bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1960: Upregulation of MCP-1 regulates invasiveness in triple negative breast cancer
Authors: Dutta Pranabananda , Paico Kimberly , Yuwanita Inez , Wu Yanyuan , Sarkissyan Marianna , Vadgama Jaydutt
Published: July 2017

Background: Triple negative breast cancer (TNBC) poses a critical problem for targeted therapy due to lack of significant expression of estrogen, progesterone receptor or Her2/neu oncogene. Hence, it is imperative to identify novel therapeutic strategies to target TNBC. Our study is aimed to examine whether Monocyte Chemoattractant Protein -1 (MCP-1) is a specific marker for TNBC metastasis. Experimental

Design: We employed ELISA to determine secreted MCP-1 in cell conditioned media, as well as Real-time PCR to determine the status of MCP-1 in TNBC cell lines. Boyden chamber assay was used to determine the effect of recombinant MCP-1 on cellular metastasis. Cellular proliferation was measured with MTT assay. Immunofluorescence staining was utilized for protein of interest in breast cancer cells. MCP-1 knockdown was performed using lentiviral vector with shRNA targeting MCP-1 coding regions.

Results: Our data show that the key inflammatory chemokine MCP-1 is upregulated in TNBC cell lines both transcriptionally as well as in terms of secretion compared to ER-positive cell line, MCF-7. MCP-1 stimulation in MDA-MB231 and MCF-7cells does not affect cellular proliferation. However, MCP-1 enhances metastatic properties of MDA-MB-231 cells along with BT-549 cells. Inhibiting Chemokine receptor 2/4 (CCR2/4), cognate receptor for MCP-1, with small molecule antagonists negatively affects invasiveness in MDA-MB-231 cells as evidenced by Boyden chamber assay. Knocking down MCP-1 by shRNA decreases cell invasion in TNBC cell line, BT-549 along with downregulation of key epithelial to mesenchymal transition markers, N-cadherin and Vimentin. MCP-1 induced cell invasion in TNBC may involve activation of p44/p42 MAPK Thr202/Tyr204.

Conclusion: Our study suggests that high MCP-1 levels in TNBC is driving up metastasis potential in cells. Thus MCP-1 and its mediated pathways could be potential therapeutic targets for the treatment of TNBC.

Citation Format: Pranabananda Dutta, Kimberly Paico, Inez Yuwanita, Yanyuan Wu, Marianna Sarkissyan, Jaydutt Vadgama. Upregulation of MCP-1 regulates invasiveness in triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1960. doi:10.1158/1538-7445.AM2017-1960
Abstract 1997: Charting the DNA methylation landscape of prostate cancer associated fibroblasts
Authors: Lawrence Mitchell G. , Pidsley Ruth , Zotenko Elena , Niranjan Birunthi , Statham Aaron , Song Jenny , Armstrong Nicola , Pedersen John , Frydenberg Mark , Taylor Renea , Stirzaker Clare , Risbridger Gail , Clark Susan
Published: July 2017

Most studies of the genome and epigenome landscape of tumors focus on the aberrations in epithelial cancer cells. However, the growth and progression of solid tumors involves dynamic cross-talk between cancer epithelium and the surrounding microenvironment, including cancer-associated fibroblasts (CAFs). The molecular landscape of these cells is still poorly understood, so it is important to define the epigenetic features that underpin the functional differences between CAFs and non-malignant prostate fibroblasts (NPFs) and influence cancer progression. Therefore, we used whole-genome bisulphite sequencing to chart the epigenome landscape of CAFs at single base resolution. Our data revealed that NPFs and CAFs from localized prostate cancer have remarkably different epigenome profiles characterized by locus-specific rather than global changes in DNA methylation. CAFs exhibited more than 7500 differentially methylated regions compared to NPFs. In contrast, no recurrent genomic aberrations were detected in CAFs, emphasising the importance of epigenome changes in reinforcing their pro-tumorigenic phenotype. Many differentially methylated regions occurred at known regulatory loci and were associated with differentially expressed genes measured using RNAseq. The methylation and gene expression changes were highly consistent across patients when validated using an independent cohort of patient-matched NPFs and CAFs. Remarkably, a subset of differentially methylated regions in CAFs is shared with prostate cancer epithelial cells, revealing the new concept of tumour-specific epigenome modifications in the tumour and its microenvironment. Collectively, these data demonstrate that CAFs are defined by an enduring epigenetic signature that comprises a distinct set of locus-specific DNA methylation alterations in cancer-related regulatory loci. The distinct methylome of CAFs provides a novel epigenetic hallmark of the cancer microenvironment and promises new biomarkers to improve interpretation of diagnostic samples.

