bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 120: Detection of acrolein-derived cyclic DNA adducts in human cells by monoclonal antibodies.
Authors: Pan Jishen , Awoyemi Bisola , Xuan Zhuoli , Vohra Priya , Wang Hsiang-Tsui , Dyba Marcin , Greenspan Emily , Fu Ying , Creswell Karen , Zhang Lihua , Berry Deborah , Tang Moon-Shong , Chung Fung-Lung
Published: August 2014

Acrolein (Acr) is a ubiquitous environmental pollutant found in cigarette smoke and automobile exhaust. It can also be produced endogenously by oxidation of polyunsaturated fatty acids. The Acr-derived 1,N2-propanodeoxyguanosine (Acr-dG) adducts in DNA are mutagenic lesions that are potentially involved in human cancers. In this study, monoclonal antibodies were raised against Acr-dG adducts and characterized using ELISA. They showed strong reactivity and specificity towards Acr-dG, weaker reactivity towards crotonaldehyde- and trans-4-hydroxy-2-nonenal-derived 1,N2-propanodeoxyguanosines, and weak or no reactivity towards 1,N6-ethenodeoxyadenosine, 8-oxo-deoxyguanosine, and benzo(a)pyrene- and malondialdehdye-derived adducts. Using these novel antibodies, we developed assays to detect Acr-dG in vivo: First, a simple and quick FACS-based assay for detecting these adducts directly in cells; Second, a highly sensitive direct ELISA assay for measuring Acr-dG in cells and tissues using only one μg DNA; And third, a competitive ELISA for better quantitative measurement of Acr-dG levels in DNA. The assays were validated using Acr-treated HT29 cell DNA samples or calf thymus DNA and the results were confirmed by LC-MS/MS-MRM. An immunohistochemical assay was also developed to detect and visualize Acr-dG in HT29 cells as well as in human oral cells. These antibody-based methods provide useful tools for the studies of Acr-dG as a cancer biomarker and of the molecular mechanisms by which cells respond to Acr-dG as a ubiquitous DNA lesion. (This work was supported by NCI grant CA134892)

Citation Format: Jishen Pan, Bisola Awoyemi, Zhuoli Xuan, Priya Vohra, Hsiang-Tsui Wang, Marcin Dyba, Emily Greenspan, Ying Fu, Karen Creswell, Lihua Zhang, Deborah Berry, Moon-Shong Tang, Fung-Lung Chung. Detection of acrolein-derived cyclic DNA adducts in human cells by monoclonal antibodies. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 120. doi:10.1158/1538-7445.AM2013-120
Abstract 121: Markers of cell proliferation and growth as potential preneoplastic biomarkers of risk for colorectal neoplasms.
Authors: Tu Huakang , Woodard Jill , Daniel Carrie , Gonzalez-Feliciano Amparo , Ahearn Thomas , Bostick Roberd M.
Published: August 2014

Background: Increased colorectal epithelial cell proliferation and growth are early, common events in sporadic colorectal cancer. To assess the potential of cell proliferation and growth markers as pre-neoplastic biomarkers of risk for colorectal neoplasms, we conducted a pilot, colonoscopy-based case-control study (n = 51 cases and 154 controls) of incident, sporadic colorectal adenoma.

Methods: We evaluated transforming growth factor alpha (TGFα), TGFβ1, and telomerase reverse transcriptase (hTERT) expression in normal-appearing mucosa from the rectum and ascending and sigmoid colon using automated immunohistochemistry and quantitative image analysis in a subset of cases and controls (n = 40 and 46, respectively). Mean differences in staining batch-standardized biomarker labeling optical density between cases and controls were assessed using analysis of covariance, and associations of the biomarkers with adenomas were assessed using unconditional logistic regression.

Results: In the normal rectal mucosa, hTERT expression was 13% (P = 0.06) higher in cases than in controls, and higher hTERT expression (above the median in controls) was associated with a three-fold increased risk of colorectal adenoma (OR = 3.28, 95% confidence interval [CI], 1.04 – 10.35). TGFα expression was 31% (P = 0.25) higher in cases than in controls, and higher TGFα expression was associated with a 44% increased risk of colorectal adenoma (OR = 1.44, 95% CI 0.50 – 4.16). TGFβ1 expression alone and the ratio of TGFα to TGFβ1 expression were 3% (P = 0.86, OR = 0.91, 95% CI 0.39 – 2.11) and 61% (P = 0.30, OR = 1.23, 95% CI 0.45 – 3.36), respectively, higher in cases than in controls.

Conclusions: These preliminary data suggest that hTERT and TGFα expression and, perhaps especially, the balance of TGFα and TGFβ1 expression in the normal rectal mucosa may be directly associated with risk for incident, sporadic colorectal neoplasms, providing support for further investigation of these markers as potential biomarkers of risk for colorectal neoplasms.

Citation Format: Huakang Tu, Jill Woodard, Carrie Daniel, Amparo Gonzalez-Feliciano, Thomas Ahearn, Roberd M. Bostick. Markers of cell proliferation and growth as potential preneoplastic biomarkers of risk for colorectal neoplasms. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 121. doi:10.1158/1538-7445.AM2013-121
Abstract 1214: RET fusion gene: translation to the therapy of lung adenocarcinoma.
Authors: Kohno Takashi , Ichikawa Hitoshi , Tsuta Koji , Mizukami Tatsuji , Shimada Yoko , Kato Mamoru , Sakamoto Hiromi , Tsuchihara Katsuya , Watanabe Shun-ichi , Nokihara Hiroshi , Goto Koichi , Yokota Jun , Yoshida Teruhiko , Shibata Tatsuhiro
Published: August 2014

