bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2730: Novel animal models of pancreatic cancer induced by smoking and chronic pancreatitis.
Authors: Xu Shiping , Grippo Paul J. , Khayrutdinova Oxana M. , French Samuel W. , Pandol Stephen J. , Gukovskaya Anna S. , Edderkaoui Mouad
Published: August 2014

Background and Aims: Smoking and pancreatitis are two major risk factors for pancreatic cancer. We developed novel mouse models of pancreatic cancer induced by smoking and chronic pancreatitis. We used two pancreatic cancer mouse models induced by overexpression of Kras with two different degrees of disease complexity. Methods: Wild type (WT), EL-Kras and Pdx1-Cre;LSL-Kras transgenic mice were subjected to a major cigarette smoke compound nitrosamine 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) at doses close to those accumulated with smoking (100mg/kg) or exposed to tobacco smoke (80mg/m3) in tobacco smoke chambers for five days a week during 6 weeks. Standard model of chronic pancreatitis induced by cerulein injections (50μg/kg) for 3 weeks was applied to WT and Pdx1-Cre;LSL-Kras mice. Mice were then sacrificed and pancreatic tissue analyzed. Apoptosis was assessed by TUNEL. Autophagy was assessed by measuring autophagic markers LC3-II and p62. Pancreatic lesions, proliferation, fibrosis and inflammation were analyzed by immunohistochemistry. Epithelial to mesenchymal transition (EMT) and expression of stem cell markers were analyzed by Western. Results: NNK and exposure to tobacco smoke significantly increased the number and stage of pancreatic neoplastic (PanIN) lesions in Kras transgenic mice. Smoking stimulated fibrosis and stellate cells activation but had no significant effect on inflammation. EMT markers (decrease in E-Cadherin and increase in vimentin protein levels) were stimulated by NNK and tobacco smoke exposure in both WT and transgenic mice. NNK decreased apoptosis and LC3-II level in Kras mice. In addition, smoking stimulated expression of stem cell markers sox2 and CD133. Cerulein stimulated fibrosis, stellate cells activation, inflammation, and slightly stimulated EMT in WT and transgenic mice. Combination of smoking and cerulein had additive effect on stimulation of PanIN lesion formation, fibrosis, EMT and stem cell markers induction. Conclusion: The results indicate that both tobacco smoke and chronic cerulein pancreatitis promote pancreatic cancer progression in both EL-Kras and Pdx1-Cre;LSL-Kras mice models. Smoking acts through enhancing fibrosis and EMT; whereas pancreatitis stimulates mainly inflammation and fibrosis. Smoking and pancreatitis have additive effect on fibrosis, EMT and acquiring of stem cell characteristics.

Citation Format: Shiping Xu, Paul J. Grippo, Oxana M. Khayrutdinova, Samuel W. French, Stephen J. Pandol, Anna S. Gukovskaya, Mouad Edderkaoui. Novel animal models of pancreatic cancer induced by smoking and chronic pancreatitis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2730. doi:10.1158/1538-7445.AM2013-2730
Abstract 2733: K14-GFP-miR-31 transgenic mice confer high susceptibility to squamous cell tumorigenesis on oral tongue and esophagus.
Authors: Tseng Ssu-Hsueh , Lee Yong-Syu , Chang Kuo-Wei , Lin Shu-Chun
Published: August 2014

Squamous cell carcinoma (SCC) occurring on head and neck region and esophagus causes tremendous cancer mortality around the world. Effects from tobacco and/or alcoholic carcinogens and other factors are highly associated with the prevalence of these malignancies. MiRNAs are small non-coding RNAs mediating the RNA interference on targeted mRNAs. They play important roles in the pathogenesis of malignancies. miR-31 is among the most eminently up-regulated miRNAs in SCC occurring in head and neck region and esophagus. To elucidate the molecular pathogenesis of SCC, we established miR-31 transgenic mouse lines, in which miR-31 is under the control of K14 promoter and co-expressed with GFP. GFP and miR-31 were detected in basal and suprabasal squamous cells of skin, oral cavity and esophagus in these transgenic mice. The transgenic mice exhibited higher potential for tumor induction than wild type mice on oral tongue and esophagus by 4-nitroquinoline 1-oxide (4NQO) treatment. The precancerous lesions on oral tongues and esophagus in transgenic mice were also in more advanced stages in contrast to counterparts of wild type mice. Homozygous transgenic mice had the worst prognosis than the others. 4NQO induction also up-regulated miR-31 expression in tumors of wild type mice. The tumors induced in transgenic mice expressed more abundant miR-31 and VEGF, and less abundance of factor inhibiting HIF (FIH) than tumors in wild type mice. Base excision repair genes were validated as targets of miR-31 in human SCC cell lines, and the expression of these target genes were also decreased in the squamous epithelium of transgenic mice. This study identifies the high sensitivity of K14-GFP-miR-31 transgenic mice for chemical carcinogen-induced squamous cell tumorigenesis, and this could be associated with the down-regulation of tumor suppressor genes and the impairment of repair activity to counteract carcinogenic stimuli in squamous cells.

