bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5662: MEK inhibitors potentiate tumor immune surveillance by enhancing cancer cell response of IFNγ
Authors: Xie Mengyu , Zheng Hong , Beg Amer
Published: July 2017

FDA approved mitogen-activated protein kinase (MAPK) kinase (MEK) targeting inhibitors such as trametinib have recently been reported to enhance anti-tumor immunity in multiple preclinical models. It is hypothesized that the combination of such MEK inhibitors with immune checkpoint blockade will have synergistic effects and cause further tumor regression. Immune checkpoint inhibitors such as anti-PD-1 overcome immunosuppressive signaling in the activated T cells, yielding higher levels of Interferon-gamma (IFNγ) release. IFNγ plays a key role in a wide range of immune responses by direct activation of JAK-STAT1 pathway through the binding of IFNγ receptor. JAK-STAT1 regulates transcription of downstream effector genes that are involved in tumor immune surveillance. Many studies have shown that in cancer cells, transcription of T cell chemokines CXCL9,10,11, as well as PD-L1 and MHC-I are induced upon stimulation with IFNγ. While there are some recent studies indicating MEK inhibitors enhance T cell activity and function, here we report for the first time that targeted inhibition of MEK increases tumor cells response to IFNγ and results in significant upregulation of target genes including T cell chemokines, MHC-I, as well as PD-L1.

We treated a panel of 20 human lung cancer cell lines with MEK inhibitor U0126 followed by IFNγ stimulation, and showed that the expression of CXCL10 is significantly increased as compared to IFNγ stimulation alone. Furthermore, we found that instead of MEK inhibition, knock down of KRAS in the KRAS mutant cell lines such as A549 and H2122 results in more dramatic CXCL10 upregulation upon IFNγ stimulation. This indicates the potential of increasing tumor T cell infiltration upon MEK targeted therapy. Next, to elucidate the underlying molecular mechanism, we analyzed JAK-STAT1 pathway in mouse lung cancer cell lines LKR, 344, and colon cancer cell line CT26, in which trametinib mediated synergistic expression of CXCL9,10, 11 and MHC-I, PD-L1 were observed. The phosphorylation as well as total protein of JAK2, STAT1 are increased in cells that are pre-treated with trametinib, as compared to IFNγ stimulation alone. Treatment of JAK inhibitor Ruxolitinib as well as knockout of IFNγR1 in LKR cells completely abolishes CXCL10 expression. Importantly, trametinib enhanced response to PD-1 blockade in the lung tumor models. In conclusion, our study demonstrates that targeting MEK signaling in cancer cells increases the response of IFNγ through JAK-STAT1 pathway, which may potentially enhance immune surveillance and response to immune checkpoint blockade.

Citation Format: Mengyu Xie, Hong Zheng, Amer Beg. MEK inhibitors potentiate tumor immune surveillance by enhancing cancer cell response of IFNγ [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 5662. doi:10.1158/1538-7445.AM2017-5662
Abstract 5663: A potential role for NKG2A to influence post-transplant outcomes in multiple myeloma
Authors: Cooperrider Jennifer , Ciarlariello Paul D. , Hughes Tiffany , Williams Nita , Hofmeister Craig , Baiocchi Robert , Benson Don M.
Published: July 2017

While there is currently no clear consensus regarding optimal maintenance therapy after autologous stem cell transplant (autoSCT), the early post-transplant setting is a potential site of augmentation to improve post-transplant outcomes. We previously characterized stem cell graft lymphocytes in association with progression free survival (PFS) after autoSCT in multiple myeloma (MM), and showed that there was an inverse correlation between NKG2A expression on CD56dim NK cells and PFS. Therefore, we aimed to investigate a potential role for NKG2A modulation, especially in the post-transplant setting with maintenance chemotherapy. We performed immunophenotypic analyses of NK cells from patients in the immediate post-transplant setting as well as from patients at various time points after transplant on lenalidomide maintenance therapy. In the immediate post-transplant setting (mean time since transplant=13 days) NKG2A was upregulated on NK cells when compared with KIR2DL1 (p=0.001) and KIR2DL2 (p=0.002). Additionally, in immunophenotypic analyses of PBMC from MM patients on various treatments, patients receiving lenalidomide maintenance therapy that recently underwent stem cell transplant (mean time since transplant=0.53 years) had elevated NKG2A expression compared to patients receiving no treatment (p=0.029). Conversely, patients receiving lenalidomide maintenance therapy over years (mean time since transplant=4.21 years) after transplant showed reduced NKG2A expression compared to patients receiving no treatment (p=0.0161). In bone marrow aspirates from patients with MM, primary CD138-positive myeloma cells in the bone marrow express HLA-E, the cognate antigen for NKG2A, which makes the overexpression of NKG2A biologically relevant. These data suggest that the NKG2A/HLA-E checkpoint axis may be more biologically relevant than KIR in myeloma, particularly in the post-transplant setting. This has important potential implications for treatment, as it demonstrates that NKG2A could serve as a potential target for modulation in the immediate post-transplant setting to improve survival, especially in the setting of maintenance chemotherapy. Further investigation should be aimed delineating the potential effect of NKG2A modulation on immune recovery and tumor surveillance to increase survival in the post-transplant setting.

