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Changes in Gene Expression Pattern of Human Primary Macrophages Induced by Carbosilane Dendrimer 2G-NN16

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An Erratum to this article was published on 30 November 2010

Abstract

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.

Materials and Methods

Genomic technologies allow us to identify global gene expression profile changes in macrophages exposed to a non-toxic concentration (5 µM) 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 which are involved in autoimmune disease, by this particular dendrimer suggests a potential pharmacological application.

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Abbreviations

2G-NN16:

\(2G - \left[ {Si\left\{ {O\left( {CH_2 } \right)_2 N(Me)_2^ + \left( {CH_2 } \right)_2 NMe_3^ + \left( {I^ - } \right)2} \right\}} \right]_8 \)

ALDOA:

aldolase A, fructose-bisphosphate

AOC3:

amine oxidase, copper containing 3

C18ORF10:

chromosome 18 open reading frame 10

CCND3:

cyclin D3

CCR1:

chemokine (C–C motif) receptor 1

CCR2:

chemokine (C–C motif) receptor 2

CD74:

CD74 antigen

CR2:

complement component (3d/Epstein Barr virus) receptor 2

CXCL1:

chemokine (C-X-C motif) ligand 1

CXCL2:

chemokine (C–X–C motif) ligand 2

G-CSF:

granulocyte colony-stimulating factor

H3F3B:

H3 histone, family 3B

HIST1H4B:

histone cluster 1, H4b

IL17F:

Interleukin 17F

IL1B:

Interleukin 1B

IL2:

Interleukin 2

IL23A:

Interleukin 23A

IL23R:

Interleukin 23 Receptor

IL6:

Interleukin 6

IL8:

Interleukin 8

INF-γ:

Interferon gamma

IPA:

Ingenuity Pathways Analysis

LDH:

Lactate dehydrogenase

MDK:

midkine (neurite growth-promoting factor 2)

MIP1-a:

Macrophage inflammatory protein 1a

MIP1-b:

Macrophage inflammatory protein 1b

MLLT6:

myeloid/lymphoid or mixed-lineage leukemia (trithorax homolog, Drosophila); translocated to, 6

NFKB1A:

nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha

ODNs:

Oligonucleotide antisense

PAMAM:

polyamidoamine dendrimers

PBMCs:

peripheral blood mononuclear cells

PI:

propide iodide

PRDX2:

peroxiredoxin 2

PTMA:

prothymosin, alpha

qRT-PCR:

quantitative reverse transcription polymerase chain reaction

siRNA:

small interference RNA

SLC1A2:

arrier family 1 (glial high affinity glutamate transporter), member 2

SYVN1:

synovial apoptosis inhibitor 1, synoviolin

TIA1:

TIA1 cytotoxic granule-associated RNA binding protein

TNF-α:

tumor necrosis factor, alpha

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Acknowledgments

The authors are grateful to Alberto Benguria for Pathways analyses, Laura Diaz for flow cytometer assays, Raquel Llorente for technical assistance and Keith Harshman for english corrections and critical reading of the manuscript. This work has been supported by grants from Fondo de Investigación Sanitaria (FIS) del Ministerio de Sanidad y Consumo (PI061479); Red RIS RD06-0006-0035; FIPSE (36514/05, 24534/05, 24632/07), Fundación Caja Navarra and Comunidad de Madrid (S-SAL-0159-2006) to MAMF and grant from Fondo Investigaciones Sanitarias del Ministerio de Sanidad y Consumo (CP06/0267) to LALF.

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Authors and Affiliations

  1. Laboratory of Pharmacogenetics and Pharmacogenomics, Hospital General Universitario Gregorio Marañon, 28007, Madrid, Spain

    Rafael Gras & Luis A. Lopez-Fernandez

  2. Laboratory of Molecular Immunobiology, Hospital General Universitario Gregrorio Marañon, 28007, Madrid, Spain

    Rafael Gras, Maria J. Serramia, Luis A. Lopez-Fernandez & M. Angeles Muñoz-Fernandez

  3. National Center for Biotechnology, 28049, Madrid, Spain

    Luis Almonacid

  4. Departamento de Quimica Inorganica, Universidad de Alcala, Campus Universitario, 28871, Alcala de Henares, Spain

    Paula Ortega, Rafael Gomez & F. Javier de la Mata

  5. Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Barcelona, Spain

    Paula Ortega, Maria J. Serramia, Rafael Gomez, F. Javier de la Mata & M. Angeles Muñoz-Fernandez

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  1. Rafael Gras
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  2. Luis Almonacid
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Correspondence to Luis A. Lopez-Fernandez.

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L. A. Lopez-Fernandez and M. A. Muñoz-Fernández have equally contributed to this work.

An erratum to this article is available at http://dx.doi.org/10.1007/s11095-010-0328-y.

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Gras, R., Almonacid, L., Ortega, P. et al. Changes in Gene Expression Pattern of Human Primary Macrophages Induced by Carbosilane Dendrimer 2G-NN16. Pharm Res 26, 577–586 (2009). https://doi.org/10.1007/s11095-008-9776-z

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