Citation Format: Ssu-Hsueh Tseng, Yong-Syu Lee, Kuo-Wei Chang, Shu-Chun Lin. K14-GFP-miR-31 transgenic mice confer high susceptibility to squamous cell tumorigenesis on oral tongue and esophagus. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2733. doi:10.1158/1538-7445.AM2013-2733
Abstract 2734: Residual embryonic cells as precursors of a Barrett's-like metaplasia.
Authors: Wang Xia
Published: August 2014

Barrett's esophagus is an intestine-like metaplasia and precursor of esophageal adenocarcinoma. Trig- gered by gastroesophageal reflux disease, the origin of this metaplasia remains unknown. p63-deficient mice, which lack squamous epithelia, may model acid-reflux damage. We show here that p63 null embryos rapidly develop intestine-like metaplasia with gene expression profiles similar to Barrett's metaplasia. We track its source to a unique embry- onic epithelium that is normally undermined and re- placed by p63-expressing cells. Significantly, we show that a discrete population of these embryonic cells persists in adult mice and humans at the squa- mocolumnar junction, the source of Barrett's meta- plasia. We show that upon programmed damage to the squamous epithelium, these embryonic cells migrate toward adjacent, specialized squamous cells in a process that may recapitulate early Barrett's. Our findings suggest that certain precancerous lesions, such as Barrett's, initiate not from genetic alterations but from competitive interactions between cell lineages driven by opportunity.

Citation Format: Xia Wang. Residual embryonic cells as precursors of a Barrett's-like metaplasia. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2734. doi:10.1158/1538-7445.AM2013-2734
Abstract 2735: Inactivation of activin signaling pathway accelerates the development of pancreatic intraductal papillary mucinous neoplasms in vivo.
Authors: Qiu Wanglong , Tang Sophia , Lee Sohyae , Turk Andrew T. , Sireci Anthony , Qiu Anne , Hruban Ralph H. , Remotti Helen E. , Su Gloria H.
Published: August 2014

The association of TGF-β pathway with human tumorigenesis has been rigorously demonstrated. However, little is known about activin signaling, part of the TGF-β family, in pancreatic tumorigenesis. We have previously reported sporadic mutations of the Activin receptor type 1B (ACVR1B) in human pancreatic ductal adenocarcinoma (PDAC). To investigate the significance of ACVR1B in pancreatic tumorigenesis, Acvr1bflox/flox; Pdx1-Cre mice were generated and examined. Chronic pancreatitis-like histological changes such as inflammatory cell infiltration, acinar to ductal metaplasia, and fibrosis were observed in Acvr1b mutant alone mice older than 8 months of age. In combination with mutant KrasG12D in the pancreas, Acvr1b deletion accelerated the development of pancreatic intraductal papillary mucious neoplasms (IPMN), but not the pancreatic intraepithelial neoplasias (PanINs). The IPMN progressed to invasive and metastatic cancer was observed in this model. The expressions of Cox-2 and phospho-PDK1 are dramatically increased in preneoplastic and neoplastic lesions, but activated Notch4 expression is exclusively observed in the IPMN lesions by immunohistochemistry, suggesting that Notch4 signaling may play a role in the formation or maintenance of the mucin-rich ductal phenotype in this model. Interestingly, the progression of IPMN to invasive cancer appears to require additional p16 loss in this double mutant mouse model. In human IPMN, loss of p16 expression is associated with increasing grade. Our data provide the first evidence that Acvr1b acts as a tumor-suppressor in vivo and that activin signaling plays a predominant role in the development of pancreatic IPMNs.

Significance: PDAC can arise from PanINs or IPMN. It has been reported previously that disruption of TGF-β signaling promotes the progression of PanIN to

PDAC in the TGFβRII knockout background. In contrast, here we present that activin signaling deficiency favors the development of IPMNs and enhances their progression to PDAC. This phenotype overlaps with those observed in the Smad4 knockout background, suggesting that the involvement of activin signaling in the IPMN to PDAC sequence is Smad4-dependent. Together these data demonstrate that TGF-β superfamily ligands play critical roles in regulating the IPMN-derived versus PanIN-derived carcinogenesis.

Citation Format: Wanglong Qiu, Sophia Tang, Sohyae Lee, Andrew T. Turk, Anthony Sireci, Anne Qiu, Ralph H. Hruban, Helen E. Remotti, Gloria H. Su. Inactivation of activin signaling pathway accelerates the development of pancreatic intraductal papillary mucinous neoplasms in vivo. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2735. doi:10.1158/1538-7445.AM2013-2735
Abstract 2750: Targeted therapy of childhood ALL and Ewing's sarcoma using antiCD19 (antiCD99)-Hybrid Polymerized Liposomal Nanoparticles (HPLN).
Authors: Kang HyungGyoo , Shahbazian Violette , Nagy Jon , Triche Timothy
Published: August 2014

Current success in childhood cancer therapy improves the survival rate of patients but unfortunately not all childhood cancers have shown successful survival rates and still many types of childhood cancer continue to have a poor prognosis. Furthermore due to the poor specificity of cancer therapies almost two-third of childhood cancer survivals experience side effects such as secondary cancers, heart or lung damage, infertility or chronic hepatitis. Therefore a reliable, non-toxic high therapeutic window targeted therapy is urgently needed to treat childhood cancer patients. In our study a novel nanoparticle (HPLN) was used for the treatment of childhood ALL and Ewing's sarcoma. The tumor specific monoclonal antibody was conjugated to the PLN into which Doxorubicin was uptaken using active loading method. AntiCD19 and antiCD99 antibodies were used for targeting of ALL and Ewing tumor, respectively. These tumor specific HPLNs effectively inhibit tumor growth in a murine model of ALL and Ewing's sarcoma. Removal of targeting antibody or drug eliminates the antitumor effects, which proves this anticancer effect of HPLN is very specific to the target cancer cells and dependent upon drug. Additionally, no abnormalities in liver and kidney function tests, complete blood counts or pathology of major organs are observed from tail-vein administrations. These data provide strong evidence for the safety and efficacy of this targeted HPLN delivery system of anticancer drugs to childhood cancer.

