bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1159: Downregulation of TMPRSS11B in squamous cell carcinoma
Authors: Amano Suburu , Iwaya Takeshi , Nishizuka Satoshi , Kume Kohei , Ito Chie , Akiyama Yuji , Shioi Yoshihiro , Endo Fumitaka , Sasaki Akira
Published: July 2016

[Background] Esophageal squamous cell carcinoma (ESCC) is one of the most malignant cancers. The molecular mechanism of ESCC development has not been fully elucidated. [Aim] The aim of this study is to identify responsible genes for ESCC development. [Materials and Methods] Firstly, we performed transcriptome sequence (RNA-seq) analysis of surgically resected tumor and corresponding normal tissues form 3 ESCC patients with different tumor locus, histrogical differentiation, and TNM stage.by RNA-seq analysis: (Case 1) upper thoracic esophagus, well differentiated SCC, TNM, Stage I; (Case 2) middle thoracic esophagus, poorly differentiated SCC, T3N0M0, Stage II; and (Case 3) lower thoracic esophagus, moderately differentiated SCC, T3N1M0, Stage III. Next, we validated the expression status of the candidate gene in 64 clinical samples and 10 ESCC cell lines (KYSE150, KYSE270, KYSE410, KYSE450, KYSE510, TE1, TE6, TE8, TE9, and TE10) using quantitative real-time PCR (qRT-PCR). [Results] In RNA-seq analysis, 34 genes showed more than 30-fold decrease in tumor samples compared to normal tissues, and the most downregulated gene was transmembrane protease serine 11B (TMPRSS11B). Amon these 34 genes, other 5 genes of TMPRSS11 family (TMPRSS11A, TMPRSS11BNL, TMPRSS11D, TMPRSS11E, and TMPRSS11F) were included. qRT-PCR analysis revealed that TMPRSS11B expression in ESCC samples were lower than those in normal samples (p < 0.001). Downregulation of TMPRSS11B in tumor samples compared to corresponding normal samples were observed in 51 cases out of 64 (79.7%). Patients with expression levels of TMPRSS11B that were below the median value were assigned to the low expression group (n = 32), whereas those with expression values above the median were assigned to the high expression group (n = 32). Significant difference was not observed between two groups regarding clinicopathological data, including age, gender, TNM stage, and lymphatic/venous invasion. TMPRSS11B expression was not detected in all of ESCC cell lines and noncancerous esophageal epithelial cell line, HET1A. [Conclusion] Our results suggest that downreguration of TMPRSS11B expression occurs in early stage of the ESCC oncogenesis and that TMPRSS11B has tumor suppressive roles in ESCC development.

Citation Format: Suburu Amano, Takeshi Iwaya, Satoshi Nishizuka, Kohei Kume, Chie Ito, Yuji Akiyama, Yoshihiro Shioi, Fumitaka Endo, Akira Sasaki. Downregulation of TMPRSS11B in squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1159.
Abstract 1215: Utilization of podoplanin as a chemotherapeutic target for oral squamous cell carcinoma
Authors: Retzbach Edward P. , Krishnan Harini , Ochoa-Alvarez Jhon A. , Shen Yongquan , Nevel Evan , Kephart David J. , Kalyoussef Evelyne , Baredes Soly , Fatahzadeh Mahnaz , Rameriz Maria I. , Yin Kingsley , Young Mary Ann , Deluca-Rapone Lisa , Shienbaum Alan J. , Jensen Lasse D. , Goldberg Gary S.
Published: July 2016

Oral cancer is diagnosed in over 300 thousand people, and kills over 100 thousand people, around the world each year. Current treatments rely on radiation and surgery procedures that often decrease the quality of life for oral cancer survivors. There is a clear need to improve treatments for these patients. Over 90% of oral cancers are formed by oral squamous cell carcinoma (OSCC). Most OSCC cells express the transmembrane receptor podoplanin (PDPN), which has emerged as a promising target for OSCC treatment. The PDPN receptor promotes tumor cell invasion and metastasis which leads to the vast majority of cancer deaths. Here, we describe efforts to target PDPN intracellularly and extracellularly in order to prevent and treat oral cancer. PDPN contains intracellular serine residues that can be phosphorylated by protein kinases including CDK5 and PKA to decrease cell migration. Additionally, the extracellular portion of PDPN can be targeted with Maackia amurensis seed lectin (MASL) to inhibit tumor cell migration and viability. Previous studies suggest that PDPN induces RhoA GTPase activity to promote cell migration. However, we show here that MASL dynamically binds PDPN to downregulate Cdc42 GTPase activity, but not RhoA or Rac1 GTPase activity. These data suggest that MASL suppresses Cdc42 GTPase activity to disrupt cell polarity and inhibit cell migration. Taken together, these data indicate that PDPN can serve as a functionally relevant target to prevent and combat oral cancer.

