bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 4014: Combination of high-dose irradiation and local interleukin-12 treatment enhance tumor killing and have less toxicities than either treatment alone
Authors: Hong Ji-Hong , Yu Ching-Fang , Chen Fang-Hsin , Chiang Chi-Shiun 
Published: July 2016

Abstract
Immunotherapy such as anti-PD-1/PD-L1 is effective in several types of tumors, but in general the response rate is less than 30%. Systemic administration of cytokine such as IL-12 could create anti-tumor immunity but has unacceptable side effects. This study is to test our hypothesis that combination of high-dose local radiotherapy (RT) and local production of IL-12 inside tumors within short period could have potent anti-tumor effects but low systemic toxicity. TRAMP-C1 tumors derived from a murine prostate cancer were either no treatment, or irradiated with 25 Gy, or intra-tumorally injected with Ad-sc-IL12 virus (1×10sup8 pfu), or treated with both modalities at the tumor size of 4mm. RT alone and Ad-sc-IL12 alone could not shrink tumors and only caused tumor growth delay around 7 and 12 days, respectively. The combination of RT with Ad-sc-IL12 virus could shrink the tumors to the sizes of &lt; 2mm and further extended the tumor growth delay more than 20 days. The percentage of lymphocytes and IFN-γproduction were evaluated by flow cytometry and ELISA. The percentages of CD4 cells in tumors were too low to be reliably measured in all groups, but the percentages of CD8 cells were increased in Ad-sc-IL12 virus-treated groups and even much higher in those combined with RT. For mice treated with Ad-sc-IL12 virus injection, the serum IL-12 levels were around 4.5 times increase at day7, but those combined with with RT had significantly lower levels than those treated with virus alone. The serum IFN-γ(Th1 response) levels were around 12 times higher in Ad-sc-IL12 groups at day7 as compared to control or RT only group, but had no significant differences between those treated with combined modality and with Ad-sc-IL12 virus alone. Liver damage is an index for IL-12 toxicity and the levels of GOT and GTP were also lower in those treated IL-12 combined with RT than those treated with IL-12 alone. This study supported our hypothesis and showed that combination of local IL-12 production and local RT are effective in increasing CD8 cells, exerting Th1 response, and enhancing tumor growth delay. The most important is that this combination has less systemic toxicity than local IL-12 production alone. This study provides evidence to develop new strategy for producing a strong anti-tumor and low toxicity for radioresistant tumors by combining RT and IL-12 immunotherapy.
Citation Format: Ji-Hong Hong, Ching-Fang Yu, Fang-Hsin Chen, Chi-Shiun Chiang. Combination of high-dose irradiation and local interleukin-12 treatment enhance tumor killing and have less toxicities than either treatment alone. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4014.
Abstract 4023: Combination therapy with immune checkpoint inhibitor and CCK-receptor blockade increases survival of pancreatic cancer
Authors: Smith Jill P. , Wang Shangzi , Reham Ajina , Jablonski Sandra A. , Weiner Louis M. 
Published: July 2016

