Preclinical Models Of Ovarian Cancer Research Articles

Targeting Aurora Kinase with MK-0457 Inhibits Ovarian Cancer Growth

The Aurora kinase family plays pivotal roles in mitotic integrity and cell cycle. We sought to determine the effects of inhibiting Aurora kinase on ovarian cancer growth in an orthotopic mouse model using a small molecule pan-Aurora kinase inhibitor, MK-0457. We examined cell cycle regulatory effects and ascertained the therapeutic efficacy of Aurora kinase inhibition both alone and combined with docetaxel using both in vitro and in vivo ovarian cancer models. In vitro cytotoxicity assays with HeyA8 and SKOV3ip1 cells revealed >10-fold greater docetaxel cytotoxicity in combination with MK-0457. After in vivo dose kinetics were determined using phospho-histone H3 status, therapy experiments with the chemosensitive HeyA8 and SKOV3ip1 as well as the chemoresistant HeyA8-MDR and A2780-CP20 models showed that Aurora kinase inhibition alone significantly reduced tumor burden compared with controls (P values<0.01). Combination treatment with docetaxel resulted in significantly improved reduction in tumor growth beyond that afforded by docetaxel alone (P <or= 0.03). Proliferating cell nuclear antigen immunohistochemistry revealed that MK-0457 alone and in combination with docetaxel significantly reduced cellular proliferation (P values<0.001). Compared with controls, treatment with MK-0457 alone and in combination with docetaxel also significantly increased tumor cell apoptosis by approximately 3-fold (P<0.01). Remarkably, compared with docetaxel monotherapy, MK-0457 combined with docetaxel resulted in significantly increased tumor cell apoptosis. Aurora kinase inhibition significantly reduces tumor burden and cell proliferation and increases tumor cell apoptosis in this preclinical orthotopic model of ovarian cancer. The role of Aurora kinase inhibition in ovarian cancer merits further investigation in clinical trials.

Marked synergism in a preclinical model of ovarian cancer with the combination of a histone deacetylase inhibitor and either topotecan or gemcitabine: Potential clinical implications

16573 Background: Gemcitabine is an active agent in ovarian cancer but relapses are common.How to combine this agent with other drugs remains unclear. Histone deacetylase inhibitors demonstrate cli...

A phase I trial of intraperitoneal EGEN-001, a novel IL-12 gene therapeutic, administered alone or in combination with chemotherapy in patients with recurrent ovarian cancer

5572 Background: EGEN-001 (E1), an IL-12 expressing plasmid formulated with a novel gene delivery system, stimulates natural killer cells, IFN-γ secretion, and T-helper 1 response, inhibits tumor neovascularization, and has potent antitumor activity in preclinical models of ovarian cancer. The activity of E1 was evaluated in two early phase I clinical trials. Methods: Patients with recurrent ovarian cancer were treated intraperitoneally (IP) with escalating dosages and cycles of E1 alone or in combination with intravenous docetaxel and carboplatin. Study endpoints included the identification of clinical toxicity, the MTD, and antitumor activity. Results: In the initial phase 1A study, 13 patients were treated IP with 0.6–24 mg/m2 of E1. All patients except one in the 24 mg/m2 dose cohort completed all 4 weekly E1 treatments. The most commonly experienced adverse events included pyrexia (69%) and abdominal pain (54%). Three patients experienced peritonitis; one of these patients in the 24 mg/m2 cohort experienced a possibly reagent related DLT. The MTD was not identified. The overall clinical response was 31% stable disease and 69% progressive disease. Ten evaluable patients had measurable IL-12 plasmid DNA and IFN-γ levels in their peritoneal fluid. In the first cohort of patients enrolled in the phase 1B study, three patients have been treated with 12 mg/m2 of E1 IP every 10 days for 4 cycles in combination with docetaxel (75 mg/m2) and carboplatin (AUC 5) at 3 week intervals for a planned 6 cycles. All patients completed therapy. Two patients experienced mild pyrexia during E1 administration. One patient developed G3 anemia following the 4th cycle of E1 and required transfusion. No other G3/4 toxicities were noted. All patients demonstrated a partial response by CT evaluation and reductions in CA- 125 levels following the completion of E1 and two cycles of chemotherapy. Conclusions: IP administration novel IL-12 gene therapeutic E1 is feasable and demonstrates potential antitumor activity when given alone or in combination with chemotherapy in patients with recurrent ovarian cancer. Further development of E1 as a potential therapy for ovarian cancer is warranted. No significant financial relationships to disclose.

