Hydra-Elastin-like Polypeptides Increase Rapamycin Potency When Targeting Cell Surface GRP78.

Autor:	Avila H; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Yu J; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Boddu G; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Phan A; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Truong A; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Peddi S; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Guo H; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Lee SJ; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States.; Department of Biomedical Engineering, USC Viterbi School of Engineering, Los Angeles, California 90089, United States., Alba M; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States., Canfield E; Mass Spectrometry Core, USC School of Pharmacy, Los Angeles, California 90089, United States., Yamamoto V; Department of Biochemistry and Molecular Medicine, USC Keck School of Medicine, Los Angeles, California 90033, United States., Paton JC; Research Centre for Infectious Diseases, Department of Molecular and Biomedical Science, University of Adelaide, Adelaide 5005, Australia., Paton AW; Research Centre for Infectious Diseases, Department of Molecular and Biomedical Science, University of Adelaide, Adelaide 5005, Australia., Lee AS; Department of Biochemistry and Molecular Medicine, USC Keck School of Medicine, Los Angeles, California 90033, United States., MacKay JA; Department of Pharmacology and Pharmaceutical Sciences, USC School of Pharmacy, Los Angeles, California 90089, United States.; Department of Biomedical Engineering, USC Viterbi School of Engineering, Los Angeles, California 90089, United States.; Department of Ophthalmology, USC Keck School of Medicine, Los Angeles, California 90033, United States.
Jazyk:	angličtina
Zdroj:	Biomacromolecules [Biomacromolecules] 2022 Aug 08; Vol. 23 (8), pp. 3116-3129. Date of Electronic Publication: 2022 Jul 05.
DOI:	10.1021/acs.biomac.2c00048
Abstrakt:	Rapalogues are powerful therapeutic modalities for breast cancer; however, they suffer from low solubility and dose-limiting side effects. To overcome these challenges, we developed a long-circulating multiheaded drug carrier called 5FA, which contains rapamycin-binding domains linked with elastin-like polypeptides (ELPs). To target these "Hydra-ELPs" toward breast cancer, we here linked 5FA with four distinct peptides which are reported to engage the cell surface form of the 78 kDa glucose-regulated protein (csGRP78). To determine if these peptides affected the carrier solubility, this library was characterized by light scattering and mass spectrometry. To guide in vitro selection of the most potent functional carrier for rapamycin, its uptake and inhibition of mTORC1 were monitored in a ductal breast cancer model (BT474). Using flow cytometry to track cellular association, it was found that only the targeted carriers enhanced cellular uptake and were susceptible to proteolysis by SubA, which specifically targets csGRP78. The functional inhibition of mTOR was monitored by Western blot for pS6K, whereby the best carrier L-5FA reduced mTOR activity by 3-fold compared to 5FA or free rapamycin. L-5FA was further visualized using super-resolution confocal laser scanning microscopy, which revealed that targeting increased exposure to the carrier by ∼8-fold. This study demonstrates how peptide ligands for GRP78, such as the L peptide (RLLDTNRPLLPY), may be incorporated into protein-based drug carriers to enhance targeting.
Databáze:	MEDLINE
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