Citation Format: Jennifer Cooperrider, Paul D. Ciarlariello, Tiffany Hughes, Nita Williams, Craig Hofmeister, Robert Baiocchi, Don M. Benson. A potential role for NKG2A to influence post-transplant outcomes 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 5663. doi:10.1158/1538-7445.AM2017-5663
Abstract 5667: Exposure of tumor-associated macrophages to apoptotic pancreatic cancer cells promotes cancer stem cell chemoresistance
Authors: D'Errico Gabriele , Vallespinos Mireia , Alcala Sonia , Valle Sandra , Martin-Hijano Laura , Sainz Bruno
Published: July 2017

Pancreatic ductal adenocarcinoma (PDAC) is the fourth leading cause of cancer related deaths worldwide. This is largely due to the existence of a subpopulation of stem-like cells present within the tumor, known as cancer stem cells (CSCs) that drive metastasis and chemoresistance. In addition, we have now come to realize that the tumor-associated microenvironment not only provides structural support for tumor development, but more importantly the microenvironment provides cues to CSCs that regulate their biological properties. Chemotherapy often leads to apoptosis of cancer cells, and in previous studies we have shown that tumor-associated macrophages (TAMs) exponentially increase following chemotherapy. We hypothesized that TAMs, in response to chemotherapy-induced apoptosis, secrete factors that potentiate PDAC chemoresistance. In line with this hypothesis, we show that monocyte-derived macrophages cultured in the presence of apoptotic PDAC cells polarize towards an M2 pro-tumor phenotype and secrete factors that render naïve PDAC cells, specifically CSCs, resistant to Gemcitabine- or Abraxane-induced apoptosis, irrespective of mutations in p53. Importantly, chemoresistant cells showed increased sphere formation capacity and increased tumorigenesis as measured in an extreme limiting dilution assay in nude mice, confirming an enrichment in CSCs. Moreover, using a syngeneic orthotropic in vivo model of PDAC, we were able to significantly augment the anti-tumor potential of Gemcitabine by pharmacologically eliminating TAMs using clodronate liposomes. To determine the mechanism by which TAMs promote PDAC chemoresistance, we performed proteomic analyses on macrophage conditioned media and identified several proteins specifically induced and secreted when macrophages were co-cultured with apoptotic PDAC cells, including 14-3-3 protein zeta/delta (14-3-3ζ), a major regulator of apoptotic cellular pathways. We present additional data to show that TAM-seceretd 14-3-3ζ promotes CSC chemoresistance. Taken together, the sum of these data highlight a unique regulatory mechanism by which chemotherapy-induced apoptosis acts as a switch to initiate a pro-tumor/anti-apoptotic mechanism in PDAC, challenging the idea that apoptosis of tumor cell is therapeutically beneficial, at least when immune sensor cells, such as macropahges, are present.

Citation Format: Gabriele D'Errico, Mireia Vallespinos, Sonia Alcala, Sandra Valle, Laura Martin-Hijano, Bruno Sainz. Exposure of tumor-associated macrophages to apoptotic pancreatic cancer cells promotes cancer stem cell chemoresistance [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 5667. doi:10.1158/1538-7445.AM2017-5667
Abstract 5676: Functional pairing of immunomodulatory targets in anaplastic thyroid cancer
Authors: Chakraborty Sanjukta , Maniyar Rachana R. , Tuli Neha Y. , Rahoma Ghada Ben , Budenz Cameron , Singh Sarnath , Geliebter Jan , Tiwari Raj
Published: July 2017

Thyroid cancer is the most common type of endocrine malignancy that has an escalating global frequency. Although most well differentiated thyroid cancers (WDTC) are manageable and respond to current therapeutic modalities, undifferentiated anaplastic thyroid cancers (ATC) exhibit a dramatically different clinical behavior and poor prognosis. With the recent development of immunotherapies, targeted, well-defined treatment plans can demonstrate promising treatment outcomes in ATC patients. Precise immunological targets in ATCs with potential clinical relevance are unknown. Major progress has been made in last 5 years toward development of immune checkpoint inhibitors using anti-CTLA-4 and anti-PD-1/PD-L1 antibodies for cancer treatment which has made immunotherapies one of the mainstream treatment choices. Few additional members of the immunoglobulin superfamily of receptors, like LAG3, TIM3 and VISTA have recently been identified as potential checkpoint targets. Interestingly some of these molecules including TIM3 and PD-L1 promote tumor progression and immune escape. Identification of specific immunotherapeutic targets requirs a better understanding of the immune microenvironment in ATC. To this end we evaluated the expression of prominent co-stimulatory and co-inhibitory cell surface molecules by RT-PCR in three thyroid cancer cell lines - TPC-1 (papillary), CGTH-W-1 (follicular) and 8505C (anaplastic). We observed that many co-inhibitory molecules were upregulated in all three tumor cell lines. CTLA4, interestingly, had the highest expression in 8505C. Additionally we observed differential expression of BTLA, LAIR1, TIM3 and VISTA between TPC-1 and 8505C. LAG3, PD-1 and PD-L1 were also upregulated in 8505C compared to TPC-1. Similar pattern was observed with the expression of co-stimulatory molecules, CD40L and GITR . GITR has been shown to have a tumor suppressor function in multiple myeloma. Another co-stimulatory molecule OX40, which has shown promise in tumor recession when targeted, was upregulated in all three cell lines and 8505C showed the highest expression. Our findings suggest that the aggressive and less immunogenic phenotype of ATC might be attributed to the differential expression of these molecules. Targeting these immunomodulatory molecules in ATC warrants a better understanding of the crosstalk between them and it might provide an efficient means for the disease management.

