Extracellular Vesicles as a Neprilysin Delivery System Memory Improvement in Alzheimer’s Disease

Document Type : Research article

Authors

1 Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

2 NeuroBiology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

3 Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine,Tehran University of Medical Sciences, Tehran.Iran.

4 Department of Neuroscience and Addiction Studies, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

5 Department of Genetics, Faculty of Basic Sciences, Tarbiat Modarres University, Tehran, Iran.

6 Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

Abstract

Alzheimer’s disease (AD) is a neurodegenerative brain disorder which has no effective treatment yet due to the blood barrier in the brain that limits the drugs with the potential of disease improvement. Extracellular vesicles (EVs) are biocompatible nanoparticles with a lipid membrane. These vesicles are secreted from various cells such as mesenchymal stem cells (MSCs) and can pass through biological barriers for transfer of information such as signals or be used as carriers for various proteins like Neprilysin (NEP). NEP is an active enzyme in the clearance of abnormal aggregated beta-amyloid sheets in the brain. In the present study, we used EVs to carry NEP for memory improvement in Alzheimer’s disease. For this purpose, bone marrow MSCs were isolated from rat femur. Stemness evaluation of established cells was characterized by differentiation potency and specific markers with flowcytometry. EVs were isolated from MSCs supernatant by ultracentrifugation and analyzed by scanning electron microscopy(SEM), dynamic light scattering(DLS) and western blotting. EVs were loaded with NEP by freeze-thaw cycle and then administrated intranasally in a rat model of the AD for 14 days. Our findings showed EV-loaded NEP caused a decrease in IL-1beta and also BAX but an increase in BCL2 expression level in the rat brain. Altogether, these data showed that EV-loaded NEP can improve brain-related behavioural functional which may be mediated through the regulation of inflammation and apoptosis. These findings suggest that EV-loaded NEP can be considered as a potential drug delivery system for the improvement of AD.

Graphical Abstract

Extracellular Vesicles as a Neprilysin Delivery System Memory Improvement in Alzheimer’s Disease