Citation Format: Edward P. Retzbach, Harini Krishnan, Jhon A. Ochoa-Alvarez, Yongquan Shen, Evan Nevel, David J. Kephart, Evelyne Kalyoussef, Soly Baredes, Mahnaz Fatahzadeh, Maria I. Rameriz, Kingsley Yin, Mary Ann Young, Lisa Deluca-Rapone, Alan J. Shienbaum, Lasse D. Jensen, Gary S. Goldberg. Utilization of podoplanin as a chemotherapeutic target for oral squamous cell carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1215.
Abstract 1239: NVP-HDM201: Biochemical and biophysical profile of a novel highly potent and selective PPI inhibitor of p53-Mdm2
Authors: Stachyra-Valat Thérèse , Baysang Frédéric , D’Alessandro Anne-Cécile , Dirk Erdmann , Furet Pascal , Guagnano Vito , Kallen Joerg , Leder Lukas , Mah Robert , Masuya Keiichi , Stutz Stefan , Vaupel Andrea , Hofmann Francesco , Chène Patrick , Jeay Sébastien , Holzer Philipp
Published: July 2016

An effective strategy to restore p53 activity in cancer cells containing wild type p53 is to inhibit the Mdm2-p53 protein-protein interaction (PPI). NVP-HDM201 is a novel PPI inhibitor currently under evaluation in a Phase I clinical trial. It binds to the p53 binding-site of the Mdm2 protein, disrupting the interaction of the two proteins and leading to the activation of the p53 pathway.

NVP-HDM201 belongs to a novel chemical series with a distinct biophysical and biochemical profile. Affinity constant of NVP-HDM201 for Mdm2 is in the picomolar range, with a selectivity ratio greater than a 10000-fold vs. Mdm4. Analysis of its binding mode provides evidence for a distinct set of critical interactions between the small molecule and its target, as compared with our other Mdm2 inhibitor NVP-CGM097, and explains as to why NVP-HDM201 binds equally to human, mouse, rat and dog Mdm2. Characterization of its binding kinetics indicates that the optimized interactions of NVP-HDM201 with Mdm2 protein are responsible for the increased stabilization of the complex resulting in high potency against Mdm2. This feature, together with favorable physicochemical and drug-like properties, supported the selection of NVP-HDM201 for clinical development.

Citation Format: Thérèse Stachyra-Valat, Frédéric Baysang, Anne-Cécile D’Alessandro, Erdmann Dirk, Pascal Furet, Vito Guagnano, Joerg Kallen, Lukas Leder, Robert Mah, Keiichi Masuya, Stefan Stutz, Andrea Vaupel, Francesco Hofmann, Patrick Chène, Sébastien Jeay, Philipp Holzer. NVP-HDM201: Biochemical and biophysical profile of a novel highly potent and selective PPI inhibitor of p53-Mdm2. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1239.
Abstract 1259: MutT homolog 1 (MTH1) is upregulated in glioblastoma multiforme: its inhibition by siRNA or crizotinib results in impaired cell migration and tumor growth in vivo
Authors: Timmer Marco , Kannampuzha Sara Gerdi , Röhn Gabriele , Goldbrunner Roland
Published: July 2016

Introduction:

MutT homolog1 (MTH1) is required for the survival of cancer cells. These are characterized by irregular redox formation resulting in the production of reactive oxygen species which damage the deoxynucleoside triphosphate pool and subsequently the DNA. MTH1 inhibits the incorporation of oxidized bases into the DNA thus avoiding apoptosis of cancer cells. Our aim was to analyze the pathophysiological role of MTH1 in glioma.

Materials and Methods:

MTH1 expression was analyzed using quantitative PCR and Western Blot analysis in human glioma tissue (n = 80). We compared peritumoral tissue with grade II, III, and IV gliomas. Moreover, we compared primary to secondary GBM and initial tumors with recurrent tumors after temozolomide treatment. U87 cell migration was tested after siRNA knock-down of MTH1 and treatment with the MTH1 inhibitor crizotinib by scratch assays. After orthotopic implantation of MTH1-siRNA treated U87 into rats, tumor size was measured after 8 days.

Results:

In RT-PCR and Western blot we found a significantly higher expression of MTH1 in primary and secondary glioblastoma than in lower grade brain tumor. Immunofluorescence staining of histological sections of glioblastoma showed also a higher level of MTH1 than in lower grade tumors. By nucleofection, we transfected siRNA MTH1 into the U-87 glioblastoma cells and a slower migration was investigated applying migration assay. Furthermore, treatment with the drug crizotinib inhibited MTH1 and decreased U-87 cell migration. The xenograft study

showed a slower tumor growth when U87 cells were transfected with MTH1 siRNA compared to normal U87.

Conclusion:

These results show that MTH1 plays an essential role in migration and proliferation of glioma cells resulting in clinically observed malignization of glioma and disease progression in recurrent glioblastoma. Moreover, MTH1 can be targeted by Crizotinib indicating a role for potential future therapies.

Citation Format: Marco Timmer, Sara Gerdi Kannampuzha, Gabriele Röhn, Roland Goldbrunner. MutT homolog 1 (MTH1) is upregulated in glioblastoma multiforme: its inhibition by siRNA or crizotinib results in impaired cell migration and tumor growth in vivo. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1259.
Abstract 1284: Lurbinectedin reduces tumor-associated macrophages and the production of inflammatory cytokines, chemokines, and angiogenic factors in preclinical models
Authors: Allavena Paola , Belgiovine Cristina , Liguori Manuela , Bello Ezia , Frapolli Roberta , Galmarini Carlos M. , D’Incalci Maurizio
Published: July 2016

