Folate receptor mediated genetic modification of human mesenchymal stem cells via folic acid-polyethylenimine-grafted poly(N-3-hydroxypropyl)aspartamide
Issue title: Selected papers of the 36th Conference of the German Society for Clinical Microcirculation and Hemorheology, 5–8. June, 2017, Greifswald, Germany
Guest editors: M. Jünger, A. Krüger-Genge and F. Jung
Article type: Research Article
Authors: Wang, Weiweia; b; 1 | Li, Wenzhongb; 1 | Wang, Jinleic | Hu, Qinglianc | Balk, Mariaa | Bieback, Karend | Stamm, Christofe | Jung, Friedricha | Tang, Gupingc; * | Lendlein, Andreasa; f; * | Ma, Nana; f; *
Affiliations: [a] Institute of Biomaterial Science and Berlin-Brandenburg Center for Regenerative Therapies, Helmholtz-Zentrum Geesthacht, Teltow, Germany | [b] Reference and Translation Center for Cardiac Stem Cell Therapy (RTC), Department of Cardiac Surgery, University of Rostock, Rostock, Germany | [c] Institute of Chemical Biology and Pharmaceutical Chemistry, Zhejiang University, Hangzhou, P. R. China | [d] Medical Faculty Mannheim, Institute of Transfusion Medicine and Immunology, German Red Cross Blood Service of Baden-Württemberg-Hessen, Heidelberg University, Mannheim, Germany | [e] Charité – Universitätsmedizin Berlin, Deutsches Herzzentrum Berlin, Berlin-Brandenburg Center for Regenerative Therapies, Germany | [f] Institute of Chemistry and Biochemistry, Department of Biology, Chemistry and Pharmacy, Free University of Berlin, Berlin, Germany
Correspondence: [*] Corresponding authors: Prof. Guping Tang, Tel./Fax: +86 571 88273284; E-mail: tangguping@zju.edu.cn; Prof. Nan Ma, Tel.: +49 3328 352483; Fax: +49 3328 352452; E-mail: nan.ma@hzg.de; Prof. Andreas Lendlein, Tel.: +49 3328 352235; Fax: +49 3328 352452; E-mail: andreas.lendlein@hzg.de.
Note: [1] These authors contributed equally to this work.
Abstract: Mesenchymal stem cells (MSCs) are targeted as vehicles for cell mediated gene therapy. Here we report on a macromolecular carrier, which was designed aiming at successful targeted gene delivery into MSCs through the mediation of folate receptor and reduced cytotoxicity compared to established cationic polymer vector – polyethylenimine with a weight average molecular weight (Mw) of 25,000 Dalton (PEI25K). The carrier PHPA-PEI1800-FA was synthesized in a two-step procedure. PHPA-PEI1800 was prepared by grafting polyethylenimine with a Mw of 1800 Dalton (PEI1800) onto the α,β-poly(N-3-hydroxypropyl)-D,L-aspartamide (PHPA) backbone. PHPA-PEI1800-FA was obtained by chemically conjugating folic acid onto PHPA-PEI1800. The grafting degree of PEI1800 was 3.9±0.2% in relation to the CH groups of PHPA and the molar ratio of folic acid conjugated to PEI1800 (χFA) was 1.8±0.1 as calculated by NMR spectroscopy. The copolymers were biodegradable and exhibited lower cytotoxicity than PEI25K. Compared to PHPA-PEI1800, PHPA-PEI1800-FA led to a significantly higher transfection efficiency in human MSCs, which could be attributed to the mediation of folate receptor during the transfection process as confirmed by folic acid competition assay. Both marker gene (GFP) and therapeutic gene (VEGF) were delivered into human MSCs from multi-donors using PHPA-PEI1800-FA. The percentage of GFP+ MSCs showed an average value of 2.85±1.60% but a large variation for different samples. The VEGF expression level of the PHPA-PEI1800-FA transfected cells was significantly higher than that of either untransfected or naked DNA transfected cells. Conclusively, PHPA-PEI1800-FA is a suitable vector to deliver genes into human MSCs through the interaction with folate receptor.
Keywords: Non-viral gene delivery, mesenchymal stem cells, folate receptor, poly(N-3-hydroxypropyl)aspartamide, polyethylenimine
DOI: 10.3233/CH-179209
Journal: Clinical Hemorheology and Microcirculation, vol. 67, no. 3-4, pp. 279-295, 2017