Gene Therapy

Main interests

Our main research interest is focused on the development of non-viral vectors for ocular gene therapy, in particular for retinopathies (diabetic, prematurity, Lebers’ Congenital Amaurosis and Retinitis Pigmentosa and other pathologies in which neovascularization is involved).

For this purpose, our approach is three-fold:

1) Develop non-viral vectors based on natural and synthetic polymers such as chitosan, hyaluronic acid, and PDMAEMA.

2) Optimize of the delivery and expression of genes of interest, using more efficient plasmid-based expression systems and linkage of nuclear localization signals.

3) Contribute to the definition of the etiology and identification of new therapeutic approaches for treating retinal diseases, with emphasis on RAS (Renin Angiotensin-System), PEDF and anti-angiogenic VEGF isoforms.

Research Areas

Projects

- PTDC/BTM-ORG/28121/2017. TARGET - TArgeting diabetic Retinopathy with GEne Therapy. Role: Investigador Principal | Principal Investigator. Funding | Financiamento: 239 550€. 2018-2021. Fundação para a Ciência e Tecnologia (FCT), Portugal e Programa Operacional Regional de Lisboa.

- PTDC/SAU-BEB/098475/2008 - "Development of non-viral vectors for ocular gene therapy". Principal Investigador. 150 000 Euro. Fundação para a Ciência e Tecnologia (FCT), Portugal.

- PTDC/SAU-FCF/100291/2008 - "EncapsDevice: Development of polymeric nanostructures for nasal and pulmonary delivery of peptides and proteins". Team member. 200 000 Euro. Fundação para a Ciência e Tecnologia (FCT), Portugal.

- PIRG05-GA-2009-249314 - Marie Curie Reintegration Grant. "EyeSee - Development of new gene therapy approaches for the treatment of ocular neovascularization", Principal Investigador. 100 000 Euro. EU, FP7.

- EXPL/BIM-MEC/1433/2013 - “Targeting the renin-angiotensin system for the treatment of retinal diseases“. Principal Investigador. 38 880 Euro. Fundação para a Ciência e Tecnologia (FCT), Portugal.

Publications

Carvalho FR, Calado SM, Silva GA, Diogo GS, Moreira da Silva J, Reis RL, Cancela ML, Gavaia PJ. (2019) Altered bone microarchitecture in a type 1 diabetes mouse model Ins2^Akita. Journal of Cellular Physiology, Volume 234, Issue 6, Pages 9338-9350, doi: 10.1002/jcp.27617.

Filipa Pires, Jeniffer Farias Santos, Diogo Bitoque, Gabriela Araújo Silva, Alexandre Marletta, Viviane Abreu Nunes, Paulo A. Ribeiro, Jorge Carvalho Silva, Maria Raposo. (2019) Polycaprolactone/Gelatin Nanofiber Membranes Containing EGCG-Loaded Liposomes and Their Potential Use for Skin Regeneration; ACS Applied Bio Materials 2019, 2 (11), 4790-4800, doi: 10.1021/acsabm.9b00524.

Mónica Machado, Gabriela A. Silva, Diogo B. Bitoque, Joana Ferreira, Luís A. Pinto, Jorge Morgado, Quirina Ferreira. (2019) Self-Assembled Multilayer Films for Time-Controlled Ocular Drug Delivery; ACS Applied Bio Materials 2019, 2 (10), 4173-4180, doi: 10.1021/acsabm.9b00417.

Araújo RS, Santos DF, Silva GA. (2018) The role of the retinal pigment epithelium and Müller cells secretome in neovascular retinal pathologies. Biochimie, Volume 155, December 2018, Pages 104-108, doi: 10.1016/j.biochi.2018.06.019.

Diogo B. Bitoque, Gabriela A. Silva. (2018) Molecular biology tools for the study and therapy of Retinitis Pigmentosa; Journal of Biotechnology, Volume 284, 20 October 2018, Pages 1-5, doi: 10.1016/j.jbiotec.2018.07.030.

Diogo B. Bitoque, Ana M. Rosa da Costa, Gabriela A. Silva. (2018) Insights on the intracellular trafficking of PDMAEMA gene therapy vectors; Materials Science & Engineering C- Materials for Biological Applications, Volume 93, 1 December 2018, Pages 277-288, doi: 10.1016/j.msec.2018.07.071.