Citation Format: Mitchell G. Lawrence, Ruth Pidsley, Elena Zotenko, Birunthi Niranjan, Aaron Statham, Jenny Song, Nicola Armstrong, John Pedersen, Mark Frydenberg, Renea Taylor, Clare Stirzaker, Gail Risbridger, Susan Clark. Charting the DNA methylation landscape of prostate cancer associated fibroblasts [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1997. doi:10.1158/1538-7445.AM2017-1997
Abstract 1998: Transcriptome analysis of hypoxic head and neck cancer cells uncovers intron retention in EIF2B5 as a mechanism to reduce protein synthesis
Authors: Brady Lauren K. , Wang Hejia , Radens Caleb , Radovich Milan , Maity Amit , Ivan Cristina , Ivan Mircea , Barash Yoseph , Koumenis Constantinos
Published: July 2017

Hypoxia is a key feature of solid tumors that contributes to resistance to therapy and reduced overall survival. Cellular adaptation to the hypoxic tumor microenvironment involves attenuation of energy-consuming processes such as macromolecular synthesis. We have identified RNA processing as another major process which is globally downregulated under hypoxic stress. By sequencing RNA of normoxic and hypoxic head and neck cancer cells, we observed widespread repression of genes that regulate RNA splicing and processing. As a result, we observed over 1,000 changes in relative mRNA isoform expression, including a significant increase in hypoxia-induced intron retention in nearly 100 genes. Genes observed to undergo intron retention in hypoxia included major regulators of RNA processing and protein synthesis, such as the translation initiation factor EIF2B5.

Surprisingly, hypoxia-induced intron retention in EIF2B5 creates a premature termination codon, which results in a truncated protein isoform of eIF2Bϵ. We provide evidence that this isoform acts in opposition to the full-length protein to inhibit translation under stringent hypoxic conditions. Moreover, this intron is expressed in solid cancers known to contain hypoxic fractions and is overexpressed in head and neck cancer in a stage-dependent manner compared to normal tissue.

To investigate how this intron is retained, we are examining the role of hypoxia-mediated changes in phosphorylation of the C-terminal domain (CTD) of RNA polymerase II. Our data suggest that increased phosphorylation of the CTD may lead to transcriptional pausing and subsequent retention of intron 12 in EIF2B5. Furthermore, distinct sequence features at this locus, such as a weak splice site at the intron12:exon13 junction, may explain how this intron is selectively retained in hypoxic cells. These results implicate mRNA splicing as a key mechanism to induce alternate transcripts and subsequent protein isoforms under hypoxia that influence cellular adaptation to stress.

Citation Format: Lauren K. Brady, Hejia Wang, Caleb Radens, Milan Radovich, Amit Maity, Cristina Ivan, Mircea Ivan, Yoseph Barash, Constantinos Koumenis. Transcriptome analysis of hypoxic head and neck cancer cells uncovers intron retention in EIF2B5 as a mechanism to reduce protein synthesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1998. doi:10.1158/1538-7445.AM2017-1998
Abstract 2: Functional and mechanistic interrogation of BET bromodomain degraders for the treatment of metastatic castration resistant prostate cancer
Authors: Kregel Steven , Malik Rohit , Asangani Irfan A. , Wilder-Romans Kari , Jiang Xia , Apel Ingrid J. , Ravi Gautham , Tien Jean , Cao Xuhong , Feng Felix Y. , Speers Corey , Wang Shaomeng , Chinnaiyan Arul M.
Published: July 2017

Metastatic castration-resistant prostate cancer (mCRPC) is a lethal disease with about 30,000 estimated annual deaths in U.S, with a vast majority of CRPC driven by androgen receptor (AR) signaling. AR-signaling is critical for the development and progression of prostate cancer, and AR also the main therapeutic clinical target. AR-targeted therapies, such as AR-antagonists, provide substantial benefits in the treatment of mCRPC; however, majority of patients fail these therapies and succumb to the disease. Therefore, there is a clear and pressing need to develop new therapeutics against the AR axis in CRPC. One such novel strategy for targeting the AR-pathway and inhibiting the growth of CRPC has been the use of bromodomain and extraterminal (BET) protein inhibitors such a JQ1. However, a new class of molecules that target BET bromodomain proteins through their proteasomal degradation can improve efficacy and specificity of these inhibitors. Based on our findings, we hypothesized that pharmacologic BET degradation represents an important advance in the treatment of CRPC, and may provide a novel therapeutic strategy for advanced prostate cancer. These inhibitors affect AR-positive prostate cancer cells preferentially over AR-negative and benign prostate epithelial cells, and proteomic and genomic mechanistic studies confirm disruption of oncogenic AR and MYC signaling, both in vivo and in vitro. The overall goal is to develop very potent small molecule that leads to the proteasomal-degradation of BET proteins, with optimized in vivo properties, and to provide a compelling scientific rationale, including detailed mechanistic insight, to facilitate advancement of BET degraders as a novel potential therapeutic strategy for patients with mCRPC.

Citation Format: Steven Kregel, Rohit Malik, Irfan A. Asangani, Kari Wilder-Romans, Xia Jiang, Ingrid J. Apel, Gautham Ravi, Jean Tien, Xuhong Cao, Felix Y. Feng, Corey Speers, Shaomeng Wang, Arul M. Chinnaiyan. Functional and mechanistic interrogation of BET bromodomain degraders for the treatment of metastatic castration resistant prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2. doi:10.1158/1538-7445.AM2017-2
Abstract 2054: Cerivastatin overcomes the acquired resistance to crizotinib in EML4-ALK positive non-small cell lung cancer by controlling YAP activation
Authors: Yun Miran , Choi Hun Mi , Pyo Kyoung Ho , Cho Byoung Chul
Published: July 2017

Crizotinib is highly effective in patients with non-small cell lung cancer (NSCLC) harboring the echinoderm microtubule-associated protein-like 4 (EML4)-anaplastic lymphoma kinase (ALK) fusion. However, its efficacy has been limited by the development of acquired resistance, and the mechanisms of such resistance remain largely unknown. Herein, we investigated a possible candidate for circumventing the acquired resistance to crizotinib.

