bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 369: Tumor detection and surgical navigation with fluorescence laparoscopy is enhanced with combinations of differently labeled fluorophore-conjugated antibodies and fluorescent proteins
Authors: Raimo Cristina A. Metildi , Kauhsal Sharmeela , Snyder Cynthia S. , Luiken George A. , Hoffman Robert M. , Bouvet Michael
Published: August 2012

The aim of this study was to improve detection of primary and metastatic lesions of human pancreatic cancer in mouse models with fluorescence laparoscopy by enhancing the fluorescence signal of tumor with different combinations of fluorophores without compromising background illumination. Human pancreatic cancer models were established with combinations of FG-RFP, MiaPaca2-GFP, BxPC-3-RFP, or BxPC-3 cancer cells implanted in 6-week-old female athymic mice. Two weeks post-implantation, diagnostic laparoscopy was performed with a Stryker L9000 LED light source 24 hours after tail vein injection of CEA antibodies conjugated with Alexa 488- and/or Alexa 555. Cancer lesions were detected and localized under the fluorescence light mode. Intravital images were obtained with the Maestro CRI Small Animal Imaging System, serving as a positive control. Tumors were collected for histologic review. Fluorescence laparoscopy with a 495-nm emission filter and an LED light source enabled real-time visualization of differently fluorescence-labeled tumors in the peritoneal cavity simultaneously. The combination of BxPC-3 RFP labeled with anti-CEA-Alexa 488 afforded the brightest signal of all established mouse models. We were able to reproduce this enhanced fluorescence signaling by doubly labeling non-color BxPC-3 tumor with anti-CEA-Alexa 488 and anti-CEA-Alexa 555. The doubly labeled tumor provided a greater signal intensity profile compared to tumors labeled with either anti-CEA-Alexa 488 or -Alexa 555 alone. This enhanced fluorescence signal permitted improved detection of sub-millimeter lesions without compromising background illumination. The ability to visualize tumor deposits expressing fluorescent proteins of different wavelengths simultaneously with an LED light source enabled us to establish the optimal combination of fluorophores that enhanced the signal intensity of fluorescence. With this optimal fluorophore combination, we improved the detection of sub-millimeter tumor lesions without impairing the visualization of surrounding structures necessary for surgical navigation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 369. doi:1538-7445.AM2012-369
Abstract 3690: 19-nor-1α-25-dihydroxyvitamin D2 induces gastric cancer cell apoptosis and cell cycle arrest and inhibits STAT3 mediated inflammation
Authors: Bae Woo Kyun , Lee Ji Hee , Hwang Jun Eul , Shim Hyun Jeong , Cho Sang Hee , Chung Ik-Joo
Published: August 2012

The active metabolite of vitamin D, 1,25-dihydroxyvitamin D3 (calcitriol), inhibits the growth of several types of human cancer cells in vitro, but its therapeutic use is limited because it causes hypercalcemia. Among its analogs, 19-nor-1,25-dihydroxyvitamin D2 has fewer calcemic effects and exhibits an activity that is equipotent to that of calcitriol in several in vivo and in vitro systems. This study demonstrated that paricalcitol has antitumor activity in gastric cancer cells. Treatment with paricalcitol inhibited gastric cancer cell growth, induced G0/G1 cell cycle arrest, and decreased the expression of cycle-dependent kinase 2 (CDK2). Additionally, paricalcitol led to apoptosis via induction of caspase-3 cleavage and decreased expression of anti-apoptotic Bcl-XL. Inhibition of STAT3, COX-2, and NF-κB activities was also observed. Moreover, growth of peritoneal metastases in vivo was significantly reduced in mice treated with19-nor-1,25-dihydroxyvitamin D2. These results suggest that the vitamin D analog (19-nor-1,25-dihydroxyvitamin D2) has anti-cancer and anti-inflammatory activity in gastric cancer cells, suggesting that it holds promise as a novel therapy for the prevention and treatment of gastric cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3690. doi:1538-7445.AM2012-3690
Abstract 3697: Phase II study of therapy selected by molecular profiling in patients with previously treated metastatic pancreatic cancer - SU2C-001
Authors: Barrett Michael T. , Lenkiewicz Elizabeth , Evers Lisa , Holley Tara , Aziz Meraj , Kiefer Jeffrey , Maitra Anirban , Hidalgo Manuel , Kumar Rajesh , Hlavacek William , Stites Edward , Posner Richard G. , Ramanathan Ramesh , Von Hoff Dan D.
Published: August 2012