Citation Format: HyungGyoo Kang, Violette Shahbazian, Jon Nagy, Timothy Triche. Targeted therapy of childhood ALL and Ewing's sarcoma using antiCD19 (antiCD99)-Hybrid Polymerized Liposomal Nanoparticles (HPLN). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2750. doi:10.1158/1538-7445.AM2013-2750
Abstract 2756: Targeted inhibition of Chromosomal Maintenance Region Protein (CRM1) potently suppresses growth of human neuroblastoma cell line models.
Authors: Attiyeh Edward F. , McKeon-Fish Aaron , Landesman Yosef , Senapedis William , McCauley Dilara , Kashyap Trinayan , Shacham Sharon , Kauffman Michael , Maris John M.
Published: August 2014

Background: Neuroblastoma is a childhood malignancy of the peripheral nervous system, and most patients with high-risk disease are not cured. New therapies are needed, but recurrent somatic mutations indicative of a tractable therapeutic target are rare. Translocation of p53, FOXO, IκB and other critical tumor suppressor proteins (TSPs) from the nucleus to the cytosol is mediated by exportin 1 (CRM1, XPO1), suggesting that inhibition of CRM1 is a rational anti-cancer strategy. KPT-330 is a Selective Inhibitor of Nuclear Export (SINE) that irreversibly binds CRM1 and is currently in Phase 1 clinical trials. SINE inhibits CRM1-mediated nuclear export across multiple tumor types leading to activation of TSPs and cancer cell death.

Methods: A panel of 14 well-characterized neuroblastoma cell lines was treated with KPT-330 across a 5-log range. Growth inhibition was measured using CellTiter-Glo (Promega) viability assays and a real time growth monitoring system (xCELLigence, Roche). NOD/SCID mice with neuroblastoma xenografts were treated orally with KPT-330 and tumor size was monitored.

Results: KPT-330 showed cytotoxicity against 14 neuroblastoma cell lines with a median IC50 of 289±47.5 nM. Sensitivity to KPT-330 was independent of MYCN amplification status. Combination of KPT-330 with commonly used cytotoxic agents (irinotecan, topotecan, cisplatin, doxorubicin) showed additive effects. In xenograft models using the neuroblastoma cell lines IMR-32 and SH-SY5Y, oral administration of KPT-330 (10 or 20 mg/kg, twice or thrice weekly) was well tolerated and showed significant tumor growth inhibition (p<0.001). KPT-330 treatment resulted in decreased Myc family protein levels as well as activation of apoptotic pathways.

Conclusions: Neuroblastomas show sensitivity to CRM1 inhibition both in vitro and in vivo. Ongoing work is focused on discovering the cellular and genomic factors responsible for increased sensitivity to nuclear export inhibition. With the expected completion of the first in human phase I trials by mid 2013, treatment with KPT-330 has the potential to be rapidly translated into a clinical trial for children with neuroblastoma.

Citation Format: Edward F. Attiyeh, Aaron McKeon-Fish, Yosef Landesman, William Senapedis, Dilara McCauley, Trinayan Kashyap, Sharon Shacham, Michael Kauffman, John M. Maris. Targeted inhibition of Chromosomal Maintenance Region Protein (CRM1) potently suppresses growth of human neuroblastoma cell line models. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2756. doi:10.1158/1538-7445.AM2013-2756
Abstract 2766: YM155 inhibits survivin-mediated survival, migration, and tumor growth in neuroblastoma.
Authors: McClung Heather M. , Zhao Ping , Smith Lauren , Eckardt Alexandra , Sholler Giselle L.
Published: August 2014

Background: Neuroblastoma is the most common extracranial solid tumor in children, with poor prognosis in children diagnosed after 12-15 months of age. Even with current multimodal therapies, the 5-year event-free survival is below 50%, necessitating the development of new, targeted therapies. Survivin is a developmentally regulated gene that is highly up-regulated in many cancers. In neuroblastoma, survivin is up-regulated by PI3Kinase-AKT signaling and its expression correlates with poor prognosis. Survivin is a member of the Inhibitor of Apoptosis family of proteins and is involved in cell survival through caspase dependent and independent pathways. Knockdown of survivin in melanoma cells reduces cell migration in vitro and its expression correlates with metastasis in Ewing's Sarcoma and Schwann cell tumors. YM155 is a transcriptional suppressor of survivin and has shown efficacy in several cancer model systems both in vivo and in vitro and is currently under clinical investigation for adult cancers. In this study, we investigate the effects of YM155 on neuroblastoma cell apoptosis and migration in vitro and on tumor growth in vivo.

Methods: Four neuroblastoma cell lines were treated with YM155 and subject to cell viability and migration assays. Gene expression was investigated by microarray and signaling by Western Blotting. In vivo, effects on tumor volume were determined by xenograft implant of neuroblastoma cell lines followed by YM155 treatment and measurement by calipers.

