br Results br Discussion Our finding that AXL overexpression

Results
Discussion Our finding that AXL overexpression is significantly associated with poor survival in patients with high-grade serous OC is consistent with previous reports indicating that AXL regulates tumor growth in some cancers, as well as reports showing that AXL activity is involved in the regulation of epithelial-to-mesenchymal transition and metastasis.10, 12, 13, 23 These data indicate AXL as a potentially useful therapeutic target in metastatic OC. There have been some efforts to develop inhibitors for AXL-RTK via small non-coding RNAs and small molecule inhibitors.32, 33, 34 Although small molecule inhibitors lead to increased therapeutic efficacy and sensitivity in targeting these kinases, they can inhibit additional kinases potentially leading to unexpected toxicities. For instance, AXL inhibitor BGB324 has been shown to inactivate the increased cell proliferation, survival, and chemoresistance of acute myeloid leukemia cells. AXL inhibitors based on small non-coding RNAs have also been shown to successfully reduce proliferation and metastasis of various cancer cells.23, 32, 36, 37, 38 However, clinical development of RNAi-based therapeutics is hampered by the lack of nanodelivery systems that can efficiently and safely deliver these small molecules into tumors. Aptamers have been shown to provide favorable specificity and stability in in vivo studies. The key features of the aptamers as therapeutics are that they can easily be modified to optimize their characteristic features, such as stability, affinity, and specificity. Nucleic Cholesterol aptamers have several advantages over other therapeutic options, including (1) high binding affinity40, 41 and (2) decreased sensitivity to endonucleases by chemical modifications, such as fluoro modifications on the 2′ moiety of the ribose and phosphorothioate modifications.42, 43, 44 In this study, we used a previously characterized RNA aptamer (GL21.T), which is known to bind AXL tyrosine kinase receptor with high affinity, as the base sequence to synthesize the corresponding DNA aptamer. Since RNA oligonucleotides are more susceptible to hydrolysis than their DNA counterparts and DNA aptamers are more cost effective in terms of synthesis compared to RNA aptamers, in this study we synthesized and used a DNA aptamer with 2′-fluoro and monothiophosphate chemical modifications to further enhance the stability against endonucleases. AXL has been further shown to be a downstream effector of the epithelial-to-mesenchymal transition, which promotes mobility, invasion, and metastasis. In OC, AXL was shown to regulate cancer cell invasion and progression via altering MMPs and PI3K/AKT signaling. Furthermore AXL inhibition led to the inhibition of FAK activity, which is one of the key regulatory mechanisms of the invasion pathway. Overall, our data suggest that inhibition of FAK phosphorylation and MT1-MMP1 and MMP2 by the AXL aptamer is a key underlying mechanism of the inhibition of migration and invasion of OC cells. We demonstrate that AXL-APTAMER treatment reduced the tumor burden and number of metastatic nodules in mice. When combined with paclitaxel, the AXL aptamer significantly enhanced antitumor efficacy of paclitaxel chemotherapy. Given the fact that AXL confers drug resistance in many cancers,48, 49, 50, 51 these results suggest that AXL-APTAMER not only provides additional antitumor effect but also, by inhibiting downstream signaling such as PI3K, ERK-MAPK, and nuclear factor κB (NF-κB), overcomes drug resistance in cancer cells. Therefore, AXL-APTAMER plus chemotherapy combinations may be a potential strategy for drug-resistant OC, and also it may serve as a tool for targeted delivery of chemotherapy to tumor while sparing healthy cells from chemotherapy-induced damage. Whether AXL-APTAMER conjugated to a chemotherapy agent provides efficient targeted therapy for OC patients can be proposed as a future scope of this study. Aptamer in combination with paclitaxel therapy opens up the possibility of conjugating the aptamer chemically to paclitaxel for more efficient and targeted therapy. Our data indicate AXL-APTAMER’s potential as a therapeutic agent either alone or in combination with chemotherapy.