We have completed a second line Phase II study of therapy to be selected by molecular profiling in 35 patients with previously treated metastatic pancreatic cancer. On this study all patients have a biopsy of a metastatic lesion (usually liver). Therapeutic targets are identified by profiling the metastatic biopsies with the combined use of immunohistochemistry (IHC), comparative genomic hybridization (CGH) on sorted tumor populations, and correlating gene expression microarray data to that of a panel of xenografts for which drug sensitivity has been previously determined. The IHC assays are performed in a CLIA certified laboratory and include the following therapeutic targets: MRP1, TOPO1, MGMT, PGP, Her2/Neu, PTEN, c-kit, RRM1, EGFR, BCRP, PDGFR, TOP2A, TS, ERCC1, SPARC, ER, PR, and AR. In addition each sample is screened by PCR for the presence of the most common K-RAS mutations. Gene expression microarray analysis is performed at John Hopkins University using Affymetrix U133 Plus 2.0 gene arrays. For CGH analysis flow sorted nuclei of diploid and aneuploid tumor cell populations are processed and hybridized to 400,000 feature CGH arrays. We have identified distinct high level focal amplicons targeting AKT2, LYN, RET, CDK6, K-RAS, MYCBP, BCL11A, RASA1 and WNT6, and gene specific homozygous deletions including CDKN2A, PTEN, MAP2K4, RASA1 and PARK2 in the sorted aneuploid tumor nuclei. In an effort to elucidate new targets and potential contexts of vulnerability based on the results of this trial, we have extracted combined gene level data for each patient and present these mapped to 32 pancreatic specific pathways and processes.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3697. doi:1538-7445.AM2012-3697
Abstract 37: CCN6 downregulation in invasive breast carcinomas: In situ characterization of changes in EMT proteins and metastatic disease
Authors: Lorenzatti Guadalupe , Gonzalez María Elena , Toy Kathy A. , Cabanillas Ana M. , Kleer Celina G.
Published: August 2012

Background: The recognized heterogeneity of invasive breast carcinomas is a result of intrinsic characteristics of the tumor cells and their interaction with the microenvironment. CCN6, an extracellular matricellular protein involved in cell communication and epithelial-stromal interactions has a tumor suppressor role in breast cancer. CCN6 downregulation in benign breast cells induces an epithelial to mesenchymal transition marked by downregulation of E-cadherin and increased levels of the transcriptional regulators ZEB1 and Snai1. However, the relevance of these studies to human breast cancer needs further investigation. We hypothesized that CCN6 mRNA and protein levels in samples of breast cancer patients may be associated with metastasis and features of EMT. Methods: Forty clinical samples of invasive carcinoma and surrounding normal breast, frozen at the time of operation were collected. We extracted mRNA and protein from each sample and performed real time RT-PCR and Western blots for CCN6, respectively. All frozen samples were immunostained with antibodies against E-cadherin (BD Biosciences, 1:4000), ZEB1 (Santa Cruz Biotechnology, 1:400) and Snai1(Cell Signaling, 1:800). E-cadherin was analyzed as aberrant or normal. ZEB1 and Snai1 proteins were scored as positive or negative, either in the cancer or stromal cells. Results: Of the 40 frozen samples of breast cancer, 23 had high quality RNA and protein for further study. CCN6 mRNA and protein were both decreased in 18/23 (78%). Five of 23 (22%) samples exhibited normal or high expression. Low CCN6 was associated with the presence of distant metastasis. Four of 18 (22%) invasive carcinomas with low CCN6 had distant metastasis compared to none of the normal CCN6 expressing tumors (p<0.05). Seventeen of 18 (94%) of invasive carcinomas with low CCN6 had aberrant E-cadherin protein expression. We noted that in the tumors with aberrant E-cadherin protein, the expression of ZEB1 or Snai1 was mutually exclusive in all cases (p<0.05). We noted that 80% of the tumors exhibited ZEB1protein in the stromal cells. Conclusions: Our results show a direct link between levels of CCN6 mRNA and protein in breast carcinoma patient samples. A decrease in CCN6 expression in the primary breast cancer is associated with distant metastasis and decreased E-cadherin. We uncover a novel inverse relationship between ZEB1 and Snai1 proteins in breast cancer, which is under further investigation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 37. doi:1538-7445.AM2012-37
Abstract 371: In vivo monitoring of metabolic inhibition in anaplastic thyroid carcinoma
Authors: Sandulache Vlad C. , Bankson James A. , Skinner Heath , Wang Yuan , Chen Yunyun , Dodge Cristina T. , Ow Thomas , Hazle John D. , Myers Jeffrey N. , Lai Stephen Y.
Published: August 2012

