_str_mv | AT carolinsophiedombrowsky tumorspecificcytosolpenetratingantibodiesforantigenandtmedependentintracellularcargodelivery AT felixklausgeyer tumorspecificcytosolpenetratingantibodiesforantigenandtmedependentintracellularcargodelivery AT dianazakharchuk tumorspecificcytosolpenetratingantibodiesforantigenandtmedependentintracellularcargodelivery AT haraldkolmar tumorspecificcytosolpenetratingantibodiesforantigenandtmedependentintracellularcargodelivery |