bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5762: Role of BMP7 in dormancy and recurrence of prostatic tumor stem cell in the bone metastasis
Authors: Kobayashi Aya , Okuda Hiroshi , Furuta Eiji , Pandey Puspa R. , Watabe Misako , Pai Sudha K. , Xing Fei , Liu Wen , Watabe Kounosuke
Published: February 2011

Prostate cancer is becoming a curable disease when it is diagnosed at an early stage. However, even those patients who have localized cancer and have been “successfully” treated with surgery often experience recurrent disease after many years of dormancy. In the case of prostate cancer, bone is the most common metastatic site which affects approximately 70% of patients with advanced disease. The growth of the tumor cells in the bone is generally slow and they often become dormant until an appropriate microenviroment is established for their re-growth. How these metastatic tumor cells become dormant and how they recur at the target organs is virtually unknown. To address this critical question, we have studied an interaction of prostate cancer cell (PC3mm) and bone stromal cell (HS5). Our data indicate that the conditioned medium of HS5 cells was able to induce growth arrest and senescence to prostate cancer cells through activation of p38 MAP kinase. Western blot, qRT-PCR and reporter assay showed that this effect was mediated by induction of the tumor metastasis suppressor gene, N-myc downstream regulated gene 1 (NDRG1). We found that this secretory factor is bone morphogenetic protein 7 (BMP7) and that a specific inhibitor of p38 significantly abrogated up-regulation of NDRG1 by BMP7, suggesting that BMP7 induces NDRG1 through activation of p38. We also found that BMP7 significantly activated p21 and that this effect was inhibited by siRNA to NDRG1. Importantly, our results indicate that BMP7 was able to induce senescence to PC3mm cell; however, this senescence was reversible and the tumor cells re-gained the ability of their growth upon withdrawal of BMP7, suggesting that BMP7-induced senescence is reversible. Notably, our results of the existing database analysis indicate that the expression level of BMPR2, one of the BMP7 receptors, showed inverse correlation with bone metastasis and recurrence of prostate cancer. Furthermore, we isolated tumor stem population from PC3mm, and examined the effect of BMP7 on these cells. BMP7 indeed induced p38, NDRG1 and p21 in the prostate tumor stem cells. BMP7 also significantly suppressed sphere forming ability of the tumor stem cells followed by activation of NDRG1 and this effect was reversed by withdrawal of BMP7. These results suggest that the BMP7-NDRG1axis plays a critical role in the regulation of dormancy and recurrence of prostate tumor cells in bone metastasis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5762.
Abstract 5763: VCAM1 mediates aggressive bone metastasis outbreak from tumor dormancy by engaging α4β1-positive osteoclast progenitor cells
Authors: Lu Xin , Yang Qifeng , Yuan Min , Mu Euphemia , Reiss Michael , Haffty Bruce , Massagué Joan , Kang Yibin
Published: February 2011

In breast cancer patients, disseminated tumor cells can maintain in dormancy at secondary organs for a long period before relapse into life-threatening metastases. However, the molecular understanding of this process is still very limited. Here, we show that Vascular Cell Adhesion Molecule-1 (VCAM1) is essential for the conversion from dormancy to macrometastasis in a xenograft mouse model of breast cancer bone metastasis. A weakly bone-metastatic subline (SCP6) of the human breast cancer cell MDA-MB-231 was found to give rise to strongly metastatic derivatives after six-month in vivo dormancy in nude mice. Whereas no additional chromosomal alterations were detected by array-CGH, the derivatives significantly upregulated a handful of genes based on microarray analysis. Five candidate genes were evaluated by silencing their expression using short-hairpin RNA in the highly metastatic derivatives and tested by in vivo bone metastasis assay. Only the loss of expression of VCAM1 dramatically reduced the metastasis ability. Ectopic overexpression of VCAM1 in dormancy-prone sublines allowed the outgrowth of bone metastasis. We successfully blocked the initiation and the progression of bone metastasis using purified monoclonal antibodies against VCAM1 or its receptor VLA4 (integrin α4β1). Biochemical analysis showed activated NF-κB pathway is essential for the upregulation of VCAM1. Bone metastases formed by VCAM1-overexpressed tumor cells harbored numerous TRAP-positive osteoclasts and their number was dramatically attenuated with VCAM1 or VLA4 antibodies. Expression analysis using cell lines and fresh murine bone marrow showed progenitor and mature osteoclasts express α4β1. Functional assays found VCAM1 promoted osteoclast differentiation by directly tethering the α4β1-positive osteoclast progenitors. Mice with VCAM1-positive bone metastases showed enriched CD11b-positive α4β1-positive monocytic progenitor cells of osteoclasts in the bone marrow compared with mice free of bone metastases. Finally, tissue microarray analysis showed VCAM1 is upregulated in breast tumor epithelial cells in a subgroup of patients and is associated with early relapse. Overall, our study (1) established a novel mouse model for bone metastasis dormancy of breast cancer; (2) demonstrated VCAM1 as the key mediator for the conversion from dormancy to overt metastasis through engaging α4β1-positive osteoclast progenitors; (3) developed monocloncal antibodies against VCAM1 or VLA4 as promising therapeutic agents for preventing and treating bone metastasis of breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5763.
Abstract 5774: Identification of novel targeted therapies for human triple-negative breast cancer
Authors: Tomlinson Christine C. , Bennett Christina N. , Hollinghead Melinda , Green Jeffrey E.
Published: February 2011

