bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 465: Downregulation of miR-193a and its correlation with clinical and pathological behavior of colorectal cancer
Authors: Mamoori Afraa M. , Wahab Riajul , Islam Farhadul , Lee Katherine , Smith Robert A. , Gopalan Vinod , Lam Alfred Ky
Published: July 2017

Aim: Deregulation of miR-193a expression was reported to have significant roles in cancer development and progression. This study aimed to investigate the expression pattern and clinicopathological implication of miR-193a-3p in patients with colorectal cancer. Also, the bio-physiological effects of miR-193a in colon cancer cells were examined.

Methodology: Fresh frozen tissues from 70 matched colorectal cancers (adenocarcinomas) and adjacent noncancer tissue were prospectively collected with no selection bias. Expression level of miR193a-3p was measured by quantitative real-time polymerase chain reaction (qRT-PCR). Immunohistochemical staining was used to detect the expression level of K-ras protein as a predicted downstream target for miR-193a. Restoration of the miR-193a level in the cell lines was achieved by permanent transfection. Multiple functional and immunological assays were performed to analyze miR-193’s cellular implications and target affinity in colon cancer cell lines.

Results: Downregulation of miR-193a-3p was noted in the majority of the colorectal cancer tissues when compared to nontumor colorectal tissues (70%, n= 49/70). In addition, high prevalence of miR-193a downregulation was noted in early stage colon cancer tissues and cell lines (SW480) when compared to advanced staged colorectal carcinomas. Immunohistochemical analysis revealed inverse correlation between miR-193a and K-ras expression in which high K-ras protein expression was reported in 28/49 cases with miR-193a downregulation. Analogues findings were also observed in cell lines using Western blot (P value< 0.05). Overexpression of miR-193a in colon cancer cell lines resulted in reduced cell proliferation, increased apoptosis and a less accumulation of cells in the G0-G1 phase cell cycle.

Conclusion: The current study confirms the downregulation of miR-193a-3P in colorectal cancer and its tumour suppressor effects in vitro plays a key role in the modulation of K-ras regulated molecular signalling pathways in colon cancers

Note: This abstract was not presented at the meeting.

Citation Format: Afraa M. Mamoori, Riajul Wahab, Farhadul Islam, Katherine Lee, Robert A. Smith, Vinod Gopalan, Alfred Ky Lam. Downregulation of miR-193a and its correlation with clinical and pathological behavior of 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 465. doi:10.1158/1538-7445.AM2017-465
Abstract 4659: Detection of autoantibodies against tumor-associated antigens (TAAs) improving immunodiagnosis in human osteosarcoma by serological proteome analysis
Authors: Li Jitian , Huang Manyu , Luo Manli , Dai Liping , Xie Wen , Qin Xiaofei , Han Zongchang , Li Wuyin , Zhang Jianying
Published: July 2017

Osteosarcoma (OS) is the most common malignant primary solid bone-tumor. Despite its relatively low incidence rate among overall cancers, it remains one of the most harmful primary malignant tumors in childhood and adolescence. Thus, a critical need in the diagnosis and management of OS is to determine an optimal combination of clinical biomarkers that could detect tumors early with high specificity/sensitivity and with limited invasiveness. The objective of this study is to identify and characterize the targeted tumor-associated antigens (TAAs) as biomarkers in OS by serological proteome analysis (SERPA) approach, and further to analyze the frequency and specificity of anti-TAA antibodies in OS patients. In this initial study, autoantibodies to 29 TAAs were detected by ELISA and Western blotting in 90 sera from patients with OS, Osteochondroma (OC), and age matched normal individuals. Only 8 protein antigens including DSF70, HMGB1, HCC1, RalA, c-myc, AnnexinA1, IMP1, PBP, can induce significantly higher antibody responses in OS patients compared to normal individuals, achieving the highest sensitivity and specificity, 66% and 95%, respectively. The cumulative positive rate of autoantibodies against these eight selected TAAs in OS reached 70.7% with an observed AUC of 0.972 (95% CI: 0.867-0.988), significantly higher than that in normal control sera. Positive results were also confirmed by Western blotting. These preliminary data extensively evidence that not all proteins identified in cancer can be used as TAA biomarkers in OS, and only some of these proteins can induce immune responses, which could be potential TAAs in OS immunodiagnosis or prognosis, and further studies of novel targeted proteins in OS are currently in progress by SERPA approach. In addition, it supports the hypothesis that a customized TAA array can be used for enhancing anti-TAA antibodies detection, and it may constitute a promising and powerful tool for immunodiagnosis of OS.

Citation Format: Jitian Li, Manyu Huang, Manli Luo, Liping Dai, Wen Xie, Xiaofei Qin, Zongchang Han, Wuyin Li, Jianying Zhang. Detection of autoantibodies against tumor-associated antigens (TAAs) improving immunodiagnosis in human osteosarcoma by serological proteome analysis [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 4659. doi:10.1158/1538-7445.AM2017-4659
Abstract 466: Role of miR-15b-3p in mitoquinone induced autophagy of breast cancer cells
Authors: Mascia Francesca , Pokrzywinski Kaytee , Rao Ashutosh
Published: July 2017