A considerable proportion of lung adenocarcinomas, the most common histological type of lung cancer that comprises 40% of the total cases, develops through activation of oncogenes, for example, somatic mutations in EGFR or KRAS or fusion of ALK, in a mutually exclusive manner. Tyrosine kinase inhibitors targeting the EGFR and ALK proteins are effective in the treatment of lung adenocarcinomas that carry EGFR mutations and ALK fusions, respectively. We performed whole-transcriptome sequencing (RNA sequencing) of 30 lung adenocarcinoma specimens from Japanese individuals to identify new chimeric fusion transcripts that could be targets for therapy. Then, we identified KIF5B (kinesin family 5B)-RET fusion gene, as another druggable oncogene that is present in 2% (6/319) of lung adenocarcinomas (Kohno et al., Nat Med, 2012). The KIF5B-RET fusion leads to aberrant activation of RET kinase and is considered to be a new driver mutation segregating from mutations or fusions in EGFR, KRAS, HER2 and ALK. The RET fusion has preferentially been detected in well or moderately differentiated tumors of never smokers. The RET fusion in lung adenocarcinoma was also identified by three other groups: Dr. Ju et al of Seoul National University, Dr. Takeuchi et al of Japanese Foundation for Cancer Research, and Dr. Lipson et al of Foundation Medicine, USA. A clinical trial to address the therapeutic efficacy of a RET tyrosine kinase inhibitor, vandetanib, will be started in 2013 by Dr. Koichi Gotoh (kgoto@east.ncc.go.jp) of National Cancer Center Hospital East, Japan.

Citation Format: Takashi Kohno, Hitoshi Ichikawa, Koji Tsuta, Tatsuji Mizukami, Yoko Shimada, Mamoru Kato, Hiromi Sakamoto, Katsuya Tsuchihara, Shun-ichi Watanabe, Hiroshi Nokihara, Koichi Goto, Jun Yokota, Teruhiko Yoshida, Tatsuhiro Shibata. RET fusion gene: translation to the therapy of lung adenocarcinoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1214. doi:10.1158/1538-7445.AM2013-1214
Abstract 1221: Targeting interferon-λ1 with a stabilized dimer of anti-tumor Fab .
Authors: Liu Donglin , Rossi Edmund A. , Cardillo Thomas M. , Goldenberg David M. , Chang Chien-Hsing
Published: August 2014

Background: The type-III interferons (IFNs), comprising IFN-λ1, IFN-λ2, and IFN-λ3, are functionally similar to the type-I IFN-α in eliciting anti-viral, anti-tumor, and immune-modulating activities. Due to its more restricted cellular targets, IFN-λ is being considered as a potential alternative to existing therapeutic regimens based on IFN-α, whose adverse effects in patients limit its use. The development of antibody-targeted IFN-λ may further improve its safety, potency and pharmacokinetics. Methods: We generated three DOC-AND-LOCKTM (DNLTM) complexes, designated (E1)-λ1, (15)-λ1, and (C2)-λ1, by conjugating IFN-λ1 site-specifically to a stabilized dimer of humanized Fab derived from hRS7 (anti-Trop-2), hMN-15 (anti-CEACAM6), and hL243 (anti-HLA-DR), respectively, which were affinity-purified and evaluated in human malignant cell lines of cervix (ME-180), colon (HT-29), esophagus (TE-11, T.Tn), lung (SK-MES-1, A549), liver (HepG2, Huh-7), and skin (A375). Results: As determined by flow cytometry, Trop-2 was highly expressed on the surface of ME-180, TE-11, and T.Tn; moderately on SK-MES-1 and HT-29; and negligibly on HepG2, Huh-7, A549, and A375. In comparison, CEACAM6 was undetectable on SK-MES-1 and A375, but positive on the remaining 7 cell lines, yet at a considerably lower level than Trop-2. Targeting of these immunocytokines to antigen-expressing cells markedly increased the amount of IFN-λ1 localized at the cell surface, as demonstrated by (E1)-λ1 on ME-180 (Trop-2+), (15)-λ1 on HepG2 (CEACAM6+), and (C2)-λ1 on A375 (HLA-DR+), respectively. In MTS assays, (E1)-λ1 inhibited the in vitro proliferation of ME-180, SK-MES-1, and TE-11, with EC50 <1 pM, which was 1,000-fold more potent than commercial IFN-λ1. Likewise, the anti-proliferation activity of (15)-λ1 was enhanced ∼100-fold in ME-180 and TE-11, but not in CEACAM6-negative SK-MES-1. The observed potency was further substantiated by enhanced cell signaling, including STAT phosphorylation and MHC-I up-regulation induced by (15)-λ1 in HepG2 cells. In the Pk study, (E1)-λ1 showed a mean residence time of 12 h with a T1/2 of 8.6 h, and clearance of 2.2 ml/h in mouse serum, which is comparable to PEG-IFN-λ1. Conclusions: These promising results, attributable to increased localization and stronger binding to antibody-targeted cells, warrant further exploration of these novel immunocytokines as potential cancer therapeutics.

Citation Format: Donglin Liu, Edmund A. Rossi, Thomas M. Cardillo, David M. Goldenberg, Chien-Hsing Chang. Targeting interferon-λ1 with a stabilized dimer of anti-tumor Fab . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1221. doi:10.1158/1538-7445.AM2013-1221
Abstract 1225: Serum cytokine profiling in metastatic colorectal cancer patients: IL-8, MCP-1, and MCP-3 are theranostic targets.
Authors: Chen Zhi-Yuan , He Wen-Zhuo , Peng Li-Xia , Guo Rong-Ping , Xia Liang-Ping , Qian Chao-Nan
Published: August 2014

Background: The prognostic value of many circulating cytokines in metastatic colorectal cancer (CRC) patients is undetermined. By using high through-put approach, this study was to evaluate the prognostic values of circulating cytokines in patients with metastatic CRC and identify potential therapeutic targets.

