bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4685: Immune checkpoint protein VISTA is a critical regulator of the IL-23/IL-17 inflammatory axis
Authors: Li Na , Xu Wenwen , Yuan Ying , Ayithan Natarajan , Imai Yasutomo , Wu Xuesong , Miller Halli , Olson Michael , Hwang Samuel T. , Malarkannan Subramaniam , Wang Li
Published: July 2017

V-domain Immunoglobulin Suppressor of T cell Activation (VISTA) is an inhibitory immune-checkpoint molecule that suppresses CD4+ and CD8+ T cell activation when expressed on antigen-presenting cells. Vsir-/- mice developed loss of peripheral tolerance and multi-organ chronic inflammatory phenotypes. Vsir-/- CD4+ and CD8+ T cells were hyper-responsive towards self- and foreign antigens. Whether or not VISTA regulates innate immunity has not been demonstrated. Our current study shows that VISTA-blocking monoclonal antibody (mAb) enhanced anti-tumor T cell response, and synergized with a peptide vaccine and TLR7 agonist imiquimod as adjuvant to suppress tumor growth in the B16 melanoma model. Surprisingly, the therapeutic effect of this combination therapy was abolished in the IL17R knockout hosts, indicating that the IL-17-mediated inflammation is regulated by VISTA and is required for anti-tumor immunity in the context of VISTA blockade. To better define the regulatory role of VISTA in inflammation, we employed a murine model of psoriasis induced by topical treatment of IMQ. Our data show that VISTA deficiency exacerbated the psoriasiform inflammation. Enhanced TLR7 signaling in Vsir-/- dendritic cells (DCs) led to the hyper-activation of Erk1/2 signaling and augmented the production of IL-23. IL-23, in turn, promoted the expression of IL-17A in both TCRgd+ T cells and CD4+ Th17 cells. Furthermore, VISTA regulates the peripheral homeostasis of CD27- gd T cells, and their activation upon TCR-mediated or cytokine-mediated stimulation. IL-17A-producing CD27- gd T cells were expanded in the Vsir-/- mice and amplified the inflammatory cascade. Together, these results indicate that VISTA is a critical regulator of the IL-23/IL-17-mediated inflammatory axis. Ongoing studies will define how IL-17 regulates tumor-reactive immunity in the context of VISTA blockade and TLR vaccine adjuvant.

Citation Format: Na Li, Wenwen Xu, Ying Yuan, Natarajan Ayithan, Yasutomo Imai, Xuesong Wu, Halli Miller, Michael Olson, Samuel T. Hwang, Subramaniam Malarkannan, Li Wang. Immune checkpoint protein VISTA is a critical regulator of the IL-23/IL-17 inflammatory axis [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 4685. doi:10.1158/1538-7445.AM2017-4685
Abstract 4689: Anti-proliferative effect and tumor targeting properties of SurVaxM-derived monoclonal antibodies
Authors: Ciesielski Michael J. , Figel Sheila , Wiltsie Laura , Galbo Phillip , Frank Cheryl , Fenstermaker Robert A.
Published: July 2017

Background: Survivin is an inhibitor of apoptosis protein (IAP) which is upregulated in many cancers. SurVaxM (active anti-survivin immunotherapy) is currently being evaluated in a multi-center phase II clinical trial. Circulating anti-survivin antibodies were recently detected in recurrent glioblastoma patients who received SurVaxM immunizations in a previous phase I study. Though primarily functioning as an intracellular molecule, survivin has also been identified in patient serum, and, recently, in tumor-derived exosomes.

Methods: Patients in the completed SurVaxM phase I clinical trial produced both antibody and CD8+ T cell responses against survivin. In order to gauge whether antibodies to the vaccine provided any therapeutic benefit, or might serve as potential biomarkers for SurVaxM responsiveness, we tested these antibodies in pre-clinical tumor models. Murine hybridomas derived from the SurVaxM peptide were established and purified yielding several high-affinity IgG antibodies. These antibodies were characterized for target recognition and therapeutic potential.

Results: Two of the clones with the highest affinities, 2C2 (IgG2a) and H30 (IgG1), recognized both SurVaxM peptide and the wild type survivin counterpart (ELISA assay), as well as endogenous, full-length survivin (western blotting). Survivin was detected on cells with these antibodies using both immunofluorescence and flow cytometry. Following s.c. implantation of glioblastoma (GL261) and melanoma (B16) tumors in immuno-competent C57BL/6 mice, administration of 2C2 and H30 antibodies (i.p. injection) reduced tumor growth in both models compared to an irrelevant IgG. An orthotopic intracranial glioma study showed similar anti-tumor effects when treated with SurVaxM-derived antibodies. Tumor growth was also inhibited in immuno-compromised (nude) animals treated with 2C2 or H30 antibodies, although to a lesser extent.

Conclusions: Antibodies generated in response to the SurVaxM vaccination are highly cross-reactive to survivin and provide therapeutic benefit in immuno-competent mouse tumor models. These antibodies retain some efficacy in immuno-compromised models, indicative of a direct T-cell independent effect. An anti-tumor response through anti-survivin targeted antibodies is unexpected as surface-accessible survivin expression has not yet been well described in the literature. Data presented here highlight possible avenues for investigation of mechanism(s) of action.