Abstract
Introduction: Advanced pancreatic ductal adenocarcinoma (PDAC) has typically been resistant to chemotherapy and immune therapy; therefore, novel strategies are needed to enhance therapeutic response. Cholecystokinin receptors (CCKRs) are present on pancreatic cancer epithelial cells, fibroblasts of the microenvironment, and lymphocytes. We hypothesized that CCK receptor blockade would improve response to immune checkpoint blockade and survival by promoting influx of tumor infiltrating lymphocytes (TILs) and reducing fibrosis.
Methods: Subcutaneous tumors were established using Panc02 murine pancreatic cancer cells (1.0- or 2.0 x10E6) in a syngeneic immune competent mouse model. Three separate experiments were performed with 40 mice each. Mice (C57BL/6) were divided into 4 groups (N = 10 mice each) and treated with PBS (control), a CCKR antagonist (L364,718 or proglumide), an immune checkpoint blockade antibody (PD-1 or CTLA-4; Im-Ab), or the combination of CCKR antagonist and immune checkpoint blockade antibody (combination). Tumor growth and animal survival were evaluated. Tumors were subjected to immunohistochemical staining for TILs, trichrome staining, and flow cytometry.
Results: On day 50 after inoculation of 10E6 cells all control mice had died while 42%, 42% and 71% of L364,718, CTLA-4 mice, or combination-treated mice, respectively were alive (p = 0.02). By day 90 all mice died except one in the combination-treated group. In the experiment where mice were injected with 2×10E6 Panc02 cells, survival was also increased in mice receiving a proglumide and PD-1 antibody combination (p = 0.0009). CD8+TILs increased by 3-,6.3-,and 7.5-fold in CCKR antagonist, Im-Ab, and combination therapy, respectively compared to controls (p&lt;0.0001). CD4+ TILs also significantly increased (p = 0.001) in the tumors of mice treated with the combination. Trichrome stain analysis revealed 50% less fibrosis in mice treated with CCKR antagonist compared to PBS controls (p = 0.017).
Conclusions: The combination of CCKR antagonism and immune checkpoint blockade antibodies significantly improve survival in a murine model of PDAC. The mechanism is associated with decreased fibrosis in the microenvironment and an accompanying influx of CD8+ and CD4+ TILs.
Supported by NCI CA50633, CA51008, and CA194745
Citation Format: Jill P. Smith, Shangzi Wang, Ajina Reham, Sandra A. Jablonski, Louis M. Weiner. Combination therapy with immune checkpoint inhibitor and CCK-receptor blockade increases survival of pancreatic cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4023.
Abstract 4025: Impact of Kras mutant subtypes on PD-L1 expression in lung adenocarcinoma
Authors: Ilie Marius , Fazzalari Laetitia , Guibert Nicolas , Yazbeck Nathalie , Signetti Laurie , Hofman Véronique , Long Elodie , Zahaf Katia , Fayada Julien , Lespinet Virginie , Bordone Olivier , Tanga Virginie , Washetine Kevin , Vénissac Nicolas , Mouroux Jérôme , Marquette Charles Hugo , Brest Patrick , Hofman Paul 
Published: July 2016

Abstract
Clinical responses to immune checkpoint blockade by anti-PD-1/PD-L1 monoclonal antibodies in non-small-cell lung cancer (NSCLC) may be associated with PD-L1 expression. This study was undertaken to determine the expression profile of PD-L1 in patients with Kras-mutant lung adenocarcinoma (LUAD) and to investigate the activation of Kras codon subtypes as a mechanism of PD-L1 regulation.
Immunohistochemistry analysis of PD-L1 expression (SP142 clone) on 117 LUAD (KrasWT, n = 51; KrasG12D, n = 25; KrasG12V, n = 14; KrasG12C, n = 27), showed significantly greater expression of PD-L1 in both tumor and immune cells compartments in Kras mutant LUAD compared with KrasWT wild-type tumors (37% vs. 18%; P = 0.005). PD-L1+ tumors had greater CD66b+ neutrophil infiltrate and lower CD8+ T-cell infiltrate than PD-L1− tumors, notably in KrasG12D tumors.
To determine the effect of Kras codon subtypes on PD-L1 expression, stable cell lines were generated by transfection of KrasG12D, KrasG12V, KrasG12C and KrasWT plasmids into Beas2B bronchial cells. At basal level, mutant Kras led to significantly higher cell-surface PD-L1 expression and PD-L1 transcripts, notably in KrasG12C and KrasG12V cells, suggesting transcriptional regulation. Moreover, there was differential activation of NF-kB, ERK and Pi3k/Akt pathways between Kras mutant subtypes. In addition, PD-L1 was upregulated 3-fold by stimulation with IFNγ, independently of the Kras codon subtypes. Instead, hypoxia significantly increased PD-L1 expression in KrasG12C and KrasG12D cells. Co-culture experiments with human PBMCs from healthy patients were performed to determine the functional effect of altered PD-L1 expression. Increased PD-L1 expression by tumor cells induced by Kras mutations led to decreased PBMCs proliferation and increased apoptosis.
PD-L1 is expressed in 37% of Kras mutant LUAD, suggesting PD-L1 as a therapeutic target in this subset. According to the Kras mutation subtype, potential drugs targeting the NF-kB, ERK or Pi3k/Akt pathways may increase the antitumor adaptive immune responses.
Citation Format: Marius Ilie, Laetitia Fazzalari, Nicolas Guibert, Nathalie Yazbeck, Laurie Signetti, Véronique Hofman, Elodie Long, Katia Zahaf, Julien Fayada, Virginie Lespinet, Olivier Bordone, Virginie Tanga, Kevin Washetine, Nicolas Vénissac, Jérôme Mouroux, Charles Hugo Marquette, Patrick Brest, Paul Hofman. Impact of Kras mutant subtypes on PD-L1 expression in lung adenocarcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4025.
Abstract 403: Performance assessment of highly sensitive NGS assay for TP53
Authors: Fredebohm Johannes , Mehnert Daniel , Löber Ann-Kathrin , Shimizu Hiroyuki , Holtrup Frank , Diehl Frank 
Published: July 2016