Mitoquinone, a redox-active mitochondrially targeted ubiquinone, activates autophagy more potently in breast cancer cells (MDA-MB-231) than in normal breast cells (MCF-12A). Mitoquinone induces the production of reactive oxygen species (ROS) and a concomitant activation of the Nrf2-dependent antioxidant response. MicroRNA are endogenous small double stranded RNA that bind to the 3’ UTR of several target mRNAs and function through translational repression or mRNA degradation. The goal of this study was to evaluate microRNA as a link between oxidative stress and autophagy signaling. We performed RNA sequencing analysis of the microRNAs modulated by mitoquinone treatment of MDA-MB-231 cells. miRNAs were identified using Illumina RNA sequencing. DEseq analysis revealed 97 miRNAs that were differentially expressed at an α < 0.05. Within this set, 15 miRNAs were down-regulated by stimulation with mitoquinone making them attractive as potential autophagy inhibitors. We characterized the role of a miR-15b-3p by transfecting the cells with miR-mimic and anti-miR before inducing an autophagy response. Western blot analysis showed a reduction in the expression of the autophagy marker LC3, the autophagosome chaperon p62 and the transcription factor Nrf2 in presence of the miR-15b-3p mimic. Conversely, miR-15b-3p mimic pre-treatment increased the expression of the cell cycle inhibitor p27 both at the basal level and after mitoquinone stimulation for 48 hrs. The administration of the anti-miR-15b-3p did not change the expression level of these proteins suggesting that miR-15b-3p may work with other miRs toward regulating autophagy. Combining the action of miR-15b-3p mimic with redox active chemotherapeutic drugs could represent a novel strategy for breast cancer therapy by reducing survival signaling through autophagy.

Citation Format: Francesca Mascia, Kaytee Pokrzywinski, Ashutosh Rao. Role of miR-15b-3p in mitoquinone induced autophagy of breast 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 466. doi:10.1158/1538-7445.AM2017-466
Abstract 4673: EZH2 as a therapeutic target in glioblastoma: a cellular and molecular study
Authors: Rosa Javier de la , Iraburu Marta , Gallo-Oller Gabriel , Urdiciain Alejandro , Fan Xing , Shahi Mehdi H. , Meléndez Bárbara , Rey Juan A. , Idoate Miguel A. , Castresana Javier S.
Published: July 2017

Glioblastoma is the most common and deadliest malignant brain tumor in adults. It is currently treated with a combination of surgery, radiotherapy and chemotherapy with temozolomide (TMZ). Resistance to TMZ is a challenge in the treatment of this type of brain cancer. Epigenetic modulation is a new possibility to be tested against glioblastoma. Inhibition of EZH2, a histone methyltransferase (H3K27met3) which is overexpressed in cancer cells, leading to angiogenesis and metastasis, might be a new strategy to treat this tumor. In this work, the EZH2 inhibitor DZNeP was tested in A172 glioblastoma cells and in A172-TMZ-resistant glioblastoma cells. Inhibition of cell proliferation, cell adhesion, colony formation, and cell migration was noted in A172 and A172-TMZ-resistant glioblastoma cells after DZNeP treatment. DZNeP also decreased EZH2 expression, and increased the expression of its target genes (E-cadherin and TIMP3), which might probably contribute to inhibiting the development of a cancer metastatic phenotype. Finally, DZNeP negatively regulated the TGFβ pathway: the expression of receptors involved in this pathway (TGFβR1 and TGFβR2) was decreased, and the expression of inhibitors of this pathway (BAMBI, SMAD6 and SMAD7) was increased. In order to support the results obtained by qRT-PCR about the TGFβ1 pathway, protein expression of SMAD2 and PSMAD2 was analyzed by Western blot. The ratio between PSMAD2/SMAD2 decreased in cells treated with DZNeP, which supports the possible therapeutic effect of this drug against the TGFβ pathway, as PSMAD2 is a negative prognostic marker in glioblastoma. In conclusion, we propose that inhibition of EZH2 might be considered as a therapeutic strategy against glioblastoma.

Citation Format: Javier de la Rosa, Marta Iraburu, Gabriel Gallo-Oller, Alejandro Urdiciain, Xing Fan, Mehdi H. Shahi, Bárbara Meléndez, Juan A. Rey, Miguel A. Idoate, Javier S. Castresana. EZH2 as a therapeutic target in glioblastoma: a cellular and molecular study [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 4673. doi:10.1158/1538-7445.AM2017-4673
Abstract 468: Evaluation of microRNA inhibition in medulloblastoma
Authors: Vavra Alexander , Stoops Sydney , White Jonathan
Published: July 2017

Medulloblastoma is the most common pediatric malignant brain tumor. The current treatment plan consists of a combination of surgery, radiation, and chemotherapy which results in an 80% five year survival rate. However, the survival rate does not take into account the developmental delays and learning disabilities inherited as a result of the aggressive treatment plan on a developing brain. As such the development of new treatment options is needed. While many protein targets are currently being explored for therapeutic intervention, the role of non-coding RNAs is of increasing interest. Specifically microRNA profiling in medulloblastoma has been performed by others to highlight expression profiles which can be mapped to the disease state and also to potentially identify associated protein targets. Moreover, it is particularly intriguing that microRNAs themselves may be directly targeted as therapeutic points of intervention. With this in mind, our lab is currently investigating methods for directly targeting microRNAs in medulloblastoma and we have recently validated several upregulated miRNAs for further investigation. Specifically, we evaluated the expression profile eighteen selected miRNAs in DAOY and D341 cell lines. Our results indicate that four miRNAs (miR-196a, miR-183, miR-96, and miR-182) are statistically overexpressed in both cell lines. Results of these findings and our initial efforts to target these selected miRNAs will be described.

