Cathepsin B Is Dispensable for Cellular Processing of Cathepsin B-Cleavable Antibody–Drug Conjugates
| العنوان: | Cathepsin B Is Dispensable for Cellular Processing of Cathepsin B-Cleavable Antibody–Drug Conjugates |
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| المؤلفون: | Tommy K. Cheung, Josefa Chuh, Katherine R. Kozak, Donglu Zhang, Yong Ma, Jack Sadowsky, Qui Phung, Robert W. Akita, Niña G. Caculitan, Byoung Chul Lee, Benjamin Haley, Yichin Liu, Andrew Polson, Mark X. Sliwkowski, Thomas H. Pillow |
| المصدر: | Cancer Research. 77:7027-7037 |
| بيانات النشر: | American Association for Cancer Research (AACR), 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, Cancer Research, Immunoconjugates, Antineoplastic Agents, Cathepsin B, 03 medical and health sciences, chemistry.chemical_compound, 0302 clinical medicine, Cell Line, Tumor, Humans, Prodrugs, Cells, Cultured, Cathepsin, Oligopeptide, Chemistry, Antibodies, Monoclonal, Valine, Cysteine protease, body regions, HEK293 Cells, 030104 developmental biology, Cell killing, Oncology, Monomethyl auristatin E, Biochemistry, 030220 oncology & carcinogenesis, Proteolysis, Citrulline, Drug Screening Assays, Antitumor, Oligopeptides, Linker, Cysteine |
| الوصف: | Antibody–drug conjugates (ADC) are designed to selectively bind to tumor antigens via the antibody and release their cytotoxic payload upon internalization. Controllable payload release through judicious design of the linker has been an early technological milestone. Here, we examine the effect of the protease-cleavable valine-citrulline [VC(S)] linker on ADC efficacy. The VC(S) linker was designed to be cleaved by cathepsin B, a lysosomal cysteine protease. Surprisingly, suppression of cathepsin B expression via CRISPR-Cas9 gene deletion or shRNA knockdown had no effect on the efficacy of ADCs with VC(S) linkers armed with a monomethyl auristatin E (MMAE) payload. Mass spectrometry studies of payload release suggested that other cysteine cathepsins can cleave the VC(S) linker. Also, ADCs with a nonprotease-cleavable enantiomer, the VC(R) isomer, mediated effective cell killing with a cysteine-VC(R)-MMAE catabolite generated by lysosomal catabolism. Based on these observations, we altered the payload to a pyrrolo[2,1-c][1,4]benzodiazepine dimer (PBD) conjugate that requires linker cleavage in order to bind its DNA target. Unlike the VC-MMAE ADCs, the VC(S)-PBD ADC is at least 20-fold more cytotoxic than the VC(R)-PBD ADC. Our findings reveal that the VC(S) linker has multiple paths to produce active catabolites and that antibody and intracellular targets are more critical to ADC efficacy. These results suggest that protease-cleavable linkers are unlikely to increase the therapeutic index of ADCs and that resistance based on linker processing is improbable. Cancer Res; 77(24); 7027–37. ©2017 AACR. |
| تدمد: | 1538-7445 0008-5472 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::97963072f0241a139d445ff8921bb822 https://doi.org/10.1158/0008-5472.can-17-2391 |
| رقم الأكسشن: | edsair.doi.dedup.....97963072f0241a139d445ff8921bb822 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15387445 00085472 |
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