Focal Adhesion Kinase Targeting Using In vivo Short Interfering RNA Delivery in Neutral Liposomes for Ovarian Carcinoma Therapy

Abstract

Purpose: Focal adhesion kinase (FAK) plays a critical role in ovarian cancer cell survival and in various steps in the metastatic cascade. Based on encouraging in vitro results with FAK silencing, we examined the in vivo therapeutic potential of this approach using short interfering RNA (siRNA) in the neutral liposome 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine (DOPC). Experimental Design:Therapy experiments of FAK siRNA with or without docetaxel were done using human ovarian cancer cell lines SKOV3ip1, HeyA8, and HeyA8MDR in nude mice. Additional experiments with a cisplatin-resistant cell line (A2780-CP20) were also done. Assessments of angiogenesis (CD31), cell proliferation (proliferating cell nuclear antigen), and apoptosis (terminal deoxynucleotidyl transferase^mediated dUTP nick end labeling) were done using immunohistochemical analysis.Results: A single dose of FAK siRNA-DOPC was highly effective in reducing in vivo FAK expression for up to 4 days as assayed by Western blot and immunohistochemical analysis. Therapy experiments were started 1 week after injection of the ovarian cancer cells. Treatment with FAK siRNA-DOPC (150 Ag/kg twice weekly) reduced mean tumor weight by 44% to 72% in the three cell lines compared with the control group (Ps < 0.05 for HeyA8, A2780-CP20, and SKOV3ip1).When FAK siRNA-DOPC was combined with docetaxel, there was even greater reduction in mean tumor weight in all models (all Ps < 0.05). Similar results were observed in combination with cisplatin. Treatment with FAK siRNA-DOPC plus docetaxel resulted in decreased microvessel density, decreased expression of vascular endothelial growth factor and matrix metalloproteinase-9, and increased apoptosis of tumor-associated endothelial cells and tumor cells. Conclusions: Taken together, these findings suggest that FAK siRNA-DOPC plus docetaxel or platinum might be a novel therapeutic approach against ovarian cancer.

Phosphoproteomic Profiling Identifies Focal Adhesion Kinase as a Mediator of Docetaxel Resistance in Castrate-Resistant Prostate Cancer

“…Early studies in ovarian cancer cell lines and xenografts demonstrated that knockdown of FAK expression enhanced docetaxel efficacy in docetaxelsensitive and docetaxel-resistant models in vitro and in vivo (31,32). Subsequently, TAE226, a TKI that targets FAK and IGF-1R, was demonstrated to enhance docetaxel cytotoxicity (33); however, at this point, development stalled due to the drug failing clinical trials.…”

Focal adhesion kinase

“…These approaches include the various small molecule inhibitors and FAK silencing using FAK targeted short interfering RNA (siRNA). 34,35 Recently, VS-6062 was the first-in-class FAK inhibitor to enter phase I dose-escalation trials in advanced, terminal solid malignancies. Disease stability (≥ six 21 d cycles) was observed in 15 of 91 patients (16%) with measurable disease, which is noteworthy for a heavily pre-treated population.…”

“…Whole cell lysate for western blot analysis of FAK and pFAK Y397 expression was prepared as previously reported. 35 Proteins were separated by 7.5% SDS-PAGE and transferred to nitrocellulose membrane. Membranes were blocked with 5% nonfat milk and incubated with anti-pFAK Y397 antibody (1:1000, BD Transduction Laboratories) for 1 h at room temperature.…”

Multifunctional nanoparticles for cancer immunotherapy

“…In ovarian cancer patients, FAK overexpression is associated with aggressive tumor features resulting to poor overall survival. 79 In this regard, systemic targeting of FAK with liposomal NPs or small molecule inhibitors has shown reduction in tumor growth and metastasis, 77,80 but such approaches are not tumor-specific and could result in undesired side effects. This highlights the need for targeted delivery of these therapeutic molecules.…”