Citation Format: Sanjukta Chakraborty, Rachana R. Maniyar, Neha Y. Tuli, Ghada Ben Rahoma, Cameron Budenz, Sarnath Singh, Jan Geliebter, Raj Tiwari. Functional pairing of immunomodulatory targets in anaplastic thyroid 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 5676. doi:10.1158/1538-7445.AM2017-5676
Abstract 5682: Circulating tumour DNA following decompression using selfexpandable metallic stent
Authors: Takahashi Goro , Yamada Takeshi , Iwai Takuma , Takeda Kohki , Koizumi Michihiro , Shinji Seiichi , Yokoyama Yasuyuki , Hara Keisuke , Furuki Hiroyasu , Watanabe Atsushi , Ohta Keiichro , Uchida Eiji
Published: July 2017

Background: Decompression using selfexpandable metallic stent (SEMS) has been reported to improve short-term outcomes in patients with malignant large bowel obstruction (MLBO). However, some reports suggest that SEMS worsen long-term survival. Conversely, trans-anal drainage tubes (TDT), which are similar to the ileus tubes widely used in Japan for MLBO, have had no reports suggesting reduced survival. SEMS worsen long-term survival by compressing tumor continuously and induce tumor cell death which can lead production of various types of mediator which could negatively affect prognosis. In this study, we investigated tumor cell death induced by decompression procedure by using circulating tumour DNA (ctDNA).

Methods: Plasma samples were obtained before and 1, 3 and 7 days after decompression. First, we purified circulating cell free DNA (ccfDNA) from 1 mL of plasma and measured the concentration using fluorometer. Second, we measured mutated fragment of ccfDNA (circulating tumour DNA) which is derived from tumor cell. Mutational analysis from the main tumour was performed using Ion PGM™ Sequencer following library preparation with Ampliseq CHPver.2 (Thermo Fisher Scientific). Each mutant fragment in ccfDNA was quantified using digital PCR (QuantStudio 3D® digital PCR system).

Results: We enrolled 30 patients (22 SEMS and 8 TDT). CcfDNA levels on day7 was significantly increased in the SEMS group (before; 623ng/ml, day7; 1455ng/ml, p=0.006) but TDT group. Sequencing result revealed that the average number of mutation per tumor was 3.7 (e.g. APC, TP53, and KRAS) out of 50 genes. Mutant fragments (ctDNA) were detected in the ccfDNA in all cases. Mutant fragment also increased after decompression in SEMS group(before; 6.2copies/μl, day3; 22.6copies/μl, p=0.03)but in TDT group.

Conclusion: SEMS insertions increased plasma levels of not only ccfDNA but also ctDNA in patients with MLBO. This indicated that SEMS would induce massive cancer cell death. This tumor cell death can drain various mediators, including DAMPs, into blood flow and may worsen prognosis of patients with MLBO. Conversely, TDT may not induce massive tumor cell death.

Citation Format: Goro Takahashi, Takeshi Yamada, Takuma Iwai, Kohki Takeda, Michihiro Koizumi, Seiichi Shinji, Yasuyuki Yokoyama, Keisuke Hara, Hiroyasu Furuki, Atsushi Watanabe, Keiichro Ohta, Eiji Uchida. Circulating tumour DNA following decompression using selfexpandable metallic stent [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 5682. doi:10.1158/1538-7445.AM2017-5682
Abstract 57: Generation and characterization of novel anti-glycan monoclonal antibody against acute myeloid leukemia
Authors: Teo Chong Kok , Chng Wee Joo , Choo Andre Boon Hwa
Published: July 2017

Hematological malignancies encompass a wide spectrum of cancers from hematopoietic and lymphoid tissues. Under this grouping, the various forms of leukemia, lymphoma and myeloma have been classified into more than 60 distinct disease types, each having particular clinical features and disease outcomes. Among these diseases, curative treatments with high long-term survival rate have been developed for some, e.g. Chronic Myeloid Leukemia, while such effective regimens are still unavailable for many others, e.g. Multiple Myeloma and Acute Myeloid Leukemia. In this study, a panel of antibodies was raised against SLAMF7, a cell surface marker that was associated to Multiple Myeloma. Among the antibodies, TAG-HC2 was surprisingly found to have preferential binding to SLAMF7 expressed on Acute Myeloid Leukemia (AML) cell lines instead. Characterization of TAG-HC2 revealed that the binding of TAG-HC2 to SLAMF7 requires the N-glycosylation of the antigen. Functionally, TAG-HC2 does not have non-specific binding to normal cell lines when tested via flow cytometry and immunohistochemistry. TAG-HC2, when complexed with saporin-conjugated secondary mAb, was able to internalize into and kill AML cell lines. These results suggest that TAG-HC2 has the potential to be developed into an Antibody Drug Conjugate against AML.