Citation Format: Michael J. Ciesielski, Sheila Figel, Laura Wiltsie, Phillip Galbo, Cheryl Frank, Robert A. Fenstermaker. Anti-proliferative effect and tumor targeting properties of SurVaxM-derived monoclonal antibodies [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 4689. doi:10.1158/1538-7445.AM2017-4689
Abstract 469: Microrna signature as a potential biomarker for predicting survival in colon cancer
Authors: Jacob Havjin , Stanisavljevic Luka , Storli Kristian Eeg , Dahl Olav , Myklebust Mette Pernille
Published: July 2017

Background: Colon cancer is one of the most common cancers with increasing incidence and high mortality worldwide. Prognosis and choice of treatment is largely based on the tumor stage at presentation. Thus, finding novel biomarkers for predicting survival is highly desirable. Lately, several studies have been looking at microRNAs (miRNAs) in several cancers, including colon cancer. MicroRNAs are conserved, non-coding RNA molecules that play an important role in the regulation of post-transcriptional gene expression.

Material and Methods: In the present study, we have profiled miRNA in one hundred and seventy two TNM stage I-IV colon cancer patients and 10 corresponding normal colon tissue samples. Total RNA was extracted from freshly frozen tissues, and the expression of miRNA profile were assessed using Pick and Mix focus panels from Exiqon containing 84 miRNAs that have been linked to cancer.

Results: The results were visualized in a heatmap (Qlucore omics Software) and more than 20 miRNAs were found to be differentially expressed in tumors compared to the normal colon. Further, twelve miRNAs were found to discriminate between relapse and no-relapse patients in TNM- stage II and III, and four of these miRNAs (miR-23a, miR-25, miR-30d and miR-31) were found to be statistically significant in binary logistic regression with relapse as outcome variable. In univariate analysis, low expression of the four-miRNA signature was associated with better 3-year disease-free survival (DFS), 88 % versus 63% in low versus high signature, respectively (P=0.001). Moreover, the signature was a predictor of poor relapse-free survival in multivariate analyses (P=0.001; HR 31; 95% CI: 3.8-248.9). Another regression analyses method (LASSO) identified a 16-miRNA signature, and the four miRNAs found earlier were among them. The 16-miRNA signature was associated with better survival (P<0.001).

Conclusion: The present study has identified a four-miRNA signature predicting relapse in colon cancer stage II and III patients.

Citation Format: Havjin Jacob, Luka Stanisavljevic, Kristian Eeg Storli, Olav Dahl, Mette Pernille Myklebust. Microrna signature as a potential biomarker for predicting survival in colon 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 469. doi:10.1158/1538-7445.AM2017-469
Abstract 4693: Fc effector bioassays enable faster and quantitative measurement of ADCC and ADCP mechanisms of action
Authors: Cheng Zhi-Jie Jey , Moravec Rich , Paguio Aileen , Garvin Denise , Krishnan Gopal B. , Fan Frank , Cong Mei
Published: July 2017

Antibody-dependent cell-mediated cytotoxicity (ADCC) and antibody-dependent cell-mediated phagocytosis (ADCP) are recognized as important mechanisms of action (MOA) of therapeutic antibodies. Primary peripheral blood mononuclear cells (PBMCs) are routinely used in traditional bioassays to measure potency, stability of antibody drugs in ADCC and ADCP. However these methods are labor intensive and highly variable. Here we report the development of a luciferase-based reporter assay that measures activation of effector cells via cross-linking of Fc receptors with target cell-bound antibodies. We will discuss functional cell-based Fc effector reporter bioassays developed to measure Human FcgRIIIa (V158 and F158 variants), Human FcgRIIa (H131 and R131 variants), Human FcgRI as well as Mouse FcgRIV and FcgRIII. Compared to primary cell-based assays these reporter bioassays are less variable, easier to use and provide more consistent analysis of the important MOAs that are critical to for drug development programs. In qualification studies, performed in accordance with ICH guidelines for antibody screening and characterization, we tested the bioassays for specificity, accuracy, precision and linearity.

Citation Format: Zhi-Jie Jey Cheng, Rich Moravec, Aileen Paguio, Denise Garvin, Gopal B. Krishnan, Frank Fan, Mei Cong. Fc effector bioassays enable faster and quantitative measurement of ADCC and ADCP mechanisms of action [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 4693. doi:10.1158/1538-7445.AM2017-4693
Abstract 4701: Blocking FSTL1 ameliorates immunity against osteosarcoma
Authors: Kudo-Saito Chie , Ogiwara Yamato , Aoki Kazunori
Published: July 2017

Purposes: Osteosarcoma is a rare malignant tumor of bone in children and young adults, and it is very hard to successfully control the systemic metastases and life-threatening health problems more than a decade after the completion of the care due to the intensiveness of the conventional treatments including surgical resection of the primary tumor and chemotherapy. Therefore, non/low-toxic but advanced treatments have been expected for a long while. We recently found that a BMP family FSTL1 is highly and functionally expressed in murine and human osteosarcoma cell lines, and systemically expands pluripotent mesenchymal stem/stromal cells (MSCs), which play a central role in amplification of tumor heterogeneity that is consequence of reciprocal evolution together with the tumor microenvironment, including induction of immune suppression and dysfunction, desmoplasia, and angiogenesis. FSTL1 and its induced MSCs also directly confer higher invasive and metastatic properties on tumor cells leading to further cancer progression. In this study, we evaluated antitumor efficacy induced by anti-FSTL1 mAb on murine osteosarcoma models, and also compared the therapeutic efficacy with those induced by immune checkpoint inhibitory mAbs (ICIs).