Citation Format: Alexander Vavra, Sydney Stoops, Jonathan White. Evaluation of microRNA inhibition in medulloblastoma [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 468. doi:10.1158/1538-7445.AM2017-468
Abstract 4681: Histone deacetylase 3 inhibition overcomes BIM deletion polymorphism-mediated osimertinib resistance in EGFR-mutant lung cancer
Authors: Tanimoto Azusa
Published: July 2017

Purpose:The BIM deletion polymorphism is associated with apoptosis resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), such as gefitinib and erlotinib, in non-small cell lung cancer (NSCLC) harboring EGFR mutations. Here, we investigated whether the BIM deletion polymorphism contributes to resistance against osimertinib, a third-generation EGFR-TKI. In addition, we determined the efficacy of a histone deacetylase (HDAC) inhibitor, vorinostat, against this form of resistance and elucidated the underlying mechanism.

Experimental Design:We used EGFR-mutated NSCLC cell lines which were either heterozygous or homozygous for the BIM deletion polymorphism to evaluate the effect of osimertinib in vitro and in vivo. Protein expression was examined by western blotting. Alternative splicing of BIM mRNA was analyzed by RT-PCR.

Results:EGFR-mutated NSCLC cell lines with the BIM deletion polymorphism exhibited apoptosis resistance to osimertinib in a polymorphism dosage-dependent manner, and this resistance was overcome by combined use with vorinostat. Experiments with homozygous BIM deletion-positive cells revealed that vorinostat affected the alternative splicing of BIM mRNA in the deletion allele, increased the expression of active BIM protein, and thereby induced apoptosis in osimertinib-treated cells. These effects were mediated predominantly by HDAC3 inhibition. In xenograft models, combined use of vorinostat with osimertinib could regress tumors in EGFR-mutated NSCLC cells homozygous for the BIM deletion polymorphism. Moreover, this combination could induce apoptosis even when tumor cells acquired EGFR-T790M mutations.

Conclusions:These findings indicate the importance of developing HDAC3-selective inhibitors, and their combined use with osimertinib, for treating EGFR-mutated lung cancers carrying the BIM deletion polymorphism.

Note: This abstract was not presented at the meeting.

Citation Format: Azusa Tanimoto. Histone deacetylase 3 inhibition overcomes BIM deletion polymorphism-mediated osimertinib resistance in EGFR-mutant 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 4681. doi:10.1158/1538-7445.AM2017-4681
Abstract 4684: MYC inhibition in human malignancy by BET-Bromodomain inhibitor JQ1, using a 3-D human tumor primary culture micro-spheroid platform to explore clinically relevant observations
Authors: Nagourney Robert Alan , Evans Steven S. , Nagourney Alexander J. , Bernard Paula J. , Francisco Federico R. , Sheth Milan , Vora Nilesh , Deo Eknath
Published: July 2017

The transcription factor MYC is associated with the regulation of over 15% of human gene expression. Deregulation of MYC is found in up to 70% of human cancers. As MYC is responsible for the reprogramming of cell metabolism, growth and survival, it is an attractive therapeutic target, yet effective drug development has been slow. As the Bromodomain/BET proteins epigenetically regulate MYC, BET inhibitors like JQ1 have been developed as MYC therapeutics. We used the Ex Vivo Analysis of Programmed Cell Death (EVA/PCD) previously shown to correlate with response, time to progression and survival, to examine JQ1 activity in 46 human tumor specimens isolated from surgical biopsies.

Methods: Surgical biopsies were mechanically and enzymatically disaggregated, with micro spheroids of desired size isolated by density centrifugation. Five-Point dose response curves were interpolated to provide LC50 values for comparison by Z-score. Synergy was conducted by median-effect.

Results: JQ1 revealed activity in hematologic, breast, glial & gastrointestinal (GI) tumors. By rank order, JQ1 activity favored hematologic (AML & NHL) & breast tumors over Lung, Ovary and GI with inter-patient differences observed within tumor types. MYC association with histone acetylation and aurora kinase (AURK) activity, led to an exploration of JQ1 synergy with SAHA (HDACi), MLN 8054 & Tozasertib (both AURKi). JQ1 synergy analysis revealed SAHA > Tozasertib > MLN 8054.

Conclusions: Based upon JQ1 activity, several tumor types appear to be candidates for agents that target MYC-mediated cell survival, offering potential treatment options for patients with advanced malignancies. JQ1 synergy with HDACi & AURKi suggests novel drug combinations for future development. As MYC signaling influences thousands of gene targets, phenotypic analyses like the EVA/PCD platform can offer insights into cellular cell death events that serve as surrogate markers for clinical response to MYC inhibition. Supported in part by the Vanguard Cancer Foundation, the Nagourney Institute and the Malcolm C. Todd Cancer Institute.

