TY - JOUR
T1 - Evaluation of AAV-mediated delivery of shRNA to target basal-like breast cancer genetic vulnerabilities
AU - Pinto, Catarina
AU - Silva, Gabriela
AU - Ribeiro, Ana S.
AU - Oliveira, Mónica
AU - Garrido, Manuel
AU - Bandeira, Vanessa S.
AU - Nascimento, André
AU - Coroadinha, Ana Sofia
AU - Peixoto, Cristina
AU - Barbas, Ana
AU - Paredes, Joana
AU - Brito, Catarina
AU - Alves, Paula M.
PY - 2019/7/20
Y1 - 2019/7/20
N2 - Adeno-associated viral vectors (AAV) for gene therapy applications are gaining momentum, with more therapies moving into later stages of clinical development and towards market approval, namely for cancer therapy. The development of cytotoxic vectors is often hampered by side effects arising when non-target cells are infected, and their production can be hindered by toxic effects of the transgene on the producing cell lines. In this study, we evaluated the potential of rAAV-mediated delivery of short hairpin RNAs (shRNA) to target basal-like breast cancer genetic vulnerabilities. Our results show that by optimizing the stoichiometry of the plasmids upon transfection and time of harvest, it is possible to increase the viral titers and quality. All rAAV-shRNA vectors obtained efficiently transduced the BLBC cell lines MDA-MB-468 and HCC1954. In MDA-MB-468, transduction with rAAV-shRNA vector targeting PSMA2 was associated with significant decrease in cell viability and apoptosis induction. Importantly, rAAV2-PSMA2 also slowed tumor growth in a BLBC mouse xenograft model, thus potentially representing a therapeutic strategy against this type of cancer.
AB - Adeno-associated viral vectors (AAV) for gene therapy applications are gaining momentum, with more therapies moving into later stages of clinical development and towards market approval, namely for cancer therapy. The development of cytotoxic vectors is often hampered by side effects arising when non-target cells are infected, and their production can be hindered by toxic effects of the transgene on the producing cell lines. In this study, we evaluated the potential of rAAV-mediated delivery of short hairpin RNAs (shRNA) to target basal-like breast cancer genetic vulnerabilities. Our results show that by optimizing the stoichiometry of the plasmids upon transfection and time of harvest, it is possible to increase the viral titers and quality. All rAAV-shRNA vectors obtained efficiently transduced the BLBC cell lines MDA-MB-468 and HCC1954. In MDA-MB-468, transduction with rAAV-shRNA vector targeting PSMA2 was associated with significant decrease in cell viability and apoptosis induction. Importantly, rAAV2-PSMA2 also slowed tumor growth in a BLBC mouse xenograft model, thus potentially representing a therapeutic strategy against this type of cancer.
KW - AAV production
KW - Adeno-associated viral vectors
KW - Basal-like breast cancer
KW - Gene therapy
UR - http://www.scopus.com/inward/record.url?scp=85066232199&partnerID=8YFLogxK
U2 - 10.1016/j.jbiotec.2019.05.016
DO - 10.1016/j.jbiotec.2019.05.016
M3 - Article
C2 - 31150679
AN - SCOPUS:85066232199
SN - 0168-1656
VL - 300
SP - 70
EP - 77
JO - Journal of Biotechnology
JF - Journal of Biotechnology
ER -