We established a model of acquired resistance to crizotinib (H3122-CR) by exposing EML4-ALK-positive H3122 lung cancer cells to increasing doses of crizotinib, and performed MTT screening using a library of FDA (Food and Drug Administration)-approved drugs composed of a collection of 640 clinically used compounds.

Our MTT screening identified that cerivastatin, a drug targeting 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase, significantly inhibited the cell growth in H3122-CR cells, which was completely restored by addition of geranylgeranyl pyrophosphate (GGPP), a key metabolite of mevalonate pathway. We also found that yes-associated protein (YAP), a major effector of the Hippo tumor suppressor pathway, was robustly accumulated in nucleus with a concomitant decrement of YAP phosphorylation in H3122-CR cells compared to parental cells. Importantly, inhibition of geranylgeranylation (GGylation) by GGTI-298 or cerivastatin markedly increased YAP phosphorylation, led to cytoplasmic translocation of YAP, and subsequently induced YAP inactivation. Moreover, the enrichment of EGFR signaling pathway and cell cycle signature including transcriptional targets of YAP was enhanced in H3122-CR cells, whose induction contributed YAP-mediated resistance to crizotinib. Inhibition of YAP function with siRNA or verteporfin as a YAP inhibitor greatly abrogated cell growth of H3122-CR cells by modulating cell cycle regulators and EGFR activation. Finally, we confirmed up-regulation of nuclear YAP expression in crizotinib-acquired resistant patient derived tumor xenograft (PDTX) models established from EML4-ALK positive NSCLC patients. Collectively, our findings define the GGylation-mediated YAP transcriptional activation as a new mechanism of resistance to crizotinib, providing a rationale for further exploring statins as a possible anticancer agent to overcome the acquired resistance to crizotinib in EML4-ALK positive NSCLC cells.

Citation Format: Miran Yun, Hun Mi Choi, Kyoung Ho Pyo, Byoung Chul Cho. Cerivastatin overcomes the acquired resistance to crizotinib in EML4-ALK positive non-small cell lung cancer by controlling YAP activation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2054. doi:10.1158/1538-7445.AM2017-2054
Abstract 2107: Tetra-arsenic hexoxide induces G2/M cell cycle arrest, apoptosis, and autophagy via p38 MAPK- and AKT-mediated pathways in SW620 human colon cancer cells
Authors: Lee Won Sup , Yun Jeong Won , Kim Min Jeong , Nagappan Arulkumar , Lu Jing Nan , Chang Seong-Hwan , Jeong Jae-Hoon , Kim GonSup , Jung Jin-Myung , Hong Soon Chan
Published: July 2017

Tetraarsenic hexoxide (As4O6) has been used in Korean folk remedy for the treatment of cancer since the late 1980’s. Evidence suggests that As4O6 show anti-cancer effects, whose mechanisms are different from those for As2O3. However, the detailed anticancer mechanism is still unclear. Here, we investigated the anticancer effects of As4O6 on SW620 human colon cancer cells. As4O6 induced cell death in a dose-dependent manner. Flow cytometry analysis revealed that As4O6 increased the sub-G1 (apoptotic cell population) and G2/M phase population in a dose-dependent manner. Further, nuclear condensation and cleaved nuclei were also observed upon staining with Hoechst 33342 in As4O6-treated SW620 cells. Western blot revealed that As4O6 significantly down-regulated the cyclin B1, cdc 2, pro-caspases -3, -8 and -9, and up-regulated p21 and cleavage of PARP in SW620 cells. In addition, As4O6 increased the expression of death receptor 5 (DR5), suppressed Bcl-2 and XIAP family proteins, and promoted the conversion of LC3-I to LC3-II in a Beclin-1 independent manner. Interestingly, As4O6 dephosphorylated Akt and JNK and phosphorylated p38 MAPK, and the cell death was inhibited by p38 MAPK inhibitor; whereas Akt inhibitor augmented the As4O6 induced cell death, suggesting p38 MAPK and AKT were associated with As4O6-induced cell death. Taken together, these findings suggest that As4O6 induced G2/M arrest, apoptosis and autophagic cell death at least in part through p38 MAPK and AKT pathways in SW620 human colon cancer cells. This study may explain the anecdotal anticancer effects showing central necrosis with dormant status of cancers when treated by As4O6 as folk remedy.

Citation Format: Won Sup Lee, Jeong Won Yun, Min Jeong Kim, Arulkumar Nagappan, Jing Nan Lu, Seong-Hwan Chang, Jae-Hoon Jeong, GonSup Kim, Jin-Myung Jung, Soon Chan Hong. Tetra-arsenic hexoxide induces G2/M cell cycle arrest, apoptosis, and autophagy via p38 MAPK- and AKT-mediated pathways in SW620 human colon cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2107. doi:10.1158/1538-7445.AM2017-2107
Abstract 2116: Combining the power of different profiling approaches to better understand the activity of kinase inhibitor drugs
Authors: King Alastair J. , Parry Jesse J. , Burkart Samantha C. , Cavedine Lee R. , Cracchiolo Alyssa M. , Epkins Brogan A. , Garner Steven M. , Sindelar Timothy J. , Wageman Charles R. , Davies Stephen P. , Plater Andrew , Woodward Anna M. , Warrior Usha
Published: July 2017