Results: We used two syngeneic tumor models that were subcutaneously implanted with murine osteosarcoma NHOS cells in BALB/c mice, or LM3 cells in C3H mice. These mice were intraperitoneally injected with anti-FSTL1 mAbs (10mg/kg) twice on day 5 and day 8 after tumor implantation, and were sacrificed for immunological analysis of tumor-infiltrating cells, spleen cells and bone marrow cells on day 15. In both osteosarcoma models, CD45- MSCs as well as other immunosuppressive CD4+Foxp3+ Tregs and CD11b+Gr1+ MDSCs systemically increased, indicating impairment of antitumor immunity in the mice. Treatment with the anti-FSTL1 mAb significantly induced potent tumor-specific CTLs through reduction of MSCs, resulting in tumor-free in some of the treated mice. Although the antitumor efficacy was similar to those induced by ICIs, the mode of action was totally distinct because MSCs and the MSC-inducible exhausted CD8+PD1+Tim3+ T cells were reduced only in the anti-FSTL1-treated mice, but not in the ICI-treated mice.

Conclusions: These results suggest that blocking FSTL1 properly reprograms immunity toward elimination of osteosarcoma by reducing the tumor-supportive MSCs. Anti-FSTL1 mAb may be a promising drug for treating osteosarcoma more moderately but effectively in clinical settings.

Citation Format: Chie Kudo-Saito, Yamato Ogiwara, Kazunori Aoki. Blocking FSTL1 ameliorates immunity against osteosarcoma [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 4701. doi:10.1158/1538-7445.AM2017-4701
Abstract 4709: Induction of enhanced tumor-specific immunity by Hsp90 targeted photodynamic therapy (Hsp90-PDT) combined with immune checkpoint inhibition
Authors: Kaneko Kensuke , Osada Takuya , Haystead Timothy A. , Morse Michael A. , Lyerly Herbert K.
Published: July 2017

Background: Immunotherapy has become an emerging anti-cancer therapy, as immune checkpoint blockade with PD-1 or PD-L1 inhibition have been active against multiple cancer types. Nonetheless, there remain many non-responders, suggesting that combinations may improve activity, particularly combinations which can target tumors and have low toxicity. Methods & Results: We have developed a novel Hsp90 targeted photodynamic therapy (Hsp90-PDT), which causes local anti-tumor responses. Hsp90-PDT can be combined with immune checkpoint blockade to generate potent anti-tumor effects. We synthesized well characterized PDT agent, verteporfin (VP) tethered to a HSP90 small molecule inhibitor, to create a Hsp90-PDT agent (Hsp90i-VP). We first compared the uptake of VP and Hsp90i-VP into the breast cancer cell lines (E0771-OVA) in vitro. Hsp90i-VP showed stronger incorporation than VP when the compounds were added 3μM or less in concentration. We next performed in vitro and vivo PDT with Hsp90i-VP and laser exposure (690 nm wavelength). Killing efficiency of VP and Hsp90i-VP was analyzed by MTT assay, which showed increased killing in a dose dependent manner for both photosensitizers (0-30µM) and laser (0-120J/cm2). Next, we treated E0771-OVA tumor-bearing C57BL/6 mice with Hsp90i-VP (25nmol/mouse) administration alone, laser irradiation (240J/cm2) alone, or the combination of Hsp90i-VP and laser. Suppression of the tumor growth was confirmed only in Hsp90i-VP and laser group. In addition, induction of tumor-specific immune response by PDT was confirmed by ELISA, where the elevation of anti-OVA antibody was seen only in PDT group. Furthermore, combination treatment of Hsp90-PDT and anti-PD-L1 mAbs was able to enhance cytotoxic T-cell response and suppress tumor growth significantly even with relatively PDT-resistant cell line. Conclusions: Hsp90-PDT showed a significant ability to induce antigen-specific immune response in both in vitro and in vivo. These results suggest that Hsp90-PDT can play an important role in anti-cancer immune therapy. Our future plan is to investigate the effect of local photodynamic therapy combined with various types of checkpoint inhibitor in breast cancer models.

Citation Format: Kensuke Kaneko, Takuya Osada, Timothy A. Haystead, Michael A. Morse, Herbert K. Lyerly. Induction of enhanced tumor-specific immunity by Hsp90 targeted photodynamic therapy (Hsp90-PDT) combined with immune checkpoint inhibition [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 4709. doi:10.1158/1538-7445.AM2017-4709
Abstract 472: Role of miRNAome deregulation in the pathogenesis of non-alcoholic steatohepatitis (NASH)-derived hepatocellular carcinoma
Authors: Ortega Juliana F. , DeConti Aline , Dreval Kostiantyn , Moreno Fernando S. , Beland Frederick A. , Pogribny Igor P.
Published: July 2017

Recent epidemiological studies have attributed the growing incidence of liver cancer, including hepatocellular carcinoma (HCC), in the United States to NASH, an advanced form of non-alcoholic fatty liver disease. Elucidating the molecular pathways that lead to the development of NASH-derived HCC is critical not only for identifying early diagnostic biomarkers of the disease, but also for treatment and prevention. We have investigated the role of microRNAs (miRNAs) in the development of NASH-derived HCC by using a Stelic Animal Model (STAM) of liver carcinogenesis. Using Liver miFinder PCR arrays, we examined the miRNA profiles in the livers of STAM mice and identified 25 and 35 miRNAs that were differentially expressed at a NASH-fibrotic stage (12 weeks) and in full-fledged HCC (20 weeks), respectively. Among these differentially expressed miRNAs, 18 miRNAs were in common. Multi-algorithm target prediction analysis of the common differentially expressed miRNAs revealed the involvement of these miRNAs in the deregulation of major molecular processes associated with the development of HCC, including epithelial-mesenchymal transition, HCC stem cell activation, and the induction of the β-catenin/Wnt and Hippo signaling pathways. These findings were confirmed by a marked up-regulation of Yes-associated protein 1 (YAP1), the main effector of the Hippo-pathway, the activation of epithelial-mesenchymal transition, and an increased-expression of hepatic progenitor cell markers. In addition to the common differentially expressed miRNAs, 10 miRNAs, including 5 members of the polycistronic oncomir miR-17-92 cluster, were found to be over-expressed in only HCC. These results indicate the fundamental role of miRNAs in the development of NASH-associated HCC. This is evidenced by the early occurrence of miRNA alterations at the preneoplastic stage of liver carcinogenesis, the persistence of these changes in HCC, and the accumulation of additional miRNA alterations in HCC, and by the existence of a mechanistic link between miRNA alterations and deregulation key cancer-related pathways.