Lurbinectedin, currently undergoing clinical evaluation in ovarian, breast and small-cell lung cancer patients, inhibits active transcription. The drug is structurally related to trabectedin containing the same pentacyclic skeleton of the fused tetrahydroisoquinoline rings, but differing by the presence of a tetrahydro-B-carboline replacing the additional tetrahydroisoquinoline of trabectedin. We investigated whether lurbinectedin has the ability to regulate the inflammatory tumor microenvironment in vitro and in vivo. Human purified monocytes were highly sensitive to lurbinectedin (IC50: 5-10 nM) and, after drug treatment, underwent caspase-8-dependent apoptosis. Furthermore, in vitro, lurbinectedin significantly inhibited the production of selected inflammatory chemokines (CCL2, CXCL8) and VEGF by stimulated monocytes and liposarcoma cell lines. Administration of lurbinectedin to mice bearing a murine fibrosarcoma -resistant to this compound in vitro- resulted in significant anti-tumor activity (T/C value around 50%). The analysis of immune cells of blood spleen and tumor tissues during treatment with lurbinectedin revealed a significant and selective decrease in the subset of monocytes and macrophages, including tumor-associated macrophages (TAM). A gene expression analysis of monocytes treated with lurbinectedin indicated a modulation of the transcriptional program in LPS-stimulated human monocytes. Overall, these results indicate that lurbinectedin affects the inflammatory micro-environment, with a selective apoptotic-inducing effect on mononuclear phagocytes and a specific inhibition of production of inflammatory cytokines.

Citation Format: Paola Allavena, Cristina Belgiovine, Manuela Liguori, Ezia Bello, Roberta Frapolli, Carlos M. Galmarini, Maurizio D’Incalci. Lurbinectedin reduces tumor-associated macrophages and the production of inflammatory cytokines, chemokines, and angiogenic factors in preclinical models. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1284.
Abstract 1287: The dual IAP antagonist, ASTX660, increases the anti-tumor activity of paclitaxel in preclinical models of triple-negative breast cancer in vivo
Authors: Smyth Tomoko , Nakatsuru Yoko , Fujita Ryoto , Ward George , Bevan Luke , Lewis Jon , Wallis Nicola , Lyons John
Published: July 2016

Background: Paclitaxel-mediated secretion of inflammatory mediators, including TNFα, potentially creates paracrine and autocrine signaling loops that can reduce paclitaxel-induced apoptosis with resultant cancer cell survival (paclitaxel resistance). One mechanism of cancer cell survival is the expression of inhibitor of apoptosis proteins (IAPs): Cellular IAP (cIAP) is involved in inflammatory pro-survival NF-κB activation, blocking the activation of effector caspases 3 and 7, while X-linked IAP (XIAP) directly binds the effector caspases 3, 7 and 9, inhibiting the full activation of the apoptotic pathway. Antagonism of IAPs in the setting of paclitaxel treatment may enhance the cancer cell death by switching the chemotherapy-induced inflammatory TNFα signaling from pro-survival to apoptosis. ASTX660 is an orally bioavailable dual antagonist of cIAP and XIAP, currently being investigated in a single-agent Phase 1/2 clinical trial in patients with advanced solid tumors and lymphomas (NCT02503423). Here, we characterize the activity of ASTX660 in triple-negative breast cancer (TNBC) preclinical models as a single agent and in combination with paclitaxel.

Results: We first investigated the viability of 21 TNBC cell lines treated with ASTX660 in vitro and found that 43% were sensitive either to ASTX660 alone (MDA-MB-231 and HCC38) or in the presence of exogenous TNFα (HCC1806, Hs578T, BT549, HCC1395, DU4475, MDA-MB-453 and mouse EMT6). In HCC1806 xenografts in mice, both ASTX660 (daily oral treatment) and paclitaxel (weekly intravenous treatment) as single agents caused moderate tumor growth inhibition but not regression. ASTX660 and paclitaxel combination treatment, however, caused regression and all tumors achieved a partial response by Day 19 of treatment. An increase in circulating acute-phase-proteins in animals was also observed on paclitaxel treatment.

Conclusions: Our study suggests that paclitaxel treatment causes an inflammatory response which could qualitatively change the tumor environment. Inhibition of IAPs in such inflammatory environment induced by paclitaxel may lead to rapid apoptotic cell death. Thus, addition of IAP antagonist may be a promising approach to enhance the TNBC response to paclitaxel therapy.

Citation Format: Tomoko Smyth, Yoko Nakatsuru, Ryoto Fujita, George Ward, Luke Bevan, Jon Lewis, Nicola Wallis, John Lyons. The dual IAP antagonist, ASTX660, increases the anti-tumor activity of paclitaxel in preclinical models of triple-negative breast cancer in vivo. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1287.
Abstract 144: Gene expression analysis of HPV negative and HPV positive penile cancer
Authors: Sánchez-Vázquez María , Rohena-Rivera Krizia , Pérez-Ruiz Carlos , Puras-Báez Antonio , Cadilla Carmen , Martínez-Ferrer Magaly
Published: July 2016

Penile cancer is a disease with a high morbidity and mortality among developing countries. Although penile cancer is a relatively uncommon cancer in developed countries, its incidence is nearly four times higher in Puerto Rico when compared with other racial and ethnic groups in the United States. Approximately 40% to 45% of penile cancers are HPV-related. The etiology of penile cancers is not completely understood and additional research is necessary to fully delineate the sequence of molecular events involved in both HPV mediated and non-HPV mediated pathways leading to penile cancer. The objective of this study is to determine the genome-wide expression profiles in HPV mediated and non-HPV mediated penile cancer. Penile cancer fresh tissue was obtained from surgically intervened cases. The global gene expression profile was identified and analyzed using the Affymetrix GeneChip® Human Gene 2.0 array. Microarray analysis showed a total of 158 genes differentially expressed between HPV mediated and non-HPV mediated penile cancer. Ingenuity Pathway Analysis (IPA) revealed alterations in the celular movement, cancer, lipid metabolism, carbohydrate metabolism, interleukin, TGF-B, integrin, PI3-kinase and MMP signaling pathways. The microarray results were confirmed by quantitative real-time PCR for some representative genes. Functional and pathway annotation using bioinformatics tools helped to identify specific pathways. Knowledge of the biological pathways and networks of the genes that are significantly expressed or altered are pertinent to understanding the molecular basis of the penile cancer.

