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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 *** 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 *** 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 *** 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 *** the spleen the groups of EGFP-positive cells located in the lymphoid follicles were *** vitro flow cytometry and fluorescent microscopy studies of the gene-modified leucoconcentrate samples also confirmed the production of EGFP by ***,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 *** paper presents a proof-of-concept of simple,safe and effective approach for personalized ex vivo gene therapy based on gene-modified leucoconcentrate *** 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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Molecular and cellular changes in the post-traumatic spinal cord remodeling after autoinfusion of a genetically-enriched leucoconcentrate in a mini-pig model

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Neural Regeneration Research 2023年第7期18卷 1505-1511页

作者： maria Aleksandrovna.davleeva Ravil Rasimovich Garifulin Farid Vagizovich Bashirov Andrei Aleksandrovich Izmailov Leniz Faritovich Nurullin Ilnur Ildusovich Salafutdinov Dilara Zilbarovna Gatina Dmitrij Nikolaevich Shcherbinin Andrei Aleksandrovich Lysenko Irina Leonidovna Tutykhina Maksim Mikhailovich Shmarov Rustem Robertovich Islamov Department of Histology Cytology and EmbryologyKazan State Medical UniversityKazanRussia Кazan Institute of Biochemistry and Biophysics Federal Research Center of Kazan Scientific Center of Russian Academy of SciencesKazanRussia Institute of Fundamental Medicine and Biology Kazan Federal UniversityKazanRussia The National Research Center for Epidemiology and Microbiology named after Honorary Academician N.F.Gamaleya of the Ministry of Health of the Russian Federation MoscowRussia

Post-traumatic spinal cord remodeling includes both degenerating and regenerating processes,which affect the potency of the functional recovery after spinal cord injury(SCI).Gene therapy for spinal cord injury is proposed as a promising therapeutic strategy to induce positive changes in remodeling of the affected neural *** our previous studies for delivering the therapeutic genes at the site of spinal cord injury,we developed a new approach using an autologous leucoconcentrate transduced ex vivo with chimeric adenoviruses(Ad5/35)carrying recombinant *** the present study,the efficacy of the intravenous infusion of an autologous genetically-enriched leucoconcentrate simultaneously producing recombinant vascular endothelial growth factor(VEGF),glial cell line-derived neurotrophic factor(GDNF),and neural cell adhesion molecule(NCAM)was evaluated with regard to the molecular and cellular changes in remodeling of the spinal cord tissue at the site of damage in a model of mini-pigs with moderate spinal cord *** animals were randomly divided into two groups of 4 pigs each:the therapeutic(infused with the leucoconcentrate simultaneously transduced with a combination of the three chimeric adenoviral vectors Ad5/35‐VEGF165,Ad5/35‐GDNF,and Ad5/35‐NCAM1)and control groups(infused with intact leucoconcentrate).The morphometric and immunofluorescence analysis of the spinal cord regeneration in the rostral and caudal segments according to the epicenter of the injury in the treated animals compared to the control mini-pigs showed:(1)higher sparing of the grey matter and increased survivability of the spinal cord cells(lower number of Caspase-3-positive cells and decreased expression of Hsp27);(2)recovery of synaptophysin expression;(3)prevention of astrogliosis(lower area of glial fibrillary acidic protein-positive astrocytes and ionized calcium binding adaptor molecule 1-positive microglial cells);(4)higher growth rates of regeneratingβIII-tubulin-positi

关键词： autologous genetically-enriched leucoconcentrate chimeric adenoviral vector gene therapy glial cell line-derived neurotrophic factor mini-pig neural cell adhesion molecule spinal cord contusion injury vascular endothelial growth factor

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