Citation Format: Chong Kok Teo, Wee Joo Chng, Andre Boon Hwa Choo. Generation and characterization of novel anti-glycan monoclonal antibody against acute myeloid leukemia [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 57. doi:10.1158/1538-7445.AM2017-57
Abstract 5706: Gene expression profile of peripheral blood mononuclear cells in breast cancer patients may be contribute to the identification and the immunological classification of breast cancer patients by blood test
Authors: Suzuki Eiji , Kawaguchi Kosuke , Sugimoto Masahiro , Pu Fengling , Uozumi Ryuji , Yamaguchi Ayane , Nishie Mariko , Tsuda Moe , Kotake Takeshi , Morita Satoshi , Toi Masakazu
Published: July 2017

It is reported that gene expression profile of peripheral blood mononuclear cells (PBMCs) exhibits unique gene expression signature in cancer patients including renal cell carcinoma, pancreatic cancer and lung cancer. Since pancreatic cancer diagnosis is difficult in not only early detection of the disease but also in the diagnosis itself, development of novel diagnostic tools in addition to conventional diagnostic strategy has been awaited. On the other hand, in breast cancer, because early diagnosis by mammography and ultra sound examination on breast is established successfully, exploration of gene expression profile of PBMCs may be important in terms of insight to host antitumor immune response aspects.

In the current study, we performed RNA sequencing (RNA-seq) analysis on RNA of PBMCs from 3 healthy volunteers, 6 early and 7 metastatic breast cancer patients including all phenotypes defined by ER, PgR and HER2. Genes that showed FDR<0.05 and fold change of <0.5 and 2.0< in comparison of healthy volunteers and breast cancer patients were applied to clustering analysis by Ward method. We found significant unique gene expression signature between healthy volunteers and breast cancer patients. Furthermore, it was shown that HER2 overexpressing breast cancer patients, which breast cancer patients were divided into 3 clusters including HER2 type and other 2 different groups, seemed to be classified into healthy group different from other phenotypes of breast cancer patients.

These findings suggested that evaluation of gene expression patterns of PBMCs of breast cancer patients might distinguish breast cancer patients from healthy subjects and the gene expression signature of PBMCs which divided breast cancer patients into 3 groups might reveal immunologically important biologic properties such as response prediction of cancer immunotherapy including immune checkpoint inhibition treatment.

Citation Format: Eiji Suzuki, Kosuke Kawaguchi, Masahiro Sugimoto, Fengling Pu, Ryuji Uozumi, Ayane Yamaguchi, Mariko Nishie, Moe Tsuda, Takeshi Kotake, Satoshi Morita, Masakazu Toi. Gene expression profile of peripheral blood mononuclear cells in breast cancer patients may be contribute to the identification and the immunological classification of breast cancer patients by blood test [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 5706. doi:10.1158/1538-7445.AM2017-5706
Abstract 5715: Gene body methylation and expression of SHOX2 gene are potent prognostic markers for survival in intermediate grade gliomas and renal cancers
Authors: Zhang Yu-An , Zhou Yunyun , Luo Xin , Xiao Guanghua , Gazdar Adi F.
Published: July 2017

Background: Previous data indicated that low level of promoter methylation of the SHOX2 gene was a prognostic marker for worse progression-free survival for non-small cell lung cancer.

Purpose: (a) to determine the relationship between SHOX2 gene methylation and expression; b) to determine whether SHOX2 gene methylation and expression were prognostic markers for other cancers.

Methods: We examined multiple methylation and expression datasets available from The Cancer Genome Atlas (TCGA) and other previously published studies. Data from a total of more than 1000 intermediate grade gliomas and 1100 renal cancer cases were examined, along with smaller number of corresponding non-malignant tissues. We performed Kaplan-Meier survival analyses and univariate and multivariate Cox regression model analyses.

Results: SHOX2 gene contains three CpG islands encompassing the promoter region and body. Methylation of various regions of SHOX2 gene body was significantly positively correlated with its expression in gliomas and renal cancer. Aberrantly high methylation and expression of SHOX2 gene, which were significantly higher than the corresponding nonmalignant tissues, were identified in subsets of gliomas and renal cancers. Survival analysis of patients indicated that the methylation and expression of SHOX2 gene in gliomas and renal cancer (both clear cell and papillary cell carcinomas) were highly potent markers for poor survival. We further investigated the combination of SHOX2 with other known clinically relevant glioma markers (IDH genotype status, TERT expression, 1p/19q chromosome co-deletion, MGMT methylation, ATRX mutation and NES expression). When combined with SHOX2 expression, we identified subsets of glioma patients with significantly favorable survival outcomes, especially in the subgroup (i.e., IDH wild-type) associated with worse prognosis for each individual marker. Multivariate analysis demonstrated that SHOX2 was a potent independent survival marker for gliomas.

Conclusion: Gene body methylation and expression of SHOX2 gene are tightly positively correlated and are potent new markers for overall survival prognosis for intermediate grade gliomas and renal cancer. The combination of IDH or other relevant prognostic markers with SHOX2 identified intermediate grade glioma subsets with improved survival outcomes.