Triple Negative (TN) breast cancers are characterized by the lack of estrogen receptor, progesterone receptor, and Her2 in the tumors. Currently, unlike breast cancers that are hormone responsive or Her2+, there are no targeted therapies for this subtype of breast cancer. Additionally, TN breast cancer is frequently diagnosed in younger women, often prior to the age of routine mammography with young African-American women having a disproportionately higher incidence and mortality from TN breast cancers than Caucasians. Despite advances in the treatment of breast cancer, molecular characteristics of this subtype remain poorly understood and improved therapies are critically needed.

Gene expression profiling has identified genetically engineered mouse (GEM) models that share important correlations with human breast tumor subtypes. In particular, the C3(1)/TAg GEM model appears to best represent basal-like/triple negative human breast cancers in several ways. The C3(1)TAg GEM model develops primary and metastatic mammary tumors following a progressive course characteristic of human breast tumor development, loss of p53 and Rb functions via SV40 T-antigen expression and amplifications of syntenic regions (hu. Chr. 12p12; mus. Chr. 6, a region containing K-RAS.) We have previously identified a critical proliferation/DNA repair/cell cycle gene signature (TAg signature), in primary mammary tumors from SV40 TAg mice that is also characteristic of human basal-like breast cancers. The TAg signature was also found to be highly predictive of human breast cancer prognosis.

By targeting the up-regulated genes identified in the TAg signature, we have identified novel targets for the treatment of TN breast cancer. We have also performed synthetic lethality studies in which gene knockdown has been combined with common chemotherapy agents (cisplatin and paclitaxel) and agents currently in clinical trials (gemcitabine, Chk1 and Notch signaling inhibitors) to identify novel combination therapies. Several combinations are being tested in in vivo tumor transplant and human xenograft experiments.

These studies have shown that the TAg signature can be used for the identification of new targets through siRNA screens of up-regulated genes in the signature and help determine new targets for preclinical testing of candidate drugs in novel combinations for treatment of TN breast cancer. The results of these studies may be translated into potentially effective therapies for women with aggressive forms of TN breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5774.
Abstract 5776: Discovery of AMG 900, a highly selective, orally bioavailable inhibitor of aurora kinases with efficacy in preclinical antitumor models and activity against multidrug-resistant cells
Authors: Geuns-Meyer Stephanie , Cee Victor J. , Deak Holly L. , Du Bingfan , Hodous Brian L. , Nguyen Hanh N. , Olivieri Philip R. , Romero Karina , Schenkel Laurie B. , Patel Vinod F. , Be Xuhai , Bush Tammy L. , Chung Grace , Eden Patrick , Huang Liyue , Kendall Richard , Lin Min-Hwa Jasmine , Radinsky Robert , Ziegler Beth , Payton Marc
Published: February 2011

The aurora family of serine/threonine kinases (Aurora-A, -B, -C) regulate cell-cycle progression in mammalian cells. Whereas aurora kinase C function appears restricted to meiosis in males, aurora kinases A and B are essential for proper chromosome congression, segregation, and cytokinesis during mitosis. Aurora kinases A and B have been implicated in tumorigenesis, with overexpression levels correlating to clinical staging of cancers and poor prognosis. Thus, these mitotic kinases have become the subject of much interest as targets for anticancer therapy.

N-(4-((3-(2-amino-4-pyrimidinyl)-2-pyridinyl)oxy)phenyl)-4-phenyl-1-phthalazinamine was a key aurora kinase inhibitor lead, possessing oral bioavailability in rats that was lacking in the anthranilamide compounds from which it was derived. This phthalazine compound possessed a key feature that was deemed important to maintain in a clinical candidate: potency against a model multidrug resistant (MDR) cell line (MES-SA Dx5) commensurate with its activity against a cell line that does not overexpress P-gp (HeLa). Improved in vivo potency was desired, as measured by suppression of the phosphorylation of the aurora kinase B substrate Histone H3 on Ser10 six hours after dosing. SAR from targeting this improvement in in vivo activity uncovered a delicate balance between protein binding, pharmacokinetic parameters, and cell potency in MES-SA Dx5 cells. AMG 900 was identified as a suitable candidate for clinical development based on its low single digit nanomolar potency against MDR cell lines, robust PD response (with complete suppression of Histone H3 phosphorylation at six hours), and high selectivity against other kinases. Oral administration of AMG 900 at a well-tolerated dose of 4 mg/kg BID inhibited tumor growth (83% TGI; p < 0.0001) in an HCT116 xenograft model. AMG 900 is currently undergoing phase 1 clinical evaluation in patients with advanced solid tumors.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5776.
Abstract 5777: EMD 1214063, an exquisitely selective c-Met kinase inhibitor in clinical phase 1
Authors: Schadt Oliver , Bladt Friedhelm , Blaukat Andree , Dorsch Dieter , Friese-Hamim Manja , Fittschen Claus , Graedler Ulrich , Meyring Michael , Rautenberg Wilfried , Stieber Frank , Wilm Claudia
Published: February 2011