Purpose/Objective(s): Anaplastic thyroid carcinoma (ATC) accounts for over half of thyroid cancer related deaths and is generally refractory to conventional treatment. Development of novel therapeutic options is critical for the management of this deadly disease. Since tumor cells fulfill energy requirements primarily through aerobic glycolysis rather than mitochondrial respiration, metabolic inhibition represents an attractive treatment modality. In vivo real time imaging may be useful for evaluating the effectiveness of targeting tumor metabolism. Materials/Methods: A panel of ATC cell lines was evaluated for dependence on glucose and glutamine for cell proliferation and survival. Intracellular ATP levels and cellular reducing potential were measured in cells treated with 2-deoxyglucose (2-DG). Enhancement of the cytotoxic effects of cisplatin and radiation therapy (XRT) with 2-DG was evaluated. In a murine orthotopic xenograft model of ATC, real-time hyperpolarized magnetic resonance imaging (HP-MRI) was employed to assess the effects of 2-DG on tumor metabolism. Results: Glucose, but not glutamine starvation, resulted in significant ATC cell death, suggesting that glucose is the primary nutrient for ATC in vitro. Alterations in extracellular glucose levels affected ATP and cellular reducing potential and both were inhibited by 2-DG. Treatment of ATC cells with 2-DG potentiated the cytotoxic effects of cisplatin alone, XRT alone or cisplatin with XRT. Dynamic spectroscopy of HP-[1-13C]-pyruvate revealed differences in the rate of conversion of pyruvate to lactate in 2-DG treated animals compared to their controls (p=0.504). Analysis with a kinetic model accounting for chemical and physical exchange highlighted this difference (p<0.001). Conclusions: Generalized dependence of ATC cells on glucose catabolism provides an opportunity to utilize metabolic inhibition to enhance sensitivity to radiation therapy and chemotherapy. HP-MRI may represent an effective means to assess metabolic targeting in vivo. Continued refinement of the kinetic parameters that relate the observed signal to multiple physical and chemical pools is necessary to facilitate interpretation of the observed changes. The synchronized optimization of metabolic inhibitors with complementary imaging agents in a preclinical platform may facilitate translation of these therapeutic options for ATC to the clinical setting.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 371. doi:1538-7445.AM2012-371
Abstract 372: Disease-free survival and overall survival from colon cancer is improved with fluorescence-guided surgery in metastatic mouse models
Authors: Raimo Cristina A. Metildi , Kaushal Sharmeela , Snyder Cynthia S. , Hoffman Robert M. , Bouvet Michael
Published: August 2012

The aim of this study was to improve disease-free survival and overall survival in orthotopic nude mouse models of human colon cancer with fluorescence-guided surgery (FGS). Fluorescent orthotopic or carcinomatosis models were established in nude mice using human colon cancer cell lines HCT-116 and HT-29 expressing either green fluorescent protein (GFP) or red fluorescent protein (RFP). The tumors were later resected by bright light surgery (BLS) or FGS. In orthotopic and carcinomatosis models, pre- and post-operative images were obtained with the OV-100 Small Animal Imaging System (Olympus Corp., Tokyo, Japan) to assess the extent of surgical resection. In the orthotopic model, whole body imaging of the mice was performed in the postoperative period to assess cancer recurrence and to follow subsequent tumor progression. The mice were sacrificed when they became premorbid and their abdomens were exposed for intravital and ex vivo imaging. Tumor burden was measured in mm2 using ImageJ v1.440. A greater extent of tumor resection in mice with carcinomatosis was achieved using FGS compared to BLS (99.9% vs. 76.9%, p = 0.006). Furthermore, all mice with localized disease had a complete surgical resection with FGS. In contrast, complete resection was achieved in only 56% of the mice undergoing BLS. (p=0.001) Fewer mice in the FGS group had evidence of tumor recurrence (33%) compared to mice in the BLS group (58%), lengthening disease-free survival from 9 weeks in the BLS group to 27 weeks in the FGS group. Overall survival of the mice also increased from 17 weeks in the BLS group to 29 weeks in the FGS group. The results of the present study demonstrate that improved surgical outcomes in the treatment of colon cancer can be achieved with FGS.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 372. doi:1538-7445.AM2012-372
Abstract 3725: Combined cytotoxic effects of volasertib and cisplatin in urothelial carcinoma cells
Authors: Lin Chia-Chi , Lin Yu-Mine , Tsai Yu-Chieh , Pu Yeong-Shiau , Cheng Ann-Lii , Yang James C.
Published: August 2012

Purpose: Patients with advanced urothelial carcinoma (UC) are treated with cisplatin-based chemotherapy. Polo-like kinase 1 (PLK1) inhibitors (e.g. volasertib) have been shown to be effective in patients with refractory UC (AACR-NCI-EORTC Meeting 2008 abstr 36). We studied the combined cytotoxic effects of volasertib and cisplatin in UC cells in vitro. Materials and methods: The cytotoxicity of the two agents either alone or in combination were studied against UC cell lines with different genetic background: T24 (HRAS mutation), RT4, J82 (FGFR3 mutation), and BFTCC909 (TP53 mutation), using the sulforhodamine B colorimetric assay. Schedule-dependence of the two-drug combination was assayed using 3 different treatment schedules: concurrent and 2 sequential exposures. We examined the effects of volasertib on cell-cycle progression in unsynchronized UC cells by determining the DNA content. Results: The volasertib IC50s of T24, RT4, J82, and BFTCC909 were 13.4±0.13, 54.0±10.6, 14.1±1.3 and 62.4±4.46 nM, respectively and the cisplatin IC50s of the four cells were 2776±361, 8767±1018, 6270±1240 and >10000 nM, respectively. With the median-effect analysis, the combined effects of the two agents when given concurrently were sub-additive in the low fraction affected (Fa) range (0.1-0.3) and additive in the median to high Fa range (0.4-1.0) for the four cells. The combined cytotoxicity was more synergistic if volasertib was given 24 hours earlier than cisplatin. The effects were less synergistic if cisplatin was given prior to volasertib. This phenomenon was seen in all four cells. UC cells in media with 50 nM volasertib arrested in G2/M and showed synchronization 24 hr after release. Conclusions: In our UC cell model, the combined effects may be optimized by sequential use of the two agents, preferably volasertib given 24 hours before cisplatin. Volasertib-treated UC cells arrested in G2/M and showed synchronization which may maximally sensitize UC cells to cisplatin. Our results have clinical implications for the treatment of UC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3725. doi:1538-7445.AM2012-3725
Abstract 3727: Targeting autotaxin to reduce chemotherapy resistance in ovarian cancer
Authors: Zhai Ling , Zamani Ali , Liu Ximeng , Wang Yimin , Maxuitenko Yulia , Alvarez Ronald , Estes Jacob , Li Rongbao , Qu Zhican
Published: August 2012