In vivo restoration of RhoB expression leads to ovarian tumor regression

Ovarian cancers are very aggressive cancers most often diagnosed when metastasis has already occurred in the entire peritoneal cavity. Ovarian adenocarcinoma cells present an undetectable level of RhoB GTPase. Using preclinical ovarian cancer models, we aimed to evaluate the potential use of RhoB cDNA as a tumor suppressor gene in gene therapy. RhoB restoration in vitro, through recombinant adenovirus transduction, resulted in the apoptosis of endogenous RhoB protein low-expressing cell lines (OVCAR-3 and IGROV-1) through the activation of the intrinsic apoptotic caspase cascade. We showed that a single injection of 10(8) p.f.u. of adenoviral vector encoding a reporter gene into the peritoneal cavity of ovarian tumor bearing mice can induce the gene modification of a large quantity of cells throughout the cavity. We thereby tested the effect of AdRhoB injections to treat ovarian cancer-bearing mice. The ectopic expression of RhoB, following its introduction via viral transduction into nude mice in vivo, was highly effective in suppressing tumor growth of ovarian cancer xenografts. Therapeutic agents designed to correct defects of RhoB at the molecular level may thereby provide innovative treatment options for patients not responding to standard therapies.

In vivo and in vitro studies of anticancer actions of α‐TEA for human prostate cancer cells

Vitamin E analog, 2,5,7,8-tetramethyl-2R-(4R,8R, 12-trimethyltridecyl) chroman-6-yloxyacetic acid, referred to as alpha-TEA induces apoptosis in a variety of human cancer cells in cell culture and reduces tumor burden and metastases in preclinical animal models of breast and ovarian cancer. The goal of this study was to determine in vivo anticancer efficacy of alpha-TEA against human prostate cancer cells and identify mechanisms of action. A PC-3-GFP xenograft model was used to assess the effects of alpha-TEA formulated in liposomes and administered orally on tumor burden and metastases. Tumor tissue was examined by immunohistochemical staining for percentage of cells undergoing apoptosis by TUNEL or cell proliferation by Ki-67. In vitro analyses of mechanisms employed western immunoblotting to examine effects of alpha-TEA-treatments in LNCaP and PC-3-GFP cells on levels of pro-survival and pro-death factors. Functional significance was determined using ectopically expressed constitutively active forms, inhibitors, or siRNA. alpha-TEA significantly reduced tumor burden and metastases, increased apoptosis and decreased proliferation of tumor cells (P < 0.05). alpha-TEA treatment of both LNCaP and PC-3-GFP cells in vitro reduced levels of pAkt1, pAkt2; FOXO1, c-FLIP(L) and survivin. Constitutively active Akt1, Akt2, c-FLIP or survivin reduced alpha-TEA-induced apoptosis. PI3K inhibitor enhanced apoptosis. Constitutively active FOXO1 enhanced alpha-TEA induced Fas ligand expression; whereas, FOXO1 siRNA reduced alpha-TEA induced Fas ligand expression. alpha-TEA is an effective anticancer agent for human prostate cancer cells. Downregulation of pro-survival and upregulation of pro-death factors play roles in alpha-TEA-induced apoptosis.

Antiangiogenic and Antitumor Effects of Src Inhibition in Ovarian Carcinoma

Src, a nonreceptor tyrosine kinase, is a key mediator for multiple signaling pathways that regulate critical cellular functions and is often aberrantly activated in a number of solid tumors, including ovarian carcinoma. The purpose of this study was to determine the role of activated Src inhibition on tumor growth in an orthotopic murine model of ovarian carcinoma. In vitro studies on HeyA8 and SKOV3ip1 cell lines revealed that Src inhibition by the Src-selective inhibitor, AP23846, occurred within 1 hour and responded in a dose-dependent manner. Furthermore, Src inhibition enhanced the cytotoxicity of docetaxel in both chemosensitive and chemoresistant ovarian cancer cell lines, HeyA8 and HeyA8-MDR, respectively. In vivo, Src inhibition by AP23994, an orally bioavailable analogue of AP23846, significantly decreased tumor burden in HeyA8 (P = 0.02), SKOV3ip1 (P = 0.01), as well as HeyA8-MDR (P < 0.03) relative to the untreated controls. However, the greatest effect on tumor reduction was observed in combination therapy with docetaxel (P < 0.001, P = 0.002, and P = 0.01, for the above models, respectively). Proliferating cell nuclear antigen staining showed that Src inhibition alone (P = 0.02) and in combination with docetaxel (P = 0.007) significantly reduced tumor proliferation. In addition, Src inhibition alone and in combination with docetaxel significantly down-regulated tumoral production of vascular endothelial growth factor and interleukin 8, whereas combination therapy decreased the microvessel density (P = 0.02) and significantly affected vascular permeability (P < 0.05). In summary, Src inhibition with AP23994 has potent antiangiogenic effects and significantly reduces tumor burden in preclinical ovarian cancer models. Thus, Src inhibition may be an attractive therapeutic approach for patients with ovarian carcinoma.