The role of the receptor tyrosin kinase c-Met in tumor progression, metastasis and aggressiveness has been convincingly demonstrated in preclinical and early clinical settings. Several compounds with different selectivity profiles inhibiting c-Met are currently under preclinical/clinical investigation and might emerge as valuable cancer therapeutics in the future. During an HTS run N-(3-(3,6-Dihydro-5-(3,4-dimethoxyphenyl)-2-oxo-2H-1,3,4-thiadiazin-3-ylmethyl)-phenyl)-carbaminic acid-(3-(N,N-diethylamino)-propylester) (1) was identified as an attractive lead structure with an interesting overall profile (clogD (7.4): 2.5, S (pH 7.4): >100 µg/ml, IC50 (c-Met in vitro): 30 nM, IC50 (c-Met cellular): 800 nM) providing a valid starting point for lead optimization. The co-crystal structure of 1 revealed the binding mode and the essential structural features. The initial HTS hit was bound in a DFG-in conformation interacting namely with the main chain nitrogen atom of methionine 1160 within the hinge region and with the main chain nitrogen of aspartic acid 1222. After subsequent optimization of potency, efficacy, PK properties and the safety profile of thiadiazinone 1, EMD1214063 was identified as candidate for further development and is currently investigated in a phase 1 clinical trial. The pyridazinone EMD1214063 inhibits enzymatic and cellular c-Met kinase activity with IC50 values in the low nanomolar range. This compound displayed an exquisite selectivity when tested in vitro against a panel of more than 250 potential off-targets, including kinases, GPCRs, ion channels, transporters and various enzymes and demonstrated excellent anti-tumor activity in vivo in a variety of xenograft models. Depending on the particular model, complete regressions were observed with doses as low as 6 mg/kg/d administered per os. The overall profile of EMD1214063 including first time disclosure of the precise structure, synthesis, structure activity relationships, in vitro potency, selectivity profile, pharmacokinetic and in vivo data will be discussed.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5777.
Abstract 5780: Development of potent inhibitors of the DNA-dependent protein kinase (DNA-PK)
Authors: Cano Celine , Bailey Chris , Bardos Julia , Curtin Nicola , Frigerio Mark , Golding Bernard , Hardcastle Ian , Hummersone Marc , Menear Keith , Newell David , Saravanan Kappusamy , Shea K , Smith Graeme , Griffin Roger
Published: February 2011

The cellular response to DNA double-strand break (DSB) formation is an essential component of normal cell survival, following exposure to DNA-damaging chemicals and ionising radiation. The serine/threonine kinase DNA-dependent protein kinase (DNA-PK) is a member of the phosphatidylinositol (PI) 3-kinase related kinase (PIKK) family of enzymes, and plays an important role in DNA DSB repair via the non-homologous end-joining (NHEJ) pathway. DNA-PK inhibitors may, therefore, be useful as agents to improve the activity of radio- and chemo-therapy in the treatment of cancer.

Identification of the lead benzo[h]chromen-4-one DNA-PK inhibitor NU7026 (IC50 = 0.23 uM) guided the development of the potent and selective ATP-competitive chromenone NU7441 (DNA-PK IC50 = 30 nM). Structure-activity relationship studies for DNA-PK inhibition by chromenone-derivatives were conducted in conjunction with homology modelling. Library synthesis was undertaken employing a solution multiple-parallel approach, by O-alkylation or N-acylation of the appropriately substituted NU7441 derivatives, respectively, followed by reaction with a range of amines to afford the target compounds. These studies resulted in the identification of the highly potent inhibitor KU-0060648 (IC50 = 8.6 nM). The further development of KU-0060648 and analogues will be described.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5780.
Abstract 5781: ITRI-260, a new azaazulenone class kinase inhibitor, induces complete tumor regression and prolongs survival in AML mice
Authors: Lee On , Liu Hsingjin Eugene , Ko Ching-Huai , Tsai Yuan-Jang , Liu Chih-Peng , Chen Yen-Chun , Tseng Hsiang-Wen , Shih Ying-Chu , Chen Ting-Shou , Chen Chih-Hung , Chiou Shu-Jiau , Wang Ling-Mei , Hwang Chrong-Shiong
Published: February 2011