Results: Treatment with YM155 caused a dose-dependent decrease in survivin expression and reduced cell viability in neuroblastoma cell lines; however IC50 values vary by cell line. Cells treated with YM155 near the respective IC50 showed increased cleavage of Caspase 3 and PARP indicating apoptosis. Gene and protein expression changes include reduced CXCR4 and Mcl-1. Low levels of YM155 reduced cell migration by 30-50%; cytotoxicity was not observed at the lower concentrations. Treatment with YM155 reduced tumor growth in vivo by 25-70% and depended on cell line.

Conclusions: YM155 inhibits survivin expression, reduces cell viability, induces apoptosis, and migration in vitro, and reduces tumor volume in vivo. Further investigation is warranted in characterization of YM155 as a possible therapeutic agent for neuroblastoma.

Citation Format: Heather M. McClung, Ping Zhao, Lauren Smith, Alexandra Eckardt, Giselle L. Sholler. YM155 inhibits survivin-mediated survival, migration, and tumor growth in neuroblastoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2766. doi:10.1158/1538-7445.AM2013-2766
Abstract 2768: Molecular properties and antiproliferative activity against tumor cells of a new poly-alkylamino-bis-maltolic synthetic molecule (maltonis).
Authors: Manara Maria Cristina , Fanelli Mirco , Amatori Stefano , Guerzoni Clara , Landuzzi Lorena , Lollini Pier-Luigi , Giorgi Luca , Fusi Vieri , Scotlandi Katia
Published: August 2014

Introduction Hydroxypyrones are highly versatile molecules and several members of this family, both alone and in combination with metals, were found to exert anti-proliferative activities against a wide range of tumor cells. In this study, we show the anti-tumor potential of the prototype of new molecules belonging to the family of hydroxypyrones, named maltonis [4(N),10(N)-bis[(3- hydroxy-4-pyron-2-yl)methyl]-1,7-dimethyl-1,4,7,10-tetraazacyclododecane], in different tumor cell lines (derived from both hematopoietic and solid human tumors).

Methods and Results We examined the potential of maltonis to induce, in cell-free assays, covalent modifications of DNA structure, together with anticancer activities (cell growth inhibition, cell cycle perturbations, effects on programmed cell death). The cellular response to maltonis treatment was assessed also at the molecular level by evaluation of the transcript level of genes involved in cell cycle and programmed cell death regulation. The possible induction of a maltonis-mediated DNA damage response was investigated monitoring the histone H2AX phosphorylation (γH2AX). Sarcoma cell lines appeared to be particularly sensitive to maltonis in in vitro assays. Consistently, preliminary in vivo experiments reported a cytostatic effect of maltonis, monitoring a consistent reduction of the neoplastic lesions, when intra-tumor injected at doses of 20/40 mg/kg (for two weeks) in a xenograft model of Ewing Sarcoma. However, a range of different levels of sensitivity was observed in the different histotypes (osteosarcoma, Ewing sarcoma and rhabdomyosarcoma) and among cell lines, indicating the need of further studies to identify markers of efficacy.

Conclusions Taken together, these results show that maltonis appears to be a good candidate for sarcoma treatment. It induces complex DNA structural modifications, which need to be further analysed in order to clarify mechanisms of action and efficacy of this new drug.

Grants from: AIRC 10452 and Ministry of Health_RF2008 to KS; A.S.S.O., PRIN 2008 and PRIN 2009 to MF and VF.

Citation Format: Maria Cristina Manara, Mirco Fanelli, Stefano Amatori, Clara Guerzoni, Lorena Landuzzi, Pier-Luigi Lollini, Luca Giorgi, Vieri Fusi, Katia Scotlandi. Molecular properties and antiproliferative activity against tumor cells of a new poly-alkylamino-bis-maltolic synthetic molecule (maltonis). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2768. doi:10.1158/1538-7445.AM2013-2768
Abstract 2769: DSRCT: Establishing xenograft and tissue culture cell lines and assessing chemosensitivity.
Authors: Markides Constantine SA , Vardeman Dana M. , Luong Linh , Coil Douglas R. , Kozielski Anthony , Mendoza John , Giovanella Beppino C.
Published: August 2014

Desmoplastic small round cell tumor (DSRCT) is an extremely rare and aggressive neoplasm which mainly affects young males and generally presents as a widely disseminated tumor within the peritoneal cavity. Due to the rarity of the tumor, a younger and overall healthier patient population, and because it lacks definitive histologic and immunohistologic features, it is possible that DSRCT is frequently delayed in diagnosis or is entirely misdiagnosed as a different kind of abdominal sarcoma. This study seeks to rectify the dearth of models that exist for this rare neoplasm through the development of several DSRCT tissue culture and xenograft lines. Samples were received from surgeries and biopsies from patients around the world and were immediately processed for xenograft development in nude mice. Tumor tissues were minced and fragments were injected onto the dorsal flank of nude mice. Of the 15 samples received, 9 were established into xenograft lines and 6 of those into tissue culture lines. Xenografts displayed the microscopic histology of their parent tumors and two different growth rates among established xenograft lines. Chemosensitivity of tumors was assessed against a variety of chemotherapeutic agents and IC50 values established for each drug and cell line. Interestingly, the cell lines were split into two distinct groups in terms of morphology, growth rate and chemosensitivity. Overall, the establishment of these xenograft and tissue culture lines gives researchers tools to evaluate DSRCT response to chemotherapy and investigate DSRCT-specific signaling pathways or mechanisms.

