Evaluation of a nanotechnology-based approach to induce gene-expression in human THP-1 macrophages under inflammatory conditions.
Macrophages orchestrate the initiation and resolution of inflammation by producing pro- and anti-inflammatory products. An imbalance in these mediators may originate from a deficient or excessive immune response. Therefore, macrophages are valid therapeutic targets to restore homeostasis under inflammatory conditions. We hypothesize that a specific mannosylated nanoparticle effectively induces gene expression in human macrophages under inflammatory conditions without undesirable immunogenic responses. THP-1 macrophages were challenged with lipopolysaccharide (LPS, 5μg/mL). Polyethylenimine (PEI) nanoparticles grafted with a mannose receptor ligand (Man-PEI) were used as a gene delivery method. Nanoparticle toxicity, Man-PEI cellular uptake rate and gene induction efficiency (GFP, CD14 or CD68) were studied. Potential immunogenic responses were evaluated by measuring the production of tumor necrosis factor-alpha (TNF-α), Interleukin (IL)-6 and IL-10. Man-PEI did not produce cytotoxicity, and it was effectively up-taken by THP-1 macrophages (69%). This approach produced a significant expression of GFP (mRNA and protein), CD14 and CD68 (mRNA), and transiently and mildly reduced IL-6 and IL-10 levels in LPS-challenged macrophages. Our results indicate that Man-PEI is suitable for inducing an efficient gene overexpression in human macrophages under inflammatory conditions with limited immunogenic responses. Our promising results set the foundation to test this technology to induce functional anti-inflammatory genes.
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA; Department of Systems' Biology, School of Medicine, University of Alcala. Campus Universitario - C/ 19, Carretera Madrid-Barcelona, Km 33,600, 28871 Alcalá de Henares, Madrid, Spain. Electronic address: lau.ber91@gmail.com.
Laura
L
Bernal
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA. Electronic address: paalvarad@presby.edu.
Abigail
A
Alvarado-Vázquez
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA; Department of Pharmacology, Ribeirao Preto Medical School-University of Sao Paulo, 3900 Bandeirantes Ave., Ribeirão Preto, SP, 14049-900, Brazil. Electronic address: davidferreira@usp.br.
David Wilson
DW
Ferreira
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA. Electronic address: candler.paige@utdallas.edu.
Candler A
CA
Paige
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA; Department of Physiology, Anatomy and Cellular Biology, University Pablo de Olavide de Sevilla, Carretera de Utrera Km. 1, 41013, Sevilla, Spain. Electronic address: cristinaulecia@gmail.com.
Cristina
C
Ulecia-Morón
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA. Electronic address: bmhill@presby.edu.
Bailey
B
Hill
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA. Electronic address: myleon@presby.edu.
Marina
M
Caesar
Department of Pharmaceutical and Administrative Sciences, Presbyterian College School of Pharmacy, 307 N. Broad St, Clinton, SC 29325, USA. Electronic address: earomero.sandoval@gmail.com.
E Alfonso
EA
Romero-Sandoval
D002454:Cell Differentiation; D002460:Cell Line; D016207:Cytokines / Q000378:metabolism; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D006801:Humans; D007249:Inflammation / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D018836:Inflammation Mediators / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology; D008264:Macrophages / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D053758:Nanoparticles; D036103:Nanotechnology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014162:Transfection
D016207:Cytokines; D018836:Inflammation Mediators; D008070:Lipopolysaccharides; D012333:RNA, Messenger
Cytokines; Gene-delivery; Inflammation; Macrophages; Polyethylenimine; THP-1 cells