Profiling of inhibitors for eventual therapeutic use is an essential part of drug discovery and development. Evaluating the in vitro potency of kinase inhibitors in a wide panel of enzyme assays is critical, not only to guiding SAR knowledge for improving drug design, but also for assessing selectivity, which has long-reaching consequences for both biological activity and safety. Evaluation of the same inhibitors in cellular models yields information on the activity of these inhibitors in a more physiologically relevant environment. However, while some selective kinase inhibitors show predictably selective profiles in cell proliferation assays, others do not. Thus, the biological context in which a kinase inhibitor functions, and the signaling networks influenced by inhibition of both the desired target and off-target activities, help to define how that inhibitor will work in a cellular setting and, ultimately, in a therapeutic application. We demonstrate the power of profiling several well-known kinase inhibitors in complementary panel assays, to predict behavior that has implications for clinical activity. The in vitro profiles of selective (trametinib, vemurafenib) and less selective (dasatinib, sorafenib) kinase inhibitors are shown and compared. With careful evaluation of their in vitro enzyme activity profiles in Eurofins’ KinaseProfiler™ panel, cellular response profiles in OncoPanel™, and predictive biomarker identification by univariate genomic analysis, a fuller picture of these inhibitors’ biological activity can be generated, and may be applied to predicting the therapeutic potential of such inhibitors. While, in some cases, clinical response is seen as expected, this kind of comprehensive analysis opens up the possibility for expanding clinical utility of an asset by potential repurposing for other indications. Collectively, this demonstrates not only the need to understand an inhibitor’s fundamental activity(ies) at its designated target(s), but also the importance of considering cellular context, when assessing its maximum potential clinical utility.

Citation Format: Alastair J. King, Jesse J. Parry, Samantha C. Burkart, Lee R. Cavedine, Alyssa M. Cracchiolo, Brogan A. Epkins, Steven M. Garner, Timothy J. Sindelar, Charles R. Wageman, Stephen P. Davies, Andrew Plater, Anna M. Woodward, Usha Warrior. Combining the power of different profiling approaches to better understand the activity of kinase inhibitor drugs [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2116. doi:10.1158/1538-7445.AM2017-2116
Abstract 2141: Development of a novel miR-129 mimic with enhanced therapeutic potential for treatment of resistant colorectal cancer
Authors: Fesler Andrew T. , Wu Ning , Liu Hua , Ju Jingfang
Published: July 2017

Treatment of advanced stage colorectal cancer remains a clinical challenge associated with resistance to fluoropyrimidine based chemotherapy. There is an urgent need to discover and develop new strategies to enhance treatment efficacy in order to improve outcomes for these patients. Non-coding microRNAs (miRNAs) have important functions as oncogenes or tumor suppressor genes in the regulation of cancer development and progression. Recently, miRNAs have emerged as potential therapeutic options. We have previously identified miR-129 as a tumor suppressor miRNA and potential therapeutic candidate in colorectal cancer. The expression of miR-129 is progressively lost in colorectal cancer patients and is an important regulator of apoptosis through the targeting of genes such as BCL-2. miR-129 was also found to enhance 5-flurouracil (5-FU) cytotoxicity in vitro and in vivo. To further developing miR-129 based novel therapeutics in colorectal cancer, we have designed a modified version of miR-129 to enhance stability and efficacy. The miR-129 mimic is significantly more potent in inhibiting proliferation of a panel of colon cancer cell lines than the native miR-129 precursor, with 79% reduction by miR-129 mimic compared to 38% for native precursor. The miR-129 mimic induces profound cell cycle arrest at the G1/S checkpoint. The G1/S ratio increased 3.8 fold compared to control when cells were transfected with miR-129 mimic. We also demonstrated that the miR-129 mimic retains its target specificity to BCL-2, TS and E2F3. The therapeutic potential of miR-129 mimic was demonstrated in vivo mouse colon tumor models as a potent inhibitor of tumor growth and metastasis. As a result, miR-129 mimic has a great potential to be further developed as a novel therapeutic drug for treatment of advanced colorectal cancer.

Citation Format: Andrew T. Fesler, Ning Wu, Hua Liu, Jingfang Ju. Development of a novel miR-129 mimic with enhanced therapeutic potential for treatment of resistant colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2141. doi:10.1158/1538-7445.AM2017-2141
Abstract 2149: Unusual expression of HPRT on the surface of the colorectal cancer cell lines HT29 and SW620
Authors: Weagel Evita G. , Townsend Michelle H. , Anderson Michael D. , Velazquez Edwin J. , Weber K Scott , Robison Richard A. , O'Neill Kim L.
Published: July 2017