Citation Format: Constantine SA Markides, Dana M. Vardeman, Linh Luong, Douglas R. Coil, Anthony Kozielski, John Mendoza, Beppino C. Giovanella. DSRCT: Establishing xenograft and tissue culture cell lines and assessing chemosensitivity. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2769. doi:10.1158/1538-7445.AM2013-2769
Abstract 277: Investigation of the signaling factors involved in Ras/MAPK-promoted metastasis in the PTEN−/−TP53−/− mouse model.
Authors: Tillman Heather S. , Yin Juan Juan , Liu Yen Nien , Awan Musaddiq , Martin Philip , Kelly Kathleen
Published: August 2014

The most significant challenges for advanced prostate cancer patients are resistance to androgen-deprivation therapy and the dissemination of metastatic cancer cells. Our study begins to address how enhanced Ras signaling cooperates with combined TP53 and PTEN loss for enhanced metastatic capability. Because neither PTEN nor TP53 deletion alone is not sufficient for metastasis, we asked what other signaling pathways are required. Ras pathway activation is implicated in prostate cancer metastatic progression in comprehensive genetic analyses of human prostate tumors. We introduced a lentiviral vector expressing K-rasG12V into a non-metastatic PTEN−/−/TP53−/- adenocarcinoma cell line (clone 2). Orthotopic prostate injection of clone2/Ras expressing cells resulted in 100% pulmonary metastasis. The metastatic phenotype was paralleled by an inhibition of sarcomatoid carcinoma transformation. We hypothesized that Ras signaling modified TGFβ-mediated EMT signaling, which we have previously demonstrated leads to sarcomatoid carcinoma formation in this model. Multiple pathways downstream of TGFβ initiate signaling including SMAD4 dependent transcription and other non-SMAD mediated pathways like TAK1 induced NF-κB activation. Pathologic analysis of nuclear p65 labeling, a canonical NF-κB marker, showed enhanced labeling in clone2/Ras developed tumors and lung metastases and undetectable levels in clone 2 developed tumors. These in vivo findings suggest enhanced canonical NF-κB signaling may enable Ras transformed prostate cancer cells with metastatic capability.

Citation Format: Heather S. Tillman, Juan Juan Yin, Yen Nien Liu, Musaddiq Awan, Philip Martin, Kathleen Kelly. Investigation of the signaling factors involved in Ras/MAPK-promoted metastasis in the PTEN−/−TP53−/− mouse model. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 277. doi:10.1158/1538-7445.AM2013-277
Abstract 2771: Development of a non-thermal plasma model for neuroblastoma treatment.
Authors: Srinivasan Priya , Snyder Jason A. , Shashurin Alexey , Keidar Michael , Sandler Anthony D.
Published: August 2014

Neuroblastoma is a common extra-cranial solid tumor in children. There are several clinical categories of this disease, ranging from curable to very poor prognosis in which this cancer is treated with multi-modal therapy. We have tested a novel ablative technology, using non-thermal helium generated plasma jets, as a means to selective targeting of neuroblastoma cells. Plasma consists of partially ionized molecules, free radicals and other species. We have investigated the molecular mechanisms involved in non-thermal plasma mediated tumor cell death, as well as the differences in response between tumor and normal cells. Treatment with non-thermal plasma induced apoptosis in cancer cells, while normal cells appear to be less sensitive to the toxic effects. Levels of reactive oxygen species (ROS) such as superoxide anion and hydrogen peroxide are elevated after plasma treatment, suggesting that this may contribute to cell death. Scavengers of ROS were able to abrogate apoptosis in plasma-treated cells. Plasma therapy of tumor cells may be a novel approach to delivery of high levels of ROS, offering the ability to directly target and selectively destroy neoplastic tissue. Methods enhancing the ROS effect and/or technical delivery of the free radicals uniformly to tumor cells within a mass will be needed before this technology could be translated to ablating solid tumors in vivo.

Citation Format: Priya Srinivasan, Jason A. Snyder, Alexey Shashurin, Michael Keidar, Anthony D. Sandler. Development of a non-thermal plasma model for neuroblastoma treatment. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2771. doi:10.1158/1538-7445.AM2013-2771

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2776: Establishment of patient non-small cell lung cancer derived models for test of anticancer drugs.
Authors: Liu Changnian , Sun Connie , Li Wenwei , Zhou Wen , Liu Yong , Zhou Rui , He Fang , Bai Chang
Published: August 2014