Citation Format: María Sánchez-Vázquez, Krizia Rohena-Rivera, Carlos Pérez-Ruiz, Antonio Puras-Báez, Carmen Cadilla, Magaly Martínez-Ferrer. Gene expression analysis of HPV negative and HPV positive penile cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 144.
Abstract 1451: Analysis of expression MHC class I chain-related gene A and B (MICA/B) in normal and tumor tissue
Authors: Ghadially Hormas , Brown Lee , Lewis Arthur , Czapiga Meggan , Valge-Archer Viia , Wilkinson Robert W.
Published: July 2016

MHC class I chain-related gene A and B (MICA and MICB) are highly polymorphic proteins that are induced upon stress, damage or transformation of cells which act as a “kill me” signal through the NKG2D receptor expressed on Natural Killer, CD8+ and γδ T cells.

Experimentally, the MIC/NKG2D axis has been shown to be important for the recognition of tumour cells by cytotoxic cells of the immune system and many tumours have evolved strategies to evade the detection by NKG2D expressing cells, e.g. by shedding MIC from the cell surface.

Expression of MIC has been reported for most tumour types and in normal gastrointestinal tract epithelium but the published data is often difficult to interpret. Additionally, it is not clear how much of the protein is expressed on the cell surface as MIC cell surface expression is known to be regulated tightly on multiple levels.

A validated MICA/B IHC assay was developed using an in-house tool antibody to profile multiple frozen human normal and tumour tissue microarrays (TMA’s) by both standard and confocal microscopy techniques.

Using a stringently characterised novel antibody that detects MICA as well as MICB this study describes the expression patterns in a wide range of tumours and normal tissues. With this method we generated data with unprecedented resolution, which enabled us to analyse the expression of MICA and MICB not only on the cellular but also on the sub-cellular level.

Citation Format: Hormas Ghadially, Lee Brown, Arthur Lewis, Meggan Czapiga, Viia Valge-Archer, Robert W. Wilkinson. Analysis of expression MHC class I chain-related gene A and B (MICA/B) in normal and tumor tissue. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1451.
Abstract 1542: Identification and utilization of molecular pathways correlated with osteosarcoma metastasis to predict targeted therapies
Authors: Klippenstein Laird , Fowles Jared S. , Lear Sierra K. , Gustafson Daniel L.
Published: July 2016

Osteosarcoma (OSA) is the most prevalent primary bone tumor and accounts for approximately 2% of childhood cancers. Approximately 40% of all OSA patients succumb to metastatic disease within 5 years. Canine OSA occurs about 10 times more frequently than human OSA and has been shown to be very similar to pediatric OSA biologically, histologically, molecularly, and in treatment. This suggests that canine OSA is an excellent pre-clinical model to investigate better treatment options for metastatic OSA. We compared in-vitro markers of metastatic potential with gene expression data in eight canine OSA cell lines to identify molecular pathways that will be potential chemotherapeutic targets to treat and prevent metastatic OSA.

The metastatic potential of 8 canine OSA cell lines was characterized utilizing Incucyte Zoom based migration and invasion scratch wound assays. Gene expression analysis was performed using Affymetrix GeneChip Canine Genome 2.0 Arrays. Genes that were statistically correlated with migratory and invasive phenotypes were used to enrich molecular pathways described in several databases utilizing Enrichr software. Altered pathways and associated targeted agents that would inhibit the pathways enriched by the correlated gene sets were identified.

Chemotherapeutics targeting significantly enriched pathways were screened for their ability to inhibit metastatic potential. Dasatinib and Stattic were the only inhibitors screened to date that inhibit migration of the osteosarcoma cell lines at clinically relevant doses. The remaining significantly correlated pathways will be targeted as well as rational combinations of pathways. An orthotopic murine model is currently being tested to further explore the utility of efficacious chemotherapeutics in-vivo. Pathway Inhibition and Anti-Migratory EffectsPathwayP-ValueTargeted AgentIC50 vs MigrationMap Kinase0.0037Selumetinib>25 uM *Focal Adhesion0.0059Dasatinib18 nM - 214 nMErbb2/30.0214Lapatinib>8 uM *TubulinControlPaclitaxel7 nM - 54 nMmTOR0.0054Rapamycin>20 nM *STAT30.0004Stattic2.4 uM - 4.6 uMBMPR2<0.0005Tacrolimus>12.5 nM *ALK<0.0005Crizotinib>900 nM *VEGFR0.0226VandetanibTBDITK<0.0005IbrutinibTBDPDPK10.0011CelecoxibTBD* We were not able to reach an IC50 at clinically relevant doses

Citation Format: Laird Klippenstein, Jared S. Fowles, Sierra K. Lear, Daniel L. Gustafson. Identification and utilization of molecular pathways correlated with osteosarcoma metastasis to predict targeted therapies. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1542.
Abstract 1544: Overexpression of Rab11 and E-cadherin promotes collective cell migration and indicates a poor prognosis in colorectal carcinoma
Authors: Chung Yuan-Chiang , Wei Wan-Chen , Hung Chia-Nung , Hsu Chih-Ping , Chiu Hsiang-Ling , Chang King-Jen , Chao Wei-Ting
Published: July 2016