Methods: Thirty-nine cytokines were simultaneously analyzed in serum samples from 99 patients with metastatic CRC by using multiplex bead-based Luminex technology. Three CRC cell lines, together with three more aggressive lines derived from them, were used to validate three potential therapeutic targets by applying neutralizing antibodies.

Results: Elevated levels of 17 cytokines were found to be significant risk factors for the overall survival (OS) of patients. Only macrophage-derived chemokine (MDC) positively correlated with better prognosis. Interleukin-8 (IL-8), monocyte chemotactic protein-1 (MCP-1), MCP-3 and MDC were significant independent predictor of OS in multivariate analyses. The risk scoring system developed by combining IL-8, MCP-1, MCP-3, and MDC, was more effective than each individual factor to predictor the OS. IL-8 was secreted by three CRC cell lines, with higher expression observed in more-aggressive cells. Blocking of IL-8 with a neutralizing antibody inhibited the invasiveness of CRC cells. MCP-1 and MCP-3 were secreted by SW480 cell line, with higher expression in more-aggressive cell. Neutralizing antibodies against MCP-1 or MCP-3 significantly inhibited the cellular invasion.

Conclusions: The serum levels of 18 cytokines correlated with the prognosis of metastatic CRC patinets. Among them, IL-8, MCP-1, MCP-3 and MDC were independent prognostic factors for OS. IL-8, MCP-1, and MCP-3 are potential therapeutic targets in CRC.

Citation Format: Zhi-Yuan Chen, Wen-Zhuo He, Li-Xia Peng, Rong-Ping Guo, Liang-Ping Xia, Chao-Nan Qian. Serum cytokine profiling in metastatic colorectal cancer patients: IL-8, MCP-1, and MCP-3 are theranostic targets. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1225. doi:10.1158/1538-7445.AM2013-1225
Abstract 1226: Interleukin-6 (IL-6) as a therapeutic target in prostate cancer.
Authors: Kura Yurie , De Velasco Marco A. , Kobayashi Yasuyuki , Ando Naomi , Fukushima Emiko , Yamamoto Yutaka , Hatanaka Yuji , Shimizu Nobutaka , Yoshimura Kazuhiro , Nozawa Masahiro , Yoshikawa Kazuhiro , Nishio Kazuto , Uemura Hirotsugu
Published: August 2014

Interleukin-6 or IL-6 is a pleiotropic cytokine which is implicated in the regulation of immune response, but is also involved in regulating the growth of various malignant tumors. Mounting evidence suggests that IL-6 contributes to prostate cancer progression. Activated IL-6 has been shown to be elevated in the sera from patients with metastatic prostate cancer and it appears that the prostate tissue itself is a source of IL-6 and its receptor. Experimental data suggests that the IL-6/IL-6R signaling axis is an autocrine and paracrine growth factor in androgen independent prostate cancer cell lines and can activate a variety of signal transduction cascades, some of which may activate androgen receptor activity. In the present study we examined the role of IL6-induced Stat3 activation in the development and progression of prostate cancer and its possible role as a target for the treatment of prostate cancer using the prostate-specific conditional targeting PTEN knockout mouse model. In PTEN-mutant mice, elevated levels of IL6/IL-6R correlated with increased cellular proliferation in non-castrate and castrate prostate cancer (CRPC). Similar to human prostate cancer, mice with CPRC showed elevated levels of IL-6 and IL6-R in both serum and prostate tissue. To establish a therapeutic role for IL6/IL6R inhibition, we conducted a series of pharmacodynamic studies using an anti-mouse IL-6R antibody (MR-16) in tumor bearing homozygous PTEN-mutant mice. Our data shows that STAT3 activation was inhibited in a dose dependent manner during acute IL6-R blockade. Inhibition of STAT3 activity after chronic IL-6R blockade was transient. However, at day 45 of treatment, tumor cell proliferation was inhibited and apoptosis was induced. These findings show that this mouse model is relevant for the study of Stat3 signaling and tumor progression and inhibition of Stat3 transcriptional activation by upstream signal blockade may be a promising approach for molecularly targeted therapeutic strategies in human prostate cancer. Thus this study will serve as the foundation for future drug intervention studies designed to assess the efficacy of targeting the JAK/STAT signal pathway using targeted monotherapy and combination drug therapy models.

Citation Format: Yurie Kura, Marco A. De Velasco, Yasuyuki Kobayashi, Naomi Ando, Emiko Fukushima, Yutaka Yamamoto, Yuji Hatanaka, Nobutaka Shimizu, Kazuhiro Yoshimura, Masahiro Nozawa, Kazuhiro Yoshikawa, Kazuto Nishio, Hirotsugu Uemura. Interleukin-6 (IL-6) as a therapeutic target in prostate cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1226. doi:10.1158/1538-7445.AM2013-1226
Abstract 1227: Macrophage depletion enhances the anticancer efficacy of eribulin mesylate in mice bearing syngeneic tumors.
Authors: Albu Diana I. , Littlefield Bruce A. , Woodall-Jappe Mary
Published: August 2014

Recent reports utilizing the MMTV-PyMT spontaneous model of breast cancer established that inhibition of macrophage recruitment to the tumors resulted in increased anti-cancer activity of paclitaxel. Eribulin mesylate (Halaven®) is a non-taxane microtubule-dynamics inhibitor recently approved for third line clinical use in patients with heavily pretreated metastatic breast cancer, based upon a statistically significant increase in median overall survival (OS) compared to treatment of physician's choice. In the present study we investigated whether systemic depletion of macrophages would enhance eribulin anti-tumor efficacy. In the subcutaneous Lewis lung carcinoma (LLC) syngeneic murine tumor model, we found that macrophage depletion via liposomal clodronate enhanced eribulin inhibition of tumor growth. Current studies are investigating the mechanism(s) of this activity.