Citation Format: Juliana F. Ortega, Aline DeConti, Kostiantyn Dreval, Fernando S. Moreno, Frederick A. Beland, Igor P. Pogribny. Role of miRNAome deregulation in the pathogenesis of non-alcoholic steatohepatitis (NASH)-derived 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 472. doi:10.1158/1538-7445.AM2017-472
Abstract 4733: Dysregulation of CUL4A and CUL4B ubiquitin ligases in lung cancer
Authors: Li Hui , Jia Lei , Yan Fan , Zhou Pengbo
Published: July 2017

Background: The Cullin-RING ubiquitin ligases 4 (CRL4) is implicated in controlling cell cycle, DNA damage repair and checkpoint response based on studies employing cell lines and mouse models. CRL4 proteins, including CUL4A and CUL4B are often highly accumulated in human malignancies. Elevated CRL4 attenuates DNA damage repair and increases genome instability that is believed to facilitate tumorigenesis. However, this has yet to be evaluated in human cancer patients.

Methods: 352 lung cancer and 62 benign lung specimens of Asian origin were constructed into tissue microarrays (TMAs) of four distinct lung cancer subtypes. Expression of CUL4A, CUL4B and their substrates was detected by immunohistochemistry (IHC), and analyzed statistically for their prognostic value and association with DNA damage response and genomic instability.

Results: Both CUL4A and CUL4B are overexpressed in the majority of lung carcinomas (PCUL4A<0.001, PCUL4B<0.001), and significantly associated with tumor size (PCUL4A<0.001, PCUL4B=0.002), lymphatic invasion (PCUL4A=0.004, PCUL4B<0.001), metastasis (PCUL4A=0.019, PCUL4B=0.006), and advanced TNM stage (PCUL4A<0.001, PCUL4B<0.001), which parallels gene amplification and abnormal activation of the canonical Wnt signaling. Moreover, overexpression of CUL4A, but not CUL4B, is significantly associated with tobacco smoking (P=0.01), and inversely correlated with XPC and p21, both of which are substrates of CUL4A (PCUL4A=0.019, PCUL4B=0.006). Higher levels of CUL4A or CUL4B are significantly associated with patients’ OS (PCUL4A<0.001, PCUL4B<0.001) and PFS (PCUL4A<0.001, PCUL4B=0.001).

Conclusion: Our findings revealed that CUL4A and CUL4B are differentially associated with etiological factors for pulmonary malignancies and are independent prognostic markers for survival of distinct lung cancer subtypes.

Citation Format: Hui Li, Lei Jia, Fan Yan, Pengbo Zhou. Dysregulation of CUL4A and CUL4B ubiquitin ligases 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 4733. doi:10.1158/1538-7445.AM2017-4733
Abstract 4735: Expression of Notch1 and Numb in small cell lung cancer
Authors: Kikuchi Hajime , Sakakibara-Konishi Jun , Furuta Megumi , Yokouchi Hiroshi , Nishihara Hiroshi , Yamazaki Shigeo , Hidetaka Uramoto , Tanala Fumihiro , Harada Masao , Akie Kenji , Sugaya Fumiko , Fujita Yuka , Takamura Kei , Kojima Tetsuya , Harada Toshiyuki , Higuchi Mitsunori , Honjo Osamu , Minami Yoshinori , Watanabe Naomi , Oizumi Satoshi , Suzuki Hiroyuki , Ishida Takashi , Dosaka-Akita Hiotoshi , Isobe Hiroshi , Munakata Mitsuru , Nishimura Masaharu
Published: July 2017

Background: Notch signaling plays an important role in tumorigenesis. Numb represses intracellular Notch signaling. Previous studies have demonstrated that Notch signaling suppresses the proliferation of small cell lung cancer (SCLC) cell lines. However, in SCLC, the association between Notch1 and Numb expression and clinicopathological factors or prognosis has remained unclear. In this study, we evaluated the expression of Notch1 and Numb in SCLC.

Methods: We immunohistochemically assessed 125 SCLCs that were surgically resected at 16 institutions participating in either the Hokkaido Lung Cancer Clinical Study Group Trial (HOT) or the Fukushima Investigative Group for Healing Thoracic Malignancy (FIGHT) between 2003 and 2013. Correlations between Notch1 or Numb expression and various clinicopathological features were evaluated.

Results: Notch1 expression was associated with ECOG performance status. Numb expression was associated with age, sex, and pathological histology (SCLC or Combined SCLC). Analysis of cellular biological expression did not demonstrate a significant correlation between the expression of Notch1 and of Numb. Multivariate Cox regression analysis showed that high Notch1 expression was an independent favorable prognostic factor for SCLC (hazard ratio = 0.503, P = 0.023).

Conclusions: We demonstrate that Notch1 expression, but not Numb, is associated with prognosis in SCLC and may provide a novel prognostic marker of SCLC.