Abstract
We have designed a highly sensitive assay based on the Safe-Sequencing technology(1) to detect de novo mutations in the TP53 gene. A custom panel was designed to cover 95% of all reported mutations in TP53. To demonstrate assay performance, we assessed LoD, LoB, reproducibility, and repeatability and evaluated concordance with BEAMing(2). A customized data analysis pipeline for ultra-deep sequencing runs was developed that includes extensive quality control and advanced mutation calling. This highly sensitive NGS-based workflow can be applied to monitor recurrent or minimal residual disease in cancer patients after surgery or chemotherapy.
References:
(1) Kinde I, Wu J, Papadopoulos N, Kinzler KW, Vogelstein B. Detection and quantification of rare mutations with massively parallel sequencing. Proc Natl Acad Sci U S A. 2011 Jun 7; 108(23): 9530-9535
(2) Diehl F, Li M, He Y, Kinzler KW, Vogelstein B, Dressman D. BEAMing: single-molecule PCR on microparticles in water-in-oil emulsions. Nat Methods. 2006 Jul;3(7):551-9.
Citation Format: Johannes Fredebohm, Daniel Mehnert, Ann-Kathrin Löber, Hiroyuki Shimizu, Frank Holtrup, Frank Diehl. Performance assessment of highly sensitive NGS assay for TP53. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 403.
Abstract 4036: Prognostic value of infiltrating immune cell profiling in lung adenocarcinoma
Authors: Kurebayashi Yutaka , Emoto Katsura , Hayashi Yuichiro , Kamiyama Ikuo , Ohtsuka Takashi , Asamura Hisao , Sakamoto Michiie 
Published: July 2016

Abstract
Neoplastic cancer cells and cancer stroma (including infiltrating immune cells) determine the biology and prognosis of cancer. Various types of adaptive and innate immune cells are known to infiltrate the cancer stroma. However, the patterns and spatial distribution of immune cell infiltration as well as their association with tumor histology remain poorly understood. To address these issues, we comprehensively analyzed the infiltrating immune cells present in lung adenocarcinoma. The principal types of both adaptive and innate infiltrating immune cells were immunohistochemically evaluated in the predominant histological components of 111 lung adenocarcinomas. The same analysis was also carried out on 143 samples of histological subtypes making up more than 20% of tumors. As a result, plasma cells and B cells with interfollicular distribution were almost exclusively observed in invasive histological subtypes, while increased number of mast cells were observed in noninvasive histological subtypes. Cluster analysis revealed four distinct immunosubtypes: those characterized by high CD8/CD3 ratios with low stromal cellularity (CD8 subtype), those with increased mast cell numbers (mast cell subtype), those with increased macrophage/DC infiltration with variable amounts of T cell infiltration (mϕ/DC subtype), and those with increased plasma cell infiltration in addition to increased macrophage/DC infiltration and variable increases in other infiltrating immune cells (T cells, Tregs, pDCs, and neutrophils) (plasma cell subtype). These immunosubtypes correlated with histological subtypes. Univariate and multivariate analyses identified the plasma cell subtype as an independent negative prognostic factor. Furthermore, we found that these plasma cells may be one of the major producers of immunosuppressive cytokine IL-35 in cancer stroma. Consequently, comprehensive immunohistochemistry-based profiling of infiltrating immune cells has potential as a powerful method for analyzing the cancer stroma of various cancer tissues.
Citation Format: Yutaka Kurebayashi, Katsura Emoto, Yuichiro Hayashi, Ikuo Kamiyama, Takashi Ohtsuka, Hisao Asamura, Michiie Sakamoto. Prognostic value of infiltrating immune cell profiling in lung adenocarcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4036.
Abstract 4038: Developmentof a murine tumor immunophenotyping platform to support drug discovery anddevelopment in immuno-oncology
Authors: Belmontes Brian , Matyas Stephanie , O’Brien Sarah , Tan Hong , Ganley Kenneth , Merriam Kimberly , Rottman Jim , Egen Jackson , Beltran Pedro , Moody Gordon 
Published: July 2016

