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

Abstract

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 vegf165 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.

Neural Correlates and Molecular Mechanisms of Memory and Learning

“…Preclinical research and computational medicine are important adjuncts to human studies to understand the neurobiological basis of cognitive functions and disorders [ 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 ]. Researchers can use these models to simulate cognitive mechanisms and investigate the complex interactions between genetics, environment, pharmacology, and comorbidities [ 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 ]. This Special Issue advances our understanding of the pathomechanisms underlying normal and pathological conditions, aids in the evaluation of potential treatments, and provides insights into the efficacy of therapies [ 66 ].…”

The Impact of C-3 Side Chain Modifications on Kynurenic Acid: A Behavioral Analysis of Its Analogs in the Motor Domain

“…Preclinical research, including in vitro and in vivo studies, can provide invaluable data that are not feasible to investigate in humans [ 176 , 177 , 178 , 179 , 180 , 181 , 182 , 183 , 184 , 185 , 186 , 187 , 188 , 189 , 190 ]. This study showed that the incorporation of the C-3 side chain elicits subtle differences in curiosity and emotion in animal models of motor function.…”

Emerging Translational Research in Neurological and Psychiatric Diseases: From In Vitro to In Vivo Models

“…In vitro-and in vivo-based preclinical research serves as a vital complement to human studies in understanding neuropsychiatric conditions [51][52][53][54][55]. These models enable researchers to simulate disease conditions and explore the intricate connections among genetics, environment, pharmacology, and comorbidities [56][57][58][59][60]. This study provides insights into the pathomechanisms underlying neurological and mental disorders, facilitates the testing of potential treatments, and evaluates therapeutic efficacy [61].…”