Citation Format: Hajime Kikuchi, Jun Sakakibara-Konishi, Megumi Furuta, Hiroshi Yokouchi, Hiroshi Nishihara, Shigeo Yamazaki, Uramoto Hidetaka, Fumihiro Tanala, Masao Harada, Kenji Akie, Fumiko Sugaya, Yuka Fujita, Kei Takamura, Tetsuya Kojima, Toshiyuki Harada, Mitsunori Higuchi, Osamu Honjo, Yoshinori Minami, Naomi Watanabe, Satoshi Oizumi, Hiroyuki Suzuki, Takashi Ishida, Hiotoshi Dosaka-Akita, Hiroshi Isobe, Mitsuru Munakata, Masaharu Nishimura. Expression of Notch1 and Numb in 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 4735. doi:10.1158/1538-7445.AM2017-4735
Abstract 4737: A lung cancer risk classifier comprising genome maintenance genes measured in normal bronchial epithelial cells
Authors: Yeo Jiyoun , Crawford Erin , Zhang Xiaolu , Khuder Sadik , Chen Tian , Levin Albert , Blomquist Thomas , Willey James
Published: July 2017

Background: Annual low dose CT (LDCT) screening of individuals at high demographic risk reduces lung cancer mortality by more than 20%. However, subjects selected for screening based on demographic criteria typically have less than a 10% lifetime risk for lung cancer. Thus, there is need for a biomarker that better stratifies subjects for LDCT screening. Toward this goal, we previously reported a lung cancer risk test (LCRT) biomarker comprising 14 genome-maintenance (GM) pathway genes measured in normal bronchial epithelial cells (NBEC) that accurately classified cancer (CA) from non-cancer (NC) subjects. The primary goal of the studies reported here was to optimize the LCRT biomarker for high specificity and ease of clinical implementation.

Methods: Targeted competitive multiplex PCR amplicon libraries were prepared for next generation sequencing (NGS) analysis of transcript abundance at 68 sites among 33 GM target genes in NBEC specimens collected from a retrospective cohort of 120 subjects, including 60 CA cases and 60 NC controls. Genes were selected for analysis based on contribution to the previously reported LCRT biomarker and/or prior evidence for association with lung cancer risk. Linear discriminant analysis was used to identify the most accurate classifier suitable to stratify subjects for screening.

Results: The best classifier comprised 11 features including age and 10 transcript abundance assays representing nine genes; specifically, CDKN1A, E2F1, ERCC5, GPX1, GSTP1, KEAP1, TP63, XPA, and XRCC1. After cross-validation, the Receiver Operator Characteristic area under the curve was 0.94 (95% CI: 0.92-0.96) and overall classification accuracy was 88.3% (95% CI 82.4%-94.1%). Using a threshold set at 95% sensitivity, specificity was > 70%.

Conclusions: The LCRT biomarker reported here displayed high specificity and ease of implementation on a high throughput, quality-controlled targeted NGS platform. As such, it is optimized for clinical validation in specimens from the ongoing LCRT blinded prospective cohort study. Following validation, the biomarker is expected to have clinical utility by better stratifying subjects for annual lung cancer screening compared to current demographic criteria alone.

Citation Format: Jiyoun Yeo, Erin Crawford, Xiaolu Zhang, Sadik Khuder, Tian Chen, Albert Levin, Thomas Blomquist, James Willey. A lung cancer risk classifier comprising genome maintenance genes measured in normal bronchial epithelial 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 4737. doi:10.1158/1538-7445.AM2017-4737
Abstract 4743: Identification of metastasis-associated biomarker PLEC1 by iTRAQ-based proteomic approach and prognostic role of PLEC1 in patients with hepatocellular carcinoma
Authors: Song Lin-lan , Chen Yu , Mu Di , Jiang Xiao-Yan , Li Yu-Nong , Jiang Ling-Yu , Zhang Zhen-Fang , Yan Liang , Yang Min , Yang Yi-xuan , Ren Hong , Liao Yong
Published: July 2017

Purpose: Hepatocellular Carcinoma(HCC)is the fifth most prevalent cancer and the second most frequent cause of cancer-related death in men worldwide because of high incidence of recurrence and widespread metastasis. However, to date, there have been no direct metastesis-associated biomarker which is critical for the effective treatment of patients with HCC, we thus aimed to identify novel HCC metastasis-associated biomarkers with prognostic significance.E

xperimental Design: Two human HCC cell lines, MHCC97-L and MHCC97-H with similar genetic background and stepwise increasing metastatic potentials, were used to explore potential metastasis-associated biomarkers for HCC by isobaric tag for relative and absolute quantitation (iTRAQ) combination with 2D-LC-MS/MS. Ninety-five HCC cancerous and paracancerous tissues were further analyzed by immunohistochemistry to validate the correlations between the expression levels of PLEC1 with various clinicopathologic factors and patient survival.

Results: Using iTRAQ combination with 2D-LC-MS/MS, a total of 47 differentially expressed proteins were found in MHCC97-L and MHCC97-H cells, of which 26 proteins were up regulated and 21 proteins were down regulated. Plectin-1 (PLEC1) was identified and re-confirmed as a potential biomarker for HCC that was highly up-regulated in tumor tissues, in comparison with that of para-cancerous tissues. Over-expression of PLEC1 was also strongly correlated with tumor size (P < 0.05) and TMN stages (P < 0.05). In addition, both univariate and multivariate analyses demonstrate that PLEC1 expression is an independent prognostic factor for patients with HCC(p<0.05).

Conclusions: In this study, iTRAQ-based proteomic technology was explored to uncover novel metastasis-associated biomarkers in HCC. We identified and validated metastasis-associated protein PLEC1 as a potential prognostic marker in patients with HCC.

Note: This abstract was not presented at the meeting.