Citation Format: Robert Alan Nagourney, Steven S. Evans, Alexander J. Nagourney, Paula J. Bernard, Federico R. Francisco, Milan Sheth, Nilesh Vora, Eknath Deo. MYC inhibition in human malignancy by BET-Bromodomain inhibitor JQ1, using a 3-D human tumor primary culture micro-spheroid platform to explore clinically relevant observations [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 4684. doi:10.1158/1538-7445.AM2017-4684
Abstract 4691: Polarization of macrophages with STING agonists
Authors: Henare Kimiora L. , Tijono Sofian M. , Ching Lai-Ming
Published: July 2017

The functional plasticity of macrophages and their ability to support tumour growth or promote potent antitumor immunity make them an attractive target cancer therapy. Macrophages have been implicated in the complex antitumor activity of murine STING agonist, DMXAA, including disruption of tumor vasculature, induction of cytokine production, and promotion of durable antitumor immune responses. Many of these activities are lost in STING-/- mice, and DMXAA does not bind to human STING, providing a pertinent explanation for why the anti-tumour effects of DMXAA observed in mice were not recapitulated human clinical trials. Using PMA-differentiated THP-1 macrophages, we sought to identify human active analogues of DMXAA in vitro, comparing their capacity to induce cytokines indicative of a classically activated antitumour phenotype; the non-canonical STING agonist 2'3'-cGAMP was also included in our investigation. IP-10 (CXCL10) production, measured by ELISA, was used as an initial marker for activity, followed by multiplex cytokine analysis for broader characterisation compared to M(IFN-γ/LPS), M(IL-4/IL-13), and ‘resting’ M(media alone) macrophage phenotypes. Parallel experiments were also conducted with RAW264.7 murine macrophages to compare the DMXAA-induced cytokine profile directly with that of 2'3'-cGAMP. 2'3'-cGAMP induced 22-fold and 18-fold increases in IP-10 production in resting and M(IL-4/IL-13) THP-1 macrophages, respectively. By comparison, THP-1 macrophages were much less responsive to DMXAA and analogues tested to date. Furthermore, 2'3'-cGAMP induced a spectrum of cytokines similar to that observed in M(IFN-γ/LPS) macrophages, and markedly different to that of M(IL-4/IL-13) THP-1 macrophages. DMXAA, and analogues previously shown to have cytokine-inducing activity in human leukocytes did not produce cytokine spectra indicative of a classic M(IFN-γ/LPS) phenotype. Unlike human cells, however, cytokine spectra produced by RAW264.7 murine macrophages were comparable between DMXAA- and 2'3'-cGAMP-treated cells. These findings support previous data that suggest that STING agonists can promote macrophage polarisation towards an antitumour phenotype. While DMXAA is active in murine macrophages, only 2'3'-cGAMP can induce in human THP-1 cells, an M(IFN-γ/LPS)-like spectrum of cytokines often attributed to an antitumour macrophage phenotype.

Citation Format: Kimiora L. Henare, Sofian M. Tijono, Lai-Ming Ching. Polarization of macrophages with STING agonists [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 4691. doi:10.1158/1538-7445.AM2017-4691
Abstract 4696: Development of an inhaled TLR9 agonist for the immunotherapy of lung cancer
Authors: Gallotta Marilena , Assi Hikmat , Coffman Robert L. , Guiducci Cristiana
Published: July 2017

CpG-oligonucleotides (CpG-ODN) stimulate innate immune responses through Toll-like receptor-9 (TLR9). Intratumoral (IT) administration of CpG-ODN has been shown to be safe and clinically effective in humans and in multiple animal species. An important feature of local administration of a CpG-ODN is the ability to stimulate immune responses that control the growth of distant, uninjected tumors. Such treatment is especially effective when combined with a complementary mode of immunotherapy, such as anti-PD-1 or radiation. However, repeated IT injection is not practical for many tumor types, including most lung cancers. In mild asthmatic patients and healthy subjects, CpG-ODN have demonstrated safety and pharmacologic activity when delivered by inhalation directly to the lungs. Thus, inhaled CpG presents an alternative to IT injection for primary lung cancers and for other tumor types with prominent lung metastases. In the 4T1 mouse model of metastatic breast cancer, treatment of mice with a CpG-ODN administered intranasally led to a marked reduction in the number of lung metastases. This reduction was accompanied by infiltration of tumors with activated CD8+ and CD4+ T cells and the formation of highly organized tertiary lymphoid structures. These changes are similar to those observed after IT injection of solid tumors growing subcutaneously, and are consistent with changes observed in human tumors responsive to immunotherapy. In the 4T1 model, intranasal CpG-ODN strongly synergized with systemic anti-PD-1 treatment and induced complete clearance of lung tumors, as well as metastases in tissues not exposed to the CpG-ODN, including liver, pancreas and bone. This generation of systemic immunity resulted in long-term survival in the majority of mice. Immune cell subset depletion studies implicate both CD8 and CD4 T cells in the rejection of lung tumors treated with inhaled CpG-ODN plus anti-PD1. Dynavax has developed a CpG-ODN, DV281, that is optimized for aerosol delivery and plans to initiate clinical studies in NSCLC patients in 2017.

Citation Format: Marilena Gallotta, Hikmat Assi, Robert L. Coffman, Cristiana Guiducci. Development of an inhaled TLR9 agonist for the immunotherapy of 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 4696. doi:10.1158/1538-7445.AM2017-4696
Abstract 4707: Evaluation of efficacy and immune response to PD1 checkpoint inhibition in human immune-reconstituted mice using patient-derived xenograft models
Authors: Izadi Hooman , Broudy Tommy , Yan Deborah , Thatte Jayant
Published: July 2017

Background: Human Immune-reconstituted Mice, generated by transplanting pre-validated human CD34+ hematopoietic stem cells (HSC) into immune-compromised mice, are extremely useful in basic and applied human disease research. Many cancer therapies, including immune checkpoint inhibitors such as the anti-PD-1 monoclonal antibody, rely on an intact immune system to release immunosuppression and destroy cancer cells.

