Controlled Release of rAAV Vectors from APMA-Functionalized Contact Lenses for Corneal Gene Therapy
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Controlled Release of rAAV Vectors from APMA-Functionalized Contact Lenses for Corneal Gene TherapyAutor(es)
Fecha
2020-04-09Cita bibliográfica
Alvarez-Rivera, F.; Rey-Rico, A.; Venkatesan, J.K.; Diaz-Gomez, L.; Cucchiarini, M.; Concheiro, A.; Alvarez-Lorenzo, C. Controlled Release of rAAV Vectors from APMA-Functionalized Contact Lenses for Corneal Gene Therapy. Pharmaceutics 2020, 12, 335. https://doi.org/10.3390/pharmaceutics12040335
Resumen
[Abstract]
As an alternative to eye drops and ocular injections for gene therapy, the aim of this work
was to design for the first time hydrogel contact lenses that can act as platforms for the controlled
delivery of viral vectors (recombinant adeno-associated virus, rAAV) to the eye in an effective
way with improved patient compliance. Hydrogels of hydroxyethyl methacrylate (HEMA) with
aminopropyl methacrylamide (APMA) (H1: 40, and H2: 80 mM) or without (Hc: 0 mM) were
synthesized, sterilized by steam heat (121 ◦C, 20 min), and then tested for gene therapy using rAAV
vectors to deliver the genes to the cornea. The hydrogels showed adequate light transparency, oxygen
permeability, and swelling for use as contact lenses. Loading of viral vectors (rAAV-lacZ, rAAV-RFP,
or rAAV-hIGF-I) was carried out at 4 ◦C to maintain viral vector titer. Release in culture medium
was monitored by fluorescence with Cy3-rAAV-lacZ and AAV Titration ELISA. Transduction efficacy
was tested through reporter genes lacZ and RFP in human bone marrow derived mesenchymal stem
cells (hMSCs). lacZ was detected with X-Gal staining and quantified with Beta-Glo®, and RFP was
monitored by fluorescence. The ability of rAAV-hIGF-I-loaded hydrogels to trigger cell proliferation in
hMSCs was evaluated by immunohistochemistry. Finally, the ability of rAAV-lacZ-loaded hydrogels
to transduce bovine cornea was confirmed through detection with X-Gal staining of β-galactosidase
expressed within the tissue.
Palabras clave
Therapeutic contact lens
Gene therapy
rAAV vectors
Controlled release
Corneal diseases
Gene therapy
rAAV vectors
Controlled release
Corneal diseases
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Atribución 4.0 Internacional (CC BY 4.0)
ISSN
1999-4923