1. Transduction Enhancers Enable Efficient Human Adenovirus Type 5-Mediated Gene Transfer into Human Multipotent Mesenchymal Stromal Cells
- Author
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Philip Helge Zeplin, Lea Krutzke, Karmveer Singh, Astrid Kritzinger, Stefan Kochanek, Hubert Schrezenmeier, Robin Nilson, Wolfgang Funk, Markus Rojewski, Olivia Lübbers, Karin Scharffetter-Kochanek, Linus Weiß, European Union (EU), and Horizon 2020
- Subjects
0301 basic medicine ,Genetic therapy ,Genetic enhancement ,Cell ,Regenerative medicine ,Transduction (genetics) ,0302 clinical medicine ,DDC 570 / Life sciences ,Transduction, Genetic ,Cells, Cultured ,Chemistry ,GMP ,Adenovirus 5 ,Cell Differentiation ,adenovirus ,Genetic vectors ,gene therapy ,QR1-502 ,3. Good health ,Cell biology ,Infectious Diseases ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,mesenchymal stromal cells ,Cell type ,viral vectors ,Microbiology ,Article ,Viral vector ,Adenoviridae ,Gentherapie ,03 medical and health sciences ,Virology ,ddc:570 ,medicine ,Humans ,ddc:610 ,Enhancer ,Mesenchymzelle ,mesenchymal stem cells ,Adenoviruses, Human ,Macrophages ,Mesenchymal stem cell ,equipment and supplies ,Coculture Techniques ,030104 developmental biology ,good manufacturing practice ,Mesenchymal stem cells ,hMSC ,DDC 610 / Medicine & health ,transduction enhancer - Abstract
Human multipotent mesenchymal stromal cells (hMSCs) are currently developed as cell therapeutics for different applications, including regenerative medicine, immune modulation, and cancer treatment. The biological properties of hMSCs can be further modulated by genetic engineering. Viral vectors based on human adenovirus type 5 (HAdV-5) belong to the most frequently used vector types for genetic modification of human cells in vitro and in vivo. However, due to a lack of the primary attachment receptor coxsackievirus and adenovirus receptor (CAR) in hMSCs, HAdV-5 vectors are currently not suitable for transduction of this cell type without capsid modification. Here we present several transduction enhancers that strongly enhance HAdV-5-mediated gene transfer into both bone marrow- and adipose tissue-derived hMSCs. Polybrene, poly-l-lysine, human lactoferrin, human blood coagulation factor X, spermine, and spermidine enabled high eGFP expression levels in hMSCs. Importantly, hMSCs treated with enhancers were not affected in their migration behavior, which is a key requisite for many therapeutic applications. Exemplary, strongly increased expression of tumor necrosis factor (TNF)-stimulated gene 6 (TSG-6) (a secreted model therapeutic protein) was achieved by enhancer-facilitated HAdV-5 transduction. Thus, enhancer-mediated HAdV-5 vector transduction is a valuable method for the engineering of hMSCs, which can be further exploited for the development of innovative hMSC therapeutics., publishedVersion
- Published
- 2021