Material and methods: We evaluated the effects of anti-PD-1 in humanized mice in donor cohorts utilizing the cancer matrix of 5 HSC donor cohorts of humanized animals with PDX models from 7 small cell lung cancer (SCLC). We also tested anti-PD-1 in 1 triple negative breast cancer (TNBC) model and in 1 MDA-MB-231 study.

Results: Here, we show that HSC derived human immune cell engraftment does not have significant effects on patient derived xenograft (PDX) tumor growth. However, humanization is required for anti-tumor response to anti-PD-1. In MDA-MB-231 study, tumors showed complete regression after the first round of treatment. Re-engraftment of same animals with MDA-MB-231 cells showed minor tumor growth before complete regression again. Aged match control naïve animals engrafted with same cells showed normal tumor growth. In SCLC studies anti-PD-1 efficacy varied between the models, which clearly underscores the inherent donor to donor variability in this humanized model system. Tumors from these studies were then analyzed to determine target engagement of anti-PD-1, tumor infiltrating lymphocytes (TILs) characterization and histology comparing anti-PD-1 responders vs non-responders. As expected, in this study, we demonstrate humanization of the animals is necessary to evaluate the efficacy of anti-PD-1 therapy and can elicit an immunotherapy mediated anti-tumor effect.

Conclusion: Here we demonstrate the development of humanized mouse studies with ability to run multi point analysis to determine the outcome of treatment.

Citation Format: Hooman Izadi, Tommy Broudy, Deborah Yan, Jayant Thatte. Evaluation of efficacy and immune response to PD1 checkpoint inhibition in human immune-reconstituted mice using patient-derived xenograft models [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 4707. doi:10.1158/1538-7445.AM2017-4707
Abstract 4713: Deubiquitination and stabilization of programmed cell death-ligand 1 by CSN5
Authors: Lim Seung-Oe , Li Chia-Wei , Hung Mien-Chie
Published: July 2017

Chronic inflammation in cancer is often associated with disease aggressiveness. Pro-inflammatory cytokines produced in the tumor microenvironment lead to eradication of antitumor immunity and enhanced tumor cell survival. In the current study, we identified TNFα as a major factor triggering cancer cell immunosuppression against T cell surveillance via stabilization of programmed cell death-ligand 1 (PD-L1). We demonstrated that COP9 signalosome 5 (CSN5), induced by NF-κB p65, is required for TNFα-mediated PD-L1 stabilization in cancer cells. CSN5 inhibits the ubiquitination and degradation of PD-L1. The role of CSN5 in modulating PD-L1 stabilization prompted us to use CSN5 inhibitor, curcumin, to show that it could destabilize PD-L1. Curcumin reduced CSN5 activity to attenuate TNFα-mediated PD-L1 stabilization and enhanced anti-tumor immunity. Preclinical data demonstrated inhibition of CSN5 sensitized cancer cells to anti-CTLA4 therapy, suggesting CSN5 inhibitor may be a useful adjuvant to enhance immune-based therapies.

Note: This abstract was not presented at the meeting.

Citation Format: Seung-Oe Lim, Chia-Wei Li, Mien-Chie Hung. Deubiquitination and stabilization of programmed cell death-ligand 1 by CSN5 [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 4713. doi:10.1158/1538-7445.AM2017-4713
Abstract 4728: Plasma interleukin-6 level predicts prognosis of patients who received sorafenib for advanced hepatocellular carcinoma
Authors: Shao Yu-Yun , Lin Hang , Li Yong-Shi , Lee Ying-Hui , Chen Ho-Min , Cheng Ann-Lii , Hsu Chih-Hung
Published: July 2017

Background: No biomarker has been proven to predict the treatment outcomes of sorafenib, the only approved first-line therapy for advanced hepatocellular carcinoma (HCC). We explored predictive and prognostic values of plasma interleukin (IL)-6 levels for patients who received sorafenib as first-line therapy for advanced HCC.

Methods: This study had 2 patient cohorts. The exploration cohort consisted of patients who were previously enrolled in a phase 2 clinical trial that examined sorafenib with tegafur/uracil as first-line therapy for advanced HCC. The validation cohort consisted of patients who received sorafenib alone as first-line therapy for advanced HCC under reimbursement of National Health Insurance of Taiwan. Pretreatment plasma IL-6 levels were determined. We used the receiver operating characteristic (ROC) curve in the exploration cohort to determine the best cut point for IL-6 levels to predict overall survival (OS). We then confirmed the cut point in the validation cohort.

Results: There were 55 and 73 patients in the exploration and validation cohort, respectively. In the exploration cohort, there was no complete response but 3 (6%) partial response. The median OS was 8.1 months. In the validation cohort, there was no complete response but 7 (10%) partial response. The median OS was 10.3 months. In the exploration cohort, we found 4.28 pg/ml was the best cut point in defining high and low IL-6 levels because it could differentiate OS (p = 0.042) with the best sensitivity and specificity. Applying the cut point on the validation cohort, patients with high pretreatment plasma IL-6 levels, compared with patients with low IL-6 levels, exhibited significantly poorer OS (median, 8.0 vs. 13.9 months, p = 0.031). After adjusting for age, gender, hepatitis etiology, tumor characteristics, α-fetoprotein level, performance status, and prior treatment, a high plasma IL-6 level remained an independent predictor for poor OS (hazard ratio 2.594, p = 0.005). By contrast, the plasma IL-6 level was not associated with progression-free survival, treatment response, disease control, or other patient characteristics.