EphA2 as a target for ovarian cancer therapy

EphA2 is a receptor tyrosine kinase that is overexpressed by many human cancers, and is often associated with poor prognostic features. It is involved in many processes crucial to malignant progression, such as migration, invasion, metastasis, proliferation, survival and angiogenesis. Inducing EphA2 downregulation by any one of several mechanisms (antibody-mediated inhibition of signalling, antibody-mediated downregulation of total EphA2 expression and siRNA-mediated inhibition of expression) has been shown to decrease tumour growth, prolong survival and inhibit angiogenesis in multiple preclinical models of ovarian, breast and pancreatic cancer. Targeting EphA2 is especially attractive in ovarian cancer, in which overexpression is present in > 75% of cases. This disease is highly responsive to chemotherapy, and EphA2 inhibition is especially effective in combination with taxanes. This demonstrated efficacy, along with the low expression of EphA2 by normal adult tissues and lack of demonstrable toxicities in preclinical models, suggest that long-term treatment with EphA2-targeting agents is an attractive approach for ovarian cancer therapy.

Antitumor activity of irofulven against human ovarian cancer cell lines, human tumor colony-forming units, and xenografts.

The objective of this study was to investigate the cytotoxic activity of irofulven (HMAF, MGI 114), a unique chemotherapeutic agent currently under clinical investigation, in various preclinical models of ovarian cancer. Antiproliferative effects of irofulven in ovarian cancer cell lines and ovarian tumor specimens were characterized in vitro using sulforhodamine B and human tumor colony-forming assays, respectively. Irofulven demonstrated marked activity against a panel of ovarian tumor cell lines, including IGROV1, OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, and SK-OV-3, all of which exhibit various drug resistance mechanisms. In human tumor cloning assays, irofulven inhibited colony formation in surgically derived ovarian tumors at concentrations as low as 0.001 micro g /ml and indicated superior activity in comparison with paclitaxel when tested against the same tumor specimens. The antitumor activity of irofulven compared to that of paclitaxel was also examined using the SK-OV-3 xenograft model. In mice bearing subcutaneously implanted SK-OV-3 tumors, treatment with paclitaxel failed to inhibit tumor growth; whereas mice treated with maximum tolerated doses of irofulven had a 25% partial shrinkage rate, and the remaining animals had a mean tumor growth inhibition of 82%. The potent activity of irofulven against ovarian tumors in vitro and in vivo supports the evaluation of its clinical activity in ovarian cancer.

Antitumor activity of irofulven against human ovarian cancer cell lines, human tumor colony-forming units, and xenografts

The objective of this study was to investigate the cytotoxic activity of irofulven (HMAF, MGI 114), a unique chemotherapeutic agent currently under clinical investigation, in various preclinical models of ovarian cancer. Antiproliferative effects of irofulven in ovarian cancer cell lines and ovarian tumor specimens were characterizedin vitrousing sulforhodamine B and human tumor colony-forming assays, respectively. Irofulven demonstrated marked activity against a panel of ovarian tumor cell lines, including IGROV1, OVCAR-3, OVCAR-4, OVCAR-5, OVCAR-8, and SK-OV-3, all of which exhibit various drug resistance mechanisms. In human tumor cloning assays, irofulven inhibited colony formation in surgically derived ovarian tumors at concentrations as low as 0.001 μg /ml and indicated superior activity in comparison with paclitaxel when tested against the same tumor specimens. The antitumor activity of irofulven compared to that of paclitaxel was also examined using the SK-OV-3 xenograft model. In mice bearing subcutaneously implanted SK-OV-3 tumors, treatment with paclitaxel failed to inhibit tumor growth; whereas mice treated with maximum tolerated doses of irofulven had a 25% partial shrinkage rate, and the remaining animals had a mean tumor growth inhibition of 82%. The potent activity of irofulven against ovarian tumorsin vitroandin vivosupports the evaluation of its clinical activity in ovarian cancer.