BACKGROUND Human tumor xenograft models established by transplantation of human tumor cell lines into immunodifficient mice have been routinely used for preclinical test of anticancer agents. But tumor cell lines have a relatively low transplantability and resulted in a limited number of tumor models available for selection of right models for testing novel agents based on their antitumor mechanism. Recently, we have developed a large number of patient primary non-small cell lung cancer (NSCLC) xenograft models by transplanting patients’ fresh tumor tissues into nude mice, which have been employed for preclinical test of anticancer agents. METHODS The fresh NSCLC tissues were collected from local hospitals. The tumor fragments of 1-2 mm were subcutaneously implanted in the flanks of the nude mice by trocar needle. Sixteen tumor fragments were grafted into four mice from one patient tumor tissue (passage 0). All therapeutic efficacy experiments used female mice bearing passage 5 xenografts. The test drugs included cisplatin, carboplatin, paclitaxel, docetaxel, gemcitabine, erlotinib, gefitinib, and pemetrexed. RESULTS A total of 213 patient primary NSCLC samples were implanted into nude mice, and 107 primary tumor models have been established with a tumor taking rate of 50% for the first passage. The tumor taking rates were higher (80-100) in the later passages. The therapeutic efficacy of the test drugs in these models is consistent with their clinical findings. The histopathology and gene sequence of the established primary tumor xenografts were analyzed; their architecture, histopathological morphology, and genomic mutation status from five generations of xenografts retained the patients’ original tumor characteristics. CONCLUSIONS The patient primary NSCLC models can be established in a large number for right models selection in preclinical setting, and have been employed for test of standard of care drugs and novel anticancer agents. The primary tumor models retain a similarity in histology and genomic mutation status to their patients’ original tumors. They may predict more relevant clinical response rate and higher correlation with clinical findings than use of traditional xenograft models established from long-term cultured cancer cell lines. Especially, they have advantages for test of target-oriented therapeutics in new drugs discovery and development programs.

Citation Format: Changnian Liu, Connie Sun, Wenwei Li, Wen Zhou, Yong Liu, Rui Zhou, Fang He, Chang Bai. Establishment of patient non-small cell lung cancer derived models for test of anticancer drugs. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2776. doi:10.1158/1538-7445.AM2013-2776
Abstract 2781: Champions TumorGraft™ models possess PIK3CA mutations - A great tool to explore the PI3K/AKT pathway.
Authors: Khor Tin Oo , Tiruchinapalli Dhanrajan , Friedman Jay , Anderson Nathan , Sidransky David , Bruckheimer Elizabeth M.
Published: August 2014

Phosphatidylinositol 3-kinase (PI3K) is an important intracellular signal transducer that is involved in many cellular functions such as cell growth, proliferation, and differentiation. PI3K is a heterodimeric molecule composed of a regulatory and a catalytic subunit. PIK3CA, which encodes the p110α catalytic subunit, is often mutated in a range of human cancers, such as colorectal and non-small-cell lung cancers. Increasing our understanding of the biological implications of these mutations and the role of PI3K pathway in cancer proliferation or survival may lead to potential new therapeutic opportunities targeting PI3K signaling.

Champions Oncology focuses on the development of TumorGraft models derived from the direct implantation of patient tumors into immunocompromised mice. Compared to traditional cell line-based xenograft models, the patient derived Champions TumorGraft™ models maintain stable gene-expression patterns and mutational status and correlate to clinical predictability. Through extensively characterization, Champions has identified 19 TumorGraft models with PIK3CA mutations. Out of these 19 models, there are 11 colorectal cancers, 3 non-small cell lung cancers, 3 pancreatic cancers, 1 gastric cancer and 1 melanoma. Genomics analyses using the Ampliseq platform identified the majority of PIK3CA mutations in exon 9 and 20. Mutations in these regions of PIK3CA are known to confer resistance to some anti-cancer agents, such as HER2 antibodies. Additionally, it has been reported that PIK3CA mutations frequently coexist with RAS and BRAF mutations in patients with advanced cancers. Upon further analysis, we identified the same pattern in Champions TumorGraft models where∼70% (13 out of 19) of the PIK3CA mutant models possess either concurrent RAS or BRAF mutations. Notably, PI3K activates the downstream kinase AKT which ultimately triggers the mTOR pathway. Mutations in cancer often result in constitutive activation of this pathway. Given the role of the PI3K/AKT signaling in cancer and the presence of these specific mutations in the Champions TumorGraft bank, utilization of Champions’ TumorGraft models in oncology drug development allows for greater insight into new therapeutic opportunities targeting the inhibition of PI3K/AKT signaling and offers new promise for patients with these activated pathways.

Citation Format: Tin Oo Khor, Dhanrajan Tiruchinapalli, Jay Friedman, Nathan Anderson, David Sidransky, Elizabeth M. Bruckheimer. Champions TumorGraft™ models possess PIK3CA mutations - A great tool to explore the PI3K/AKT pathway. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2781. doi:10.1158/1538-7445.AM2013-2781
Abstract 2782: Utilization of in vivo human isogenic cancer models in the new era of targeted therapies.
Authors: Sun Yanmei , Zhang Songling , Astley Holly , Foster Rebecca , Schofield Christine , Torrance Chris , Ning Jinying , Chen Taiping , Shi Qian
Published: August 2014

The explosion of knowledge regarding the genetic underpinnings of human cancer heralds a new era of targeted therapy. To date, in vivo cell based screening has proven a useful tool in almost all drug development programs. Cells used in such screens are usually harvested from cancer patients that harbor the specific mutation of interest, but these cells almost invariably contain many other additional mutations making it difficult to ascertain the specific functions of molecules being screened. Thus the lack of true control cells hampers the development of new cancer therapeutics.

Using cell lines generated with Horizon Discovery's proprietary rAAV-based GENESIS™ gene editing platform, we have established a comprehensive range of isogenic cancer models for our in vivo compound screening program to service our clients in academia and industry, with mutations in a wide variety of genes including KRAS, PIK3CA, PTEN, IDH1 and p53. These isogenic tumor models comprise pairs of cell lines which share the same genetic background, differing only by the mutation of interest and therefore allowing definitive studies of specific genetic variances to be performed. The same isogenic pairs of lines can be used for in vitro and in vivo experiments to ensure continuity and relevance of results. We have carried out intensive validation of these models in vivo to ensure the lines generate robust tumor growth in mice and the matched tumors differ only in the genetic composition of target gene.

