Engineering Toxoplasma gondii secretion systems for intracellular delivery of multiple large therapeutic proteins to neurons.

Autor: Bracha S; Department of Neurobiology, Biochemistry and Biophysics, Wise Faculty of Life Sciences and Sagol School for Neuroscience, Tel Aviv University, Tel Aviv, Israel. shaharbr@mit.edu.; McGovern Institute for Brain Research, MIT, Cambridge, MA, USA. shaharbr@mit.edu., Johnson HJ; Neuroscience Graduate Interdisciplinary Program, University of Arizona, Tucson, AZ, USA.; Departments of Neurology and Immunobiology, College of Medicine, and BIO5 Institute, University of Arizona, Tucson, AZ, USA., Pranckevicius NA; Centre for Parasitology, School of Infection and Immunity, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK., Catto F; Institute of Neuropathology, University Hospital Zurich, University of Zurich, Zurich, Switzerland., Economides AE; Institute of Neuropathology, University Hospital Zurich, University of Zurich, Zurich, Switzerland., Litvinov S; Computational Science and Engineering Laboratory, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA., Hassi K; Centre for Parasitology, School of Infection and Immunity, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK., Rigoli MT; Human Technopole, Milan, Italy.; Department of Experimental Oncology, European Institute of Oncology IRCCS, Milan, Italy.; Department of Oncology and Hemato-oncology, University of Milan, Milan, Italy., Cheroni C; Human Technopole, Milan, Italy.; Department of Oncology and Hemato-oncology, University of Milan, Milan, Italy., Bonfanti M; Human Technopole, Milan, Italy., Valenti A; Human Technopole, Milan, Italy.; Department of Experimental Oncology, European Institute of Oncology IRCCS, Milan, Italy.; Department of Oncology and Hemato-oncology, University of Milan, Milan, Italy., Stucchi S; Human Technopole, Milan, Italy.; Department of Experimental Oncology, European Institute of Oncology IRCCS, Milan, Italy.; Department of Oncology and Hemato-oncology, University of Milan, Milan, Italy., Attreya S; Undergraduate Biology Research Program, University of Arizona, Tucson, AZ, USA., Ross PD; Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.; Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK., Walsh D; Centre for Parasitology, School of Infection and Immunity, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK., Malachi N; Epeius Pharma, Ness Ziona, Israel., Livne H; Epeius Pharma, Ness Ziona, Israel., Eshel R; Epeius Pharma, Ness Ziona, Israel., Krupalnik V; Epeius Pharma, Ness Ziona, Israel., Levin D; Epeius Pharma, Ness Ziona, Israel., Cobb S; Institute of Neuroscience and Psychology, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK.; Centre for Discovery Brain Sciences, University of Edinburgh, Edinburgh, UK., Koumoutsakos P; Computational Science and Engineering Laboratory, School of Engineering and Applied Sciences, Harvard University, Cambridge, MA, USA., Caporale N; Human Technopole, Milan, Italy.; Department of Experimental Oncology, European Institute of Oncology IRCCS, Milan, Italy.; Department of Oncology and Hemato-oncology, University of Milan, Milan, Italy., Testa G; Human Technopole, Milan, Italy. giuseppe.testa@fht.org.; Department of Experimental Oncology, European Institute of Oncology IRCCS, Milan, Italy. giuseppe.testa@fht.org.; Department of Oncology and Hemato-oncology, University of Milan, Milan, Italy. giuseppe.testa@fht.org., Aguzzi A; Institute of Neuropathology, University Hospital Zurich, University of Zurich, Zurich, Switzerland. adriano.aguzzi@usz.ch., Koshy AA; Departments of Neurology and Immunobiology, College of Medicine, and BIO5 Institute, University of Arizona, Tucson, AZ, USA. akoshy@arizona.edu., Sheiner L; Centre for Parasitology, School of Infection and Immunity, College of Medical, Veterinary and Life Sciences, University of Glasgow, Glasgow, UK. lilach.sheiner@glasgow.ac.uk., Rechavi O; Department of Neurobiology, Biochemistry and Biophysics, Wise Faculty of Life Sciences and Sagol School for Neuroscience, Tel Aviv University, Tel Aviv, Israel. odedrechavi@gmail.com.
Jazyk: angličtina
Zdroj: Nature microbiology [Nat Microbiol] 2024 Aug; Vol. 9 (8), pp. 2051-2072. Date of Electronic Publication: 2024 Jul 29.
DOI: 10.1038/s41564-024-01750-6
Abstrakt: Delivering macromolecules across biological barriers such as the blood-brain barrier limits their application in vivo. Previous work has demonstrated that Toxoplasma gondii, a parasite that naturally travels from the human gut to the central nervous system (CNS), can deliver proteins to host cells. Here we engineered T. gondii's endogenous secretion systems, the rhoptries and dense granules, to deliver multiple large (>100 kDa) therapeutic proteins into neurons via translational fusions to toxofilin and GRA16. We demonstrate delivery in cultured cells, brain organoids and in vivo, and probe protein activity using imaging, pull-down assays, scRNA-seq and fluorescent reporters. We demonstrate robust delivery after intraperitoneal administration in mice and characterize 3D distribution throughout the brain. As proof of concept, we demonstrate GRA16-mediated brain delivery of the MeCP2 protein, a putative therapeutic target for Rett syndrome. By characterizing the potential and current limitations of the system, we aim to guide future improvements that will be required for broader application.
(© 2024. The Author(s).)
Databáze: MEDLINE
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