Citation Format: Yu-An Zhang, Yunyun Zhou, Xin Luo, Guanghua Xiao, Adi F. Gazdar. Gene body methylation and expression of SHOX2 gene are potent prognostic markers for survival in intermediate grade gliomas and renal cancers [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 5715. doi:10.1158/1538-7445.AM2017-5715
Abstract 5717: PKR promotes EphA2 and other proteins degradation and is predictor of survival in lung cancer
Authors: Pataer Apar , Shao Ruping , Ozpolat Bulent , Fang Bingliang , Swisher Stephen G.
Published: July 2017

Relative expression of PKR and EphA2 was a significant predictor of survival in non-small cell lung cancer. To study the PKR-regulated protein, we evaluated expression of 149 proteins in lung cancer cells by reverse-phase protein assay (RPPA) after induction of PKR. Expression of several proteins (EphA2, IRS1, Chk1 and c-Raf and paxillin) was inhibited by PKR. In validating the downregulated proteins by western blot analysis, we observed that PKR reduced EphA2, IRS1, Chk1 and paxillin protein expression. We next investigated the involvement of the proteasomes and lysosomes in PKR-induced protein degradation. Human lung cancer H1299 and A549 cells were transfected with Ad-PKR for 48 h in the presence or absence of the proteasome inhibitor MG132 or lysosome inhibitor (3MA: 3-methyladenine). We observed that MG132 prevented PKR-induced IRS1 and Chk1 degradation, but not PKR-induced EphA2 and paxillin degradation in these cells. However, we found that 3MA prevented PKR-induced EphA2 and paxillin degradation in these cells. These data suggest that PKR is involved in proteasome as well as lysosome function. To study the involvement of autophagy pathway in PKR mediated protein degradation, we evaluated proteins which known involved in autophagy by Western blotting after induction of PKR on these cancer cells. We observed slightly decrease of LC-3, ATG5 and Beclin1 in these cancer cells after induction of PKR. Our data suggest that PKR may directly involve in in lysosome function.

Citation Format: Apar Pataer, Ruping Shao, Bulent Ozpolat, Bingliang Fang, Stephen G. Swisher. PKR promotes EphA2 and other proteins degradation and is predictor of survival in 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 5717. doi:10.1158/1538-7445.AM2017-5717
Abstract 5727: A functional polymorphism in DTX1 gene of Notch pathway predicts the prognosis of surgically resected non-small cell lung cancer
Authors: Lee Shin Yup , Lee Janghyuck , Shin Kyung Min , Son Ji Woong , Park Jae Yong
Published: July 2017

Notch signaling pathway has been implicated in the prognosis of non-small cell lung cancer (NSCLC). This study was conducted to determine the association between single nucleotide polymorphisms (SNPs) in the Notch pathway genes and survival outcomes of surgically resected NSCLC. Among the 64 SNPs analyzed in the discovery set, 8 SNPs were significantly associated with overall survival (OS) or disease-free survival (DFS). Among the 8 SNPs, the association was consistently observed only for DTX1 rs1732786A>G in the validation sets. In combined analysis, the rs1732786A>G was significantly associated with better OS and DFS (adjusted HR aHR for OS, 0.81; 95% CI, 0.68-0.95; P = 0.01; aHR for DFS, 0.82; 95% CI, 0.72-0.94; P = 0.003; under codominant model). To verify the functional relevance of the rs1732786A>G in DTX1 promoter region, we conducted dual-luciferase reporter assays. The G allele showed significantly higher luciferase activity than the A allele (P < 0.005) in lung cancer cell lines. The relative expression level of DTX1 mRNA was significantly lower in tumor tissues than in nonmalignant lung tissue (P = 0.003). Moreover, the level of DTX1 mRNA expression in tumor tissue increased as the number of G allele increased (P trend = 0.02). Higher levels of DTX1 mRNA expression were significantly correlated with a better OS and DFS (P = 0.03 and P = 0.002). Our results suggest that the DTX1 rs1732786 A>G polymorphism may affect DTX1 expression and may be a useful marker for predicting the prognosis of patients with early stage NSCLC.

Citation Format: Shin Yup Lee, Janghyuck Lee, Kyung Min Shin, Ji Woong Son, Jae Yong Park. A functional polymorphism in DTX1 gene of Notch pathway predicts the prognosis of surgically resected 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 5727. doi:10.1158/1538-7445.AM2017-5727
Abstract 5732: PI3K/Akt activity regulates androgen receptor expression and predicts poor clinical outcome in non-metastatic hormone-naïve prostate cancer
Authors: Blom Sami , Mäki-Teeri Petra , Erickson Andrew , Paavolainen Lassi , Mirtti Tuomas , Rannikko Antti , Potdar Swapnil , Östling Päivi , Weerden Wytske van , Kallioniemi Olli , Pellinen Teijo
Published: July 2017

