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Gene-modified leucoconcentrate for personalized ex vivo gene therapy in a mini pig model of moderate spinal cord injury

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Neural Regeneration Research 2021年第2期16卷 357-361页

作者： Rustem R.Islamov Farid V.Bashirov Mikhail E.Sokolov Andrei A.Izmailov Filip O.Fadeev Vage A.Markosyan Maria A.Davleeva Olga V.Zubkova Maxim M.Smarov Denis Yu.Logunov Boris S.Naroditskyi Ilnur I.Salafutdinov Albert A.Rizvanov Ramil G.Turaev Kazan State Medical University KazanRussia Gamaleya Research Institute of Epidemiology and Microbiology MoscowRussia Kazan Federal University KazanRussia The Republican blood Center of the Ministry of Health of the Republic of Tatarstan KazanRussia

We previously demonstrated that gene-modified umbilical cord blood mononuclear cells overexpressing a combination of recombinant neurotrophic factors are a promising therapeutic approach for cell-mediated gene therapy for neurodegenerative diseases,neurotrauma,and stroke.In this study,using a mini pig model of spinal cord injury,we proposed for the first time the use of gene-modified leucoconcentrate prepared from peripheral blood in the plastic blood bag for personalized ex vivo gene therapy.Leucoconcentrate obtained from mini pig peripheral blood was transduced with a chimeric adenoviral vector(Ad5/35 F)that carried an enhanced green fluorescent protein(EGFP)reporter gene in the plastic blood bag.The day after blood donation,the mini pigs were subjected to moderate SCI and four hours post-surgery they were intravenously autoinfused with gene-modified leucoconcentrate.A week after gene-modified leucoconcentrate therapy,fluorescent microscopy revealed EGFP-expressing leucocytes in spinal cord at the site of contusion injury.In the spleen the groups of EGFP-positive cells located in the lymphoid follicles were observed.In vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by leucocytes.Thus,the efficacy of leucocytes transduction in the plastic blood bag and their migratory potential suggest their use for temporary production of recombinant biologically active molecules to correct certain pathological conditions.This paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate autoinfusion.The animal protocols were approved by the Kazan State Medical University Animal Care and Use Committee(approval No.5)on May 27,2014.

关键词： chimeric Ad5/35F virus enhanced green fluorescent protein gene-modified leucoconcentrate mini pig peripheral blood personalized ex vivo gene therapy plastic blood bag spinal cord injury

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