Conclusion: High pretreatment plasma IL-6 level predicted prognosis of patients who received sorafenib as first-line therapy for advanced HCC.

Citation Format: Yu-Yun Shao, Hang Lin, Yong-Shi Li, Ying-Hui Lee, Ho-Min Chen, Ann-Lii Cheng, Chih-Hung Hsu. Plasma interleukin-6 level predicts prognosis of patients who received sorafenib for advanced hepatocellular carcinoma [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 4728. doi:10.1158/1538-7445.AM2017-4728
Abstract 4747: Subcutaneous fat predict outcome in head and neck cancer
Authors: Tsang Ngan M.
Published: July 2017

Background: Previous studies have shown that body mass index (BMI) is linked to survival in cancer patients. Our objective was to analyze whether body composition before treatment can predict outcomes in patients with head-and neck cancer.

Methods: All 881 patients with loco-regional head-and -neck cancer treated with curative-intent RT during 2005 to 2012 were retrospectively studied. Body composition analyses by using pre-RT CT images acquired for RT simulation. We compared subcutaneous adipose tissue (SAT) index and muscle index by measuring cross-sectional area of tissues at level of the third thoracic vertebrae (T3) in both genders. Overall survival (OS), loco-regional control (LRC), and distant metastasis-free survival (DFS) were analyzed by body composition index that divided into two groups based on values higher and lower than the median. The median follow-up was 4.68 years.

Results: SAT index in female patients was significantly higher than that in male (p <0.001). The median SAT index and muscle index for female were 15.25 cm2/m2 (range, 0.93-88.09 cm2/m2) and 34.3 cm2/m2 (range, 14.78-73.05 cm2/m2), and for male were 6.19 cm2/m2 (range, 0.21-40.48 cm2/m2) and 51.74 cm2/m2 (range, 8.03-89.06 cm2/m2). The 5-year and 10-year OS, LRC and DFS were 66.3%, 57.6%, 73.4%, 71.4%, 83%, and 82.1%. Higher pre-treatment SAT index was associated with longer OS (HR = 0.616; P = 0.001) and DFS (HR = 0.649; P = 0.015) and a trend towards longer LRC (HR = 0.772; P = 0.066). Pre-treatment muscle index was also a significant predictor of OS (HR = 0.662; P = 0.006) and LRC (HR = 0.684; P = 0.029).

Conclusion: SAT index and muscle index of T3 level obtained from CT images appear to predict outcomes in patients with curative head-and-neck cancers.

Note: This abstract was not presented at the meeting.

Citation Format: Ngan M. Tsang. Subcutaneous fat predict outcome in head and neck 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 4747. doi:10.1158/1538-7445.AM2017-4747
Abstract 4751: Correlating the genomic and transcriptomic profiles of pre-treatment cervical tumor biopsies with patient recurrence after definitive chemoradiation
Authors: Ruiz Fiona J. , Schwarz Julie K.
Published: July 2017

Purpose: The purpose of this study was to identify genomic mutations that correlate with patient recurrence after curative intent chemoradiation therapy in advanced stage cervical cancer patients.

Methods: Pre-treatment cervical tumor biopsies and normal blood were prospectively collected from patients treated at Washington University in St. Louis. DNA and RNA was isolated from the fresh frozen tumor biopsies and sent for next-generation sequencing and RNAseq respectively. Targeted exome sequencing was done using a panel of 127 known cancer associated genes, and an additional 8 genes previously reported to be mutated in cervical cancer. Gene tiling was done to probe for 30 different HPV subtypes.

Results: The mutational signatures of single nucleotide polymorphisms were an APOBEC cytidine deaminase and CpG site cytidine deamination mutagenesis pattern. The non-silent somatic mutations were analyzed by online prediction algorithms to assess the mutation’s putative effect on protein function. Univariate COX regression was used to correlate patient recurrence with non-silent putatively functional mutations. In our patient cohort 88% of samples were HPV positive, with 74% being the high-risk HPV 16 and 18 subtypes. Patient’s with HPV positive tumors had increased progression-free survival when compared to the HPV negative patient tumors; and the HPV positive patients with HPV 16 had increased progression-free survival over the other HPV subtypes. Complete gene tiling of the HPV 16 and 18 subtypes allowed us to determine whether the HPV DNA was integrated in the host genome or retained episomally. In our patient cohort less than 30% of samples had integrated HPV DNA, and there was no significant difference in progression-free survival between patients with integrated or episomal HPV.

Conclusions: The genomic analysis of our advanced staged cervical cancer patient cohort found that epigenetic modifiers were frequently mutated and that cytidine deamination mutagenesis was primarily responsible for the generation of the SNP mutations. Our study identified NSD1 mutations as being correlated with tumor recurrence, indicating that histone modifications and chromatin structure may play an important role in tumor response to ionizing radiation therapy. Gene expression analysis is currently ongoing.