Citation Format: Diana I. Albu, Bruce A. Littlefield, Mary Woodall-Jappe. Macrophage depletion enhances the anticancer efficacy of eribulin mesylate in mice bearing syngeneic tumors. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1227. doi:10.1158/1538-7445.AM2013-1227
Abstract 1230: Impact of apheresis (Aph) on number and function of lymphocytes in cancer patients (pts).
Authors: Gulley James L. , Heery Christopher R. , Steinberg Seth , Marte Jenn , Madan Ravi A. , Schlom Jeffrey
Published: August 2014

It has been suggested that Aph, used for Sipuleucel-T, may negatively affect the OS of cancer pts by removing circulating lymphocytes and diminishing host immunity. We explored data from prior clinical trials in which pts had Aph or simple phlebotomy for evidence to support or contradict this.

We examined data from cancer pts on clinical trials who had Aph (n=64) or only peripheral blood draws (n=90) as controls for immune analysis. As the 1° functional analysis, we evaluated the impact of Aph on the flu specific T-cells using ELISPOT. We also evaluated the impact on absolute lymphocyte count (ALC), T cells (CD3, CD4, CD8, and CD4:CD8 ratio). ELISPOT was expressed as number of cells/spot (i.e. 1/10,000 would be expressed as 10,000). For all parameters, baseline (pre) values were compared to post-treatment (post) values using a Wilcoxon signed rank test. A large two-tailed p-value would indicate no significant change in values. Findings from smaller groups who underwent Aph multiple times are described.

In the Aph group, the median age was 63 (range 38-82); 92% male. Comparing the pre and post values within the groups, with each pt as its own control, there was no significant difference when comparing ELISPOT, ALC, and T cells (Table). 13 pts had a 3rd Aph with ELISPOT; 7 had improved responses to flu with an even distribution around no change. 29 pts underwent >2 Aph (range 3-19, median 4), 20 with matched ALC and 20 with T cell values. There was a trend toward a slight decrease (<10%) in ALC and T cell subsets. These findings supported the statistical analysis, which identified no consistently positive or negative change in pts who received Aph.

This analysis indicates there is no difference in the number and function of immune cells post Aph in cancer pts. This data provides evidence that Aph inflicts negligible effects on a pts immune system, rejecting the hypothesis that pts immune competence is impacted by the procedure in a meaningful way.

Post minus pre valuesELISPOT (n = 64)ALC (n = 52)CD3 (n = 36)CD4 (n = 36)CD8 (n = 36)CD4:CD8 ratio (n = 15)Aph groupDifference Pre vs PostMean500.906−0.02133.72102.7219.420.51Median0.000.0097.5063.5020.000.23P value1.000.780.0360.0240.560.0045InterpretationNo changeNo changeTrend IncreaseTrend IncreaseNo changeSignificant IncreaseNon-Aph group N = 90Difference Pre vs PostMean500.600Median0.00P value1.00

Citation Format: James L. Gulley, Christopher R. Heery, Seth Steinberg, Jenn Marte, Ravi A. Madan, Jeffrey Schlom. Impact of apheresis (Aph) on number and function of lymphocytes in cancer patients (pts). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1230. doi:10.1158/1538-7445.AM2013-1230
Abstract 1241: Targeting the IL-13 receptor alpha 2 with novel antibody-drug conjugates for the treatment of cancer.
Authors: Ma Dangshe , Zhang Haige , Jin Fang , Charati Manoj , Khandke Kiran , Lucas Judy , Follettie Max , Tumey L. Nathan , Gerber Hans-Peter , Tchistiakova Lioudmila , Sapra Puja
Published: August 2014

The IL-13 receptor alpha 2 (IL-13Rα2) is a transmembrane protein that is highly expressed on several cancers including glioblastoma, ovarian, pancreatic cancers. It can internalize after binding to its ligand IL-13 or in response to ligation with an antibody. In addition, we have discovered that IL-13Rα2 is up-regulated in tumor cell lines that are resistant to chemotherapeutic drugs. These properties make IL-13Rα2 an attractive target for antibody-drug conjugate (ADC), an emerging modality of molecularly targeted cancer therapies. A humanized monoclonal antibody, hAB08 with an affinity of 0.17 nM to the soluble recombined hIL-13Rα2, was conjugated with two novel microtubule inhibitors (MTIs), MTI1 and MTI2, developed by Pfizer. In vitro cytotoxicity of the conjugates was tested against IL-13Rα2 positive and negative cell lines. These conjugates were effective against the IL-13Rα2 positive cell lines (PC3MM2 and A375), having an IC50 ranging from 1.1 to 3.8 ng Ab/mL. However, both of the ADCs were not active against the IL-13Rα2 negative H460 cell line. When treating mice with established human PC3MM2 and A375 tumors, these conjugates were efficacious at 3 mg/kg in reducing tumor growth. In the PC3MM2 model, treatment with hAB08-MTI1 and hAB08-MTI2 resulted in 5 out of 8 or 3 out of 8 animals without measurable tumors at Day 76, respectively. In contrast, the vehicle control group and control ADC (hIgG8.8-MTI1 and hIgG8.8-MTI2) groups were terminated at Day 15, 15, 19 due to the large size of tumors, respectively. These in vitro and in vivo results demonstrate potent antitumor activities of hAB08-MTI1 and hAB08-MTI2 against IL-13Rα2 positive tumors. The findings support the development of novel hAB08-MTI1 or hAB08-MTI2 ADCs as a molecularly targeted therapy for cancer patients that are resistant to standard chemotherapy.