Development of resistance to chemotherapy presents the biggest challenge in the treatment of ovarian cancer. Autotaxin (ATX) is a secreted enzyme that catalyzes lysophosphatidic acid (LPA) production and is responsible for the up-regulation of LPA in ovarian cancer. The ATX-LPA axis has been identified to be one of the mechanisms of chemotherapy resistance in ovarian cancer. Thus, inhibition of autotaxin may be a potential strategy to increase the chemotherapy efficacy in this disease context. At Southern Research, we previously identified a known anti-parasitic small molecule, Bithionol as a potent antiangiogenic agent, which inhibits endothelial cell proliferation, migration and tubular morphogenesis in vitro and directly inhibits autotaxin enzyme activity. Our recent results have shown that Bithionol not only directly inhibits the enzyme activity; it also reduces autotaxin secretion from human endothelial and ovarian cancer cells. Recently, using a human ovarian cancer xenograft mouse model, Biothionol was shown to have in vivo anti-tumor activity as a single drug treatment. In addition, in combination therapy studies in mice, Bithionol significantly increased the efficacy of Paclitaxel and Cisplatin against ovarian tumor growth. These results suggest that Bithionol may provide a promising approach for reducing chemotherapy associated resistance in ovarian cancer. Additional preclinical studies are in progress to assess the potential clinical utility of Bithionol in combination with current ovarian cancer therapy. (This work is supported by a pilot grant from Norma Livingston Foundation and SRI SIP fund).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3727. doi:1538-7445.AM2012-3727
Abstract 3733: Combined effect of linalool as component of Humulus lupulus on doxorubicin-induced antitumor activity
Authors: Miyashita Michiko , Sadzuka Yasuyuki
Published: August 2012

[Purpose] Cancer chemotherapy has been applied to advanced tumors and intractable tumors, playing an important role in their treatments. However, there are many problems including the appearance of adverse reactions and acquirement of drug resistance. The increased numbers of medications resulted in a decreased quality of life for the patients. However, food and food components can be used to increase the activity of antitumor agents without increasing the number of medicines. In this study, the effects of some components in Humulus lupulus (Hop) were examined on the doxorubicin (DOX) induced antitumor activity in vitro and in vivo. [Methods] The effects of Hop components (linalool, myrcene, α-humulene, α-caryophyllene) on DOX permeability and DOX cytotoxicity in P388 leukemia cells in vitro were examined. P388 leukemia cells (5 x 105 cells per animal) were transplanted onto the backs of BDF1 mice, and DOX (2.0mg/kg/day x 4 days, i.p.) and linalool (1.0mg/kg/day x 4 days, p.o.) were administered. The solid tumor and normal tissues were removed and measured DOX concentrations in each tissue. [Results and Discussion] In vitro, myrcene, α-humulene and α-caryophyllene did not have beneficial effects on DOX permeability in tumor cells. In contrast, the addition of linalool was shown to significantly promote on DOX influx and increased DOX concentration in tumor cells. The combination of DOX with linalool (0.1 μM) showed 3.5-fold (P<0.05) cytotoxicity, compared to treatment with DOX alone. Therefore, the combined linalool with DOX enhanced DOX cytotoxicity by the increase of DOX concentration in tumor cells. In vivo, the tumor weight in the DOX alone group decreased to 77% of the control level, whereas the combination of linalool with DOX decreased the tumor weight by 53% (P<0.05) of that in DOX alone group. In the tumor, the DOX concentration in the linalool combined group increased by 1.8-fold (P<0.05) compared to DOX alone group. In the normal tissue, the DOX concentration after combined linalool and DOX treatment tended to decrease. In particular, the DOX concentration in the heart decreased by 49% (P<0.05) of that in DOX alone group, expecting that linalool may decrease DOX-induced cardiotoxicity. Thus, the combination of linalool with DOX increased DOX concentration in the tumor, consequently enhanced antitumor activity of DOX. In conclusion, linalool is expected effectiveness as DOX modulator.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3733. doi:1538-7445.AM2012-3733
Abstract 3739: TAK-733, an investigational, selective MEK1/2 inhibitor, in combination with alisertib (MLN8237), an investigational, selective Aurora A kinase inhibitor is tolerated and results in additive to synergistic antitumor activity: Results from In Vivo Studies
Authors: Fabrey Robyn , O'Connell Shawn , Stanton Andrew , Chakravarty Arijit , Gangolli Esha , Ecsedy Jeffery , Zarycki Jodi , Vincent Patrick
Published: August 2012