Keywords


(1)        Joshi P, Benussi L, Furlan R, Ghidoni R, and Verderio C. Extracellular vesicles in Alzheimer's disease: friends or foes? Focus on abeta-vesicle interaction. Int J Mol Sci. (2015) 16: 4800-13.
(2)        Deane R, Bell R, Sagare A, and Zlokovic B. Clearance of amyloid-β peptide across the blood-brain barrier: implication for therapies in Alzheimer's disease. CNS & Neurological Disorders-Drug Targets (Formerly Current Drug Targets-CNS & Neurological Disorders). (2009) 8: 16-30.
(3)        Serrano-Pozo A, Frosch MP, Masliah E, and Hyman BT. Neuropathological alterations in Alzheimer disease. Cold Spring Harbor perspectives in medicine. (2011) 1: a006189.
(4)        Grill JD and Cummings JL. Novel targets for Alzheimer's disease treatment. Expert review of neurotherapeutics. (2010) 10: 711.
(5)        Karran E, Mercken M, and De Strooper B. The amyloid cascade hypothesis for Alzheimer's disease: an appraisal for the development of therapeutics. Nature reviews Drug discovery. (2011) 10: 698.
(6)        Wang Y-J, Zhou H-D, and Zhou X-F. Clearance of amyloid-beta in Alzheimer's disease: progress, problems and perspectives. Drug discovery today. (2006) 11: 931-8.
(7)        Patel N and Gluck J. Is Entresto good for the brain? World journal of cardiology. (2017) 9: 594.
(8)        Ozudogru S and Lippa C. Disease modifying drugs targeting β-amyloid. American Journal of Alzheimer's Disease & Other Dementias®. (2012) 27: 296-300.
(9)        Huang S-M, Mouri A, Kokubo H, Nakajima R, Suemoto T, Higuchi M, Staufenbiel M, Noda Y, Yamaguchi H, and Nabeshima T. Neprilysin-sensitive synapse-associated Aβ oligomers impair neuronal plasticity and cognitive function. Journal of Biological Chemistry. (2006)
(10)      El-Amouri SS, Zhu H, Yu J, Marr R, Verma IM, and Kindy MS. Neprilysin: an enzyme candidate to slow the progression of Alzheimer's disease. The American journal of pathology. (2008) 172: 1342-54.
(11)      Wang D-S, Iwata N, Hama E, Saido TC, and Dickson DW. Oxidized neprilysin in aging and Alzheimer’s disease brains. Biochemical and biophysical research communications. (2003) 310: 236-41.
(12)      Nalivaeva NN, Fisk LR, Belyaev ND, and Turner AJ. Amyloid-degrading enzymes as therapeutic targets in Alzheimer's disease. Current Alzheimer Research. (2008) 5: 212-24.
(13)      Webster CI, Burrell M, Olsson L-L, Fowler SB, Digby S, Sandercock A, Snijder A, Tebbe J, Haupts U, and Grudzinska J. Engineering neprilysin activity and specificity to create a novel therapeutic for Alzheimer’s disease. PLoS One. (2014) 9: e104001.
(14)      Saraiva C, Praça C, Ferreira R, Santos T, Ferreira L, and Bernardino L. Nanoparticle-mediated brain drug delivery: overcoming blood–brain barrier to treat neurodegenerative diseases. Journal of Controlled Release. (2016) 235: 34-47.
(15)      Antimisiaris S, Mourtas S, and Marazioti A. Exosomes and Exosome-Inspired Vesicles for Targeted Drug Delivery. Pharmaceutics. (2018) 10: 218.
(16)      Lee Y, El Andaloussi S, and Wood MJ. Exosomes and microvesicles: extracellular vesicles for genetic information transfer and gene therapy. Human molecular genetics. (2012) 21: R125-R34.
(17)      Lai CP-K and Breakefield XO. Role of exosomes/microvesicles in the nervous system and use in emerging therapies. Frontiers in physiology. (2012) 3: 228.
(18)      Agrawal M, Tripathi DK, Saraf S, Saraf S, Antimisiaris SG, Mourtas S, Hammarlund-Udenaes M, and Alexander A. Recent advancements in liposomes targeting strategies to cross blood-brain barrier (BBB) for the treatment of Alzheimer's disease. Journal of Controlled Release. (2017) 260: 61-77.
(19)      Nooshabadi VT, Mardpour S, Yousefi‐Ahmadipour A, Allahverdi A, Izadpanah M, Daneshimehr F, Ai J, Banafshe HR, and Ebrahimi‐Barough S. The extracellular vesicles‐derived from mesenchymal stromal cells: A new therapeutic option in regenerative medicine. Journal of cellular biochemistry. (2018)
(20)      Gholizadeh S, Draz MS, Zarghooni M, Sanati-Nezhad A, Ghavami S, Shafiee H, and Akbari M. Microfluidic approaches for isolation, detection, and characterization of extracellular vesicles: current status and future directions. Biosensors and Bioelectronics. (2017) 91: 588-605.
(21)      Izadpanah M, Seddigh A, Barough SE, Fazeli SAS, and Ai J. Potential of Extracellular Vesicles in Neurodegenerative Diseases: Diagnostic and Therapeutic Indications. Journal of Molecular Neuroscience. (2018) 66: 172-9.
(22)      Fuhrmann G, Neuer AL, and Herrmann IK. Extracellular vesicles–A promising avenue for the detection and treatment of infectious diseases? European Journal of Pharmaceutics and Biopharmaceutics. (2017) 118: 56-61.