The FMS-like tyrosine kinase-3 (FLT3) is a receptor tyrosine kinase with important roles in hematopoietic stem/progenitor cell survival and proliferation. FLT3 is over-expressed in the majority of acute myeloid leukemia (AML) and the presence of FLT3 active mutations is associated with poor prognosis, implicating FLT3 as a potential target for AML treatment. ITRI-260 is a new azaazulenone class kinase inhibitor that inhibited FLT3-ITD-dependent MV4-11 cell growth with IC50 of 21 nM and cellular phosphorylation with IC50 of 17 nM. In in vivo PK study, ITRI-260 showed favorable oral PK profiles. The maximum concentration of ITRI-260 was 700 nM after a single 100 mg/kg oral dosing in mice, with dose proportional exposure from 10 to 150 mg/kg. ITRI-260 showed therapeutic efficacy in a human AML orthotopic SCID mouse model, following a 100 mg/kg/qd oral dosing regimen. The treatment group showed 100% survival of animals at day 90 after tumor cell inoculation, while the vehicle control group showed 100% mortality. In the other study, ITRI-260 also increased survival in primary tumor cell engraftment model inoculated with cells isolated from sunitinib- and sorafenib-resistant AML patient. Through hERG assay with patch clamp technology, we found higher IC50 of ITIR-260 compared with sunitinib and MLN-518, which indicates that ITRI-260 might arise less cardiotoxic concern. In a 14-day subchronic toxicology study in mice, ITRI-260 was generally tolerated at 300 mg/kg/qd. The most prominent adverse effect appeared at 600 mg/kg/qd group is starry sky appearance in spleen, which is moderate and reversible. No specific clinical signs or body weight loss were attributed to the test article. In conclusion, the in vitro/in vivo PK/PD/toxicology studies have provided the basis for the clinical developmental potential of ITRI-260 in AML. The bridge studies from discovery to development such as CMC, GLP toxicology/safety pharmacology and pre-formulation are now in progress.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5781.
Abstract 5783: In vitro ADME properties of ARQ 621: A specific Eg5 inhibitor
Authors: Savage Ronald E. , Zhong Caiyun , Hall Terence , Bresciano Karen , Song Pengfei , Wang Yunxia , Chan Thomas C.K.
Published: February 2011

ARQ 621 is a potent and selective allosteric inhibitor of Eg5, a microtubule-based ATPase motor protein involved in cell division. Eg5 inhibition is recognized as a potential therapeutic strategy in cancer, supported by the observation that over-expression of Eg5 causes genomic instability and tumor formation in mice. ARQ 621 is currently being tested in a Phase I clinical trial in cancer patients. Prior to its entry into the clinic, the in vitro ADME properties of ARQ 621 were studied. Data from these studies showed that ARQ 621 had a t1/2 of 53 min in human liver microsomes. The t1/2 of ARQ 621 in male and female mouse, rat, dog and monkey liver microsomes was similar to that of human liver microsomes with t1/2 values of 43, 53, 56, 53, 47, 44, 36, and 32 minutes, respectively. Consistent with the microsomal stability data, in vitro metabolic studies conducted with individual human CYP P450 isoforms indicated that ARQ 621 was relatively stable with t1/2 values all greater than 27 min. IC50 values of ARQ 621 were measured for CYP 1A2, 2C9, 2D6, 3A4, 2C19, and 2C8 and determined to be >20, >20, >20, 4.1, 4.0, and 15 µM, respectively. ARQ 621 was found to modestly induce 1A2 but not 2A6 or 3A4. Data from in vitro Caco 2 studies demonstrated that ARQ 621 has poor GI absorption potential with a Papp value of 0.69 × 10−6 cm/s. Additionally, Caco 2 bi-directional experiments suggested that ARQ 621 may be a P-glycoprotein substrate with an efflux ratio of 45. These data are consistent with rat in vivo oral bioavailability studies in which the bioavailability of ARQ 621 was determined to be approximately 9%. Hence, ARQ 621 was subsequently developed for IV administration only. Protein binding studies conducted in human plasma showed that ARQ 621 is highly bound (∼96.4-99.2%) to plasma proteins.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5783.
Abstract 579: Chinese herbal mixture LQ inhibits tumor growth and metastasis and increases survival in orthotopic and other mouse models of human and mouse cancer
Authors: Zhang Lei , Wu Chengyu , Zhang Yong , Wang Xiaoen , Reynoso Jose , Liu Fang , Zhao Ming , Hoffman Robert M.
Published: February 2011

Natural products have been used as sources of first-line cancer chemotherapeutic agents, including vinca alkaloids, taxanes, podophyllotoxin and camptothecin. However, Traditional Chinese Medicine (TCM) combinations (Fufang), which have been widely used in China for over 5000 years, are still poorly understood with regard to the types of cancer they are effective against as well as their mechanism of action. We report here on the use of patient-like orthotopic nude-mouse models of metastatic cancer and other mouse tumor models to obtain proof-of-concept that TCM (Fufang) combinations have anti-tumor and anti-metastatic activity. In the present study, human and mouse lung, pancreatic and prostate cancer cell lines (H460, Lewis lung carcinoma [LLC) MIA-PaCa-2 and PC-3, respectively), labeled with red fluorescent protein (RFP) or green fluorescent protein (GFP), were orthotopically implanted or injected subcutaneously, or injected in the tail vein in nude mice to evaluate anti-tumor and anti-metastatic efficacy of the Fufang LQ, which contains a mixture of herbs. Results showed that LQ (gavage, 600 mg/kg/day) significantly inhibited prostate, pancreatic and lung cancer tumor growth as measured by imaging, tumor size and weight, in both subcutaneous and metastatic models. LQ was efficacious against primary and metastatic cancer without weight loss in contrast to drugs such as doxorubicin and cisplatin, which caused significant weight loss. Survival of tumor-bearing mice was also prolonged by LQ treatment. Furthermore, in vitro experiments demonstrated that LQ inhibited proliferation of tumor cells (LLC, H460, PC-3 and MIA-PaCa-2) in a dose-dependent manner. Those results indicate that LQ has efficacy against metastatic pancreatic and lung and prostate cancer which demonstrates its clinical potential. The results also suggest that GFP- and RFP-expressing tumors in orthotopic and other nude mouse models can be widely used for in vivo evaluation of TCM.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 579.
Abstract 58: Hyaluronan metabolism enhances the Warburg effect and provides an alternate energy source in the breast cancer microenvironment
Authors: Thomas Natalie K. , Waltham Mark , Udabage Lishanthi , Brown Tracey J.
Published: February 2011