Collective cell migration, whereby the cell-cell contacts such as E-cadherin are maintained during migration, has recently emerged, and the detailed mechanisms of this process are still unclear. The present study identified the role of Rab11 which functions in recycling endosome and the relation to E-cadherin in colorectal carcinoma and clarified the mechanism of Rab11 in the collective cell migration of cancer cells. The relationships between the overexpression of Rab11 and E-cadherin and survival were evaluated from colorectal carcinoma patients. A total of 107 patients with surgically resected colorectal carcinoma were enrolled in immunohistochemical study. The relationships between the overexpression of Rab11 and E-cadherin and survival were evaluated. GFP tagged Rab11 or Rab11 shRNA was introduced into HT-29 colon cancer cells for overexpression or knockdown. The interaction between E-cadherin and Rab11 was determined by immunoprecipitation. Rac1 and matrix metalloproteinase (MMP) activities were evaluated as functional effectors of collective cell migration. In the results, the expression of Rab11 and E-cadherin was not correlated with the stages of cancer or lymph node metastasis. However, the overall survival was poor in the group of 60 patients with duo-positive Rab11 and E-cadherin expression compared to the group (47 patients) without duo-positive expression (p = 0.038). In the cell biology assay, Rab11 was demonstrated through its physical interaction with E-cadherin, and overexpression of Rab11 was found to promote collective cell migration through the increased distribution of E-cadherin, which enhanced cell-cell connections. In addition, Rac1 activation and MMP2 expression were up-regulated upon Rab11 expression. This study demonstrated that Rab11and E-cadherin expression are poor indicators of survival time in colorectal carcinoma, but that Rab11 overexpression may contribute to increased collective cell invasion in colorectal carcinoma.

Citation Format: Yuan-Chiang Chung, Wan-Chen Wei, Chia-Nung Hung, Chih-Ping Hsu, Hsiang-Ling Chiu, King-Jen Chang, Wei-Ting Chao. Overexpression of Rab11 and E-cadherin promotes collective cell migration and indicates a poor prognosis in colorectal carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1544.
Abstract 1573: The progesterone analogue, Norgestrel, impairs tumor immunity and promotes metastatic breast cancer progression
Authors: Dalotto Moreno Tomas , Rabinovich Gabriel Adrian , Salatino Mariana
Published: July 2016

The immune system is responsible for recognition and elimination of cancer cells. Tumors, however, can elicit different evasive mechanisms that contribute to immune escape Thus, understanding the immunosuppressive pathways that hamper tumor immunity is critical for the identification of novel therapeutic targets. Progestin-containing hormonal supplementation (as hormone replacement therapy or contraceptive) has been associated with an increase in the risk of breast cancer incidence and recurrence. Moreover, progesterone, as a natural hormone, generally impairs cellular immune responses responsible of preventingfetal rejection during pregnancy. We postulated that progestins and in particular the widely used contraceptive Norgestrel may impair tumor immunity and foster immunosuppressive tumor microenvironments. We injected Balb/c mice with the triple-negative 4T1breast tumor,, and treated them with a Norgestrel releasing depot for 30 days. Norgestrel-treated mice displayed a higher frequency of suppressive regulatory T cells (Tregs) in draining lymph nodes and tumors which displayed a CD4+CD25+Foxp3+CD44+ phenotype. Accordingly, norgestrel-treated mice showed greater number of exhausted PD1+ Tim-3+ CD8 T cells in the tumor microenvironment. Furthermore, in vitro exposure to Norgestrel also promoted differentiation and expansion of Tregs which activated the mTOR pathway assessed by phosphorylation of S6 ribonucleoprotein. These effects could not be prevented by the progesterone nuclear receptor antagonist, Zk 230211, suggesting that Norgestrel acts preferentially through membrane progesterone receptors. Although no significant difference regarding tumor growth could be observed, Norgestrel-treated mice showed greater number of lung metastasis. Mechanistically, Norgestrel-driven Treg cells showed increased expression of RANKL, a protein involved in breast cancer development and progression. Our results identify a novel progestin-mediated tumor-promoting mechanism and indicate caution onthe use of Norgestrel-supplementation therapies. Our findings suggest that Norgestrel may impair tumor immunity, thereby promoting metastatic dissemination of breast cancer cells.

Citation Format: Tomas Dalotto Moreno, Gabriel Adrian Rabinovich, Mariana Salatino. The progesterone analogue, Norgestrel, impairs tumor immunity and promotes metastatic breast cancer progression. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1573.
Abstract 1585: Molecular subtypes of breast cancers and normal mammary epithelial cells show hierarchical heterogeneity and complexity at single-cell resolution
Authors: Azizi Ebrahim , Colacino Justin , Fouladdel Shamileh , Wicha Max S.
Published: July 2016