Colorectal cancer is one of the most common cancers in the world, affecting nearly 1.2 million people in the United States alone. The aim of this study is to investigate the salvage pathway enzyme HPRT as a possible biomarker in two colorectal cancer cell lines: HT29 and SW620. HPRT is a transferase in the purine salvage pathway that functions primarily by catalyzing the conversion of hypoxanthine to inosine monophosphate and guanine to guanosine monophosphate. Because of its role in proliferation and cell cycle regulation, we hypothesized an increase in HPRT expression within cancer cells, which could potentially lead to presentation on the surface of the cell. HPRT surface localization was assessed utilizing confocal microscopy, flow cytometry, and scanning electron microscopy. These techniques allowed us to visualize HPRT on the plasma membrane and quantify expression. There was statistically significant expression of HPRT on the surface of both HT29 and SW620 cells with a 28% and 58% fluorescent shift in the population, respectively. Confocal microscopy images revealed overlap between cells stained with a membrane dye and anti-HPRT FITC antibody, which indicates a direct relationship between HPRT and the plasma membrane of SW620 cells. To visualize the location of HPRT on the plasma membrane, anti-HPRT antibodies were labelled with gold and protein presence was quantified using an electron microscope, which measured the gold elemental weight percentage of each sample. When exposed to anti-HPRT antibody, the gold weight percentage of the samples significantly increased with 12.4% gold in SW620 cells and 11.2% gold in HT29 cells, indicating significant HPRT surface presence. The observed localization of HPRT to the plasma membrane was variable between the two different cancer cell lines. SW620 cells, which are faster growing and more aggressive, had significantly higher HPRT presentation that HT29 cells. These data collectively suggest that HPRT may be a possible biomarker target for the identification and possible treatment of colorectal cancer cells.

Citation Format: Evita G. Weagel, Michelle H. Townsend, Michael D. Anderson, Edwin J. Velazquez, K Scott Weber, Richard A. Robison, Kim L. O'Neill. Unusual expression of HPRT on the surface of the colorectal cancer cell lines HT29 and SW620 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2149. doi:10.1158/1538-7445.AM2017-2149
Abstract 2155: Small molecule NSC59984 suppresses cancer cell growth under hypoxia
Authors: Zhang Shengliang , Zhou Lanlan , El-Deiry Wafik S.
Published: July 2017

Hypoxia is one of the main features of solid tumors, contributes to drug resistance and is associated with poor prognosis. We have reported that a small-molecule NSC59984 restores p53 signaling and degrades mutant p53 protein in cancer cells. We find that NSC59984-mediated p53 signaling restoration and mutant p53 degradation is significantly blocked by NAC, an inhibitor of reactive oxygen species (ROS), while, further enhanced by BSO, a chemical compound increasing ROS in cells. These results suggest that ROS is a factor required for the effect of NSC59984 on cancer cells. Cellular ROS is produced by hypoxia in solid tumors. We investigated the effect of NSC59984 on p53 pathway and cell death in p53 mutant colorectal cancer (CRC) cells cultured under hypoxia. NSC59984 increases p53 signaling in p53 mutant cancer cells under hypoxia, as it does under normoxia. The effect of NSC59984 under hypoxia is blocked by NAC. Moreover, NSC59984 decreases cancer cell viability in a dose-dependent manner under hypoxia similar to normoxia. Our results suggest that NSC59984 inhibits cancer cell growth under hypoxia. Hypoxia has been known to increase cancer stem cells in tumors, which is one of the reasons causing drug resistance. We then evaluate the effect of NSC59984 on the stem cell growth. NSC59984 significantly reduces colonosphere formation of SW480, DLD-1 and HT29 cells. CRC stem cell markers such as CD44 and ALDH are decreased in colonospheres upon NSC59984 treatment, suggesting that NSC59984 inhibits stem cell self-renewal or/and growth. Our results provide the rationale for administration of NSC59984 in solid tumors with or without hypoxia and in combination treatment with ROS activators.

Citation Format: Shengliang Zhang, Lanlan Zhou, Wafik S. El-Deiry. Small molecule NSC59984 suppresses cancer cell growth under hypoxia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2155. doi:10.1158/1538-7445.AM2017-2155
Abstract 2179: Targeted delivery of nanoparticulate cytochrome c into GL261 glioma cells through the proton-coupled folate transporter
Authors: Kucheryavykh Yuriy , Ortiz-Rivera Jescelica , Inyushin Michael , Cubano Luis , Morales-Cruz Moraima , Cruz-Montañez Alejandra , Griebenow Kai , Kucheryavykh Lilia
Published: July 2017

In this study we identified the proton-coupled folate transporter (PCFT) as a route for targeted delivery of drugs to glioma cells. Using the techniques of confocal imaging, western blotting, and siRNA knockdown against the PCFT (encoded by the SLC46A1 gene), we demonstrated that Gl261 glioma cells, but not primary cultured astrocytes, express the PCFT, which provides selective internalization of folic acid (FA)-conjugated cytochrome c-containing nanoparticles (FA-Cyt c NPs), followed by glioma cell death. FA-Cyt c NPs, used at a concentration of 100 µg/ml, had no cytotoxic effects in astrocytes but caused 40% death in GL261 cells. Whole-cell patch clamp recording revealed FA-induced membrane currents, with maximum activity at pH 6.0, which is specific for the PCFT. Currents were reduced by siRNA PCFT knockdown in a similar manner as by application of FA-Cyt c NPs, indicating that the PCFT is a major carrier of FA and a route for internalization of FA-conjugated NPs in GL261 cells. We conclude that the PCFT provides a mechanism for targeted delivery of FA-conjugated nanodrugs to glioma cells with high specificity for GL261 cells, effectively causing apoptosis in these cells, and provides a starting point for the development of efficient methods for treating gliomas. The research was supported by NIH grants SC2GM102040, SC2GM095410, G12MD007583, R25GM110513, and US Department of Education Grant P031S130068.