Introduction: A hallmark of cancer is a shift in cellular metabolism away from oxidative phosphorylation of glucose towards aerobic glycolysis thereby facilitating the rapid generation of energy required for proliferation. In breast cancer, the overproduction of hyaluronan (HA) is associated with poor disease outcomes where its metabolism is a finely regulated equilibrium between rapid synthesis and degradation. Synthetic products are involved in the structural integrity of the tumoral architecture while catabolic intermediates contribute to cell proliferation, neovascularisation and metastasis but little is known about their role in the generation of intratumoral energy.

Methods: A hyaluronanhigh producing cell line was generated by the stable transfection of HAS2 into the ZR-75-1 breast cancer cell line. The HA metabolism and expression of the HA receptor, CD44 was characterised in the parental, vector- and HAS2-transfected cell lines. The effect of elevated HA synthesis on the glycolytic phenotype was determined by quantitation of phosphoinositide 3-kinase/Akt pathway activation, transketolase-like protein 1 (TKTL1), pyruvate kinase M2 (PKM2), glucose transporter 1 (GLUT1), aerobic glycolysis, cell proliferation and metastasis both in vitro and in vivo.

Results: Compared to the controls, HAS2-transfected cells liberated up to 10-fold more HA and had increased expression of CD44s and CD44v9. The resultant increase in CD44-HA interactions mediated in vitro activation of the PI3K/Akt pathway, enhanced proliferation and metastatic capacity, increased propensity for aerobic glycolysis and a higher HA metabolic index. High levels of HA production conferred a characteristic glycolytic phenotype to HAS2 transfectants where Akt and TKTL1 were up-regulated and PKM2 reduced. Unlike the classic glycolytic phenotype, GLUT1 was not increased, but down-regulated 3-fold in the HAS2 cells. HA-induced enhancement of the glycolytic phenotype was confirmed in vivo by a >6 fold increase in endpoint tumor volume and reduced overall survival in an animal model of metastases.

Conclusions: Elevated HA production confers an aggressive phenotype to breast cancer where CD44-HA interactions alter the metabolic fate of the cell driving it into an aerobic glycolytic pathway. This study has provided preliminary evidence that in highly proliferative breast cancer glucose may not be the sole catabolic substrate and hyaluronan may contribute to this process.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 58.
Abstract 581: Development of RNA interference-based therapeutics for bladder cancer and hepatocellular carcinoma
Authors: Fosnaugh Kathy L. , Seth Shaguna , Matsui Yoshiyuki , Adami Roger , Vaish Narendra , Chen Yan , Liu Yan , Harvie Pierrot , Johns Rachel , Severson Gregory , Bell Susan , Granger Brian , Zhu Tianying , Charmley Pat , So Alan , Templin Michael V. , Polisky Barry
Published: February 2011

Bladder cancer and hepatocellular carcinoma are two of many cancers refractory to current treatments. Small interfering RNAs (siRNAs) are a new therapeutic modality able to specifically silence expression of targets not accessible via current small molecule and antibody options. MDRNA is developing UsiRNAs, a novel siRNA construct containing unlocked nucleobase analogs, with improved specificity for RNA interference (RNAi). Delivery of UsiRNAs to target tissues is achieved using proprietary Di-alkylated Amino Acid (DiLA2)-based liposomes. Survivin, overexpressed in many cancers, is involved in cell division and inhibition of apoptosis. In orthotopic and xenograft models of liver cancer, systemic administration of survivin UsiRNA-DiLA2 liposomes resulted in approximately 60% and 70% reductions in survivin mRNA, respectively, and > 50% decreases in tumor weight. Local intravesical administration in an orthotopic bladder cancer model resulted in 90% inhibition of mRNA expression and substantial tumor growth inhibition. Polo-like kinase 1 (PLK-1), also elevated in many tumors, regulates cell cycle progression and mitosis. Treatment of bladder cancer and liver cancer cell lines with PLK-1 UsiRNA/DiLA2 liposomes leads to significant caspase activation and corresponding apoptotic cell death. In vivo studies with PLK-1 UsiRNA are in progress. These UsiRNAs and UsiRNAs directed against additional targets are being evaluated in vitro and in orthotopic and xenograft models of cancer as single agents, in combinations of target-specific UsiRNAs, and with existing small molecule and antibody therapeutics.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 581.
Abstract 583: Treatment of human liver cancer in nude mice with a tumor-targeting Salmonella typhimurium auxotroph
Authors: Zhang Yong , Zhao Ming , Liu Fang , Hoffman Robert M.
Published: February 2011

