Abstract
The antioxidant enzyme catalase represents an important therapeutic target due to its role in mitigating cellular reactive oxygen species that contribute to the pathogenesis of many disease states. Catalase-SKL (CAT-SKL), a genetically engineered, peroxisome-targeted, catalase derivative, was developed in order to increase the therapeutic potential of the enzyme, and has previously been shown to be effective in combating oxidative stress in a variety of in vitro and in vivo models, thereby mitigating cellular degeneration and death. In the present study we addressed important considerations for the development of an extracellular vesicle-packaged version of CAT-SKL (evCAT-SKL) as a therapeutic for neurodegenerative diseases by investigating its delivery potential to the brain when administered intranasally, and safety by assessing off-target toxicity in a mouse model. Mice received weekly intranasal administrations of evCAT-SKL or empty extracellular vesicles for 4 weeks. Fluorescent labeling for CAT-SKL was observed throughout all sections of the brain in evCAT-SKL-treated mice, but not in empty extracellular vesicle-treated mice. Furthermore, we found no evidence of gross or histological abnormalities following evCAT-SKL or empty extracellular vesicle treatment in a full-body toxicological analysis. Combined, the successful brain targeting and the lack of off-target toxicity demonstrates that intranasal delivery of extracellular vesicle-packaged CAT-SKL holds promise as a therapeutic for addressing neurological disorders.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Ambani LM, Van Woert MH, Murphy S (1975) Brain peroxidase and catalase in Parkinson disease. Arch Neurol
Batrakova EV, Kim MS (2015) Using exosomes, naturally-equipped nanocarriers, for drug delivery. J Control Release
Birben E, Sahiner UM, Sackesen C, Erzurum S, Kalayci O (2012) Oxidative stress and antioxidant defense. World Allergy Organ J
Cipolla CM, Lodhi IJ (2017) Peroxisomal dysfunction in age-related diseases. Trends Endocrinol Metab
Crowe TP, Greenlee MHW, Kanthasamy AG, Hsu WH (2018) Mechanism of intranasal drug delivery directly to the brain. Life Sci
Fan Y, Chen M, Zhang J, Maincent P, Xia X, Wu W (2018) Updated progress of nanocarrier-based intranasal drug delivery systems for treatment of brain diseases. Crit Rev Ther Drug Carrier Syst
Giordano CR, Terlecky LJ, Bollig-Fischer A, Walton PA, Terlecky SR (2014) Amyloid-beta neuroprotection mediated by a targeted antioxidant. Sci Rep
Giordano CR, Terlecky SR (2012) Peroxisomes, cell senescence, and rates of aging. Biochim Biophys Acta - Mol Basis Dis
Gsell W, Conrad R, Hickethier M, Sofic E, Frölich L, Wichart I, Jellinger K, Moll G, Ransmayr G, Beckmann H, Riederer P (1995) Decreased catalase activity but unchanged superoxide dismutase activity in brains of patients with dementia of Alzheimer type. J Neurochem
Haney MJ, Klyachko NL, Zhao Y, Gupta R, Plotnikova EG, He Z, Patel T, Piroyan A, Sokolsky M, Kabanov AV, Batrakova EV (2015) Exosomes as drug delivery vehicles for Parkinson’s disease therapy. J Control Release
Haney MJ, Zhao Y, Jin YS, Batrakova EV (2020) Extracellular vesicles as drug carriers for enzyme replacement therapy to treat CLN2 Batten disease: optimization of drug administration routes. Cells
Hanson LR, Fine JM, Svitak AL, Faltesek KA (2013) Intranasal administration of CNS therapeutics to awake mice. J Vis Exp
Iraci N, Leonardi T, Gessler F, Vega B, Pluchino S (2016) Focus on extracellular vesicles: physiological role and signalling properties of extracellular membrane vesicles. Int J Mol Sci
Johnsen KB, Gudbergsson JM, Skov MN, Pilgaard L, Moos T, Duroux M (2014) A comprehensive overview of exosomes as drug delivery vehicles—endogenous nanocarriers for targeted cancer therapy. Biochim Biophys Acta - Rev Cancer
Kalani A, Chaturvedi P, Kamat PK, Maldonado C, Bauer P, Joshua IG, Tyagi SC, Tyagi N (2016) Curcumin-loaded embryonic stem cell exosomes restored neurovascular unit following ischemia-reperfusion injury. Int J Biochem Cell Biol
Kalani A, Tyagi N (2015) Exosomes in neurological disease, neuroprotection, repair and therapeutics: problems and perspectives. Neural Regen Res
Khan AR, Liu M, Khan MW, Zhai G (2017) Progress in brain targeting drug delivery system by nasal route. J Control Release
Koepke JI, Nakrieko KA, Wood CS, Boucher KK, Terlecky LJ, Walton PA, Terlecky SR (2007) Restoration of peroxisomal catalase import in a model of human cellular aging. Traffic