The analysis of average gene expression in cell populations may obscure the heterogeneity of rare cells such as cancer stem cells (CSCs). The use of CSC markers to enrich for these cell populations has proven valuable in their molecular characterization. However, conventional gene expression analysis of these CSC enriched populations is still not conducive to elucidating their heterogeneity. We exploited single-cell resolution methods using C1 and BioMark HD platform and TaqMan assays in the analysis of CSCs and non-CSCs/differentiated tumor cells. The expression of 96 target genes was studied at single cell level in a multiplex RT-qPCR setting. Breast cancer cell lines including MCF7 (luminal), BT474 (HER2+), SUM149 (Basal), and MDA-MB-231 (Basal-Claudin low) that represent major different subtypes of breast cancers and normal mammary epithelial cells (NM) were studied at single cell resolution. Despite similarities, expression profiles of 96 target genes were distinctly different between subtypes of breast cancers and also with that of NM cells. Interestingly, results of single cell studies revealed significant levels of hierarchical heterogeneities within and between each analyzed population of cells. The observed heterogeneous signatures were due to both different expression levels as well as presence/absence of certain genes. We could identify CSCs with EMT (i.e. CD44+/CD24-/ALDH-/CDH2+/Vimentin+), MET (i.e. ALDH+/CD44-/CD24+/CDH1+/Vimentin-), and dual EMT-MET (CD44+/CD24-/ALDH+/CDH1+ or CDH2+/Vimentin+) characteristics. This methodology enabled us to also identify a range of several different non-CSCs/differentiated single cells with heterogeneous expression patterns for CD44, CD24, ALDH, ESR, PGR, HER2, Vimentin, CDH1, CDH2, EpCAM, and Cytokeratins that were similar to luminal, HER2 overexpressed, basal, and basal-Claudin low breast cancer cells. Additionally, NM single cells also showed significant heterogeneity that was similar to the wide spectrum patterns of some single breast cancer cells described above. This study highlights the use of single cell technologies to redefine tumor cell characterization, revealing heterogeneity of single tumor cells with respect to stem cell properties, metastatic and drug resistance characteristics. This will further improve the identification of new markers/pathways that can be used to develop new target therapies for better therapeutic responses in cancer patients.

Citation Format: Ebrahim Azizi, Justin Colacino, Shamileh Fouladdel, Max S. Wicha. Molecular subtypes of breast cancers and normal mammary epithelial cells show hierarchical heterogeneity and complexity at single-cell resolution. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1585.
Abstract 162: Genomic analysis of uterine aspirates improves diagnosis and captures the intratumor heterogeneity in endometrial cancer
Authors: Mota Alba , Colas Eva , Garcia-Sanz Pablo , Campoy Irene , Rojo-Sebastian Alejandro , Chiva Luis , Alonso Sonsoles , Gatius Sonia , Gil-Moreno Antonio , González-Tallada Xavier , Díaz-Feijoo Berta , Ziober Malinwska Patrycja , Bobiński Marcin , Reventos Jaume , Matias-Guiu Xavier , Moreno-Bueno Gema
Published: July 2016

Background: Endometrial cancer (EC) is the most common cancer in female genital tract in developed countries. Although the majority of ECs are diagnosed in early stages, with a 5-year overall survival of around 80%, an early detection of these tumors is crucial to increase survival of patients given that advanced tumors are associated with poorer outcome. Furthermore, the correct assessment of pre-clinical diagnosis is also decisive to guide the surgical management and treatment of the patient.

Results: Genetic analysis of uterine aspirates identifies the mutational profile of endometrial cancers and represents the intra-tumor heterogeneity found in the analysis of multiple tumor regions from hysterectomy.

Conclusions: The current study reveals the potential of the genetic analysis of uterine aspirates as a pre-operative tool to diagnosis, even in biopsies with insufficient material, with a sensitivity of 93.7% and a specificity of 91.7%. Additionally, genetic analysis of uterine aspirates allows to capture the intra-tumor heterogeneity identified in ECs.

Citation Format: Alba Mota, Eva Colas, Pablo Garcia-Sanz, Irene Campoy, Alejandro Rojo-Sebastian, Luis Chiva, Sonsoles Alonso, Sonia Gatius, Antonio Gil-Moreno, Xavier González-Tallada, Berta Díaz-Feijoo, Patrycja Ziober Malinwska, Marcin Bobiński, Jaume Reventos, Xavier Matias-Guiu, Gema Moreno-Bueno. Genomic analysis of uterine aspirates improves diagnosis and captures the intratumor heterogeneity in endometrial cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 162.
Abstract 1620: Elucidating metastatic signaling networks in TNBC by investigating RKIP-regulated kinome
Authors: Yesilkanal Ali E. , Frankenberger Casey , Rabe Daniel C. , Johnson Gary L. , Rosner Marsha
Published: July 2016

Metastatic progression of tumors is the major cause of death in patients with triple negative breast cancer (TNBC), the most aggressive subtype of breast cancer. However, since metastasis is a multi-step process, unraveling its complexity is a major challenge. One effective way of tackling this question is to study natural blockers of the metastatic process, metastasis suppressors, and identify the mechanisms by which they regulate metastasis. Raf kinase inhibitory protein (RKIP), a protein that regulates kinase activity, is a suppressor of TNBC metastasis. Although RKIP inhibits the activity of key kinases such as Raf-1, GRK2, NIK/IKK in cultured cells, the kinase targets of RKIP in tumors are not known. To address this question, we used a mass spectrometry approach involving inhibitor-conjugated beads to identify kinases that are down-regulated by RKIP in human TNBC xenograft tumors. Our results identified novel targets of RKIP whose kinase activity is either down-regulated or up-regulated by RKIP. We used bioinformatics analysis to build RKIP-regulated signaling networks based upon RKIP-induced changes in kinase activity and related gene expression. Elucidating RKIP function at a systems level reveals the interplay between key metastatic signaling cascades, particularly in relation to cell migration and invasion, and can potentially identify novel anti-metastatic target combinations in TNBCs.

