A Biosafety Study of Human Umbilical Cord Blood Mononuclear Cells Transduced with Adenoviral Vector Carrying Human Vascular Endothelial Growth Factor cDNA In Vitro

• Published in: Biomedicines Vol. 11; no. 7; p. 2020
• Main Authors: Salafutdinov, Ilnur I, Gatina, Dilara Z, Markelova, Maria I, Garanina, Ekaterina E, Malanin, Sergey Yu, Gazizov, Ilnaz M, Izmailov, Andrei A, Rizvanov, Albert A, Islamov, Rustem R, Palotás, András, Safiullov, Zufar Z
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 01.07.2023; MDPI
• Subjects: Adenosine; Amyotrophic lateral sclerosis; Analysis; Biological activity; Biosafety; Blood & organ donations; Bone marrow; Brain research; Chemokines; Clinical medicine; Cord blood; cord blood stem cell transplantation; Disease; Enzymes; Fluorescence; Gene expression; Gene therapy; Genetic disorders; genetic vectors; Green fluorescent protein; Health aspects; Ischemia; Leukocytes (mononuclear); Medical research; Neurological disorders; Patients; Secretome; Spinal cord injuries; Stem cells; Stroke; transcriptome; Transcriptomes; Transgenes; Umbilical cord; Vascular endothelial growth factor; vascular endothelial growth factor A; United States > US; genetic vectors; genetic therapy; transcriptome; secretome; cord blood stem cell transplantation; vascular endothelial growth factor A; biosafety
• ISSN: 2227-9059; 2227-9059
• DOI: 10.3390/biomedicines11072020

The biosafety of gene therapy remains a crucial issue for both the direct and cell-mediated delivery of recombinant cDNA encoding biologically active molecules for the pathogenetic correction of congenital or acquired disorders. The diversity of vector systems and cell carriers for the delivery of therapeutic genes revealed the difficulty of developing and implementing a safe and effective drug containing artificial genetic material for the treatment of human diseases in practical medicine. Therefore, in this study we assessed changes in the transcriptome and secretome of umbilical cord blood mononuclear cells (UCB-MCs) genetically modified using adenoviral vector (Ad5) carrying cDNA encoding human vascular endothelial growth factor (VEGF165) or reporter green fluorescent protein (GFP). A preliminary analysis of UCB-MCs transduced with Ad5-VEGF165 and Ad5-GFP with MOI of 10 showed efficient transgene expression in gene-modified UCB-MCs at mRNA and protein levels. The whole transcriptome sequencing of native UCB-MCs, UCB-MC+Ad5-VEGF165, and UCB-MC+Ad5-GFP demonstrated individual sample variability rather than the effect of Ad5 or the expression of recombinant on UCB-MC transcriptomes. A multiplex secretome analysis indicated that neither the transduction of UCB-MCs with Ad5-GFP nor with Ad5-VEGF165 affects the secretion of the studied cytokines, chemokines, and growth factors by gene-modified cells. Here, we show that UCB-MCs transduced with Ad5 carrying cDNA encoding human VEGF165 efficiently express transgenes and preserve transcriptome and secretome patterns. This data demonstrates the biosafety of using UCB-MCs as cell carriers of therapeutic genes.