We have previously developed a tumor-targeting Salmonella typhimurium double-amino acid auxotrophic strain termed A1-R (Cancer Research 66, 7647-7652, 2006; Proc. Natl. Acad. Sci. USA 104, 10170-10174, 2007). In the present study, we evaluated the efficacy of A1-R against a nude mouse model of human liver cancer. GFP-expressing A1-R bacteria targeted, colonized, and replicated in the human liver tumor growing s.c. in transgenic nude mice expressing RFP as observed by fluorescence imaging on day 3 after i.v. injection of A1-R. Subsequently, the bacteria grew throughout most of the tumor by day 10 after i.v. injection. Bacterial growth within the tumor was observed by fluorescence imaging of the infected tumor tissue in transverse sections. For efficacy experiments, bacteria (5 × 107 CFU/100 μl) were inoculated i.v. in HepG2-bearing nude mice. Tumor growth was monitored by caliper measurement in two dimensions. Treatment with A1-R resulted in tumor shrinkage. The difference in tumor volume between the treated and control group was statistically significant. There were no obvious toxic effects on the host even with 5 × 107 A1-R injected i.v. The mice tolerated A1-R treatment and could survive as long as untreated mice. A1-R only replicated and persisted within the microenvironment of the tumor and not outside the tumor site. The results suggest the potential of bacterial therapy of liver cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 583.
Abstract 584: Identification of mutations in HSV-1 envelope glycoprotein B that enhance retargeted infection
Authors: Uchida Hiroaki , Chan Janet , Goins William F. , Grandi Paola , Kumagai Izumi , Cohen Justus B. , Glorioso Joseph C.
Published: February 2011

Conditionally-replicating herpes simplex virus type 1 (HSV-1) vectors, including mutants deleted for the neurovirulence factor ICP34.5 currently in clinical trials, are promising agents for oncolytic virotherapy. As an alternative strategy to accomplish cytocidal HSV replication exclusively in cancer cells, we have designed modified viral envelope glycoproteins to limit virus entry uniquely to EGF receptor (EGFR)-bearing cells. HSV-1 entry is initiated by the interaction of glycoprotein D (gD) with one of its receptors, herpesvirus entry mediator (HVEM) or nectin-1. Receptor binding results in a conformational change in gD triggering activation of gB and gH as the effectors of fusion between the viral envelope and cell membranes. To target virus entry solely to EGFR-expressing cells, we mutated or deleted gD residues essential for binding to the natural entry receptors and inserted an EGFR-specific single-chain antibody (scFv). Although ablation of virus entry through HVEM and nectin-1 was successful, entry through the EGFR was detectable but inefficient. To address this flaw, we carried out in vitro evolution experiments to isolate gain-of-function mutations that could compensate for defective gD-receptor interactions. We passaged a nectin-1-binding impaired HSV-1 mutant repeatedly through cells expressing a gD-binding impaired nectin-1 until visible plaques appeared. Plaque purification and sequence analysis revealed that each isolate contained the same double mutation in gB. We transferred the gB double mutation into wild-type virus and observed entry into cells that are normally resistant to HSV infection due to the absence of authentic gD receptors. In addition, we found that the mutant-gB virus was capable of entering cells through nectins other than nectin-1, and that entry into nectin-1-bearing cells was markedly accelerated compared to wild-type virus. We combined the gB double mutation with the EGFR-retargeted gD allele and found that this combination yielded increased entry into target cells without raising off-target infection. We are currently expanding this strategy to target other cancer-specific cell-surface markers. By enhancing the efficiency of retargeted infection, the gB double mutation is likely to reduce the need for strong mutations in current oncolytic vectors that not only prevent virus replication in normal cells, but also diminish replication in tumor cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 584.
Abstract 59: Impact of omega-3 fatty acids on obesity-induced DNA oxidation and prostate cancer progression
Authors: Cavazos David A. , deGraffenried Linda A.
Published: February 2011