MEK and Aurora A kinase inhibitors (AAK) have profound impacts on cell cycle progression. Inhibition of MEK leads to defects in cell-cycle progression followed by cell cycle arrest and/or apoptosis. These cell-cycle progression defects may in part be due to the role of MEK in the DNA replication or damage checkpoint responses through positive regulation of DNA repair mechanisms. AAK inhibition leads to chromosome congression and segregation defects leading to DNA damage followed by apoptosis or senesence. The overlapping biological effects of inhibition of AAK and MEK kinases in cell cycle progression and chromosomal integrity raise the possibility that inhibiting both targets would provide added benefit over the inhibition of either target alone. The in vivo antitumor activity of TAK-733, an investigational potent, selective, non-ATP-competitive allosteric inhibitor of MEK, in combination with alisertib, an investigational potent, selective, reversible, ATP-competitive inhibitor of Aurora A kinase, was examined in experimental human solid tumor xenograft models including NSCLC (NCI H23 [KRAS and LKB1 mutations]), CRC (SW620 [KRAS, APC, p53 mutations]), and pancreatic cancer (Panc 1 and Capan 1 [KRAS mutations] and BxPC-3 [No MAPK mutations]) models in immunocompromised mice. The previously established maximally efficacious doses for the single agents dosed once daily (QD) orally (PO) (10 mg/kg TAK-733 and 30-mg/kg alisertib) were examined. A lower dose of 20 mg/kg alisertib was also examined in the event the maximally efficacious dose of each agent was not tolerated in combination. The once daily (QD) concurrent oral administration of TAK-733 and alisertib resulted in additive to synergistic antitumor activity and in prolonged inhibition of tumor regrowth after terminating treatment compared to single agent treatment in all xenograft models examined. Concurrent administration of TAK-733 and alisertib daily for 21 days was well tolerated. Treatment with TAK-733 alone and in combination with alisertib clearly inhibited phosphorylated ERK and produced a slight increase in phosphorylated histone H3. The results from the nonclinical models examined demonstrate considerable improvement in nonclinical antitumor activity over either single agent alone and provide a biological rationale for clinical evaluation of a TAK-733/alisertib combination in patients with advanced malignancies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3739. doi:1538-7445.AM2012-3739
Abstract 3756: Structure activity relationship studies of novel FAK inhibitor C4
Authors: Agharkar Priyanka A. , Ethirajan Manivannan , Liao Jianqun , Yemma Michael , Cance William , Pandey Ravindra , Kurenova Elena
Published: August 2012

Introduction: FAK and VEGFR3 have stirred considerable interest in recent years as attractive anti-cancer targets, owing to their high expression across several tumor types and relatively low levels in normal tissues. Our lab has shown that VEGFR3 binds to FAK and the physical interaction between the two proteins confers a significant survival advantage to tumor cells. We discovered small molecule C4 that specifically targets and disrupts FAK-VEGFR3 interaction to exert anti-tumor effects, but does so at moderate concentrations. This prompted us to design C4 analogs that display enhanced potency, minimum toxicity and high specificity for the target site. Here, we have shown that some C4 analogs disrupt FAK-VEGFR3 interaction and dramatically inhibit pancreatic tumor growth at low concentrations as single agents as well as in combination with Gemcitabine. Methods: The structure of C4 was modified at various positions by substituting alkyl and aryl moieties. The resulting analogs were screened for anti-cancer inhibitory effects in different cancer cell lines by MTS assay. Effects of C4 analogs on pancreatic cancer cell lines were determined by MTS assay (viability), Clonogenecity assay, Western blot (phosphorylation, apoptosis) and TUNEL assay. Target specificity (immunoprecipitation) and binding confirmation (Forte-Bio's Octet Red assay) of all analogs were performed. In vivo anti-tumor activity of lead analogs was done using a subcutaneous xenograft mouse model of pancreatic cancer. Results: Twenty C4 analogs were screened and four were selected for further analyses. These four analogs strikingly decreased survival of six pancreatic cancer cell lines. Analog C9 showed highest cytotoxic effects, inhibited colony formation of Panc-1 cells at 1µM, induced dose dependent cleavage of caspase-3 and PARP and decreased phosphorylated forms of Akt and Erk within 24h treatment. Disruption of FAK-VEGFR3 interaction was seen with 1µM C9. Octet method revealed that 20µM C9, KD (M) = 1.29×10-7. In vivo, 30 days treatment with low doses of the four analogs led to massive reduction in tumor growth when treated as single agents (60%) and in combination with Gemcitabine (80%)(P<0.05). Conclusions: Targeting the FAK-VEGFR3 protein-protein interaction site highlights a unique approach for molecular-targeted highly specific cancer therapeutics. The outcomes from this project could potentially widen the spectrum of oral-based treatment options for pancreatic cancer, which yet remains a formidable disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3756. doi:1538-7445.AM2012-3756
Abstract 3770: PK/PD/Efficacy characterization of AZ285, a selective CK2 kinase inhibitor
Authors: Schroeder Patricia , Alimzhanov Marat , Brassil Patrick , Denz Christopher R. , Dowling James E. , Lyne Paul D. , Cooke Emma L.
Published: August 2012

