Laser-induced choroidal neovascularization model to study age-related macular degeneration in mice

Developers

Vincent Lambert, Julie Lecomte, Jean-Marie Rakic, Agnès Noel, etc.

Description of the technology

Standardized protocol developed in the frames of this technology can be applied to transgenic mice. It includes procedures with using of drugs, recombinant proteins, antibodies, adenoviruses and pre-microRNAs to aid in the search for new molecular regulators and the identification of novel targets for innovative treatments of age-related macular degeneration. The protocol is based on the mouse model of laser-induced choroidal neovascularization, which is extensively used in studies of the exudative form of age-related macular degeneration. This experimental in vivo model relies on laser injury to perforate Bruch’s membrane, resulting in subretinal blood vessel recruitment from the choroid. This method simulates the main features of the exudative form of human age-related macular degeneration, so it is used as the backbone for testing antiangiogenic therapies.

The assay with using of this model excels at robustness and requires 7–14 d to complete, depending on the treatment applied and whether immunostaining is performed. Besides, the protocol includes the method of induction of choroidal neovascularization, including laser induction, lesion excision, various types of processing and the approaches to quantify neoformed vasculature.

Practical application

The technology has clear advantages over other currently used in vivo models. The model recapitulates the complex biological processes involved in the exudative form of age-related macular degeneration disease (inflammation and angiogenesis) and is relatively rapid to develop on an easily accessible biological tissue that can be flat-mounted or prepared for histological sectioning or on protein or nucleic-acid extractions.

In contrast to the transgenic models, based on overexpression apolipoprotein E31 or deletion of superoxide dismutase 1, which are long-term assays requiring senescent animals, the laser-induced choroidal neovascularization model can be applied to a panel of young wild-type or transgenic mice (knockout or knockin). In addition, viruses, cells or compounds, including neutralizing antibodies, siRNA or shRNA, pre-miRNA, recombinant proteins, nanoparticles and drugs, can be administered via different pathways (i.p. injection, i.v. injection, per os, drinking water, intravitreous injection, subretinal injection, bone marrow engraftment and others) and combined or not with genetic manipulation.

Laboratories

• Department of Ophthalmology, University Hospital (CHU), Liège (Belgium)
• Laboratory of Tumor and Development Biology, GIGA-Cancer, University of Liège, Liège (Belgium)
• Unit of Molecular Biology and Genetic Engineering, GIGA-Cancer, University of Liège, Liège (Belgium)
• Drug Research Center (CIRM), University of Liège, Liège (Belgium)

Links

Publications

• Lambert, v. et al. «Laser-induced choroidal neovascularization model to study age-related macular degeneration in mice." 8 Nature Protocols (2013): 2197–2211.
• Lecomte, J. et al. ‘Bone marrow-derived mesenchymal cells and MMP13 contribute to experimental choroidal neovascularization." 68 Cell Mol. Life Sci. (2011): 677–686.
• Noel, A., et al. «Anti-angiogenic therapy of exudative age-related macular degeneration: current progress and emerging concepts." 13 Trends Mol. Med. (2007): 345–352.