Citation Format: Lin-lan Song, Yu Chen, Di Mu, Xiao-Yan Jiang, Yu-Nong Li, Ling-Yu Jiang, Zhen-Fang Zhang, Liang Yan, Min Yang, Yi-xuan Yang, Hong Ren, Yong Liao. Identification of metastasis-associated biomarker PLEC1 by iTRAQ-based proteomic approach and prognostic role of PLEC1 in patients with 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 4743. doi:10.1158/1538-7445.AM2017-4743
Abstract 4748: Novel predictive biomarker for monitoring adverse reactions to radiation therapy
Authors: OKunieff Paul , Swarts Steven , Peletskya Elena , Vallerga Anne , Chuang Rachel , Powell Michael J. , Zhang Aiguo
Published: July 2017

Introduction: Radiation treatment is required by 70% of cancer patients, however there is currently no clinical method for determining the therapeutic response or radiation induced toxicity that can be used during a course of radiation therapy to personalize the dose for individual patients. The only standard method is CT/PET and/or MRI. This is a major clinical concern for radiation oncologists with so many new agents being approved in combination with radiation therapy.

Methodology: Herein we describe a highly sensitive clinically validated assay that measures the extent of normal tissue damage induced by radiation by quantitation of circulating free DNA (cfDNA) derived from cellular apoptosis detected in plasma of patients undergoing radiation therapy. The assay employs DNA capture probes and SuperbDNATM signal amplification technology with alkaline phosphatase labelled signaling probes coupled with dioxetane phosphate chemiluminescence detection. The assay can be performed directly on patient plasma samples and can be readily automated.

Conclusion: RadTox TM can be used both for research and clinical testing of plasma samples for patients undergoing radiation therapy for optimization and personalization of treatment.

Citation Format: Paul OKunieff, Steven Swarts, Elena Peletskya, Anne Vallerga, Rachel Chuang, Michael J. Powell, Aiguo Zhang. Novel predictive biomarker for monitoring adverse reactions to radiation therapy [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 4748. doi:10.1158/1538-7445.AM2017-4748
Abstract 475: Role of microRNA-34a in regulating oncoprotein STMN1
Authors: Chakravarthi Balabhadrapatruni V.S.K. , Mehra Rohit , Wang Rui , Chandrashekar Darshan Shimoga , Balasubramanya Sai Akshaya Hodigere , Asangani Irfan A. , Lonigro Robert J. , Chinnaiyan Arul M. , Varambally Sooryanarayana
Published: July 2017

MicroRNAs, as negative regulators of gene expression, play a major role in cellular homeostasis. Downregulation of miRs is common in cancer which results in upregulation of multiple oncogenes in cancer. Here we show that stathmin (STMN1), which is over-expressed and oncogenic in wide variety of cancers, is regulated by microRNA-34 in prostate cancer. Earlier studies suggest that stathmin is involved in vital cellular processes such as cell proliferation, motility and metastasis. In the present study, we show elevated expression of stathmin in metastatic prostate cancer. Functionally, knockdown of stathmin in prostate cancer cells resulted in reduced cell proliferation and invasion in vitro, and tumor growth and metastasis in vivo. We also show that tumor suppressor microRNA miR-34a targets stathmin and mir-34 is regulated by transcriptional corepressor CtBP1 (C-Terminal Binding Protein 1). Further, Microarray analyses using stathmin modulated prostate cancer cell line RNA revealed multiple molecular alterations which may play a critical role in STMN1-mediated oncogenic activity. Thus, our study suggests that CtBP1-regulated miR-34a modulates stathmin expression in prostate cancer. This study therefore demonstrates functional role of CtBP1-miR-34a-Stathmin axis in prostate cancer progression.

Citation Format: Balabhadrapatruni V.S.K. Chakravarthi, Rohit Mehra, Rui Wang, Darshan Shimoga Chandrashekar, Sai Akshaya Hodigere Balasubramanya, Irfan A. Asangani, Robert J. Lonigro, Arul M. Chinnaiyan, Sooryanarayana Varambally. Role of microRNA-34a in regulating oncoprotein STMN1 [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 475. doi:10.1158/1538-7445.AM2017-475
Abstract 4750: Predicting severe hematologic toxicity from extended-field chemoradiation of para-aortic nodal metastases from cervical cancer
Authors: Yan Kevin , Ramirez Ezequiel , Gu Xuejun , Albuquerque Kevin
Published: July 2017

Background and Purpose: To determine significant factors predictive for severe hematologic toxicity (HT) in cervical cancer patients with para-aortic lymph node (PALN) metastasis treated with concurrent chemoradiation with a specific focus on radiation dose to total bone marrow (BMTOT) and active bone marrow (BMACT). To create a nomogram using significant factors to predict HT in these patients.

Material and Methods: 38 Patients with cervical cancer and PALN metastasis who underwent 18F-FDG-PET / CT before treatment with extended field radiation therapy (EFRT) and concurrent cisplatin were analyzed. BMACT was defined as the region within BMTOT with a standardized uptake value (SUV) greater than or equal to the mean for the individual. Blood counts were collected weekly from the beginning of radiation treatment to the end of radiation treatment. HT was graded based on the guidelines set by the National Cancer Institute Common Terminology Criteria for Adverse Events Version 4.0.