Activation of PI3K/Akt pathway is associated with adverse outcome and aggressive disease in many cancers. In prostate cancer (PCa), the activity of this pathway has been shown to promote disease progression and metastasis. However, it is still controversial how PI3K/Akt regulates androgen receptor (AR), a central signaling molecule in prostate pathophysiology, and whether it has an active role in hormone naïve non-metastatic PCa. Here, we show using immunohistochemistry (IHC) and advanced quantitative multiplexed IHC that the expression of phosphorylated-Akt(S473) and AR are highly correlated in clinical PCa, even at the cellular level. Furthermore, we found that high expression of p-Akt(S473) predicts poor clinical outcome in two independent hormone-naïve non-metastatic PCa cohorts. To study whether PI3K/Akt regulates AR expression, we performed an in vitro drug screen with 32 PI3K/Akt/mTOR inhibitors in PC346C, an AR expressing cell line derived from a hormone-naïve primary tumor of prostate. We observed a strong correlation between p-Akt(S473) and AR also in vitro in individual cells independent of the inhibitor used. Although both PI3K and Akt specific inhibition reduced cell viability, the response in nuclear expression of AR was highly dependent on the target of inhibition: Akt specific inhibition reduced AR nuclear expression and resulted in large, spindle-shaped cells, whereas PI3K specific inhibition increased AR nuclear expression and resulted in smaller, round-shaped cells. These data suggest that PI3K and Akt have different roles in sustaining AR activity in PCa as perturbations of the two components leads to differential responses in terms of AR nuclear expression and cell morphology. In conclusion, activated Akt associates with AR expression and predicts poor clinical outcome in hormone-naïve non-metastatic PCa. Furthermore, the differing roles of PI3K and Akt in AR regulation warrants for further studies as it may have implications in the design of PCa therapy targeting PI3K/Akt, especially when the inhibitors are administered in combination with anti-androgens.

Citation Format: Sami Blom, Petra Mäki-Teeri, Andrew Erickson, Lassi Paavolainen, Tuomas Mirtti, Antti Rannikko, Swapnil Potdar, Päivi Östling, Wytske van Weerden, Olli Kallioniemi, Teijo Pellinen. PI3K/Akt activity regulates androgen receptor expression and predicts poor clinical outcome in non-metastatic hormone-naïve 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 5732. doi:10.1158/1538-7445.AM2017-5732
Abstract 5734: The influence of body mass index on overall survival following surgical resection of non-small cell lung cancer
Authors: Liu Xi , Sepesi Boris , Gold Kathryn A. , Correa Arlene M. , Heymach John V. , Vaporciyan Ara A. , Roszik Jason , Dmitrovsky Ethan
Published: July 2017

Prior work implicated an association between increased body mass index (BMI) and lower risk of mortality from lung cancer. The aim of our study was to comprehensively evaluate the influence of BMI on long-term overall survival in surgical patients with non-small cell lung cancer. This study investigated 1935 patients who underwent surgical resection for lung cancer at MD Anderson Cancer Center (from 2000 - 2014). Study variables included both patient and treatment related characteristics. Univariate and multivariate Cox regression analyses were performed to identify variables associated with overall survival. By univariate analysis, significant predictors of better survival were higher BMI, pathologic tumor stage (stage I versus stages II, III, or IV), type of surgery (lobectomy/pneumonectomy versus wedge resection/segmentectomy), younger age, female gender, and adenocarcinoma histology (versus squamous) (all p < 0.05). Patients considered morbidly obese (BMI ≥ 35) had a trend towards better outcomes than those classified as obese (BMI ≥ 30 and <35) (p = 0.05), overweight (BMI ≥25 and <30) (p = 0.13), or healthy weight (BMI < 25) (p = 0.37) (HR 0.727, 0.848, 0.926, and 1, respectively). By multivariate analysis, BMI remained an independent predictor of survival (p = 0.02). Propensity matching analysis showed significantly better overall survival (p = 0.008) in patients with BMI ≥ 30 as compared to BMI < 25. For exploratory analysis of expressed mRNAs associated with obesity in lung cancer, the association between obesity-related species (LEP, LEPR, PCSK1, POMC, MC4R, BMIQ1, BMIQ2, BMIQ3, UCP2, BMIQ5, BMIQ6, INSIG2, FTO, TMEM18, GNPDA2, NEGR1, BDNF, KCTD15, SH2B1, MTCH2 and NPC1) and survival was explored using The Cancer Genome Atlas (TCGA). Kaplan Meier analyses demonstrated significantly improved overall survival in lung cancer patients with higher Uncoupling Protein 2 (UCP2) expression, as will be presented. In summary, this large, single center series, after controlling for disease stage and other variables found higher BMI was associated with improved overall survival following surgical resection of non-small cell lung cancer. Studies are underway to elucidate the underlying mechanisms responsible for this association between BMI and lung cancer survival.

Citation Format: Xi Liu, Boris Sepesi, Kathryn A. Gold, Arlene M. Correa, John V. Heymach, Ara A. Vaporciyan, Jason Roszik, Ethan Dmitrovsky. The influence of body mass index on overall survival following surgical resection of 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 5734. doi:10.1158/1538-7445.AM2017-5734
Abstract 5736: Clinical impact of high serum hepatocyte growth factor in advanced non-small cell lung cancer
Authors: Tsuji Takahiro , Ozasa Hiroaki , Sakamori Yuichi , Nomizo Takashi , Funazo Tomoko , Yasuda Yuto , Yoshida Hironori , Nagai Hiroki , Kim Young Hak
Published: July 2017

