In vitro fabrication of microscale secretory granules

HECTOR LOPEZ LAGUNA; ELOI PARLADE; PATRICIA ÁLAMO,; JULIETA M. SANCHEZ; ERIC VOLTA DURAN; NAROA SERNA; LAURA SÁNCHEZ-GARCIA; OLIVIA CANO-GARRIDO; ALEJANDRO SÁNCHEZ-CHARDI; ANTONIO VILLAVERDE; RAMON MANGUES; UGUTZ UNZUETA; ESTHER VÁZQUEZ

ADVANCED FUNCTIONAL MATERIALS

WILEY-V C H VERLAG GMBH

Lugar: Weinheim; Año: 2021

1616-301X

In diverse clinical settings, the application of advanced medical treatments and drug delivery approaches require the development of fully biocompatible materials with the ability to release functional drugs in a time-prolonged way. Ideally, the delivered molecules should be self-contained in the form of a chemically homogenous entity, to prevent the use of potentially toxic scaffolds or holding matrices. In nature, peptidic hormones are self-stored in protein-only secretory granules formed by the reversible coordination of Zn2+ and histidine residues. Inspired by this concept, we have developed, analyzed and comparatively applied an in vitro transversal procedure for the fabrication of protein-only secretory granules at the microscale, aimed to sustained protein drug delivery. These materials can be produced out from any polyhistidine-tagged protein by using physiological concentrations of Zn2+ as a potent and versatile glue-like agent. By the screening of granules formed by 12 engineered and non-engineered proteins at different Zn2+ concentrations, we have defined optimal fabrication conditions and determined the consequent further release kinetics. Moreover, we have assessed the functional and structural properties of the delivered protein using a drug-targeting protein platform in a mouse model of human colorectal cancer. Finally, the incorporation of short histidine tags allows the packaging of structurally and functionally dissimilar polypeptides, what validates the proposed fabrication method as a standard, generic and powerful protocol extensible to diverse scenarios in which slow protein drug delivery is required.