(23)      Johnsen KB, Gudbergsson JM, Skov MN, Pilgaard L, Moos T, and Duroux M. A comprehensive overview of exosomes as drug delivery vehicles—endogenous nanocarriers for targeted cancer therapy. Biochimica et Biophysica Acta (BBA)-Reviews on Cancer. (2014) 1846: 75-87.
(24)      Battaglia L, Panciani PP, Muntoni E, Capucchio MT, Biasibetti E, De Bonis P, Mioletti S, Fontanella M, and Swaminathan S. Lipid nanoparticles for intranasal administration: application to nose-to-brain delivery. Expert opinion on drug delivery. (2018) 15: 369-78.
(25)      Dey S, Mahanti B, Mazumder B, Malgope A, and Dasgupta S. Nasal drug delivery: An approach of drug delivery through nasal route. Der Pharmacia Sinica Journal. (2011) 2: 94-106.
(26)      Falcone JA, Salameh TS, Yi X, Cordy BJ, Mortell WG, Kabanov AV, and Banks WA. Intranasal administration as a route for drug delivery to the brain: evidence for a unique pathway for albumin. Journal of Pharmacology and Experimental Therapeutics. (2014) 351: 54-60.
(27)      Krishnan JK, Arun P, Appu AP, Vijayakumar N, Figueiredo TH, Braga MF, Baskota S, Olsen CH, Farkas N, and Dagata J. Intranasal delivery of obidoxime to the brain prevents mortality and CNS damage from organophosphate poisoning. Neurotoxicology. (2016) 53: 64-73.
(28)      Cunningham CJ, Redondo-Castro E, and Allan SM. The therapeutic potential of the mesenchymal stem cell secretome in ischaemic stroke. Journal of Cerebral Blood Flow & Metabolism. (2018) 38: 1276-92.
(29)      Vidal MA, Kilroy GE, Johnson JR, Lopez MJ, Moore RM, and Gimble JM. Cell growth characteristics and differentiation frequency of adherent equine bone marrow–derived mesenchymal stromal cells: adipogenic and osteogenic capacity. Veterinary Surgery. (2006) 35: 601-10.
(30)      Théry C, Amigorena S, Raposo G, and Clayton A. Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Current protocols in cell biology. (2006) 30: 3.22. 1-3.. 9.
(31)      Akers JC, Gonda D, Kim R, Carter BS, and Chen CC. Biogenesis of extracellular vesicles (EV): exosomes, microvesicles, retrovirus-like vesicles, and apoptotic bodies. Journal of neuro-oncology. (2013) 113: 1-11.
(32)      Haney MJ, Klyachko NL, Zhao Y, Gupta R, Plotnikova EG, He Z, Patel T, Piroyan A, Sokolsky M, and Kabanov AV. Exosomes as drug delivery vehicles for Parkinson's disease therapy. Journal of Controlled Release. (2015) 207: 18-30.
(33)      Lin Y-H, Chen C-T, Liang H-F, Kulkarni AR, Lee P-W, Chen C-H, and Sung H-W. Novel nanoparticles for oral insulin delivery via the paracellular pathway. Nanotechnology. (2007) 18: 105102.
(34)      Xu H, Hou Z, Zhang H, Kong H, Li X, Wang H, and Xie W. An efficient Trojan delivery of tetrandrine by poly (N-vinylpyrrolidone)-block-poly (ε-caprolactone)(PVP-b-PCL) nanoparticles shows enhanced apoptotic induction of lung cancer cells and inhibition of its migration and invasion. International journal of nanomedicine. (2014) 9: 231.
(35)      Parsi S, Pandamooz S, Heidari S, Naji M, Morfini G, Ahmadiani A, and Dargahi L. A novel rat model of Alzheimer’s disease based on lentiviral-mediated expression of mutant APP. Neuroscience. (2015) 284: 99-106.
(36)      Paxinos G and Watson C. Atlas of the rat brain in stereotaxic coordinates. Academic, New York. (1986)
(37)      Paxinos G, C. Watson C:“The rat brain in stereotaxic coordinates, Ed 6”. 2007, Academic Press.
(38)      Hemmati F, Dargahi L, Nasoohi S, Omidbakhsh R, Mohamed Z, Chik Z, Naidu M, and Ahmadiani A. Neurorestorative effect of FTY720 in a rat model of Alzheimer's disease: comparison with memantine. Behavioural brain research. (2013) 252: 415-21.
(39)      Bryan KJ, Lee H-g, Perry G, Smith MA, and Casadesus G. Transgenic mouse models of Alzheimer’s disease: behavioral testing and considerations. (2009)
(40)      Gao J, He H, Jiang W, Chang X, Zhu L, Luo F, Zhou R, Ma C, and Yan T. Salidroside ameliorates cognitive impairment in a d-galactose-induced rat model of Alzheimer’s disease. Behavioural brain research. (2015) 293: 27-33.
(41)      Nakajima A, Aoyama Y, Shin E-J, Nam Y, Kim H-C, Nagai T, Yokosuka A, Mimaki Y, Yokoi T, and Ohizumi Y. Nobiletin, a citrus flavonoid, improves cognitive impairment and reduces soluble Aβ levels in a triple transgenic mouse model of Alzheimer's disease (3XTg-AD). Behavioural brain research. (2015) 289: 69-77.
(42)      Onishi T, Iwashita H, Uno Y, Kunitomo J, Saitoh M, Kimura E, Fujita H, Uchiyama N, Kori M, and Takizawa M. A novel glycogen synthase kinase‐3 inhibitor 2‐methyl‐5‐(3‐{4‐[(S)‐methylsulfinyl] phenyl}‐1‐benzofuran‐5‐yl)‐1, 3, 4‐oxadiazole decreases tau phosphorylation and ameliorates cognitive deficits in a transgenic model of Alzheimer’s disease. Journal of neurochemistry. (2011) 119: 1330-40.