Activation of c-MET through hepatocyte growth factor (HGF) increases tumorigenesis, induces resistance, and is associated with poor prognosis in various solid tumors. We previously reported that lung cancer cell lines had increased expression of c-MET due to gene amplification-induced cytotoxic drug resistance, and that resistant cells paracrine HGF and promote its resistance. However, the clinical significance of sHGF in patients with advanced or recurrent NSCLC, especially in patients treated with cytotoxic chemotherapy, is yet to be identified. Here, we present sHGF may be useful to predict tumor response and PFS in patients with advanced NSCLC. A total of 81 patients with 101 treatment regimens were investigated; 53 patients received first-line therapy and 48 patients received second-line therapy during the observation period. sHGF levels were evaluated using ELISA at 4 time-points: at pre-treatment, at response-evaluation (1-2 months after treatment initiation), at the best tumor response, and at the disease progression. As a control biomarker, serum carcinoembryonic antigen (CEA) was also evaluated. sHGF at response-evaluation in patients with progressive disease was higher compared with that of disease controls in both first-line and second-line treatment (median value (MV): under the limited of detection (LOD) vs. 0.40; P=0.0086 and MV: under the LOD vs. 0.41; P=0.0035, respectively). Positive-sHGF at response-evaluation predicts poor progression-free survival (PFS) compared with negative-sHGF in both first-line (median, 153.5 vs. 288.0; P=0.047) and second-line treatment (87.0 vs. 219.5; P=0.014). Lung adenocarcinoma subgroup analysis showed that in patients receiving second cytotoxic chemotherapy, there were no significant differences in PFS or HR between patients with low-CEA compared with those with high-CEA, but positive-sHGF at pre-treatment or at response-evaluation predicts poor PFS (35 vs. 132; P=0.0045, 50 vs. 215; P=0.0047, respectively). Simple and repeatable markers for activation of the HGF/c-MET pathway would provide rationale for future research investigating the merit of sHGF as a potential clinical biomarker to indicate administration of MET inhibitors.

Citation Format: Takahiro Tsuji, Hiroaki Ozasa, Yuichi Sakamori, Takashi Nomizo, Tomoko Funazo, Yuto Yasuda, Hironori Yoshida, Hiroki Nagai, Young Hak Kim. Clinical impact of high serum hepatocyte growth factor in advanced 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 5736. doi:10.1158/1538-7445.AM2017-5736
Abstract 5739: Nicotine promotes stemness-related properties and cell migration/metastasis through IGF-1R regulation in triple negative breast cancer
Authors: Kuo Yung-Che , Chu Jan-Show , Lee Kha-Liang , Drew Victor James , Zhuang Wei-Zhan , Chen Chi-Long , Ho Yuan-Soon , Huang Yen-Hua
Published: July 2017

Purpose: To determine the role of insulin-like growth factor (IGF)-I receptor (IGF-IR) in alpha 9 nicotinic acetylcholine receptor (α9-nAChR)-induced stemness-related properties and to assess the therapeutic potential for preventing early tumor relapse and metastasis of TNBCs.

Experimental Design: Expression levels of stemness-related genes, α9-nAChR and IGF-IR in tissues (n = 67) were analyzed by real-time Q-PCR and the correlation of different gene levels were calculated by the Pearson correlation analysis. The effects of either α9-nAChR activation or IGF-1R transactivation on stemness expression in TNBC were examined using ALDEFLUOR assay with flow cytometry, RNA interference, wound closure/migration/invasion assay, and animal models. The α9-nAChR, OCT4 and IGF-1R protein, p-IGF-1R in tissues were detected by immunohistochemical staining.

Results: A high positive correlation between the gene expression levels of OCT4/NANOG/CD24/CD44 and α9-nAChR/IGF-IR in human breast cancer tissues was observed. Nicotine induced α9-nAChR activation trans-activates an IGF-IR expression and a phosphorylation of STAT3, which stimulated stemness-related properties in both the cell lines and the xenografted mouse tumors. The inhibition of either α9-nAChR or IGF-IR activation by RNA interference significantly suppressed the nicotine-induced stemness-related properties and cell metastasis both in vitro and in vivo.

Conclusions: IGF-1R signal regulates the nicotine/α9-nAChR-induced stemness properties and cell metastasis therefore convene poor prognosis in TNBCs.

Citation Format: Yung-Che Kuo, Jan-Show Chu, Kha-Liang Lee, Victor James Drew, Wei-Zhan Zhuang, Chi-Long Chen, Yuan-Soon Ho, Yen-Hua Huang. Nicotine promotes stemness-related properties and cell migration/metastasis through IGF-1R regulation 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 5739. doi:10.1158/1538-7445.AM2017-5739
Abstract 5744: The role of atypical protein kinase C in colorectal cancer cell growth and proliferation
Authors: Islam S M Anisul , Duncan Mildred Acevedo
Published: July 2017

Colorectal cancer (CRC) is the second most common cancer in the world and death from colorectal cancer accounts for 8% of all cancer deaths both in men and women in the United States. CRC is life threatening disease due to therapy resistant cancerous cells. The exact mechanisms of cell growth, survival, metastasis and inter & intracellular signaling pathways involved in colon cancer are still a major challenge for scientists. Hence, investigating the signaling pathways that lead to colon carcinogenesis may give insight into the therapeutic target. In this study, the role of atypical Protein kinase C (aPKC) on colon cancer was identified by using four inhibitors of that particular protein class: 1) ACPD (2-acetyl-1,3-cyclopentanedione and 2) DNDA (3,4-diamino-2,7 naphthalenedisulfonic acid) are non-specific inhibitors of aPKCs; 3) ζ-Stat (8-hydroxy-1, 3, 6-naphthalenetrisulfonic acid) is a specific inhibitor of PKC-ζ; and 4) ICA-I (4, 5-amino-4-carbamoylimidazol-1-yl-2, 3-dihydroxycyclopentyl) methyl dihydrogen phosphate) is a specific inhibitor of PKC-ι. The cell lines tested were HT-29 colon cancer and CCD18CO normal colon epithelial. Although PKC-ι is an oncogene in many cancers, we found that the PKC-ζ was responsible for the abnormal growth and proliferation in HT-29 colon cancer cells. Additionally, the inhibition of aPKCs did not bring any significant toxicity on CCD18CO normal colon cell line. These results suggest the potentiality of utilizing aPKC-ζ inhibitors to block colon cancer cell growth and proliferation.