Citation Format: Dangshe Ma, Haige Zhang, Fang Jin, Manoj Charati, Kiran Khandke, Judy Lucas, Max Follettie, L. Nathan Tumey, Hans-Peter Gerber, Lioudmila Tchistiakova, Puja Sapra. Targeting the IL-13 receptor alpha 2 with novel antibody-drug conjugates for the treatment of cancer. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1241. doi:10.1158/1538-7445.AM2013-1241
Abstract 1244: Phosphatidylserine-targeting antibody reactivates tumor immunity and destroys tumor vasculature in mice.
Authors: Yin Yi , Huang Xianming , Ye Dan , Thorpe Philip
Published: August 2014

The immunosuppressive lipid, phosphatidylserine (PS), becomes exposed on tumor blood vessels and tumor cells responding to therapy. The exposed PS contributes to the immunosuppressed tumor microenvironment. Bavituximab is a phosphatidylserine (PS)-targeting antibody that is being combined with chemotherapy in clinical trials in cancer patients. Here, we tested the hypothesis that innate immunity against cancer can be elicited by combining chemotherapy with a monoclonal antibody that indirectly binds exposed phosphatidylserine (PS). We used a murine version of bavituximab, 2aG4, plus docetaxel to treat prostate tumors in mice. Combination treatment markedly retarded tumor growth, prolonged survival times and delayed progression to androgen-independent disease. Antibody treatment reduced the presence of myeloid-derived suppressor cells (MDSCs) in the tumors and caused macrophages to repolarize from the immunosuppressive, proangiogenic M2-like state into a tumoricidal M1-like state. The reactivated macrophages destroyed antibody-coated tumor vasculature and tumor cells. Addition of 2aG4 to MDSCs in vitro caused them to differentiate into M1-like macrophages and DCs. Thus, PS-targeting antibodies appear to reactivate innate immunity in tumors resulting in tumor growth inhibition.

Citation Format: Yi Yin, Xianming Huang, Dan Ye, Philip Thorpe. Phosphatidylserine-targeting antibody reactivates tumor immunity and destroys tumor vasculature in mice. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1244. doi:10.1158/1538-7445.AM2013-1244
Abstract 1246: Development of an agonistic antibody against the human T-cell costimulatory receptor CD27 as a potential immunotherapeutic tool.
Authors: van Eenennaam Hans , Veraar Elise , Mulder Winfried , Bastiaanssen Ellen , Driessen Lilian , Lutje Hulsik David , Vink Paul , van der Horst Gerda , Xiao Yanling , Borst Jannie , van Elsas Andrea
Published: August 2014

Immunotherapy of cancer has reached a mature and exciting stage. Clinical activity of monoclonal antibodies (mAb) against the T-cell inhibitory receptors CTLA-4 and PD-1 indicate that cancer patients often have latent tumour-specific T cells that can be mobilized by blocking these receptors. Activation of the T-cell costimulatory receptor CD27, a member of the TNF receptor family, provides another opportunity to promote cytotoxic T cell (CTL)-based anti-tumor immunity. To generate agonistic mAb against human (h)CD27, we used our unique B-cell selection and immortalization technology. Various novel anti-hCD27 mAb were generated and tested for their agonistic properties. We used in vitro read-out systems detecting CD27 (co-)stimulatory function on the basis of NF-κB activity, T cell division and survival. One mAb hCD27.15 (KD ∼1 nM), induced CD27 receptor activation (EC50 ∼1 nM) without the need of cross-linking. Similar to its natural ligand CD70, our hCD27.15 mAb showed bona fide costimulatory properties. hCD27.15 strongly promoted CD8+ T cell expansion under conditions of suboptimal T-cell receptor (TCR) stimulation. hCD27.15 did not stimulate T cell function in absence of TCR engagement. hCD27.15 mAb demonstrated different binding kinetics and a unique epitope, compared to other antibodies that depend on cross-linking to activate CD27. Importantly, hCD27.15 retained its agonistic properties when expressed as a human IgG1 or IgG4 chimera. hCD27.15 was shown to bind and activate primate CD27 with similar potency, providing a rationale for pre-clinical safety testing. In conclusion, to our knowledge we have developed the first mAb to hCD27 that mimics ligand function without the need for cross-linking. Data from mouse models strongly support the application of CD27 agonism to enhance CTL-based anti-tumor immunity, by promoting the generation and survival of CTL, as well as their memory function. We propose therefore that hCD27.15 mAb is a promising tool for cancer immunotherapy.

Citation Format: Hans van Eenennaam, Elise Veraar, Winfried Mulder, Ellen Bastiaanssen, Lilian Driessen, David Lutje Hulsik, Paul Vink, Gerda van der Horst, Yanling Xiao, Jannie Borst, Andrea van Elsas. Development of an agonistic antibody against the human T-cell costimulatory receptor CD27 as a potential immunotherapeutic tool. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1246. doi:10.1158/1538-7445.AM2013-1246
Abstract 1248: Intralesional injection with PV-10 induces a systemic anti-tumor immune response in murine models of breast cancer and melanoma.
Authors: Pilon-Thomas Shari A. , Weber Amy , Kodumudi Krithika , Kuhn Lisa , Toomey Paul , Sarnaik Amod
Published: August 2014