Oliveira AV, Rosa da Costa AM, Silva GA (2017) Non-viral strategies for ocular gene delivery. Mater Sci Eng C Mater Biol Appl. 2017 Aug 1;77:1275-1289. doi: 10.1016/j.msec.2017.04.068. Epub 2017 Apr 18.

Simão S, Santos DF, Silva GA. (2016) Aliskiren inhibits the renin-angiotensin system in retinal pigment epithelium cells. Eur J Pharm Sci. 2016 Jun 23;92:22-27. doi: 10.1016/j.ejps.2016.06.019. [Epub ahead of print]

Simão S, Santos DF, Silva GA. (2016) Aliskiren decreases oxidative stress and angiogenic markers in retinal pigment epithelium cells. Angiogenesis. 2016 Oct 1. [Epub ahead of print]

Calado SM, Diaz-Corrales F, Silva GA (2016) pEPito-driven PEDF expression ameliorates Diabetic Retinopathy hallmarks. Hum Gene Ther Methods. 2016 Mar 4. [Epub ahead of print]

Calado SM, Alves LS, Simão S, Silva GA (2016) GLUT1 activity contributes to the impairment of PEDF secretion by the RPE. Mol Vis. 2016 Jul 14;22:761-70. eCollection 2016.

S. Simão, D. B. Bitoque, S. M. Calado, G. A. Silva.(2016) Oxidative stress modulates the expression of VEGF isoforms in the diabetic retina. New Frontiers in Ophthalmology. 2016. Accepted for publication.

A.V. Oliveira, A. Marcelo, A.M. Rosa da Costa, G.A. Silva. Evaluation of cystamine-modified hyaluronic acid/chitosan polyplex as retinal gene vector. Material Science and Engineering C, 58:264-72. 2016. DOI: 10.1016/j.msec.2015.08.047.

A.V. Oliveira, A. Marcelo, A.M. Rosa da Costa, G.A. Silva. “Modification and characterization of hyaluronic acid: a new polymer for retinal gene delivery”. Material Science and Engineering C. 2016.

A.V. Oliveira, G.A. Silva. Chitosan-based vectors mediate long-term gene expression in the retina. Journal of Bionanoscience 9(5). pp. 373-382(10). 2015. DOI: 10.1166/jbns.2015.1314

A. V. Oliveira, G. A. Silva*, D. C. Chung. “Prolonged, large gene expression with phiC31-integrase-chitosan mediated gene delivery”. Acta Biomaterialia. 2015. *Corresponding Author. DOI:10.1016/j.actbio.2015.01.013

A. V. Oliveira, G. A. Silva. “Hyaluronic acid increases transfection efficiency of chitosan gene delivery”. Journal of Nanomaterials. 2014. DOI:10.1155/2014/246347

S. Machado, S.M. Calado, D. M. Bitoque, A. V. Oliveira, C. L. Øpstad, M. Zeeshan, H.-R. Sliwka, V. Partali, M. D. Pungente, G.A. Silva. Cationic polyene phospholipids as DNA carriers for ocular gene therapy. BioMed Research International. 2014. Volume 2014 (2014), Article ID 703253. http://dx.doi.org/10.1155/2014/703253.

S. M. Calado, A. V. Oliveira, S. Machado, A. P. Silva, R. Haase, G. A. Silva, Sustained gene expression in the retina by improved episomal vectors, Tissue Engineering: Part A. 2014. DOI:10.1089/ten.TEA.2013.0672

D. M. Bitoque, S. Simao, A. V. Oliveira, S. Machado, E. Lopes, M. R. Duran, A. M. R. Costa, G. A. Silva. Efficiency of RAFT-synthesized PDMAEMA in gene transfer to the retina. Journal of Tissue Engineering and Regenerative Medicine. 2014. DOI:10.1002/term.1909.

A. V. Oliveira#, A. P. Silva#, D. B. Bitoque, G. A. Silva*, A. M. R Costa, “Transfection efficiency of chitosan and thiolated chitosan in retinal pigment epithelium cells: a comparison”. Journal of Pharmacy and Bioallied Sciences 5:111-8. 2013 [# equally contributing authors.] *Corresponding Author.

G. A. Silva, N. F. Silva, T. M. Fortunato. Stem cell and tissue engineering therapies for ocular regeneration. Current Stem Cell Research Therapy. 2011 Sep 1;6(3):255-72.