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| Status |
Public on Feb 10, 2009 |
| Title |
Gene expression of human primary macrophages induced by carbosilane dendrimer 2G-NN16 complexed with or without siRNA |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by array
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| Summary |
Purpose: The use of dendrimers for biomedical applications has emerged with promising results. 2G-NN16 is a carbosilane dendrimer with sixteen positive charges per molecule tested to be capable to bind and release antisense oligonucleotides (ODNs) and small interference RNA (siRNA) in vitro. In spite of low cytotoxicity observed for these dendrimers, little is known about cellular changes they produce in cells in general and in immune cells in particular.
Methods: Genomic technologies allow us to identify global gene expression profile changes in macrophages exposed to a non-toxic concentration (5µM) of 2G-NN16, alone or complexed with a random siRNA (dendriplex). Results were confirmed by quantitative real-time RT-PCR.
Results: Exposing macrophages to this dendrimer or dendriplex causes multiple gene expression changes, but no specific action of random siRNA was detected. Pathway analysis of differentially expressed genes shows the altered functions to be immune response, proliferation and transcription regulation. Interleukin 17F (IL17F) was the most regulated gene.
Conclusions: Global gene expression profiles are a highly sensitive method to measure the toxicity degree of a gene delivery vehicle. The strong repression of IL17F, IL23R and IL23A, all of them involved in autoimmune disease, by this particular dendrimer suggests a potential pharmacological application.
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| Overall design |
This experiment was designed to identify changes in gene expression profiles in human macrophages by exposure to carboxilane dendrimer 2G-NN16. Macrophages from three donors were isolated and cultured for 5 hours alone or in the presence of 5µM 2G-NN16 complexed with or without siRNA (Dendriplex). Control samples were labeled with Alexa Fluor 555 and dendrimer and dendriplex samples with Alexa Fluor 647. For each donor, Alexa Fluor 555 and Alexa Fluor 647 were mixed and hybridized to 4x44k whole genome human Agilent microarrays. 3 different microarrays were used and treated as biological replicates for statistical analysis.
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| Contributor(s) |
Gras R, Almonacid L, Ortega P, Gomez R, de la Mata FJ, Lopez Fernandez LA, Muñoz-Fernandez MA |
| Citation(s) |
19034630, 21116693 |
| |
| Submission date |
Aug 11, 2008 |
| Last update date |
Feb 22, 2018 |
| Contact name |
Luis A. Lopez-Fernandez |
| E-mail(s) |
llopezf.hgugm@salud.madrid.org
|
| Phone |
34 914265026
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| Organization name |
Hospital General Universitario Gregorio Marañon
|
| Department |
Pharmacy
|
| Lab |
Pharmacogenetics and Pharmacogenomics
|
| Street address |
Dr. Esquerdo 46
|
| City |
Madrid |
| ZIP/Postal code |
28007 |
| Country |
Spain |
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| Platforms (1) |
| GPL4133 |
Agilent-014850 Whole Human Genome Microarray 4x44K G4112F (Feature Number version) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA113167 |
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