Citation Format: Ali E. Yesilkanal, Casey Frankenberger, Daniel C. Rabe, Gary L. Johnson, Marsha Rosner. Elucidating metastatic signaling networks in TNBC by investigating RKIP-regulated kinome. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1620.
Abstract 1650: Whole-genome RNAi screen identified BCAS1 as a novel modifier of non-small cell lung carcinoma radioresistance
Authors: O’Leary Colin , Deraska Peter , D’Andrea Alan , Kozono David
Published: July 2016

Introduction: Despite optimal chemotherapy, radiotherapy and/or surgery, non-small cell lung carcinoma (NSCLC) remains the leading cause of cancer-related death in the United States. Therefore, there is a need for agents, such as radiosensitizers, that can improve the efficacy of current therapies. Using a whole-genome shRNA screen, we sought to identify and characterize novel gene products that may be involved in NSCLC radioresistance. We hypothesize that these proteins may represent novel targets for NSCLC radiosensitization.

Materials and Methods: A whole-genome pooled retroviral shRNA screen was performed using the Hannon-Elledge library of 74,705 distinct shRNAs directed against over 18,000 genes, to identify gene knockdowns that showed cytotoxicity only in cells treated with fractionated radiation given daily Monday-Friday. To confirm targets identified in the whole-genome screen, we treated A549 and NCI-H460 NSCLC cells with siRNAs targeted against the genes of interest and assessed cell viability following irradiation using the CellTiter-Glo assay and clonogenic survival assays.

Results: We identified six novel genes (BCAS1, c7orf24, CDC45L, KIAA0101, TCN1 and TNC) whose knockdown sensitized NSCLC cells to radiation. Of the six genes, siRNA knockdown of BCAS1 showed the most consistent increases in sensitivity to ionizing radiation. Analysis of cell viability following radiation revealed that BCAS1 knockdown resulted in a 50% additional decrease in cell viability in A549 and NCI-H460 cells following 8 and 4 Gy of ionizing radiation, respectively. We confirmed these results by clonogenic survival assays. To assess for tumor-specific expression in NSCLC, we compared expression data between matched normal tissue and NSCLC biopsy samples and found that BCAS1 is substantially more highly expressed in tumor tissues. Examination of affinity capture-mass spectroscopy data suggested that BCAS1 interacts with RUVBL2, a RuvB-like AAA ATPase, which has known functions in DNA damage and repair.

Conclusions: BCAS1 emerged from a whole-genome RNAi screen as a potential target for improving the efficacy of radiotherapy in NSCLC. We demonstrated that BCAS1 is overexpressed in tumor tissues compared to normal lung, and that its knockdown consistently sensitizes NSCLC cell lines to radiation, validating the screen hit. Ongoing work suggests that BCAS1 may modulate DNA damage repair. Further studies to understand the mechanism by which BCAS1 is involved in radioresistance are necessary.

Citation Format: Colin O’Leary, Peter Deraska, Alan D’Andrea, David Kozono. Whole-genome RNAi screen identified BCAS1 as a novel modifier of non-small cell lung carcinoma radioresistance. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1650.
Abstract 1658: M3814, a novel investigational DNA-PK inhibitor: enhancing the effect of fractionated radiotherapy leading to complete regression of tumors in mice
Authors: Zenke Frank T. , Zimmermann Astrid , Sirrenberg Christian , Dahmen Heike , Vassilev Lubo , Pehl Ulrich , Fuchss Thomas , Blaukat Andree
Published: July 2016

Physical or chemical agents that damage DNA such as ionizing radiation are among the most widely used classes of cancer therapeutics today. Double strand breaks (DSB) generated in DNA by radiation induce multitude of cellular responses, including DNA repair, cell cycle arrest or cell death if the damage is left unrepaired. A complex set of molecular events are responsible for DNA repair via two major mechanisms - homologous recombination (HR) or non-homologous end joining (NHEJ). DNA-PKcs with its regulatory protein subunits, Ku70 and Ku80, is an integral component of NHEJ and considered an attractive intervention point to inhibit DNA repair.

We have developed an orally bioavailable, highly potent, and selective inhibitor of DNA-PK, M3814, for cancer therapy in combination with DNA damaging modalities such as radiation, and radio-chemotherapy. Here, we present the preclinical characterization of M3814 using biochemical, cellular and human tumor xenograft models. M3814 sensitized multiple tumor cell lines to radiation therapy in vitro and strongly enhanced the antitumor activity of ionizing radiation in vivo with complete tumor regression applying a clinically relevant fractionated radiation regimen. These effects are due to inhibition of DNA-PK protein kinase activity as demonstrated by the levels of DNA-PK autophosphorylation in human tumor cell lines, and xenograft tumors M3814 is currently investigated in PhI clinical trials.