Citation Format: Yuriy Kucheryavykh, Jescelica Ortiz-Rivera, Michael Inyushin, Luis Cubano, Moraima Morales-Cruz, Alejandra Cruz-Montañez, Kai Griebenow, Lilia Kucheryavykh. Targeted delivery of nanoparticulate cytochrome c into GL261 glioma cells through the proton-coupled folate transporter [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2179. doi:10.1158/1538-7445.AM2017-2179
Abstract 2181: Antibody-conjugated nanoparticles for targeting metastatic triple-negative breast cancer
Authors: Khanna Vidhi , Kalscheuer Stephen , Kirtane Ameya , Panyam Jayanth
Published: July 2017

Early detection and the availability of new treatments have improved the survival rates of patients presenting with local or regional breast cancer to as high as 99% and 85%, respectively. On the contrary, patients with metastatic disease have a dismal 5-year survival rate of 17%. Thus, there is an urgent need for treatment strategies directed towards metastasis. Our lab has developed antibodies (Clone 6 and AM6) capable of recognizing tumor cells that have undergone epithelial-to-mesenchymal transition (EMT), a key step in the generation of circulating tumor cells and metastasis. The goal of the current study was to determine whether we use these antibodies as targeting ligands for directing anticancer drug-loaded polymeric nanoparticles to metastatic triple negative breast cancer cells as a novel therapeutic option. Polymeric PLGA nanoparticles loaded with paclitaxel, a chemotherapeutic agent, were functionalized with the antibodies using thiol-maleimide chemistry. We optimized the conjugation reaction in order to achieve maximal cell uptake of nanoparticles without compromising antibody binding. In vitro studies were carried out in an MDA-MB-231 derivative cell line with enhanced lung metastatic potential as well as a melanoma metastatic cell line M12. Clone 6 nanoparticles and AM6 nanoparticles showed significant improvement in cellular uptake as well as retention. A competition experiment confirmed target-mediated uptake of nanoparticles. Cytotoxicity studies showed improved cell kill using Clone 6 nanoparticles and AM6 nanoparticles. Based on these promising in vitro results, we are currently carrying out in vivo studies in mice. The development of a targeted drug delivery system for the treatment of metastatic triple negative breast cancer can significantly enhance the survival rate for patients who often have a life-expectancy of less than one year.

Citation Format: Vidhi Khanna, Stephen Kalscheuer, Ameya Kirtane, Jayanth Panyam. Antibody-conjugated nanoparticles for targeting metastatic triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2181. doi:10.1158/1538-7445.AM2017-2181
Abstract 2190: Theranostical nanoscale metal-organic frameworks-mediated rapid multiplexed microRNA detection and highly therapeutic efficacy in living oral cancer cells
Authors: Yu Pei , Zhuang Zehang , Qiu Guihua , Chen Haolin , Zhao Yuying , Chen Jinxiang , Liu Xiqiang
Published: July 2017

Oral cancer represents the sixth most frequent cancer in the world, characterized by high poor prognosis and low survival rate due to local relapse and metastasis. MicroRNAs (miRNAs) have been discovered as diagnostic biomarkers and therapeutic targets for oral cancer. However, one of the major challenges of miRNAs-based theranostical approach is to achieve efficient and safe delivery. Thus, a reliable sensing strategy that can detect miRNAs expression levels in living cancer cells is urgently needed. Herein, a novel sensing platform based on nanoscale metal-organic framework (NMOF) conjugated with the fluorophores-labeled single-stranded DNA probe (p-DNA) was fabricated to monitor and inhibit multiplexed miRNAs expression in living oral cancer cells. The flow cytometry and confocal laser scanning microscopy (CLSM) were carried out to evaluate the expression levels of miRNAs in oral cancer cells. The miRNA inhibition efficiency was examined by RT-PCR. The effects of the complexes on tumor cell invasion and migration were assessed by transwell assay and wound healing. The flow cytometry, colony formation assay and CCK8 were used to assess cell apoptosis and proliferation. The results showed that NMOF works as fluorescence quencher of the labeled p-DNA. In presence of the target miRNA (including miR-7, miR-21, and miR-155), p-DNA was able to rapidly hybridize and release from the NMOF leading to the recovery of fluorescence in living cells, which can be detected in real-time. Moreover, RT-PCR results demonstrated that the multiplexed miRNAs could be effectively downregulated after treatment of p-DNA@NMOF, resulting in the simultaneous suppression of oral cancer cell proliferation, anti-apoptosis, migration and invasion in vitro. In summary, our study suggests that p-DNA@NMOF possess great potential for combining early diagnosis and gene therapy of oral cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Pei Yu, Zehang Zhuang, Guihua Qiu, Haolin Chen, Yuying Zhao, Jinxiang Chen, Xiqiang Liu. Theranostical nanoscale metal-organic frameworks-mediated rapid multiplexed microRNA detection and highly therapeutic efficacy in living oral cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2190. doi:10.1158/1538-7445.AM2017-2190
Abstract 2200: Dual carfilzomib and doxorubicin carrying nanoparticles for synergistic efficacy in multiple myeloma
Authors: Bilgicer Basar , Kiziltepe Tanyel , Omstead David
Published: July 2017