PV-10 is a 10% solution of Rose Bengal that is currently being examined as a novel cancer therapeutic. In melanoma patients, intralesional injection (IL) of PV-10 has led to regression of injected lesions as well as distant metastases. In this study, we examined the efficacy and potential immune mechanism of PV-10 treatment in murine models of breast cancer and melanoma. In BALB/c mice bearing MT-901 breast cancer, injection of PV-10 led to regression of injected and untreated contralateral subcutaneous lesions (p<0.05 compared to IL-PBS-treated mice). A significant increase in survival was observed in mice treated with PV-10. To examine immune response, MT901-specific IFN-gamma production and cytotoxicity were measured in splenocytes collected from mice treated with IL-PBS or IL-PV-10. MT901-bearing mice treated with IL-PV-10 demonstrated enhanced IFN-gamma production (992 ± 453 pg/ml) compared to splenocytes from PBS-treated mice (174 ± 105, p<0.05). In addition, a significant increase in lysis of MT-901 cells by T cells after PV-10 treatment was observed (p<0.01 compared to PBS-treated mice). No lysis of irrelevant CT-26 cells was detected. In a murine model of melanoma, B16-F10 cells were injected into C57BL/6 mice to establish one subcutaneous tumor and multiple lung lesions. Treatment of the subcutaneous lesion with a single injection of IL-PV-10 led to regression of the injected lesion as well as distant B16 melanoma lung metastases. In B16-bearing mice, treatment with IL-PV-10 led to the induction of T cells that produced IFN-gamma in response to B16 tumors but not irrelevant tumor (p<0.05) and demonstrated specific lysis of B16 (p<0.01 compared to T cells isolated from PBS-treated mice). In total, these studies support the induction of tumor-specific T cell-mediated immunity after single treatment with IL-PV-10 in multiple histologic subtypes. The immune mechanism of PV-10 ablation in cutaneous melanoma patients is currently under investigation at our institution.

Citation Format: Shari A. Pilon-Thomas, Amy Weber, Krithika Kodumudi, Lisa Kuhn, Paul Toomey, Amod Sarnaik. Intralesional injection with PV-10 induces a systemic anti-tumor immune response in murine models of breast cancer and melanoma. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1248. doi:10.1158/1538-7445.AM2013-1248
Abstract 1249: Potent immunomodulatory effects of lenalidomide enhance the effect of a therapeutic vaccine against established lymphoma in mice .
Authors: Sakamaki Ippei , Kwak Larry W. , Cha Soung-chul , Yi Qing , Lerman Beatrisa , Chen Jian , Surapaneni Sekhar , Bateman Scott , Qin Hong
Published: August 2014

Lenalidomide is an effective therapeutic agent with direct inhibitory effects on malignant B- and plasma cells and immunomodulatory effects on the T cell activation. The dual function of lenalidomide makes it an appealing candidate for combination with other novel cancer agents. We investigated the immune stimulatory effects of lenalidomide, administrated to mice in doses which provided comparable pharmacokinetics to human patients, on the potency of a novel fusion DNA lymphoma vaccine. The combination protected mice from lethal challenge with syngeneic A20 murine lymphoma, resulting in significantly improved survival compared with vaccine or lenalidomide alone and induced immune memory. In vivo depletion experiments demonstrated a requirement for effector CD8+ and CD4+ T cells. Surprisingly, lenalidomide alone was associated with reduced numbers of systemic immune suppressive cells (MDSC/Treg) in tumor-bearing, but not naïve mice, an effect that was independent of simple tumor burden reduction. Finally, the combination of lenalidomide and vaccine produced significantly improved survival in mice with 7 day established tumors. These results demonstrated the dual effect of lenalidomide on enhancing antigen-specific T-cell immunity and a novel mechanism of action reversing tumor-induced immune suppression, which makes it ideal for combination with vaccines as an immune adjuvant.

Citation Format: Ippei Sakamaki, Larry W. Kwak, Soung-chul Cha, Qing Yi, Beatrisa Lerman, Jian Chen, Sekhar Surapaneni, Scott Bateman, Hong Qin. Potent immunomodulatory effects of lenalidomide enhance the effect of a therapeutic vaccine against established lymphoma in mice . [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1249. doi:10.1158/1538-7445.AM2013-1249
Abstract 125: Hematological cancer incidence among semiconductor workers.
Authors: Lee Kyoungho , Han Joohee , Kim Kwansick , Kim Soo-Geun
Published: August 2014

Background/objective. There has recently been such great interest in cancer incidence among the semiconductor workers. Therefore, an epidemiological study was conducted to figure out the cancer incidence rate among semiconductor workers and potential factors affecting cancer.

Methods. The study was performed for the semiconductor workers employed from 1999 to 2008, and the total number of workers in a cohort was 56,283 who employed for over one month. Cancer incidence rate was analyzed by departments, divisions, job titles, gender, age, viral hepatitis and working duration. We compared the incidence of hematological cancer in this cohort with that expected on the basis of cancer incidence in the general population.

Results. Overall, the standardized incidence rate (SIR) for workers indicates 0.76 (95% CI: 0.66-0.86) compared to general population data. In particular, the rate for leukemia was 0.86(0.50-1.47) in the total. Non-Hodgkin's lymphoma was observed high SIR which was not statistically significant among female assembly workers(2.42, 95% CI: 0.78-7.49), whereas in office workers were significant high (5.23, 95% CI: 1.31-20.92). We also found that the hepatitis virus affected in female workers (7.69, 95% CI: 1.08-54.60).

Conclusions. We concluded that the SIR for non-Hodgkin's lymphoma among female workers is high because of personal factors and additional research works are considered for some confounder factors. Epidemiological study based on a constant, long-term prospective cohort is required for the future studies.