Citation Format: Fiona J. Ruiz, Julie K. Schwarz. Correlating the genomic and transcriptomic profiles of pre-treatment cervical tumor biopsies with patient recurrence after definitive chemoradiation [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 4751. doi:10.1158/1538-7445.AM2017-4751
Abstract 476: MicroRNA mediated regulation of ATF5 contributes to homeostasis and benign to malignant transformation in breast cancer cells
Authors: Gaither Kari Ann , Madarampalli Bhanupriya , Liu David X.
Published: July 2017

The transcription factor ATF5 modulates survival, proliferation, differentiation, and homeostasis. In unstressed conditions, ATF5 has a short half-life and is rapidly degraded due to post-translational modifications. Conversely, ATF5 is upregulated in cells under cellular stress. Furthermore, we have found that ATF5 is elevated and is a survival factor in transformed C6 glioma and MCF7 breast cancer compared to non-transformed cells. Regulation of ATF5 expression is not fully understood. We hypothesized that microRNA (miRNA) play a role in regulating the expression of ATF5 at the 3’ UTR and sought to better understand the role of ATF5 in the transformation to a malignant cell phenotype. To date, no studies have examined the regulation of ATF5 by miRNA. MiRNAs are endogenous small non-coding RNAs 20-25 nucleotides in length that contribute to regulation of gene expression at the translational level. We used in silico modeling programs to identify miRNAs predicted to bind to the 3’ UTR of ATF5. We then aimed to identify the presence of specific miRNA and their ability to downregulate ATF5 during cellular stress and other physiological conditions in vitro. Luciferase reporter assays and immunoprecipitations of ATF5 3’ UTR mRNA and cell lysate were performed and microRNA quantity analyzed via qPCR analysis. Subsequently, transfections of precursor microRNA were carried out in human MCF10A breast epithelial, MCF7 mammary epithelial, and breast adenocarcinoma MDA-MB-231 cell lines, and expression levels of ATF5 were measured under varying physiological conditions via Western Blot analysis. In vitro migration and invasion assays were performed and transformation was studied via MCF10A mammary epithelial cells with inducible SRC. We demonstrate that miRNA are bound to the 3’ UTR of ATF5 and that miRNAs 433-3p and 520b help to regulate the expression of ATF5 under varying stress conditions and at steady state in MCF10A, MCF7, and MDA-MB-231 cells. Additionally, preliminary data indicate that ATF5 is upregulated during transformation of MCF10A cells with inducible SRC and is overexpressed in MDA-MB-231 cells as compared to MCF7 and MCF10A cell lines, and suggest that ATF5 may enhance migration and invasion. Further studies are necessary to elucidate the role of ATF5 in transformation, migration, and invasion. Better understanding of the regulation of ATF5 could have implications in a broad range of human malignancies.

Citation Format: Kari Ann Gaither, Bhanupriya Madarampalli, David X. Liu. MicroRNA mediated regulation of ATF5 contributes to homeostasis and benign to malignant transformation in breast 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 476. doi:10.1158/1538-7445.AM2017-476
Abstract 4761: BCL2 induced by LAMTOR3-MAPK is a druggable target of chemoradioresistance in mesenchymal lung cancer
Authors: Kwon Ok-Seon , Hong Soon-Ki , Cha Hyuk-Jin
Published: July 2017

Metastasis and chemoresistance, which are main causes of lung cancer related death, have been major interest in cancer research. Recently, Epithelial-Mesenchymal Transition (EMT) a foremost process for acquiring metastatic properties has been demonstrated to be strongly link to chemoresistance. Due to limited understanding of mechanism for EMT mediated chemoresistance, druggable targets for chemoresistance remained unidentified yet. In this study, we established the mesenchymal like cancer cells (MLCCs) from A549 lung cancer cells with chronic exposure of TGFβ. With clinicogenomics database analysis and following validation with MLCC model, we determined that BCL2, of which expression was clearly induced and indicated a poor prognosis in the mesenchymal cancer patients, was responsible for high chemoresistance in MLCCs. Therefore combined treatment of chemotherapeutic drug and BH3 mimetic such as ABT-263 or ABT-737, remarkably sensitized MLCCs to chemo and radio therapy. Furthermore, BCL2 expression was governed by sustained ERK1 activity in MLCCs, which resulted from high level of MEK partner-1 (MP1) protein expression. Therefore, combined chemotherapy with small molecules (approved by FDA or clinically tested) targeting for MEK1 or BCL2 would be a clinically feasible approach to overcome EMT related chemoresistance.

Citation Format: Ok-Seon Kwon, Soon-Ki Hong, Hyuk-Jin Cha. BCL2 induced by LAMTOR3-MAPK is a druggable target of chemoradioresistance in mesenchymal 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 4761. doi:10.1158/1538-7445.AM2017-4761
Abstract 4762: New device for radiation delivery in breast cancer patients - double balloon applicator for optimal dose distribution
Authors: Subramanian Manny R. , Cutrer Mike , Mehta Rohit
Published: July 2017

Currently available applicators for delivery of accelerated partial breast irradiation using a high dose afterloader, have treatment catheters close to the center of the balloon in fixed positions, restricting one’s ability to modify the dose to critical organs. Recently we developed a novel, dual balloon brachytherapy applicator using which one could optimize radiation dose by shaping the resection cavity with an outer balloon and allowing an inner balloon to selectively position the treatment catheters away from the central axis of the balloon based on cavity size.

