Highly efficient transfection of rat cortical neurons using carbosilane dendrimers unveils a neuroprotective role for HIF-1alpha in early chemical hypoxia-mediated neurotoxicity.

Abstract

PURPOSE:

To study the effect of a non-viral vector (carbosilane dendrimer) to efficiently deliver small interfering RNA to postmitotic neurons to study the function of hypoxia-inducible factor-1alpha (HIF1-alpha) during chemical hypoxia-mediated neurotoxicity.

METHODS:

Chemical hypoxia was induced in primary rat cortical neurons by exposure to CoCl(2). HIF1-alpha levels were determined by Western Blot and toxicity was evaluated by both MTT and LDH assays. Neurons were incubated with dendriplexes containing anti-HIF1-alpha siRNA and both uptake and HIF1-alpha knockdown efficiency were evaluated.

RESULTS:

We report that a non-viral vector (carbosilane dendrimer) can deliver specific siRNA to neurons and selectively block HIF1-alpha synthesis with similar efficiency to that achieved by viral vectors. Using this method, we have found that this transcription factor plays a neuroprotective role during the early phase of chemical hypoxia-mediated neurotoxicity.

CONCLUSION:

This work represents a proof-of-concept for the use of carbosilane dendrimers to deliver specific siRNA to postmitotic neurons to block selected protein synthesis. This indicates that this type of vector is a good alternative to viral vectors to achieve very high transfection levels in neurons. This also suggests that carbosilane dendrimers might be very useful for gene therapy.

• Authors: Inmaculada Posadas, Beatriz López-Hernández, Maria Isabel Clemente, Jose Luis Jiménez, Paula Ortega, Javier de la Mata, Rafael Gómez, María Angeles Muñoz-Fernández, Valentín Ceña
• Journal: Pharmaceutical research (Pharm Res) Vol. 26 Issue 5 Pg. 1181-91 (May 2009) ISSN: 1573-904X [Electronic] United States
• PMID: 19191011 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Dendrimers
• Hypoxia-Inducible Factor 1, alpha Subunit
• Neuroprotective Agents
• RNA, Small Interfering
• Silanes
• carbosilane
• Cobalt

Topics

• Animals
• Cell Hypoxia (drug effects)
• Cell Survival (drug effects)
• Cerebral Cortex (cytology)
• Cobalt
• Dendrimers (chemistry, toxicity)
• Gene Knockdown Techniques
• Genetic Therapy
• Hypoxia (chemically induced, genetics, therapy)
• Hypoxia-Inducible Factor 1, alpha Subunit (genetics)
• Neurons (drug effects, metabolism)
• Neuroprotective Agents (administration & dosage, pharmacology)
• RNA, Small Interfering (administration & dosage, genetics, pharmacology)
• Rats
• Rats, Sprague-Dawley
• Silanes (chemistry, toxicity)
• Transfection