In a POC study, colon cancer cell lines with the KRAS G13D mutation have been demonstrated to respond to Cetuximab treatment, consistent with recent findings in the clinic. These results challenge the current clinical practice of only using Cetuximab as a therapeutic for KRAS wild type patients, and provide the basis for expanding the usage of the drug to benefit more patients.

In summary, we have established a series of reliable in vivo isogenic models with precise and highly specific genetic modifications as predictors of clinical effect. They will provide a valuable tool in the drug discovery and development arena to drive forward personalized medicine by enabling novel target validation, expansion of the target population for existing therapeutics, and definition of patient responsive genotypes.

Citation Format: Yanmei Sun, Songling Zhang, Holly Astley, Rebecca Foster, Christine Schofield, Chris Torrance, Jinying Ning, Taiping Chen, Qian Shi. Utilization of in vivo human isogenic cancer models in the new era of targeted therapies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2782. doi:10.1158/1538-7445.AM2013-2782
Abstract 2785: Development program of patient tumor tissue bank to support the drug and target discovery.
Authors: Hillairet de Boisferon Marc , Bichat Francis , Mignard Caroline , Morgand Loïc , Blot Lydia , Venot Corinne , Merino-Trigo Ana , Vrignaud Patricia
Published: August 2014

Background: The humanization of mice with various tissues named Chi-mice® aimed to reproduce the human situation to be more predictive than conventional models. Despite significant progress in identifying malignancy of cancer cells, a more detailed understanding of tumor generation is needed. Xenograft of tumor cells into immunodeficient rodents has constituted the major preclinical screen for the development of new drugs. These models have identified efficacious agents, but their chemosensitivity, genetic drift and clonal selection induced by cell culture have been part of the high attrition rate observed in the clinical development. Patient-derived tumor xenograft (PDX) obtained in xenografting fresh patient tumor samples in mice are reported as being more predictive to the clinical situation in maintaining the histopathology and molecular diversity of the patient tumors.

Methods: The PDXs collection has been set up under ethical agreement with informed consent of patients. The patients have been screened for absence of HIV, HBV and HCV. The anonymized patient's clinical history and tissue banking (including normal tissue when available) are centralized in our internal biological resource center. Tumor samples were freshly implanted in nude or SCID mice.

Results:

PDX Passage 0 Passage 1 Take rate (%) Breast 152 10 6 Lung 18 7 39 Bladder 31 11 35 Liver 31 4 13 Stomach 17 7 41 Kidney 28 8 29 Pancreas 21 4 19 Prostate 57 5 9 Total 355 56 Xenograft Passage 0 Lymphoma Established model SCID mice 85 16 (19%) 19 (22%) Nude mice 270 0 (0%) 37 (14%)

Cryopreservation of the PDX is performed at early passages allowing using these PDX only at low passage. The histopathology, EGFR, PSA, AR, HER, ER and PR statute for lung, prostate and breast carcinoma and tumor growth characteristics of these PDX are being performed. Lymphoma characterization was performed using immunohistochemistry (hCD20, m/hKi67). Lymphoma detection, probably related to EBV infection, leads to switch to nude mice for xenografting.Conclusion: To create a highly diversified panel of PDX, we organized a global process from multiple centers. These PDX are currently being used in preclinical development of new therapies and clinical positioning including biomarkers identification.

Citation Format: Marc Hillairet de Boisferon, Francis Bichat, Caroline Mignard, Loïc Morgand, Lydia Blot, Corinne Venot, Ana Merino-Trigo, Patricia Vrignaud. Development program of patient tumor tissue bank to support the drug and target discovery. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2785. doi:10.1158/1538-7445.AM2013-2785
Abstract 2787: PrimePanel provides a high throughput in vitro drug screening platform that intimately links to in vivo pharmacological analysis in PDX models.
Authors: Zhang Yanxia , Ge Yuqiang , Liu Yanan , Zhao Jing , Li Lanhua , Qiao Meng , Ning Jinying , Zha Jiping , Liu Guizhong
Published: August 2014

Patient derived xenografts (PDX) are in vivo animal tumor models established directly from patient tumor samples without any in vitro manipulation. It has been shown that these tumor xenografts maintain essential histopathological features and genetic profiles of the original tumors, and thus are the most clinically relevant animal models for cancer drug discovery. Comparing to the traditional cell line derived xenograft (CDX) tumor models, however, one challenge encountered by the PDX in vivo studies is the lack of corresponding in vitro cell culture system for cost-effective and high throughput drug screening and model selection. Here we set out to establish such an in vitro platform, so-called PrimePanelTM, by deriving homogeneous primary cancer cell cultures from the PDX tumors in several major tumor types. These cells are early passage cultures (usually p<10), which maintain similar cellular characteristics as the corresponding tumor xenografts, including the genomic mutational status, biochemical signaling, and responses to tumor cell autonomously targeted therapeutics. Thus, PrimePanelTM offers a higher throughput, faster turnaround, more versatile and lower cost platform to assess drug efficacy prior to in vivo study, which could fulfill a critical need in early stage drug discovery.