The Double Balloon Breast Brachytherapy applicator offers a unique design with adjustable treatment catheters, allowing for both optimized dose delivery and conformal avoidance of critical structures. This applicator is a single entry (1 cm incision), multi-channel high dose rate afterloader (HDR) accessory device. The treatment catheters can be expanded to an optimal location greater than or equal to 50% of the outer balloon diameter(> 1 cm diameter) - thereby optimizing dose delivery to the patient. The Conformal Double Balloon Breast Brachytherapy Device has the ability to optimize either conformality or homogeneity by allowing expansion of an inner polyurethane balloon to selectively position multiple treatment catheters significantly away from the central axis of an expanded outer polyurethane balloon.

A model study of the Double Balloon Breast Brachytherapy Device was performed with a wide range of volumes of the lumpectomy cavity. Each case is scanned in a CT scanner and the data transported to a treatment planning system. Dosimetric parameters such as dose delivered to treatment site, different organs such as skin, ribs, etc are evaluated and compared against existing breast brachytherapy applicator devices.

Both ultrasound and CT images clearly indicate that the quality of the Double Balloon Device images are comparable to other commercial devices. All catheters are distinguishable and can be reconstructed for treatment planning, and the balloon volume can be clearly delineated. Planning Treatment Volume (PTV) based on balloon volume can be easily generated and optimized dosimetry performed. Initial patient studies also indicate that using the double balloon applicator optimized dosimetry can be obtained thereby reducing the dose to critical structures such as skin and rib while maintaining the dose to the target.

Citation Format: Manny R. Subramanian, Mike Cutrer, Rohit Mehta. New device for radiation delivery in breast cancer patients - double balloon applicator for optimal dose distribution [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 4762. doi:10.1158/1538-7445.AM2017-4762
Abstract 4763: Metronomic chemotherapy prevents therapy-induced stromal activation and induction of cancer stem cells
Authors: Tsai Kelvin K. , Chan Tze-Sian , Hsu Chung-Chi , Pai Vincent C. , Huang Shenq-Shyang , Weaver Valerie M.
Published: July 2017

Whilst traditional chemotherapy kills a fraction of tumor cells, it also activates the stroma and can promote the growth and survival of residual cancer cells to foster tumor recurrence and metastasis. Accordingly, overcoming the host response induced by chemotherapy could substantially improve therapeutic outcome and patient survival. Treatment resistance and metastasis have been attributed to expansion of cancer stem-like cells (CSCs). Molecular analysis of the tumor stroma in neoadjuvant chemotherapy-treated human desmoplastic cancers and orthotopic tumor xenografts revealed that traditional maximal tolerated dose chemotherapy, regardless of the agents used, induces persistent STAT-1 and NF-κB activity in carcinoma-associated fibroblasts that induces the expression and secretion of ELR-motif-positive (ELR+) chemokines, which signal through CXCR-2 on carcinoma cells to trigger their phenotypic conversion into CSCs and promote their invasive behaviors, leading to paradoxical tumor aggression following the therapy. By contrast, the same total accumulated dose administered as a low-dose-metronomic chemotherapy regimen largely prevented the therapy-induced stromal ELR+-chemokine paracrine signaling for CSCs, thereby substantially enhancing treatment response and extending survival of mice carrying desmoplastic cancers (Chan et al, J. Exp. Med. 2016). These studies illustrate the importance of stroma in cancer therapy and how its impact on treatment resistance could be tempered by altering the dosing schedule of systemic chemotherapy.

Note: This abstract was not presented at the meeting.

Citation Format: Kelvin K. Tsai, Tze-Sian Chan, Chung-Chi Hsu, Vincent C. Pai, Shenq-Shyang Huang, Valerie M. Weaver. Metronomic chemotherapy prevents therapy-induced stromal activation and induction of cancer stem 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 4763. doi:10.1158/1538-7445.AM2017-4763
Abstract 4766: Translation from unconventional 5' start sites drives tumor initiation
Authors: Sendoel Ataman , Dunn Joshua G. , Rodriguez Edwin H. , Naik Shruti , Gomez Nicholas C. , Hurwitz Brian , Levorse John , Dill Brian D. , Schramek Daniel , Molina Henrik , Weissman Jonathan S. , Fuchs Elaine
Published: July 2017

How translational control impacts tumor-initiation and malignancy is just beginning to unfold. Here, we devise an epidermis-specific, in vivo ribosome profiling strategy to interrogate the translational landscape during the transition from normal homeostasis to malignancy. Inducing SOX2, broadly expressed in oncogenic RAS-associated cancers, we find that despite widespread reductions in translation and protein synthesis, certain oncogenic mRNAs are spared. Seeking mechanism, we find that during tumor-initiation, the translational apparatus is redirected towards unconventional upstream initiation sites, enhancing translational efficiency of oncogenic mRNAs. An in vivo RNAi screen of translational regulators revealed that dampening conventional EIF2 complexes has dire consequences for normal but not oncogenic growth. Conversely, we identify alternative initiation factors essential for cancer progression, where they mediate initiation at these upstream sites, differentially skewing translation and protein expression. Our findings unveil a hitherto unappreciated role of 5’UTR translation in cancer, and expose new targets for therapeutic intervention.

Citation Format: Ataman Sendoel, Joshua G. Dunn, Edwin H. Rodriguez, Shruti Naik, Nicholas C. Gomez, Brian Hurwitz, John Levorse, Brian D. Dill, Daniel Schramek, Henrik Molina, Jonathan S. Weissman, Elaine Fuchs. Translation from unconventional 5' start sites drives tumor initiation [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 4766. doi:10.1158/1538-7445.AM2017-4766