Results: 19 patients (50%) had Grade 3 or higher hematologic toxicity (HT3+), not including lymphocyte toxicity. Patients who were obese (n=12) were less likely to get HT3+ compared to patients who were not obese (p=0.03) despite getting the same weight related dose of chemotherapy. Volume of BMTOT receiving 20 Gy, 30 Gy, and 45 Gy were significant predictors for HT3+ at 78.56% (p=0.01), 47.14% (p=0.00), and 20.36% (p=0.01) respectively. Volume of BMACT receiving 10 Gy, 20 Gy, 30 Gy, and 45 Gy were significant predictors for HT3+ at 95.50% (p=0.03), 80.52% (p=0.05), 59.64% (p=0.03), and 31.74% (p=0.01) respectively. Through logistic regression, the probability of developing HT3+ is given by the equation: Prob(HT3+) = 1 / (1 + exp(7.34 + 0.22*BMI - 0.44*Mean Dose to BMTOT)). Patients who had HT3+ received an average of 4 cycles of chemotherapy and 62 days of treatment time, significantly different than the 4.74 chemotherapy cycles and 53 days of treatment in patients without HT3+ (p=0.05, 0.00 respectively).

Conclusions: Both higher patient BMI and bone marrow irradiation were associated with HT3+. A simplified nomogram has been created to predict HT3+ in these patients. Radiation parameters have been identified for cervical cancer patients with PALN involvement receiving EFRT concurrently with chemotherapy. Bone marrow sparing approaches for EFRT need to be addressed to improve patient care.

Citation Format: Kevin Yan, Ezequiel Ramirez, Xuejun Gu, Kevin Albuquerque. Predicting severe hematologic toxicity from extended-field chemoradiation of para-aortic nodal metastases from cervical 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 4750. doi:10.1158/1538-7445.AM2017-4750
Abstract 4753: Prognostic significance of galectin-1 expression in patients with glioblastoma multiforme following radiotherapy
Authors: Huang Eng-Yen , Chou Shang-Yu
Published: July 2017

Purpose: Galectin-1 is a radioresistant marker and prognostic factor in our previous study for cervical cancer. The aim of current study is to determine the prognostic significance of galectin-1 expression level in patients with glioblastoma multiforme (GBM) undergoing adjuvant radiotherapy (RT).

Materials and Methods: We included 45 patients with GBM who treated with maximal safe surgical resection or biopsy only, followed by adjuvant RT of EQD2 >= 60 Gy (equivalent doses as 2-Gy fractions) for homogeneous treatment. Paraffin-embedded tissues acquired from the Department of Pathology were analyzed using immunohistochemistry staining for galectin-1 expression. Overall survival (OS) and cancer-specific survival were compared between strong and weak expression of galectin-1.

Results: According to univariate analysis for optimal cutoff of galectin-1 expression level, better median survival (27.9 months) in patients with weak level than that (10.7 months) in patients with strong level (p = 0.009) was revealed. We compared characteristics between weak and strong galectin-1 expression level, and only galectin-3 expression level was correlated. The weak galectin-1 expression group displayed a 3-year survival rate of 27.3% and 3-year cancer-specific survival (CSS) rate of 27.3%, but which are only 5.9% and 7.6% respectively in strong galectin-1 expression group (p = 0.009 and 0.020, respectively). Cox regression was used to confirm that the role of galectin-1 expression level (weak vs. strong) is a significant factor of OS (p = 0.020) and CSS (p = 0.022). Other parameters such as galectin-3 expression level, ECOG performance, gender, operation method, age > 50 years, or radiation field were not significant factors.

Conclusion: Galectin-1 expression level affects OS and CSS in patient with GBM treated with adjuvant RT.

Citation Format: Eng-Yen Huang, Shang-Yu Chou. Prognostic significance of galectin-1 expression in patients with glioblastoma multiforme following radiotherapy [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 4753. doi:10.1158/1538-7445.AM2017-4753
Abstract 4757: Radioresistance is linked with stem-like properties via activation of aryl hydrocarbon receptor
Authors: Liu SHuang , Yan Bin , Tao Yongguang
Published: July 2017

Most cancer patients receive radiotherapy in the course of their disease and the occurrence of radioresistance is associated with a poor prognosis for cancer patients. The molecular pathways that drive the enhanced tumorigenic potential during the development of radioresistance are poorly understood. Here, we demonstrate that the aryl hydrocarbon receptor (AhR) plays a vital and previously unrecognized role in the maintenance of stem-like cells that are isolated from radioresistant epithelial lung cancer and nasopharyngeal carcinoma. AhR promotes the stem-like phenotype and drives metastasis by directly targeting the promoters of ‘stemness’ genes, such as the ATP-binding cassette sub-family G member 2 (ABCG2) gene. Cancer cell populations enriched for ABCG2 expression (CD338+) show an elevated ‘stemness’ signature and are more prone to cancer progression in vitro and in vivo. Ubiquitin C-terminal hydrolase-L3 (UCHL3), a deubiquitinating enzyme, may counteract AhR, while IKKα, a bone fide chromatin modifier, may directly activate stemness related genes. The radioresistant cancer cell sublines are associated with a hybrid state in epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET), and elevated tumor associated macrophages and proinflammatory factors. Moreover, the radioresistant sublines display high levels of oncometabolites, including α-ketoglutarate, and treatment of cancer cells with α-ketoglutarate enhances their stem-like properties through activation of the AhR. Thus AhR is functionally linked with radioresistance in lung cancer and nasopharyngeal carcinoma, and it drives tumorigenesis by an epithelial and mesenchymal composition, tumor associated macrophages and oncometabolites.

Note: This abstract was not presented at the meeting.

