Engineered glycated amino dendritic polymers as specific nonviral gene delivery vectors targeting the receptor for
M Dolores Giron-Gonzalez1, Arturo Morales-Portillo, Alfonso Salinas-Castillo
1Departamento de Bioquimica y Biologia Molecular II, Facultad de Farmacia, and ‡Departamento de Quı́mica Organica and §Departamento de Quı́mica Analítica, Facultad de Ciencias, Instituto de Biotecnología, Universidad de Granada , E-18071Granada, Spain.
Abstract:
The receptor for advanced glycation end products (RAGE) is involved in diabetes or angiogenesis in tumors. Under pathological conditions, RAGE is overexpressed and upon ligand binding and internalization stimulates signaling pathways that promote cell proliferation. In this work, amino dendritic polymers PEI 25 kDa and alkylated derivatives of PAMAM-G2 were engineered by the nonenzymatic Maillard glycation reaction to generate novel AGE-containing gene delivery vectors targeting the RAGE. The glycated dendritic polymers were easily prepared and retained the capability to bind and protect DNA from endonucleases. Furthermore, while glycation decreased the transfection efficiency of the dendriplexes in CHO-k1 cells which do not express RAGE, glycated dendriplexes acted as efficient transfection reagents in CHO-k1 cells which stably express recombinant RAGE. In addition, preincubation with BSA-AGEs, a natural ligand of the RAGE, or dansyl cadaverine, an inhibitor of the RAGE internalization, blocked transfection, confirming their specificity toward RAGE. The results were confirmed in NRK and RAW264.7 cell lines, which naturally express the receptor. The glycated compounds retain their transfection efficiency in the presence of serum and promote in vivo transfection in a mouse model. Accordingly, RAGE is a suitable molecular target for the development of site-directed engineered glycated nonviral gene vectors.
Insights
Engineered glycated dendritic polymers target the receptor for advanced glycation end products (RAGE) for enhanced gene delivery. These novel vectors show specific, efficient, and in vivo transfection capabilities, highlighting RAGE as a key target.
Area of Science:
- Biomaterials Science
- Gene Therapy
- Molecular Biology
Background:
- Receptor for advanced glycation end products (RAGE) is implicated in pathological conditions like diabetes and tumor angiogenesis.
- RAGE overexpression and ligand binding stimulate signaling pathways promoting cell proliferation.
- Targeting RAGE offers a potential strategy for developing novel therapeutic vectors.
Purpose of the Study:
- To engineer novel gene delivery vectors by glycation of dendritic polymers (PEI 25 kDa and PAMAM-G2).
- To investigate the RAGE-targeting capability and transfection efficiency of these glycated vectors.
- To evaluate the specificity and in vivo performance of the RAGE-targeted gene delivery system.
Main Methods:
- Nonenzymatic Maillard glycation reaction to modify PEI 25 kDa and PAMAM-G2 dendrimers.
- Preparation and characterization of glycated dendritic polymers for DNA binding and protection.
- In vitro transfection assays in RAGE-expressing and non-expressing cell lines (CHO-k1, NRK, RAW264.7).
- Specificity studies using RAGE ligands (BSA-AGEs) and inhibitors (dansyl cadaverine).
- In vivo transfection assessment in a mouse model.
Main Results:
- Glycated dendritic polymers were successfully prepared, retaining DNA binding and protection capabilities.
- Transfection efficiency was significantly enhanced in RAGE-expressing cells compared to non-expressing cells.
- Transfection was specifically mediated by RAGE, as confirmed by blocking experiments.
- Glycated vectors demonstrated serum tolerance and successful in vivo transfection in a mouse model.
Conclusions:
- Engineered glycated dendritic polymers effectively target RAGE for gene delivery.
- RAGE is a viable molecular target for developing site-directed, engineered glycated nonviral gene vectors.
- These novel vectors show promise for targeted gene therapy applications with enhanced specificity and efficacy.