Casein Kinase 2 (CK2) is an abundant, highly conserved protein serine/threonine kinase with numerous physiological targets and participates in the maintenance of cell viability with a role in cell growth and proliferation. While the expression level of CK2 in normal cells is tightly regulated, there is an increasing body of evidence demonstrating it's over expression in cancerous cells, thereby making inhibition a viable chemotherapeutic target. Here we will describe a mathematical tumor growth inhibition model describing the relationship between systemic exposure to a potent inhibitor of CK2 (AZ285) and efficacy in the HCT116 xenograft model. Furthermore, a PK/PD model describing the onset, intensity, and duration of the pAKT and cleaved caspase 3 response as function of concentration and time is described and linked to pre-clinical xenograft efficacy outcomes.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3770. doi:1538-7445.AM2012-3770
Abstract 3790: Preclinical profile of LGX818: A potent and selective RAF kinase inhibitor
Authors: Stuart Darrin D. , Li Nanxin , Poon Daniel J. , Aardalen Kimberly , Kaufman Susan , Merritt Hanne , Salangsang Fernando , Lorenzana Edward , Li Allen , Ghoddusi Majid , Caponigro Giordano , Sun Frank , Kulkarni Swarupa , Kakar Shefali , Turner Nancy , Zang Richard , Tellew John , Pryer Nancy
Published: August 2012

Selective RAF inhibitors have significant activity in patients with metastatic melanoma whose tumors express BRAFV600E. However, not all patients respond equally well to treatment and the duration of response is often limited to less than 6 months. LGX818 was developed with the hypothesis that a more potent inhibitor with excellent pharmacological properties would maximize the degree and duration of patient response. LGX818 is a highly potent RAF inhibitor with selective anti-proliferative and apoptotic activity in cells expressing BRAFV600E. In the A375 (BRAFV600E) human melanoma cell line LGX818 suppresses phospho-ERK (EC50 = 3 nM) leading to potent inhibition of proliferation (EC50 = 4 nM). No significant activity was observed against a panel of 100 kinases (IC50 > 900 nM) and LGX818 did not inhibit proliferation of > 400 cell lines expressing wild-type BRAF. Contributing to the high potency of LGX818 is the extremely slow off-rate from BRAFV600E which is not observed with other RAF inhibitors. In biochemical assays the dissociation half-life was >24 hours which translated into sustained target inhibition in cells following drug wash-out. Single dose PK/PD studies in human melanoma xenograft models (BRAFV600E) indicated that LGX818 treatment at oral doses as low as 6 mg/kg resulted in strong (75%) and sustained (>24 hours) decrease in phospho-MEK, even following clearance of drug from circulation. Decreases in phospho-ERK were consistent with phospho-MEK but markers of downstream transcriptional output (DUSP6 and SPRY4) appeared to provide a more sensitive measure of pathway activation. LGX818 induced tumor regression in multiple BRAF mutant human tumor xenograft models grown in immune compromised mice and rats at doses as low as 1 mg/kg. Consistent with the in vitro data, LGX818 was inactive against BRAF wild-type tumors at doses up to 300 mg/kg bid, with good tolerability and linear increase in exposure. Efficacy was also achieved in a more disease-relevant spontaneous metastatic melanoma and a model of melanoma brain metastasis. LGX818 is a potent and selective RAF kinase inhibitor with unique biochemical properties that contribute to an excellent pharmacological profile. A Phase I clinical trial in patients with BRAF mutant tumors is ongoing.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3790. doi:1538-7445.AM2012-3790
Abstract 3792: Population Pharmacokinetic (PPK) modeling of ProLindac™ (AP5346), a nanopolymer prodrug of diaminocyclohexane (DACH)-platinum (Pt) in recurrent advanced ovarian cancer (AOC) patients
Authors: Rezaï Keyvan , Urien Saik , Weill Sophie , Bekradda Mohamed , Bourdel Fabrice , Nowotnik David P. , Cvitkovic Esteban , Lokiec François
Published: August 2012

Background: ProLindac™ (PL) is a 25 kDa hydroxypropylmethacrylamide copolymer linked to a DACH-Pt. The DACH-Pt complex is thought to be biologically inert while attached to the polymer, with slow in vivo Pt release accelerated in low pH environments (extracellular space of hypoxic tumor and intracellular lysosomal compartment). Methods: 9/35 patients (pts) with recurrent OAC treated with single-agent PL via 2-h IV infusion at doses of 900 mg/m*2 q2w (3 pts), 1400 mg/m*2 q3w (3 pts) and 1680 mg/m*2 q3w (3 pts) were assessable for full PK analysis. 8 time point samples were collected on cycle 1, and 2 samples at cycles 2, 3 and 4. The ultrafiltration procedure used Ultra-4 filters (Amicon) with 50 kDa for unbound PL (U-PL) and 3 kDa for unbound Pt (U-Pt) cut-off. Urine was collected every 6 h from day 1 to 5 of dosing 1. Pt levels were measured by a validated Flameless Atomic Absorption Spectrometry Method (concentration range: 12.5 - 500 ng/mL). PPK was modeled using a nonlinear mixed effect model program (Monolix version 3.1s) by computing the maximum likelihood estimation of the parameters without any approximation of the model. Results: A 3-compartment open model adequately described the total PL time-concentration curve with linear elimination while a 2-compartment model satisfactorily described U-PL and U-Pt time-concentration curves with linear elimination. Mean PPK parameters with SD are summarized below: The mean recovery of PL dose in urine was at least 60% within 5 days. Conclusion: The central Vd for total PL Pt is rather small, close to the circulating blood volume suggesting that the major part of parent drug is restrained in the circulatory space with weak tissue diffusion probably due to a high protein binding rate. However, for U-PL and U-Pt, the Vd is much higher suggesting: 1) a significant tissue diffusion of active species (free-Pt), 2) PL is bound to plasma proteins with low-energy bonds.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3792. doi:1538-7445.AM2012-3792
Abstract 3813: DNA polymerase eta and a novel anticancer selenophene compound
Authors: Chen Yih-wen , Chang Jang-yang , Chou Kai-Ming
Published: August 2012

