Exosome Delivered Anticancer Drugs Across the Blood-Brain Barrier for Brain Cancer Therapy in Danio Rerio

PurposeThe blood–brain barrier (BBB) essentially restricts therapeutic drugs from entering into the brain. This study tests the hypothesis that brain endothelial cell derived exosomes can deliver anticancer drug across the BBB for the treatment of brain cancer in a zebrafish (Danio rerio) model.Materials and MethodsFour types of exosomes were isolated from brain cell culture media and characterized by particle size, morphology, total protein, and transmembrane protein markers. Transport mechanism, cell uptake, and cytotoxicity of optimized exosome delivery system were tested. Brain distribution of exosome delivered anticancer drugs was evaluated using transgenic zebrafish TG (fli1: GFP) embryos and efficacies of optimized formations were examined in a xenotransplanted zebrafish model of brain cancer model.ResultsFour exosomes in 30–100 diameters showed different morphologies and exosomes derived from brain endothelial cells expressed more CD63 tetraspanins transmembrane proteins. Optimized exosomes increased the uptake of fluorescent marker via receptor mediated endocytosis and cytotoxicity of anticancer drugs in cancer cells. Images of the zebrafish showed exosome delivered anticancer drugs crossed the BBB and entered into the brain. In the brain cancer model, exosome delivered anticancer drugs significantly decreased fluorescent intensity of xenotransplanted cancer cells and tumor growth marker.ConclusionsBrain endothelial cell derived exosomes could be potentially used as a carrier for brain delivery of anticancer drug for the treatment of brain cancer.

[1]  Kyu-Won Kim,et al.  Functional and developmental analysis of the blood–brain barrier in zebrafish , 2008, Brain Research Bulletin.

[2]  A. M. Al-Abd,et al.  A simple HPLC method for doxorubicin in plasma and tissues of nude mice , 2009, Archives of pharmacal research.

[3]  Dongmei Sun,et al.  Treatment of brain inflammatory diseases by delivering exosome encapsulated anti-inflammatory drugs from the nasal region to the brain. , 2011, Molecular therapy : the journal of the American Society of Gene Therapy.

[4]  M. Aschner,et al.  Developmental aspects of blood-brain barrier (BBB) and rat brain endothelial (RBE4) cells as in vitro model for studies on chlorpyrifos transport. , 2003, Neurotoxicology.

[5]  Tianzhi Yang,et al.  Comparative Studies on Chitosan and Polylactic-co-glycolic Acid Incorporated Nanoparticles of Low Molecular Weight Heparin , 2012, AAPS PharmSciTech.

[6]  R. Weil,et al.  Capecitabine and lapatinib uptake in surgically resected brain metastases from metastatic breast cancer patients: a prospective study. , 2015, Neuro-oncology.

[7]  P. Steeg,et al.  Heterogeneous Blood–Tumor Barrier Permeability Determines Drug Efficacy in Experimental Brain Metastases of Breast Cancer , 2010, Clinical Cancer Research.

[8]  F. Ahsan,et al.  Evaluation of human nasal RPMI 2650 cells grown at an air-liquid interface as a model for nasal drug transport studies. , 2008, Journal of pharmaceutical sciences.

[9]  You-hong Cui,et al.  A Novel Zebrafish Xenotransplantation Model for Study of Glioma Stem Cell Invasion , 2013, PloS one.

[10]  Ç. Gerçel-Taylor,et al.  Exosomes/microvesicles: mediators of cancer-associated immunosuppressive microenvironments , 2011, Seminars in Immunopathology.

[11]  Dongmei Sun,et al.  A novel nanoparticle drug delivery system: the anti-inflammatory activity of curcumin is enhanced when encapsulated in exosomes. , 2010, Molecular therapy : the journal of the American Society of Gene Therapy.

[12]  Lihong Liu,et al.  Modern methods for delivery of drugs across the blood-brain barrier. , 2012, Advanced drug delivery reviews.

[13]  C. Coch,et al.  Exosomes as nucleic acid nanocarriers. , 2013, Advanced drug delivery reviews.

[14]  M. Wood,et al.  Delivery of siRNA to the mouse brain by systemic injection of targeted exosomes , 2011, Nature Biotechnology.

[15]  Masahiko Sugimoto,et al.  A novel transgenic zebrafish model for blood-brain and blood-retinal barrier development , 2010, BMC Developmental Biology.

[16]  Neeraj Kumar,et al.  HPLC method for the determination of carboplatin and paclitaxel with cremophorEL in an amphiphilic polymer matrix. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[17]  Tianzhi Yang,et al.  In vitro evaluation of optimized liposomes for delivery of small interfering RNA , 2014, Journal of liposome research.

[18]  M. Wood,et al.  Exosome nanotechnology: An emerging paradigm shift in drug delivery , 2011, BioEssays : news and reviews in molecular, cellular and developmental biology.

[19]  T. Dowling,et al.  Determination of Rhodamine 123 in cell lysate by HPLC with visible wavelength detection. , 2005, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[20]  Jaesung Park,et al.  Bioinspired exosome-mimetic nanovesicles for targeted delivery of chemotherapeutics to malignant tumors. , 2013, ACS nano.

[21]  W. Pardridge,et al.  Blood-brain barrier delivery. , 2007, Drug discovery today.

[22]  W. Pardridge.  The blood-brain barrier: Bottleneck in brain drug development , 2005, NeuroRx : the journal of the American Society for Experimental NeuroTherapeutics.

[23]  S. Lim,et al.  Exosomes for drug delivery - a novel application for the mesenchymal stem cell. , 2013, Biotechnology advances.

[24]  K. Hynynen,et al.  Chemotherapy delivery issues in central nervous system malignancy: a reality check. , 2007, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[25]  R. Schiffelers,et al.  Exosome mimetics: a novel class of drug delivery systems , 2012, International journal of nanomedicine.

[26]  J. Rich,et al.  New approaches to primary brain tumor treatment , 2006, Anti-cancer drugs.

[27]  A. Haqqani,et al.  Method for isolation and molecular characterization of extracellular microvesicles released from brain endothelial cells , 2013, Fluids and Barriers of the CNS.

[28]  X Yu,et al.  J.Chromatogr., B: Anal. Technol. Biomed. Life Sci. , 2004 .

[29]  M. Record,et al.  Exosomes as intercellular signalosomes and pharmacological effectors. , 2011, Biochemical pharmacology.

[30]  Anja Schneider,et al.  Exosomes: vesicular carriers for intercellular communication in neurodegenerative disorders , 2012, Cell and Tissue Research.

[31]  G. Salzano,et al.  Nanotechnologies: a strategy to overcome blood-brain barrier. , 2012, Current drug metabolism.

[32]  R. Setterquist,et al.  Exosomes: current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials. , 2012, Biochimica et biophysica acta.

[33]  Elke S. Bergmann-Leitner,et al.  Editorial [Hot Topic: Anti-Cancer Drugs Executive Editor: Elke Bergmann-Leitner] , 2005 .

[34]  R. A. Umans,et al.  Zebrafish as a Model to Study Drug Transporters at the Blood–Brain Barrier , 2012, Clinical pharmacology and therapeutics.

[35]  Trairak Pisitkun,et al.  Large-scale proteomics and phosphoproteomics of urinary exosomes. , 2009, Journal of the American Society of Nephrology : JASN.

[36]  Graça Raposo,et al.  Exosomes--vesicular carriers for intercellular communication. , 2009, Current opinion in cell biology.