Kohen R, Nyska A (2002) Oxidation of biological systems: oxidative stress phenomena, antioxidants, redox reactions, and methods for their quantification. Toxicol Pathol
Nell HJ, Au JL, Giordano CR, Terlecky SR, Walton PA, Whitehead SN, Cechetto DF (2017) Targeted antioxidant, catalase–SKL, reduces beta-amyloid toxicity in the rat brain. Brain Pathol
Niedzielska E, Smaga I, Gawlik M, Moniczewski A, Stankowicz P, Pera J, Filip M (2016) Oxidative stress in neurodegenerative diseases. Mol Neurobiol
Patel Z, Patel B, Patel S, Pardeshi C (2012) Nose to brain targeted drug delivery bypassing the blood-brain barrier: an overview. Drug Invent Today
Pires A, Fortuna A, Alves G, Falcão A (2009) Intranasal drug delivery: how, why and what for?. J Pharm Pharm Sci
Southam DS, Dolovich M, O’Byrne PM, Inman MD (2002) Distribution of intranasal instillations in mice: effects of volume, time, body position, and anesthesia. Am J Physiol - Lung Cell Mol Physiol
Sun D, Zhuang X, Xiang X, Liu Y, Zhang S, Liu C, Barnes S, Grizzle W, Miller D, Zhang HG (2010) A novel nanoparticle drug delivery system: the anti-inflammatory activity of curcumin is enhanced when encapsulated in exosomes. Mol Ther
Terlecky SR (2012) Peroxisomes, oxidative stress, and inflammation. World J Biol Chem
Undyala V, Terlecky SR, Vander Heide RS (2011) Targeted intracellular catalase delivery protects neonatal rat myocytes from hypoxia-reoxygenation and ischemia-reperfusion injury. Cardiovasc Pathol
Uttara B, Singh A, Zamboni P, Mahajan R (2009) Oxidative stress and neurodegenerative diseases: a review of upstream and downstream antioxidant therapeutic options. Curr Neuropharmacol
Valko M, Leibfritz D, Moncol J, Cronin MTD, Mazur M, Telser J (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol
Walsh KM, Courtney CL (1998) Nasal toxicity of CI-959, a novel anti-inflammatory drug, in Wistar rats and Beagle dogs. Toxicol Pathol
Wen Z, Yan Z, He R, Pang Z, Guo L, Qian Y, Jiang X, Fang L (2011) Brain targeting and toxicity study of odorranalectin-conjugated nanoparticles following intranasal administration. Drug Deliv
Wood CS, Koepke JI, Teng H, Boucher KK, Katz S, Chang P, Terlecky LJ, Papanayotou I, Walton PA, Terlecky SR (2006) Hypocatalasemic fibroblasts accumulate hydrogen peroxide and display age-associated pathologies. Traffic
Yáñez-Mó M et al (2015) Biological properties of extracellular vesicles and their physiological functions. J Extracell Vesicles
Yang T, Martin P, Fogarty B, Brown A, Schurman K, Phipps R, Yin VP, Lockman P, Bai S (2015) Exosome delivered anticancer drugs across the blood-brain barrier for brain cancer therapy in Danio Rerio. Pharm Res
Young CN, Koepke JI, Terlecky LJ, Borkin MS, Boyd SL, Terlecky SR (2008) Reactive oxygen species in tumor necrosis factor-α-activated primary human keratinocytes: implications for psoriasis and inflammatory skin disease. J Invest Dermatol
Yuan D, Zhao Y, Banks WA, Bullock KM, Haney M, Batrakova E, Kabanov AV (2017) Macrophage exosomes as natural nanocarriers for protein delivery to inflamed brain. Biomaterials
Zhu X, Badawi M, Pomeroy S, Sutaria DS, Xie Z, Baek A, Jiang J, Elgamal OA, Mo X, Perle K La, Chalmers J, Schmittgen TD, Phelps MA (2017) Comprehensive toxicity and immunogenicity studies reveal minimal effects in mice following sustained dosing of extracellular vesicles derived from HEK293T cells. J Extracell Vesicles
Zhuang X, Xiang X, Grizzle W, Sun D, Zhang S, Axtell RC, Ju S, Mu J, Zhang L, Steinman L, Miller D, Zhang HG (2011) Treatment of brain inflammatory diseases by delivering exosome encapsulated anti-inflammatory drugs from the nasal region to the brain. Mol Ther
Acknowledgements
We would like to thank Dr. Lynn Wang and Dr. Lin Zhao for their technical assistance and Kyra Keer for the help with histology.
Funding
This work was supported by the Weston Brain Institute grant to PW, PK, and SNW. SHH is a BrainsCAN Postdoctoral Fellow. QL is a CIHR and OGS scholar.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflicts of Interest
The recombinant enzyme CAT-SKL is covered by United States patent 7601366 and 8663630 to P.A. Walton and S.R. Terlecky, who have no commercial or equity interests in the CAT-SKL molecule at this time. Other authors state no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Hayes, S.H., Liu, Q., Selvakumaran, S. et al. Brain Targeting and Toxicological Assessment of the Extracellular Vesicle-Packaged Antioxidant Catalase-SKL Following Intranasal Administration in Mice. Neurotox Res 39, 1418–1429 (2021). https://doi.org/10.1007/s12640-021-00390-6
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12640-021-00390-6