Citation Format: Kyoungho Lee, Joohee Han, Kwansick Kim, Soo-Geun Kim. Hematological cancer incidence among semiconductor workers. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 125. doi:10.1158/1538-7445.AM2013-125
Abstract 1259: Murine HLA-restricted CD4+ T cell lines as source of high affinity TCRs specific for the human breast cancer-associated tumor antigen NY-BR-1.
Authors: Gardyan Adriane , Osen Wolfram , Eichmüller Stefan B. , Zoernig Inka , Jaeger Dirk
Published: August 2014

Out of all malignancies, breast cancer is the second most common cancer worldwide and the leading cause of cancer death in females. Passive immunization approaches such as adoptive transfer of TCR-transduced autologous T cells specific for tumor associated antigens, provide an innovative strategy for cancer immunotherapy. The differentiation antigen NY-BR-1 has been described to be expressed in 60% of all invasive mammary carcinomas. Since NY-BR-1 protein levels are highly elevated in malignant breast tissues compared to healthy breast tissue, NY-BR-1 might represent a suitable target antigen for T cell based immunotherapy approaches against breast cancer. Successful immunological eradication of tumors depends on the presence of activated tumor antigen-specific CD4+ effector T cells as documented by numerous reports. Therefore, NY-BR-1 was screened for the presence of CD4+ T cell epitopes using HLA-transgenic mice immunized with a NY-BR-1-encoding expression plasmid and subsequent ex vivo analysis of the NY-BR-1-specific T cell responses against a synthetic peptide library covering the entire NY-BR-1 protein. Stable CD4+ T cell lines specific for six newly identified epitopes could be established from peptide-immunized DRB1*0301 and HLA-DRB1*0401-transgenic mice and HLA-DR-restriction of the cell lines was confirmed on peptide loaded T2/DR3 and T2/DR4 target cells. Next, natural processing of the epitopes identified in HLA transgenic mice will be tested in the human system using human HLA-matched cells lines infected with NY-BR-1 encoding adenovirus as targets for recognition by the murine NY-BR-1-specific CD4+ T cell lines. Clinical relevance of the newly identified NY-BR-1-specific epitopes will be investigated by screening of blood samples from NY-BR-1+, HLA-matched breast cancer patients for the presence of cognate CD4+ T cells. High affinity TCRs from NY-BR-1 specific CD4+ T cells will be sequenced and subsequently used to generate autologous TCR-transduced T cell lines ready for adoptive transfer of breast cancer patients

Citation Format: Adriane Gardyan, Wolfram Osen, Stefan B. Eichmüller, Inka Zoernig, Dirk Jaeger. Murine HLA-restricted CD4+ T cell lines as source of high affinity TCRs specific for the human breast cancer-associated tumor antigen NY-BR-1. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1259. doi:10.1158/1538-7445.AM2013-1259
Abstract 1260: Generation of human T cells directed against an agonist epitope of a transcription factor involved in epithelial to mesenchymal transition (EMT).
Authors: Tucker Jo A. , Poole Diane J. , Palena Claudia , Jochems Caroline , Schlom Jeffrey , Tsang Kwong Y.
Published: August 2014

Purpose: The T-box family transcription factor Brachyury is overexpressed in a variety of human carcinomas, including lung, breast, colon, ovarian and prostate. Brachyury has been shown to promote epithelial to mesenchymal transition (EMT) in tumor cells, a critical step in the path to metastasis. An HLA-A2 epitope of Brachyury has been shown to expand human T cells that are capable of lysing Brachyury-expressing tumor cells in an HLA-dependent manner. This study sought to define an agonist of this epitope in order to increase T cell activation and tumor lysis.

Experimental Design: A novel agonist epitope of Brachyury was generated by residue substitution of the native epitope. Characterization of this epitope as an agonist included; comparison of HLA binding affinity and stability, interferon γ production by epitope-specific T cell lines, as well as Brachyury-and HLA-specific lysis of tumor cells. The presence of Brachyury-specific T cells that would recognize the agonist peptide, within the circulating PBMC of cancer patients, was determined by ELISPOT.

Results: The agonist epitope was shown to bind HLA-A2 with higher affinity and stability than the native. T cell lines generated from both the native and agonist epitopes produced higher levels of interferon γ in response to stimulation with the agonist epitope. The agonist-specific T cell line was able to lyse a variety of Brachyury-expressing tumor cells more efficiently than the T cell line generated with the native epitope, and specificity of lysis was confirmed by cold-target inhibition and HLA-A2 blocking. Tetramer staining revealed Brachyury agonist-specific T cells in the PBMC of a prostate cancer patient after in vitro stimulation with the agonist peptide. PBMC from colon and ovarian cancer patients reacted to the agonist peptide in an interferon γ ELISPOT assay.

Conclusions: An agonist epitope for Brachyury has been identified, which increased T cell activation and function as compared to the native epitope. T cells that react to the agonist peptide were detected in patients with carcinomas known to express high levels of Brachyury. This study supports the use of Brachyury as a cancer vaccine strategy, including the Brachyury agonist epitope.

Citation Format: Jo A. Tucker, Diane J. Poole, Claudia Palena, Caroline Jochems, Jeffrey Schlom, Kwong Y. Tsang. Generation of human T cells directed against an agonist epitope of a transcription factor involved in epithelial to mesenchymal transition (EMT). [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1260. doi:10.1158/1538-7445.AM2013-1260
Abstract 1265: Sustained PI3-kinase signaling is mediated by ICOS, not CD28, for anti-tumor T-cell responses elicited by anti-CTLA-4.
Authors: Chen Hong , Fu Tihui , Tsavachidou Dimitra , Wen Sijin , Ng Tang Derek , He Qiuming , Sun Jing Jing , Sharma Padmanee
Published: August 2014

The generation of anti-tumor T-cell responses requires at least two distinct signals consisting of T-cell receptor signaling and CD28-costimulation to mediate PI3-kinase signaling. In previous studies, we reported that cancer patients treated with anti-CTLA-4 had a significant increase in T-cells expressing inducible costimulator (ICOS). Here, we observed that PI3-kinase signaling was significantly increased in ICOShi as compared to ICOSlow CD28+ T-cells from anti-CTLA-4 treated patients. Our data also indicate that after T-cell activation, ICOS-mediated PI3-kinase signaling plays an important role in downstream T-cell responses such as expression of the T-bet transcription factor and anti-tumor Th1 cytokine production. An ICOS-specific siRNA transfected into activated human T-cells leads to diminished PI3K-signaling and T-bet expression, which controls production of anti-tumor Th1 cytokines. Furthermore, T-cells from ICOS-deficient mice have impaired PI3-kinase signaling after in vitro activation. Our data also demonstrate that T-cells from ICOS-deficient mice have impaired PI3-kinase signaling in vivo, which correlates with impaired tumor rejection after treatment with anti-CTLA-4. These data offer strong evidence to support that CD28-mediated PI3-kinase signaling is limited and ICOS provides a critical third signal to enable sustained PI3-kinase signaling for optimal anti-tumor T-cell responses in the setting of anti-CTLA-4 therapy.