Citation Format: SHuang Liu, Bin Yan, Yongguang Tao. Radioresistance is linked with stem-like properties via activation of aryl hydrocarbon receptor [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 4757. doi:10.1158/1538-7445.AM2017-4757
Abstract 4770: Novel CXCR3 allosteric modulators inhibit breast cancer stem cells and metastasis
Authors: Kundu Namita , Brox Regine , Ma Xinrong , Reader Jocelyn , Tschammer Nuska , Fulton Amy M.
Published: July 2017

There is an urgent need to identify molecular targets that are relevant to metastatic and/or stem-like cancer cells (CSC). Theoretically, surface-expressed G-protein coupled receptors on CSC are attractive therapeutic targets. There is considerable evidence that the inflammatory milieu of the tumor microenvironment drives CSC and we now show that two isoforms of the chemokine receptor CXCR3 play important but distinct roles in cancer behavior. In the bulk tumor cell population, CXCR3, isoform A is highly expressed relative to CXCR3B. CXCR3A is critical to metastatic success. CXCR3, isoform B, is highly upregulated in breast cancer CSC. When CXCR3B is gene-silenced in a model of basal-type, metastatic breast cancer, tumorsphere-forming capacity is reduced and the aldehyde dehydrogenase-positive (ALDH1+) population is correspondingly decreased. Conversely, overexpression of CXCR3B enhances these CSC properties. Thus, CXCR3B is critical to maintenance of the phenotype and function of breast CSC. CXCR3 ligands CXCL9, CXCL10 and CXCL11 all bind with high affinity to CXCR3, however, each ligand is coupled to different intracellular signaling pathways. For example, others have reported that, in HEK cells, CXCL10 strongly induces ERK activation, but CXCL11 does not. In our hands, CXCL11 more potently induces an ALDH1+ population in MDA-MB-231 cells compared to CXCL10. We have developed novel allosteric modulators of CXCR3 that demonstrate probe (ligand)-dependence. BD64 preferentially inhibits CXCL11-mediated β-arrestin 2 recruitment relative to inhibition of GPCR signaling; BD103 is significantly more effective at blocking CXCL11 versus CXCL10-mediated GPCR signaling. Neither BD64 nor BD103 affects the proliferation of non-CSC, however both compounds inhibit the ability of CSC to form tumorspheres. Furthermore, both CXCR3 antagonists reverse ligand-mediated induction of the ALDH1+ fraction. Most interesting is that both BD64 and BD103 effectively inhibit tumor cell colonization of the lung. These data suggest that optimal tumor control will be achieved when both metastatic, CXCR3Ahigh as well as CXCR3Bhigh CSC populations are inhibited by highly specific CXCR3 modulators.

Note: This abstract was not presented at the meeting.

Citation Format: Namita Kundu, Regine Brox, Xinrong Ma, Jocelyn Reader, Nuska Tschammer, Amy M. Fulton. Novel CXCR3 allosteric modulators inhibit breast cancer stem cells and metastasis [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 4770. doi:10.1158/1538-7445.AM2017-4770
Abstract 4771: The role of Hes1 in the normal and tumor stem cells of the intestine
Authors: Goto Norihiro , Fukuda Akihisa , Chiba Tsutomu , Seno Hiroshi
Published: July 2017

Notch-Hes pathway plays a crucial role in the differentiation, proliferation, and tumorigenesis of the intestine. We previously reported that prenatal deletion of Hes1 reduces intestinal tumor proliferation; however, postnatal deletion of Hes1, which recapitulates Hes1-targeted tumor therapy, has not been thoroughly investigated. Here, we elucidated the novel role of Hes1 in normal stem cells (NSCs) and tumor stem cells (TSCs) of the adult intestine. First, we deleted Hes1 in Lgr5-expressing or Bmi1-expressing NSCs and performed lineage tracing using Lgr5CreERT2/+; Hes1flox/flox; Rosa26LacZ/+ and Bmi1CreER/+; Hes1flox/flox; Rosa26LacZ/+ mice. After Hes1 deletion, Lgr5-expressing or Bmi1-expressing NSCs gave rise to progeny cells, but could not self-renew. However, Hes1 deletion in NSCs did not disrupt intestinal homeostasis, because Hes1-positive NSCs were newly generated soon after Hes1-deleted NSCs were depleted. Next, we stabilized beta-catenin for intestinal tumor formation and deleted Hes1 in Lgr5-expressing cells using Lgr5CreERT2/+; Ctnnb1lox(ex3)/+; Hes1flox/flox mice. Tumor formation was dramatically suppressed, and the animal survival was significantly improved, suggesting that Hes1 is required for the initiation of intestinal tumor formation. Finally, we deleted Hes1 in Lgr5-expressing TSCs of established intestinal tumors using Lgr5CreERT2/+; Hes1flox/flox; ApcMin/+ mice. Of note, Hes1 deletion in TSCs of established intestinal tumors induced immediate apoptosis of TSCs and significantly reduced the number of tumors. qRT-PCR analysis of sorted Lgr5-expressing cells one day after Hes1 deletion revealed upregulation of pro-apoptotic genes and downregulation of anti-apoptotic genes in Hes1-deleted TSCs, but not in Hes1-deleted NSCs. We also deleted Hes1 in Dclk1-expressing TSCs of established intestinal tumors, and confirmed that, irrespective of stem cell markers analyzed, Hes1 deletion in TSCs leads to tumor regression by inducing immediate apoptosis in TSCs. Taken together, Hes1 is required in different ways for the maintenance of NSCs and TSCs: Hes1 deletion in NSCs does not perturb homeostasis, whereas Hes1 deletion in TSCs leads to tumor regression. Furthermore, Hes1 is necessary for the initiation of intestinal tumor formation. Therefore, we propose Hes1 is a novel tumor-specific therapeutic target.

Citation Format: Norihiro Goto, Akihisa Fukuda, Tsutomu Chiba, Hiroshi Seno. The role of Hes1 in the normal and tumor stem cells of the intestine [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 4771. doi:10.1158/1538-7445.AM2017-4771