Citation Format: S M Anisul Islam, Mildred Acevedo Duncan. The role of atypical protein kinase C in colorectal cancer cell growth and proliferation [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 5744. doi:10.1158/1538-7445.AM2017-5744
Abstract 5746: TRAF1 is required for solar UV-induced skin carcinogenesis
Authors: Yamamoto Hiroyuki , Ryu Joohyun , Min Eli , Oi Naomi , Bai Ruihua , Zykova Tatyana A. , Yu Dong Hoon , Malakhova Margarita , Moriyama Kenji , Bode Ann M. , Dong Zigang
Published: July 2017

Tumor necrosis factor receptor-associated factor 1 (TRAF1) is a member of the TRAF protein family, which regulates the canonical and non-canonical NF-κB signaling cascades. Although aberrant TRAF1 expression in tumors is reported, the role of TRAF1 remains elusive. Here, we report that TRAF1 is required for skin carcinogenesis induced by chronic solar UV radiation. In vivo studies with solar UV exposure indicate that the deletion of TRAF1 results in the inhibition of AP-1 activity by down-regulating the induction of c-Fos and c-Jun by regulating ERK5 activity. Furthermore, we show that TRAF1 is required for solar UV-induced ERK5 activation. Mechanistic studies revealed that TRAF1 expression enhances the ubiquitination of ERK5 on K184, which is necessary for AP-1 activation. Altogether, our results suggest that TRAF1 mediates ERK5 activity by regulating the upstream effectors of ERK5 and also by modulating its ubiquitination status. Targeting TRAF1 function might lead to strategies for preventing and treating skin cancer.

Citation Format: Hiroyuki Yamamoto, Joohyun Ryu, Eli Min, Naomi Oi, Ruihua Bai, Tatyana A. Zykova, Dong Hoon Yu, Margarita Malakhova, Kenji Moriyama, Ann M. Bode, Zigang Dong. TRAF1 is required for solar UV-induced skin carcinogenesis [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 5746. doi:10.1158/1538-7445.AM2017-5746
Abstract 5748: Roles of miR29c-ID2 axis in tumor initiating cells of breast ductal carcinoma in situ
Authors: Liu Yin , Pandey Puspa R. , Sharma Sambad , Wu Kerui , Xing Fei , Watabe Kounosuke
Published: July 2017

Ductal carcinoma in situ (DCIS) is an early-stage form of breast cancer that accounts for over 20% of all breast cancers in the US. Therefore, prevention of DCIS is a major goal for reducing breast cancer incidence. However, the underlying molecular and pathological mechanisms of DCIS remain poorly understood. It is believed that tumor-initiating cells are formed at a very early stage of tumorigenesis, which then leads to the progression of DCIS. Our recent work that focused on tumor-initiating cells of DCIS revealed that the expression of miR29c in tumor-initiating cells was significantly down-regulated in DCIS cells compared to that in normal cells. Furthermore, we found that the ID2 gene is one of the targets of miR29c in tumor-initiating cells and that miR29c suppresses the ID2 gene by directly binding to the 3’-UTR sequence. ID2 is a well-known dedifferentiation factor in embryonic development, and the ID family is involved in progression of cancer by affecting cell differentiation and proliferation. We also found that ectopic expression of ID2 in premalignant cells accelerated tumor growth and promoted self-renewal of tumor-initiating cells in DCIS, suggesting that ID2 also controls cancer cell dedifferentiation and promotes tumor-initiating cells formation. These results indicate that the miR29c-ID2 axis plays a critical role in self-renewal of DICS in vivo and that ID2 is a potential biomarker for tumor-initiating cells in DCIS. ID2 is thought to sequester E-protein transcription factors and thus regulates a wide range of gene expression. Our data suggest that ID2 upregulates expression of several cancers stem cell genes including SOX2, which is a well- known self-renewal regulator of stem cells. ID2 also upregulates the fatty acid synthase gene (FASN), which promotes cancer cell proliferation and survival. We also found that FASN expression was significantly suppressed by pterostilbene, indicating potential utility of this natural compound for preventing DCIS formation. Taken together, these results suggest that miR29c-ID2 functions as master regulators in forming DCIS. ID2 promotes self-renewal of tumor-initiating cells by regulating expression of the SOX2 and FASN genes.

Note: This abstract was not presented at the meeting.

Citation Format: Yin Liu, Puspa R. Pandey, Sambad Sharma, Kerui Wu, Fei Xing, Kounosuke Watabe. Roles of miR29c-ID2 axis in tumor initiating cells of breast ductal carcinoma in situ [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 5748. doi:10.1158/1538-7445.AM2017-5748