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bioRxiv · 10.1101/481192

Engineering Brain Parasites for Intracellular Delivery of Therapeutic Proteins

Bracha, S.·Hassi, K.·Ross, P. D.·Cobb, S.·Sheiner, L.·Rechavi, O.
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Abstract

Protein therapy has the potential to alleviate many neurological diseases; however, delivery mechanisms for the central nervous system (CNS) are limited, and intracellular delivery poses additional hurdles. To address these challenges, we harnessed the protist parasite Toxoplasma gondii, which can migrate into the CNS and secrete proteins into cells. Using a fusion protein approach, we engineered T. gondii to secrete therapeutic proteins for human neurological disorders. We tested two secretion systems, generated fusion proteins that localized to T. gondiis secretory organelles and assessed their intracellular targeting in various mammalian cells including neurons. We show that T. gondii expressing GRA16 fused to the Rett syndrome protein MeCP2 deliver a fusion protein that mimics the endogenous MeCP2, binding heterochromatic DNA in neurons. This demonstrates the potential of T. gondii as a therapeutic protein vector, which could provide either transient or chronic, in situ synthesis and delivery of intracellular proteins to the CNS.

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Bracha, S., Hassi, K., Ross, P. D., Cobb, S., Sheiner, L., Rechavi, O.. 2018-12-03. Engineering Brain Parasites for Intracellular Delivery of Therapeutic Proteins. https://doi.org/10.1101/481192

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