Citation Format: Frank T. Zenke, Astrid Zimmermann, Christian Sirrenberg, Heike Dahmen, Lubo Vassilev, Ulrich Pehl, Thomas Fuchss, Andree Blaukat. M3814, a novel investigational DNA-PK inhibitor: enhancing the effect of fractionated radiotherapy leading to complete regression of tumors in mice. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1658.
Abstract 1709: STRAP mediates the stemness of human colorectal cancer cells by epigenetic regulation of Notch pathway
Authors: Jin Lin , Vu Trung Tam , Datta Pran K.
Published: July 2016

Human colon cancer-initiating cells (CCICs) are responsible for the unrestrained cell proliferation and chemoresistance of colorectal tumors. Notch signaling is critical for the maintenance and self-renewal of cancer-initiating cells in colon cancer. We have shown that the serine-threonine kinase receptor-associated protein (STRAP), a ubiquitous WD40 protein, is involved in promoting tumorigenesis through activated MAPK/ERK pathway and pRb phosphorylation. In addition, our previous studies uncovered that STRAP is up-regulated in 60% colon carcinomas and behaves as an oncogene in a TGF-β-dependent or -independent manner. Based on our new finding that STRAP stabilizes intracellular fragment of Notch3 (ICN3) via post-translational modification, we sought to explore the cross-talk between STRAP and Notch pathway in regulating stemness of CCICs. Here, we have generated STRAP knock down (KD) colorectal cancer cell lines (RKO, DLD1, HCT116, HT29, WiDr, SW480, SW620, and LOVO) to show how STRAP abrogates apoptosis by up-regulating CD44+/CD133+ subpopulations of CCICs. Heterozygous Strap KO inhibits tumorigenesis accompanied by the reduction of stemness phenotype (CD44+, Lgr5+) in a colitis-associated mouse model. Western blot assays and immunofluorescene staining demonstrated that the expression of stemness biomarkers (Nanog, Oct4 and Sox2) is decreased in STRAP KD cells. Furthermore, the transcriptional activation of Notch pathway related genes is downregulated in the STRAP KD cells. EpiTect Chip qPCR Arrays profiled that 17 out of 84 genes key to Notch signal transduction had higher activating (H3K4me3) and lower repressive (H3K27me3) histone modifications in WT cells compared to STRAP KD cells. Among these genes, we identified two transcription factors, Hes1 and Hes5, that are transcriptionally regulated by STRAP as demonstrated by luciferase assays and qPCR. We further demonstrated that Hes1 and Hes5 were transcriptionally activated by STRAP through Polycomb Repressive Complex 2 (PRC2)-dependent mechanism. Immunoprecipitation experiments showed that STRAP interacts with the key components, EZH2 and SUZ12 of PRC2 complex in vivo, which antagonizes the histone methyltransferase activity in H3K27 on the promoters of Hes1 and Hes5, resulting in activating these two genes. Of interest, ectopic expression of either Hes1 or Hes5 can rescue the stemness phenotype in STRAP KD cells. Together, our findings provide a novel mechanistic insight into the regulation of CCICs via STRAP-PRC2-Hes1/5 regulatory axis, which represents a potential target for CCICs, thus overcoming cancer chemoresistance and relapse.

Citation Format: Lin Jin, Trung Tam Vu, Pran K. Datta. STRAP mediates the stemness of human colorectal cancer cells by epigenetic regulation of Notch pathway. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1709.
Abstract 1723: Novel drug candidates CEP1430 and CEP1507 against cancer stem cells and circulating tumor cells in advanced stage pancreatic cancer
Authors: Sharma Cristian , Narayanan Padmini , Sharma Michael , Rodriguez Robert , Navel Miriam , Stanton Donna , Amezcua Natalee , Sharma Shaleekha , Amiri Mandana , Jani Jitesh , Sharma Sherven , Sharma Jay P.
Published: July 2016

Despite improvements in treatment modalities, pancreatic cancer remains the fourth leading cause of cancer mortality in the US. The 5-year survival rate is less than 6% because of the extent of metastases at the time of diagnosis. The treatment of pancreatic cancer remains challenging however there is much optimism with targeted strategies and trial designs. The interrogation of pathway centric targets that regulate the regenerative and metastatic activities in the tumor, adequate molecular-hypothesis driven trial designs with robust prognostic and predictive markers of response are necessary for the development of effective therapeutic strategies for long term anti-tumor benefit.

Here, we examined the efficacy of CEP1430 and CEP1507 to target cancer stem cells (CSC) and circulating tumor cells (CTC) in human pancreatic cancer. The pancreatic CSCs expressed the following markers(CD133,CD44,CXCR4,SSEA3/4,Oct4,ALDH,Telomerase,& Nestin). In vitro CEP1430 inhibited the growth of CSCs whereas CEP1507 inhibited CTCs that expressed the following markers [CK19, CEA, CA19-9,CK8 /18,h-TERT,C-MET,CK20,EpCAM, Vimentin, E- Cadherin, MUC1, CEA CAM5,CD45 neg]. Gemcitabine on the other hand suppressed the viability of non-CSCs (differentiated tumor Cells) by 70%. In accord, in vivo studies showed that CEP1507 when combined with gemcitabine eliminated the engraftment of human pancreatic cancer CTCs by 85% (n = 20,p<0.001). In contrast, CEP1430 in combination with gemcitabine selectively inhibited CSCs by 80% (n = 20,p<0.001) and was more effective than the individual agents. This data demonstrates that CEP1430 and CEP1507 that target CSCs and CTCs respectively may change the treatment paradigm and enhance the efficacy of gemcitabine in advanced pancreatic cancer. Further studies will evaluate the full potential of this approach to target the CSCs - and metastasis-initiating cells (CTCs) as combination therapy against locally advanced and metastatic pancreatic cancer.

Citation Format: Cristian Sharma, Padmini Narayanan, Michael Sharma, Robert Rodriguez, Miriam Navel, Donna Stanton, Natalee Amezcua, Shaleekha Sharma, Mandana Amiri, Jitesh Jani, Sherven Sharma, Jay P. Sharma. Novel drug candidates CEP1430 and CEP1507 against cancer stem cells and circulating tumor cells in advanced stage pancreatic cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1723.