Obesity is associated with poor prognosis and contributes significantly to prostate cancer-related mortality. As the prevalence of obesity increases among young adult males, the contribution to prostate cancer progression and mortality are expected to rise dramatically. One potential mechanism by which obesity may promote worse outcome is through the induction of functional metabolic abnormalities known to promote inflammation and oxidative stress. Pre-clinical and clinical observations indicate that a BMI >30 promotes oxidative DNA damage and is associated with shorter time to PSA failure and prostate cancer mortality. Growing evidence supports a beneficial role of omega-3 (ω-3) fatty acids in delaying time to PSA failure and reducing the aggressiveness of advanced prostate cancer, however, the mechanisms are multivariate and relatively unresolved. As oxidative burden is strongly implicated in the pathogenesis of age-related diseases, including prostate cancer tumor formation, and as omega-3 fatty acids possess known antioxidant and anti-inflammatory properties, we investigated the effects of docosahexaenoic acid (DHA-22:6n-3), one component of fish oil, in reducing the effects of obesity-induced oxidative DNA damage. Preliminary data demonstrates that prostate cancer cells (LNCaP) pretreated with ω-3 fatty acids and pulsed with physiologic levels of H2O2 were preferentially sensitive to induction of growth arrest compared to normal prostate epithelial cells (PrEC). LNCaP cells pulsed with 32 uM H2O2 exhibit DNA breaks associated with translocation of NF-κB into the nucleus, whereas pretreatment with DHA prior to H2O2 treatment prevented NF-κB translocation, selectively sensitizing LNCaP cells, but not PrEC, to cell death. Further, DHA attenuated H2O2-induced NF-κB transcriptional activity and diminished expression of the downstream target, survivin. NF-κB is heavily implicated in promoting prosurvival signaling and may be critical for resistance to the chronic oxidative stress and inflammation observed in the pathogenesis of prostate cancer. Here we describe one potential mechanism by which obesity may contribute to prostate cancer and the role of ω-3 fatty acids in mitigating these affects, ultimately setting the stage for Phase III clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 59.
Abstract 590: Preclinical biosafety test of oncolytic adenovirus-infected carrier cell for Phase I study
Authors: Hamada Katsuyuki
Published: February 2011

Although almost 1000 clinical trials of gene therapy have been tried in USA, any significant clinical effects have not yet been reported. This is due to the anti-viral neutralizing antibodies production after the induction of viral mediated gene delivery for clinical trials. To address this limitation, we developed a carrier cell system to deliver a replication-competent adenovirus. Carrier cells infected with oncolytic adenovirus were incubated with target ovarian cancer cells in high titer of antiadenovirus antibody. Carrier cells were injected into syngeneic subcutaneous ovarian tumors after immunization with adenovirus. Carrier cell-derived cell fragments containing viral particles were engulfed by proliferative target ovarian cancer cells. This engulfment-mediated transfer of adenovirus was not inhibited by antiadenovirus antibody and enabled repetitive infection. After the induction of antiadenoviral CTL responses by immunization of adenovirus, administration of carrier cells infected with a replication-competent adenovirus induced complete tumor regression. Short term toxicity and distribution test were carried out to inject carrier cells into ovarian tumor in nude mice at once and long term toxicity test was carried out to inject carrier cells into rabbits at 8 times for 4 weeks. Short term toxicity test did not show any lethal side effects in nude mice. In distribution test, single injection of carrier cells into ovarian tumor induced the peak levels of cells and oncolytic virus in tumors on the next day but did not show any significant levels of cells and oncolytic virus in tumors 14 days after injection. In long term toxicity test, 8 injections of carrier cells (1.25 × 107 cells/kg) did not show any significant toxicity in rabbits

This novel adenoviral receptor independent carrier cell-mediated viral transfection system might prove safe in this preclinical biosafety test and useful to treat solid tumors. Phase I clinical trial of this carrier cells infected with oncolytic adenovirus will be started soon to treat recurrent solid tumors which is resistant to the common treatment of surgery, radiation and chemotherapy.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 590.
Abstract 592: Downregulation of uPAR and uPA retards angiogenesis and proliferation and activates the extrinsic apoptotic pathway in pancreatic cancer cells
Authors: Gorantla Bharathi , Rao Jasti S. , Patel Jitendra , Gondi Christopher S.
Published: February 2011

Pancreatic cancer is one of the major causes of mortality in the United States. Despite well established chemotherapy and surgical resection strategies, the five-year mean survival rate is less than five percent. The activation of the uPAR/uPA system is an early event in the development of pancreatic ductal adenocarcinoma (PDAC), and uPAR gene amplification identifies a subgroup of particularly aggressive tumors. In the present study, we used two pancreatic cancer cell lines [MIA PaCa-2 (CRL1420) and PANC-1 (CRL1469)] to demonstrate the effect of uPA and uPAR downregulation. From the results, we observed that RNAi-expressing plasmids efficiently downregulated the expression of uPAR and uPA both at the mRNA and protein levels. In vitro and in vivo angiogenic assays revealed a significant decrease in angiogenic potential of CRL1420 (in vitro 26±3% and in vivo 13±2%) and CRL1469 (in vitro 23±1% and in vivo 10±2%) cells that were downregulated for both uPA and uPAR. From the antibody array analysis, we observed that the simultaneous downregulation of uPAR and uPA subsequently caused the downregulation of angiogenin and the overexpression of RANTES. The MTT proliferation assay results demonstrated that the downregulation of uPAR alone caused a decrease in proliferation (CRL1420: 24±3%; CRL1469: 38±2%) and downregulation of uPA alone caused a lesser decrease in proliferation (CRL1420: 19±2%; CRL1469: 26±1%) whereas the simultaneous downregulation of both uPA and uPAR caused the most significant decrease in proliferation in both cell lines (CRL1420: 84±4%; CRL1469: 93±3%). Further, FACS analysis demonstrated that the simultaneous downregulation of uPAR and uPA caused the accumulation of cells in the sub-G0/1 phase in both CRL1420 and CRL1469 cells. In addition, western blot analysis revealed that downregulation of uPA and uPAR caused the activation of caspase 8 and induced the cleavage of PARP, which is indicative of apoptosis; these results were confirmed by TUNEL assay. In conclusion, our results clearly demonstrate that the targeting of the uPAR-uPA system has potential clinical relevance for the treatment of pancreatic cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 592.
Abstract 593: Abusing oncogenic RAS to selectively target colorectal and pancreatic cancer cells
Authors: Naumov Inna , Lisiansky Victoria , Kazanov Dina , Shapira Shiran , Kraus Sarah , Arber Nadir
Published: February 2011