We describe the synthesis and analysis of dual carfilzomib and doxorubicin loaded nanoparticles in their ability to deliver both drugs to multiple myeloma tumor cells at their optimal synergistic ratio. First, to identify the optimal synergistic ratio, various molar ratios of carfilzomib to doxorubicin were screened against multiple myeloma cell lines using the Chou-Talalay method. Both therapeutic agents were then incorporated into liposomes at the identified optimal synergistic ratio of 1:1 to achieve dual drug loaded nanoparticles with a narrow size distribution of ~100 nm and with high reproducibility. Our results established that the dual drug loaded nanoparticles exhibited synergy in vitro and were more efficacious in inhibiting tumor growth in vivo than a combination of free drugs, while at the same time reducing systemic toxicity. In conclusion, this study achieved the preclinical evaluation of dual drug loaded liposomes containing carfilzomib and doxorubicin for enhanced therapeutic efficacy to improve patient outcome in multiple myeloma.

Citation Format: Basar Bilgicer, Tanyel Kiziltepe, David Omstead. Dual carfilzomib and doxorubicin carrying nanoparticles for synergistic efficacy in multiple myeloma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2200. doi:10.1158/1538-7445.AM2017-2200
Abstract 2205: Identification of potential biomarker related to EGFR mutation by functional proteome profiling in primary non-small cell lung cancer
Authors: Hsu Yuan-Ling , Pan Szu-Hua
Published: July 2017

Non-small-cell lung cancer (NSCLC) accounts for approximately 85% of lung cancers which the leading cause of cancer-related death worldwide. Recently, epidermal growth factor receptor (EGFR) activating mutations have been proved relevant to NSCLC and let treatment of EGFR tyrosine kinase inhibitors (EGFR-TKIs) become an alternative therapy for NSCLC patients. Despite excellent clinical response to EGFR-TKIs, the intrinsic and acquired resistance to EGFR-TKI created challenges in clinical practice. Although several mechanisms may contribute to solve EGFR-TKIs resistance, there still have 20% to 30% of these patients that how the resistance developed remain unclear. Studies indicated that the increment of membrane efflux may be associated with drug resistance. In this study, we use differential membrane proteomic analysis to identify potential biomarkers for therapeutics of EGFR-TIKs resistance in NSCLC patients.Several primary NSCLC cell lines with different EGFR status were be isolated by our laboratory. Identification of modulators on membrane of primary cancer cells with different EGFR status was analyzed by membrane proteomic assay using LC-MS/ MS. The expression patterns of EGFR mutation cells would compare with that of wild type groups and the potential candidates would be selected by cross the results of interactome profiles of EGFR mutation cell lines. The expression patterns and functional role of these candidates were be examined in NSCLC cells by several molecular, cellular and biochemical analysis; and demonstrated the clinical relevance of these specific targets with EGFR mutation. The expressions of 100 modulators up-regulated in EGFR mutation cells compared with that of wild type group. After cross reacted with the 474 interaction proteins of mutated EGFR, 8 proteins were selected as potential candidates that related to EGFR-TKI resistance, especially the expression of a protein we called SHX in EGFR mutation cells. We demonstrated the real expression levels of SHX in different lung cancer cells, and performed network analysis of SHX and EGFR in translational level.The analysis of differential membrane proteomic found several novelbiomarkers and potential regulation mechanisms that related to the development of EGFR-TKIs resistance in NSCLC. These identification may provide us new direction for exploiting therapeutic strategy for NSCLC treatment in the future.

Citation Format: Yuan-Ling Hsu, Szu-Hua Pan. Identification of potential biomarker related to EGFR mutation by functional proteome profiling in primary non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2205. doi:10.1158/1538-7445.AM2017-2205
Abstract 2229: Prevention by tolfenamic acid and characterization of the molecular targets in Pirc and FAP colorectal adenomas
Authors: Ertem Furkan U. , Zhang Wenqian , Chang Kyle , Dashwood Wan-Mohaiza , Rajendran Praveen , Sun Deqiang , Abudayyeh Ala , Abdulrahim Maen , Vilar Eduardo , Dashwood Roderick H.
Published: July 2017

A critical need exists in sporadic and high-risk colorectal cancer patients for endoscopic screening combined with safe and effective preventive agents. To prioritize molecular targets for prevention studies in vivo, RNA-seq analyses were conducted on colorectal adenomas from familial adenomatous polyposis (FAP) patients, and from the Apc-mutant polyposis in rat colon (Pirc) model. Using a new murine endoscopic polypectomy methodology (F.U. Ertem et al., Gastrointest Endosc 2016;83:1272-6), we evaluated agents that might substitute for the current standard of care, Sulindac, and identified tolfenamic acid (TA) as a promising candidate for further investigation. At clinically-relevant doses in the Pirc model, TA suppressed tumorigenesis significantly both in the small intestine and in the colon. Inhibition by TA of colonic crypt cell proliferation, and the concomitant activation of apoptosis, coincided with reduced expression in target tissues of β-catenin, cyclin D1, Survivin and matrix metalloproteinase 7. Based on the oncogene candidates defined by RNA-seq analysis, endoscopic tracking revealed the time-course of tumor suppression by TA, and the temporal changes in S100a9, Nppb, Aldh1a3 and Mmp7, highlighting their value as potential early biomarkers for prevention in the clinical setting. As an “old drug” repurposed from migraine, TA appears to offer an exciting new therapeutic avenue in FAP and other high-risk colorectal cancer patient populations. This work was supported in part by NIH grants CA090890, CA122959, ES00210, ES023512, the John S. Dunn Foundation, and a Chancellor’s Research Initiative.