Citation Format: Yanxia Zhang, Yuqiang Ge, Yanan Liu, Jing Zhao, Lanhua Li, Meng Qiao, Jinying Ning, Jiping Zha, Guizhong Liu. PrimePanel provides a high throughput in vitro drug screening platform that intimately links to in vivo pharmacological analysis in PDX models. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2787. doi:10.1158/1538-7445.AM2013-2787

Note: This abstract was not presented at the AACR Annual Meeting 2013 because the presenter was unable to attend.
Abstract 2788: Characterization of an in vivo generated subpopulation of human LnCaP prostate cancer cells as an improved testosterone-dependent in vivo mouse xenograft model for compound testing.
Authors: Lingnau Andreas , Hoffmann Steffen , Schaefer-Obodozie Cynthia , Leisegang Ulrike , Klotzbuecher Andreas , Schaechtele Christoph
Published: August 2014

Prostate cancer is one of the most frequently diagnosed cancer entities in men. It can be estimated that in 80% of all men reaching the age of 80 years prostate tumors are present. Since a depletion of testosterone in prostate cancer therapies is often performed, androgen-dependent and -independent prostate carcinoma cell lines have been widely used to establish mouse in vivo xenograft models in order to identify novel drugs helping to eliminate prostate tumors.

A commercially available LnCaP cell line was shown to be androgen-dependent since addition of synthetic androgens to androgen-free culture medium was able to restore proper cell proliferation in cell culture. However, when implanted in SCID mice in order to establish a xenograft model, the take rate turned out to be very low and tumor growth was highly heterogeneously. Therefore, we resected and recultivated one of the few well-growing tumors and reimplanted the arising subpopulation subcutaneously in mice. After performing two rounds of this “subpopulationing” procedure we could generate a new cell line, LnCaP-Z2, which was able to form tumors in mice with a take rate of nearly 100%, but still shows heterogenous tumor growth similar to other subcutaneously implanted prostate cancer cells (e.g. PC-3).

In order to demonstrate an androgen dependency of our new LnCaP subpopulation model in vivo we used castrated mice and implanted them subcutaneously with two different prostate cancer cell lines in the absence or presence of also subcutaneously inoculated testosterone pellets. Whereas no influence of testosterone was detectable on the growth behavior of subcutaneous PC-3 xenograft tumors, the development of the LnCaP-Z2 xenografts was strictly dependent on testosterone release.

Finally, treatment of Bicalutamide, a typical member of the class of anti-androgens, which is frequently used in the treatment of prostate cancer, led to significant inhibition of tumor growth using our improved subpopulation LnCaP-Z2 in vivo model. Thus, we could demonstrate the model's potency as a testosterone-dependent in vivo tool suitable for screening and development of novel anti-prostate cancer drugs.

Citation Format: Andreas Lingnau, Steffen Hoffmann, Cynthia Schaefer-Obodozie, Ulrike Leisegang, Andreas Klotzbuecher, Christoph Schaechtele. Characterization of an in vivo generated subpopulation of human LnCaP prostate cancer cells as an improved testosterone-dependent in vivo mouse xenograft model for compound testing. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 2788. doi:10.1158/1538-7445.AM2013-2788
Abstract 279: A small molecular miR-21 inhibitor inhibits glioma cell invasion and mesenchymal transition via upregulation of VHL expression.
Authors: Shi Zhendong , Zhang Junxia , Han Lei , Zhang Kailiang , Chen Luyue , Qian Xiaomin , Pu Peiyu , Kang Chunsheng
Published: August 2014

Glioblastomas (GBMs) are the most common malignant brain tumors which characterized by rapid and invasive growth. Accumulating evidence points to a critical role of EMT like events during glioma progression and malignant transformation. Cancer cells undergoing EMT can acquire invasive properties and enter the surrounding stroma, resulting in the creation of a favorable microenvironment for cancer progression and metastasis. In this study, we identified a small molecular miR-21 inhibitor through a high-throught screening approach and found it significantly repressed the U87 and LN229 glioma cell migration and invasion. Meanwhile, VHL, a direct target of miR-21, was increased expression in mRNA and protein level after this miR-21 inhibitor treatment, but the β-catenin expression was reduced. Silencing of VHL using small interefering RNA (siRNA) also reduced β-catenin expression. Furthermore, suppression of β-catenin transactivation by miR-21 inhibitor, through upregulation of VHL expression, increased E-cadherin expression and partially reversed the epithelial-mesenchymal transition (EMT). On the other hand, confocal microscopy and Western blot assay presented that N-cadherin, ZEB1, ZEB2 and MMP9 expression were reduced. Additionally, in a U87 intracranial glioma xenograft nude mice model, we found a less invading tumor edge of miR-21 inhibitor treated group in comparison with DMSO group. Moreover, there was an increased expression of E-cadherin and reduced expression of β-catenin, N-cadherin, ZEB1, ZEB2 and MMP9 in tumor samples after the inhibitor treatment. Together, these results provided insight into the specific treatment efficacy of this small molecular inhibitor in EMT and tumor metastasis, which achieved via up-regulation of VHL expression. Therefore, it might be a novel and potentially antimetastatic agent for therapeutic intervention in glioblastomas in future.

Citation Format: Zhendong Shi, Junxia Zhang, Lei Han, Kailiang Zhang, Luyue Chen, Xiaomin Qian, Peiyu Pu, Chunsheng Kang. A small molecular miR-21 inhibitor inhibits glioma cell invasion and mesenchymal transition via upregulation of VHL expression. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 279. doi:10.1158/1538-7445.AM2013-279