(43)      Galipeau J, Krampera M, Barrett J, Dazzi F, Deans RJ, DeBruijn J, Dominici M, Fibbe WE, Gee AP, and Gimble JM. International Society for Cellular Therapy perspective on immune functional assays for mesenchymal stromal cells as potency release criterion for advanced phase clinical trials. Cytotherapy. (2016) 18: 151-9.
(44)      Dominici M, Le Blanc K, Mueller I, Slaper-Cortenbach I, Marini F, Krause D, Deans R, Keating A, Prockop D, and Horwitz E. Minimal criteria for defining multipotent mesenchymal stromal cells. The International Society for Cellular Therapy position statement. Cytotherapy. (2006) 8: 315-7.
(45)      Lötvall J, Hill AF, Hochberg F, Buzás EI, Di Vizio D, Gardiner C, Gho YS, Kurochkin IV, Mathivanan S, and Quesenberry P, Minimal experimental requirements for definition of extracellular vesicles and their functions: a position statement from the International Society for Extracellular Vesicles. 2014, Taylor & Francis.
(46)      Yáñez-Mó M, Siljander PR-M, Andreu Z, Bedina Zavec A, Borràs FE, Buzas EI, Buzas K, Casal E, Cappello F, and Carvalho J. Biological properties of extracellular vesicles and their physiological functions. Journal of extracellular vesicles. (2015) 4: 27066.
(47)      Mardpour S, Hassani SN, Mardpour S, Sayahpour F, Vosough M, Ai J, Aghdami N, Hamidieh AA, and Baharvand H. Extracellular vesicles derived from human embryonic stem cell‐MSCs ameliorate cirrhosis in thioacetamide‐induced chronic liver injury. Journal of cellular physiology. (2018) 233: 9330-44.
(48)      Honary S and Zahir F. Effect of zeta potential on the properties of nano-drug delivery systems-a review (Part 2). Tropical Journal of Pharmaceutical Research. (2013) 12: 265-73.
(49)      Béduneau A, Saulnier P, and Benoit J-P. Active targeting of brain tumors using nanocarriers. Biomaterials. (2007) 28: 4947-67.
(50)      Park JS, Han TH, Lee KY, Han SS, Hwang JJ, Moon DH, Kim SY, and Cho YW. N-acetyl histidine-conjugated glycol chitosan self-assembled nanoparticles for intracytoplasmic delivery of drugs: endocytosis, exocytosis and drug release. Journal of Controlled Release. (2006) 115: 37-45.
(51)      Alvarez-Erviti L, Seow Y, Yin H, Betts C, Lakhal S, and Wood MJ. Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes. Nature biotechnology. (2011) 29: 341.
(52)      Sun D, Zhuang X, Xiang X, Liu Y, Zhang S, Liu C, Barnes S, Grizzle W, Miller D, and Zhang H-G. A novel nanoparticle drug delivery system: the anti-inflammatory activity of curcumin is enhanced when encapsulated in exosomes. Molecular Therapy. (2010) 18: 1606-14.
(53)      Yang T, Martin P, Fogarty B, Brown A, Schurman K, Phipps R, Yin VP, Lockman P, and Bai S. Exosome delivered anticancer drugs across the blood-brain barrier for brain cancer therapy in Danio rerio. Pharmaceutical research. (2015) 32: 2003-14.
(54)      Meilandt WJ, Cisse M, Ho K, Wu T, Esposito LA, Scearce-Levie K, Cheng IH, Yu G-Q, and Mucke L. Neprilysin overexpression inhibits plaque formation but fails to reduce pathogenic Aβ oligomers and associated cognitive deficits in human amyloid precursor protein transgenic mice. Journal of Neuroscience. (2009) 29: 1977-86.
(55)      Wang S-S, Jia J, and Wang Z. Mesenchymal Stem Cell-Derived Extracellular Vesicles Suppresses iNOS Expression and Ameliorates Neural Impairment in Alzheimer’s Disease Mice. Journal of Alzheimer's Disease. (2018) 1-9.
(56)      de Godoy MA, Saraiva LM, de Carvalho LR, Vasconcelos-dos-Santos A, Beiral HJ, Ramos AB, de Paula Silva LR, Leal RB, Monteiro VH, and Braga CV. Mesenchymal stem cells and cell-derived extracellular vesicles protect hippocampal neurons from oxidative stress and synapse damage induced by amyloid-β oligomers. Journal of Biological Chemistry. (2018) 293: 1957-75.
(57)      Akama KT and Van Eldik LJ. β-Amyloid stimulation of inducible nitric-oxide synthase in astrocytes is interleukin-1β-and tumor necrosis factor-α (TNFα)-dependent, and involves a TNFα receptor-associated factor-and NFκB-inducing kinase-dependent signaling mechanism. Journal of Biological Chemistry. (2000) 275: 7918-24.
(58)      Kundu M, Roy A, and Pahan K. Selective neutralization of IL-12 p40 monomer induces death in prostate cancer cells via IL-12–IFN-γ. Proceedings of the National Academy of Sciences. (2017) 114: 11482-7.
(59)      Sui A, Zhong Y, Demetriades AM, Lu Q, Cai Y, Gao Y, Zhu Y, Shen X, and Xie B. Inhibition of integrin α5β1 ameliorates VEGF-induced retinal neovascularization and leakage by suppressing NLRP3 inflammasome signaling in a mouse model. Graefe's Archive for Clinical and Experimental Ophthalmology. (2018) 256: 951-61.
(60)      McBride JD, Liu X, Berry WL, Janknecht R, Cheng R, Zhou K, Badiavas EV, and Ma J-x. Transgenic expression of a canonical Wnt inhibitor, kallistatin, is associated with decreased circulating CD19+ B lymphocytes in the peripheral blood. International journal of hematology. (2017) 105: 748-57.