Citation Format: Hong Chen, Tihui Fu, Dimitra Tsavachidou, Sijin Wen, Derek Ng Tang, Qiuming He, Jing Jing Sun, Padmanee Sharma. Sustained PI3-kinase signaling is mediated by ICOS, not CD28, for anti-tumor T-cell responses elicited by anti-CTLA-4. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1265. doi:10.1158/1538-7445.AM2013-1265
Abstract 1272: Myeloid leukemia cell lines suppress T cell activation and proliferation.
Authors: Tanimoto Kazushi , Ito Sawa , Miner Samantha , Muranski Pawel , Hensel Nancy F. , Melenhorst J. Joseph , Barrett A. John
Published: August 2014

In preliminary experiments to develop the K562 human leukemia cell line as an artificial antigen presenting cell we found that K562 cells strongly inhibited T cell proliferation. To explore this phenomenon further we tested CD4+CD25- T cells from healthy donor PBMC as targets of proliferation suppression. Targets were co-cultured with 3 chronic myeloid leukemia (CML) derived cell lines (K562, BV173 and CML1 established from a CML patient in our laboratory), two monocytic leukemia derived cell lines (THP-1 and U937), the myeloid leukemia derived cell line (KG1a) and the Burkitt lymphoma cell line (Daudi). To evaluate early activation, CD154 expression was tested by flow cytometry 16h after stimulation with CD3/CD28 beads. T cell proliferation was tested by CFSE dilution in T cells stimulated for 72h. All myeloid lines blocked CD154 expression (10.9-62.8% of control at 1:8 target: suppressor ratio). In CFSE proliferation assay, K562, KG1a, CML1, THP-1 and U937 induced comparable suppression (28.4-93.9%, at 1:8 ratio). In contrast, BV173 and Daudi cells were the least suppressive (4.96-23.0%, P<0.01). Strikingly, CML1 cell line and THP-1 showed more suppressive effect on T cell proliferation than CD4+CD25+CD127- autologous regulatory T cell (Treg). Our result showed that myeloid leukemia cell lines but not B cells strongly inhibit both T cell activation and T cell proliferation. Suppression of proliferation was greatest in acute myeloid and monocytic leukemia derived lines. These data indicate that myeloid lekemia cell lines exhibit features of myeloid-derived suppressor cells (MDSC) which can induce T cell tolerance to tumors. Our findings raise the possibility that myeloid leukemias may actively evade immune control through T cell suppression. We plan to use such cell lines to further define the mechanism of MDSC related T cell suppression.

Citation Format: Kazushi Tanimoto, Sawa Ito, Samantha Miner, Pawel Muranski, Nancy F. Hensel, J. Joseph Melenhorst, A. John Barrett. Myeloid leukemia cell lines suppress T cell activation and proliferation. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1272. doi:10.1158/1538-7445.AM2013-1272
Abstract 1276: UV radiation interacts with Endothelin 3 and loss of nucleotide excision repair pathway to promote melanomagenesis.
Authors: Benaduce Ana Paula , Batista Deannys , Grilo Gabriel , Jorge Karen , Kos Lidia
Published: August 2014

Melanomagenesis is influenced by the interaction of environmental (such as UV radiation) and genetic factors. The Xpa gene is important in the Nucleotide Excision Repair (NER) pathway. Similar to humans afflicted with Xeroderma Pigmentosum, which results from mutations in genes that play a role in NER, Xpa deficient mice show high sensitivity to UV light, leading to skin cancer development. However, Xpa deficient mice do not develop melanoma even upon UV exposure. The Endothelin 3 (Edn3) signaling pathway is essential for proliferation, survival and migration of melanocyte precursor cells. In humans, this pathway has also been implicated in melanoma progression and its metastatic potential. The purpose of this study is to develop a UV induced melanoma mouse model that combines Xpa deficiency with the over-activation of the Edn3 pathway. To this end, transgenic mice over-expressing Edn3 under the control of the keratin 5 promoter (K5-Edn3) and carrying a targeted mutation in Xpa were exposed to a single suberythemal neonatal dose of UV radiation. A subgroup of mice was also exposed to a second dose of UV radiation at 6 weeks of age. Histomorphology and immunostaining were used to confirm the melanocytic origin of primary skin tumors and metastases. Melanoma was only found in animals with the K5-Edn3 transgene. High penetrance was observed in animals exposed to one dose of UV radiation that were Xpa null (100%, n=3) when compared to Xpa heterozygous (73%, n=11) or Xpa wild type (43%, n=7). Enlarged and hyperpigmented lymph nodes were present in animals with melanoma skin lesions and these were diagnosed as local metastases. These results indicate that UV radiation exposure, in conjunction with over-activation of the Edn3 pathway is sufficient to lead to melanomagenesis in mice.

Citation Format: Ana Paula Benaduce, Deannys Batista, Gabriel Grilo, Karen Jorge, Lidia Kos. UV radiation interacts with Endothelin 3 and loss of nucleotide excision repair pathway to promote melanomagenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1276. doi:10.1158/1538-7445.AM2013-1276