1

Citation Format: Furkan U. Ertem, Wenqian Zhang, Kyle Chang, Wan-Mohaiza Dashwood, Praveen Rajendran, Deqiang Sun, Ala Abudayyeh, Maen Abdulrahim, Eduardo Vilar, Roderick H. Dashwood. Prevention by tolfenamic acid and characterization of the molecular targets in Pirc and FAP colorectal adenomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2229. doi:10.1158/1538-7445.AM2017-2229
Abstract 2235: Preventive effects of the sodium glucose cotransporter 2 inhibitor tofogliflozin on liver tumorigenesis in obese and diabetic mice
Authors: Shirakami Yohei , Obara Koki , Ohnishi Masaya , Ideta Takayasu , Sakai Hiroyasu , Tanaka Takuji , Shimizu Masahito , Seishima Mitsuru
Published: July 2017

Obesity, diabetes mellitus, and their related metabolic abnormalities are associated with increased risk of hepatocellular carcinoma (HCC). Sodium glucose cotransporter (SGLT)-2 inhibitors, recently approved anti-diabetic agents, are expected to ameliorate the abnormalities associated with metabolic syndrome including non-alcoholic fatty liver disease (NAFLD). In this study, we investigated the effects of the SGLT2 inhibitor tofogliflozin on the development of NAFLD-related liver tumorigenesis in C57BL/KsJ-+Leprdb/+Leprdb (db/db) obese and diabetic mice. The direct effects of tofogliflozin on human HCC cell proliferation were also evaluated. Male db/db mice were administered diethylnitrosamine-containing water for two weeks and were treated with tofogliflozin throughout the experiment. Tofogliflozin was kindly provided by Kowa Co., Ltd. and the chemical structure will be disclosed at the time of presentation at the meeting. In mice treated with tofogliflozin, the development of hepatic pre-neoplastic lesions was markedly suppressed, and hepatic steatosis and inflammation significantly reduced, as evaluated using the NAFLD activity score, in comparison to those in the control mice. Serum levels of glucose and free fatty acid and mRNA expression levels of pro-inflammatory markers in the liver were reduced by tofogliflozin treatment. Conversely, the proliferation of SGLT2 protein-expressing HCC cells was not inhibited by this agent. These findings suggest that tofogliflozin suppressed the early phase of obesity- and NAFLD-related liver carcinogenesis by attenuating chronic inflammation and steatosis in the liver, while the agent had no significant direct effect on the proliferation of HCC cells. Therefore, SGLT2 inhibitors may have a chemopreventive effect on obesity-related HCC.

Citation Format: Yohei Shirakami, Koki Obara, Masaya Ohnishi, Takayasu Ideta, Hiroyasu Sakai, Takuji Tanaka, Masahito Shimizu, Mitsuru Seishima. Preventive effects of the sodium glucose cotransporter 2 inhibitor tofogliflozin on liver tumorigenesis in obese and diabetic mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2235. doi:10.1158/1538-7445.AM2017-2235
Abstract 2261: Synergistic effect and VEGF/HSP70-hom haplotype analysis: Relationship to prostate cancer risk and clinical outcome
Authors: Sfar Sana , Saad Hamadi , Mosbah Faouzi , Chouchane Lotfi
Published: July 2017

Prostate cancer (PCa) is a complex disorder resulting from the combined effects of multiple environmental and genetic factors. Our previous single-locus analysis showed that VEGF and HSP70-hom polymorphisms were significantly associated with PCa susceptibility and prognosis. Both genes encoding these proteins were located on chromosome 6p21, and combining the neighboring single nucleotide polymorphisms (SNPs) into haplotypes may increase the association with the disease. Three tagging polymorphisms, the HSP70-hom 2437 T/C, the VEGF-1154 G/A, and the VEGF-634 G/C SNPs were genotyped in 101 cases and 80 controls. For the combined analysis of VEGF and HSP70-hom, we found a positive gradient in the odds ratios (ORs) related to the number of high-risk genotypes with a 3.53-fold increase of prostate carcinoma risk (OR= 3.53; p= 0.015). Furthermore, the TAG and CAG haplotypes at positions HSP70-hom, VEGF-1154 and VEGF -634 exhibited a two-fold (OR= 0.46; p= 0.014) and a seven-fold (OR= 0.14; p= 0.00005) reduction in PCa risk, respectively. Regarding PCa prognosis, the TAG haplotype had a negative association with the aggressive phenotype as defined by the histopathological grade (OR= 0.28; p= 0.006). Our findings confirm the role of at-risk haplotype across the HSP70-hom/VEGF gene cluster in determining susceptibility to PCa.

Note: This abstract was not presented at the meeting.

Citation Format: Sana Sfar, Hamadi Saad, Faouzi Mosbah, Lotfi Chouchane. Synergistic effect and VEGF/HSP70-hom haplotype analysis: Relationship to prostate cancer risk and clinical outcome [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2261. doi:10.1158/1538-7445.AM2017-2261