Human DNA polymerase ≤ (pol α) plays a key role in replicating across the UV radiation induced cyclobutane pyrimidine dimers (CPD) in DNA and mutations in pol ≤ gene result in a syndrome called Xeroderma Pigmentosum Variant (XP-V). In the absence of pol β, the DNA replication forks collapse after prolonged stalling at CPD sites, which consequently lead to DNA breakages and cell death. Therefore, pol ≤ has been considered to serve as a cellular protection mechanism to enhance cell survival and reduce mutagenesis. In addition to UV-induced CPD lesion, pol ≤ has the ability to handle DNA lesions introduced by chemotherapeutic anticancer agents, such as platinum based compounds and nucleoside analogs. Such damage tolerance ability of pol ≤ has been shown to reduce cell sensitivity to these DNA targeting agents. D-501036 [2,5-bis(5-hydroxymethyl-2-selenienyl)-3-hydroxymethyl-N-methylpyrrole] is a selenophene compounds that demonstrated promising antitumor activity against various human cancer cells and in vivo xenograft mouse model. Mechanistic studies have shown that D-501036 induced DNA adduct(s) and caused genome instability, which result in cell cycle accumulation in the S-phase, DNA breakage, and subsequently lead to cell death. Interestingly, the IC50 for human fibroblast XP30RO cells derived from XP-V patients was >50 fold higher than the XP30RO cells that stably transfected with a pol ≤ expressing vector (XP30RO-EGFP-pol α). In addition, D-501036 caused an increased amount of DNA breakage in the XP30RO-EGFP-pol ≤ cells as compared to its parental XP30RO cells in both DNA fragmentation and comet assays. These results revealed a novel role of pol ≤ in the action of D-501036 that is very different from its protective role against UV irradiation or cisplatin.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3813. doi:1538-7445.AM2012-3813
Abstract 3820: Reduced migration and enhanced killing of glioblastoma multiforme by mefloquine via alteration of sphingolipid levels
Authors: Sharma Natasha , Cho Heeyon Y. , Thomas Simmy , Chen Thomas C. , Hofman Florence M. , Petasis Nicos A. , Schonthal Axel H. , Louie Stan G.
Published: August 2012

Purpose: Glioblastoma multiforme (GBM) is one of the most aggressive and common brain tumors. Current therapy for GBM is limited to surgery, radiation therapy and chemotherapy using temozolomide (TMZ). However, the emergence of resistance to TMZ and the ability of malignant gliomas to migrate away from the primary tumor site often results in recurrence of the tumor and failure of the initial therapy. This dire scenario illustrates the urgent need to identify novel agents that will be effective in TMZ-resistant tumors. In this study, we investigated the anticancer potential of an FDA approved antimalarial drug, mefloquine (MQ), in the in vitro model of GBM. MQ's impact on glioma viability, proliferation and migration was studied in a number of wild type and TMZ-resistant cells lines. We also evaluated the effects of MQ on intracellular levels of sphingolipids, which are bioactive lipids and have been recognized as the mediators of cellular proliferation and invasion in GBM. Method: To study the effect of MQ on TMZ resistance in vitro, we generated TMZ-resistant cell lines from established GBM cells. We evaluated the effects of MQ on cellular viability and proliferation by MTT assay and colony formation assay. Moreover, cell death was determined by analyzing the effect of MQ on various markers of apoptosis (e.g. cleavage of caspase 7 and PARP) using western blot. The effect of MQ on the migration of both TMZ-sensitive and -resistant cells was evaluated using wound healing scratch assay and Boyden chamber. To evaluate the impact of MQ on sphingolipids, we employed a validated LC-MS lipidomic assay to quantify the levels of various sphingolipids. Results: MQ was able to reduce cellular viability and proliferation in both TMZ-sensitive and -resistant GBM cells. MQ exerted its antitumor activity through induction of apoptosis. Our results further demonstrate the ability of MQ to reduce the migration of both TMZ-sensitive and -resistant GBM cell lines. In addition MQ was able to reduce the intracellular levels of sphingosine-1-phosphate (S1P), a pro-survival and pro-migratory sphingolipid and increased the levels of ceramide, a pro-apoptotic sphingolipid. Conclusion: Altogether our results demonstrate the ability of MQ to reduce cellular viability and proliferation and to induce apoptotic cell death in both TMZ-sensitive and -resistant GBM cells. It is conceivable that MQ exerts these effects by altering sphingolipid metabolism. These preliminary data suggests that MQ should be further investigated for cancer therapeutic purposes for TMZ-resistant GBM.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3820. doi:1538-7445.AM2012-3820
Abstract 3823: 2-Methyl-6-substituted pyrrolo[2,3-d]pyrimidine classical antifolates as selective folate receptor substrates, glycinamide ribonucleotide formyltransferase inhibitors and antitumor agents
Authors: Gangjee Aleem , Zhao Sai , Wang Lei , Orr Steven , Etnyre Erika , Raghavan Sudhir , Matherly Larry H. , Kisliuk Roy L.
Published: August 2012