Background: RAS mutations are present in 40-60% of human colorectal (CRC) and 75-95% of pancreatic (PC) tumors leading to the activation of downstream target genes. The pro-apoptotic PUMA protein interacts with Bcl-XL and Bcl-2 to induce the release of mitochondrial cytochrome C and apoptosis.

Aim: To selectively kill RAS transformed cells by overexpressing the PUMA gene under the control of a RAS-responsive promoter.

Materials and methods: Four CRC and three PC cell lines were used. Six of them harboring KRAS mutations; SW480, HCT116, DLD1, Colo357, Panc1 and MiaPaca, whereas HT29 cells, harboring wt KRAS, served as control. In addition, normal rat enterocytes, IEC18 and RIE and their KRAS (R1), HRAS (RIE-RAS) transformed counterparts, were tested. We constructed two adenoviral vectors with the wt PUMA sequence, one containing RAS-responsive elements as an upstream promoter (PY4-PUMA) and a second lacking these elements which served as control (SV40-PUMA). Prior to the infections, RAS activity was determined by pull-down assay using GST-RBD. Inhibition of proliferation was determined on a Coulter counter; cell viability by MTT assay, apoptosis by FACS and protein expression by Western blotting. The effect of the adenoviruses was evaluated in vivo following injection of 106 cells into two flanks of nude mice (n=30).

Results: As expected, SW480, HCT116, DLD1, Colo357, Panc1, R1 and RIE-RAS showed increased RAS activity compared to HT29, IEC18 and RIE cells which had no activity. Infection with PY4-PUMA induced a marked inhibition of cell growth, up to 40-50% and spontaneous apoptosis, in all cells exhibiting high levels of RAS activity whereas HT29, IEC18 and RIE were unaffected by PY4-PUMA. The SV40-PUMA had no effect. Selective and high expression of PUMA was detected in PY4-PUMA affected cells.

In xenograft models of CRC in nude mice, PY4-PUMA injected i.p twice during a period of 4 weeks, suppressed de novo tumor formation and inhibited by ∼35% the growth of established tumors compared to the control virus.

Conclusions: Selective expression of PUMA in RAS transformed cells can induce cancer cell death. The minimal quantity of only two adenoviral injections in our in vivo model showed no toxicity and holds promise for its future application in humans that, unlike immunodeficient mice, are immunocompetent. Gene therapy may become a useful, effective and safe approach to target RAS mutated tumors while sparing the normal surrounding tissue. It might be applicable not only to GI tumors but to other tumors harboring KRAS or HRAS mutations.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 593.
Abstract 596: Oncolytic vesicular stomatitis virus selectively kills bladder cancer cells which have a low level of type I interferon receptor
Authors: Zhang Kevin , Matsui Yoshiyuki , Hadaschik Boris A. , Lee Cleo , Jia William , Bell John C. , Fazli Ladan , So Alan I. , Rennie Paul S.
Published: February 2011

Bladder cancer is the second most common genitourinary malignancy. Current treatments options are of limited efficacy since approximately 80% of patients will develop recurrent tumors of which 20-30% evolve into more aggressive, potentially lethal cancers. A new strategy to treat bladder cancer is the intra-vesical application of vesicular stomatitis virus (VSV), which is a replication competent oncolytic virus that relies on an aberrant interferon signaling pathway within cancer cells.

Using a tissue microarray composed of human bladder cancer cores and by immunohistochemistry, we observed that expression of type I interferon (IFN) receptor (IFNAR) was decreased relative to normal bladder tissue. Advanced bladder cancers had even lower expression of IFNAR. We observed that VSV preferentially targeted high grade bladder cancer cells which were resistant to type I IFN treatment. We also found that these high grade bladder cancer cells susceptible to VSV-induced lysis had low expression of IFNAR. Furthermore, siRNA knockdown of IFNAR indeed facilitated replication of VSV in cells previously resistant to VSV treatment. Consequently, intra-vesicular instillation of wildtype VSV and the Delta51M strain, which has an impaired ability to shutdown innate immunity, significantly inhibited high grade tumor growth by 98% and 90% respectively as compared to controls treated with UV-inactivated VSV. No evidence of toxicity or systemic viral infection in either group was found.

In conclusion, down-regulation of IFNAR in bladder cancer may be one of the primary molecular mechanisms for clinical IFN resistance. However, this also facilitates VSV replication and oncolysis in high risk bladder cancers and provides a basis for selecting bladder cancer patients for IFN or oncolytic VSV therapy in future clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 596.