The important role of reduced folates in one-carbon transfer reactions has rendered folate metabolism an attractive target in cancer chemotherapy for decades. Clinically used antifolates enter cells via folate uptake systems, such as the reduced folate carrier (RFC), folate receptors (FRs), and the proton-coupled folate transporter (PCFT). Toxicity of clinically used antifolates is due to their lack of selectivity for tumor cells, since these antifolates are transported by RFC which occurs in normal as well as tumor cells. FRs are not usually expressed in normal cells. However, the ≤ isoform of FR is abundantly expressed in certain tumor cells, such as ovarian, endometrial, etc. Thus, selective FR-targeted antifolates that also possess cytotoxic activity without transport by RFC would likely circumvent the major toxicities of currently used antifolates. 2-Methyl-6-substituted pyrrolo[2,3-d]pyrimidine antifolates were designed to evaluate the importance of the 2-amino group for transport by RFC, FR, and /or PCFT, as well as for inhibition of α-glycinamide ribonucleotide formyltransferase (GARFTase) in de novo purine biosynthesis, compared with the 2-amino analogs. The most active compound in our series, AG179, showed an IC50 of 1.6 nM in inhibiting proliferation of KB human tumor cells, and was ∼3-fold less potent than the corresponding 2-amino compound AG71 (0.55 nM). In Chinese hamster ovary (CHO) cells engineered to individually express the major folate transporters, AG179 also inhibited growth of FRα-expressing RT16 cells but was inactive toward RFC-expressing PC43-10 and PCFT-expressing R2/PCFT4 CHO cells. The 2-amino analog, AG71, is inert toward RFC but is avidly transported by PCFT and is cytotoxic toward PCFT-expressing cells. In KB cells, growth inhibition by AG179 like AG71 is protected by excess adenosine and 5-amino-4-imidazole carboxamide, establishing GARFTase and de novo purine biosynthesis as the likely intracellular targets. These results establish that replacement of the 2-amino group of AG71 with a methyl group has a limited impact on FRα uptake and anti-proliferative activity, however, this replacement has a profound impact on cellular uptake by PCFT. This report will present the details of the design, synthesis, biological evaluation and molecular modeling of the title compounds.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3823. doi:1538-7445.AM2012-3823
Abstract 3828: Potent curcumin analogues inhibit pancreatic cancer cell growth and angiogenesis
Authors: Nagaraju Ganji Purnachandra , Zhu Shijun , Diaz Roberto , Shoji Mamoru , El-Rayes Bassel F.
Published: August 2012

Background: Curcumin, a component of turmeric, has demonstrated anti-cancer properties mediated through inhibition of NF-κB in preclinical models. In clinical trials, curcumin failed to demonstrate activity against pancreatic cancer. The pharmacologic properties, low absorption, and low potency of curcumin may account for this discrepancy between preclinical and clinical results. Using the curcumin molecular model, our group synthesized over 100 analogs. Of these analogs, EF31 and UBS109 demonstrated higher potency in squamous cell cancer xenograft models as well as inhibition of NF-κB signaling pathways. Based on this preclinical data, we hypothesized that these analogs EF-31 and UBS-109 would be more effective in inhibiting growth of pancreatic cell lines than curcumin. Methods: MiaPaCa-2 and PANC-1 cell lines were used in the experiments. Cells were cultured and then treated with nothing (control), curcumin, UBS-109, EF-31, 5FU, or combination. Cell proliferation was evaluated using XTT assay. Western blot was used to evaluate effects on HSP90, HIF-1α, and VEGF levels. Angiogenic effects were measured by tube formation and chicken egg chorioallantoic membrane (Egg CAM) assays. Results: UBS-109 and EF-31 demonstrated significantly improved inhibition of cell proliferation as compared to curcumin. UBS-109 and EF-31 potentiated the growth inhibitory effects of 5FU. UBS-109 and EF-31 resulted in significant inhibition of expression of protein and mRNA for HSP90 and HIF-1α as compared to curcumin. In turn, UBS-109 and EF-31 significantly inhibited VEGF protein and mRNA expression resulting in inhibition of HUVEC tube formation in vitro and also inhibited angiogenesis in vivo using the Egg CAM assay. Conclusion: UBS-109 and EF-31 are more potent analogs of curcumin. Preliminary data demonstrate potentiation of chemotherapy effects as well as antiangiogenic effects. Evaluating UBS-109 and EF-31 in patients with pancreatic cancer is a rational approach to target HSP90 and HIF-1α in this disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 3828. doi:1538-7445.AM2012-3828