Abstract
Recent clinical data highlights the importance of immune cell localization, phenotype, and gene signature as it correlates to a productive anti-tumor response. Preclinically, multiple syngeneic mouse models have been used to study the effects of immunomodulation and define anti-tumor responses to transplanted “self” tumors. However, while the literature describes distinct aspects of many of these models, there is no comprehensive dataset comparing and contrasting their tumor-immune microenvironments across models. These data are critical for better understanding the role that various immune populations play in the anti-tumor response and interpreting observed changes in tumor clearance following treatment with immunomodulatory agents. We have therefore established a platform that 1) quantitates the types of immune cells within murine tumor models, and 2) describes the location of these cells within the tumor. In parallel to the immunophenotyping efforts we have benchmarked tumor models based on their response to antibodies against T cell checkpoint pathways.
We sought to use this immunophenotyping platform to identify specific immune modulation that occurs in syngeneic tumors post depletion of macrophages via CSF1R blockade. Tumor-associated macrophages (TAMs) are believed to help promote tumor survival through suppression of the adaptive immune response and the secretion of growth factors that promote tumor growth and angiogenesis. Therefore depletion of TAMs should lead to T-cell recruitment and bolster the antitumor T-cell response. Here we show that treatment of CT-26 and RENCA syngeneic tumors with a CSF1R antagonist leads to depletion of MHCII+ and F4/80+ expressing cells. Future experiments will seek to understand if CSF1R blockade improves the response to T-cell checkpoint immunotherapies. In summary, the development of a murine tumor immunophenotyping platform has allowed insight and evaluation of immune cells in the tumor microenvironment that can ultimately be leveraged to understand the synergistic effects of immunotherapeutics.
Citation Format: Brian Belmontes, Stephanie Matyas, Sarah O’Brien, Hong Tan, Kenneth Ganley, Kimberly Merriam, Jim Rottman, Jackson Egen, Pedro Beltran, Gordon Moody. Developmentof a murine tumor immunophenotyping platform to support drug discovery anddevelopment in immuno-oncology. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4038.
Abstract 4039: Mimotope variance analysis: A novel immunoprofiling method to monitor progression of cervical cancer
Authors: Enerly Espen , Nygård Mari , Orumaa Madleen , Pihlak Arno , Pihelgas Susan , Langseth Hilde , Neuman Toomas , Palm Kaia 
Published: July 2016

Abstract
The purpose of the study was to identify and describe cervical cancer specific changes in humoral immune response by using a novel technology platform Mimotope Variation Analysis (MVA) developed by Protobios.
The Janus Serum Bank is one of the world's oldest and largest population-based research biobanks established in 1973. The cohort is annually linked to the Cancer Registry of Norway using personal identification numbers. More than 1000 women have developed cervical cancer after donating sera. We identified sets of successive sera samples from ten patients with invasive cervical cancer, ten patients with pre-invasive cervical neoplasia and twenty cancer-free individuals (matched for age, gender, length of sample storage +3 months). For each subject we identified 4-10 samples donated to Janus over a period of up to 18 years, in total 213 samples. We applied MVA which combines phage display technology and high-throughput sequencing analysis to generate quantitative serologic profiles of millions of 12-mer peptide antigens called mimotopes from 2 μl of blood serum per analysis.
Hierarchical clustering analysis of the top 5000 mimotopes from each sample resulted in individual-specific immunoprofiles. Multiple samples from the same person clustered together. Moreover, clustering reflected the time the sera was drawn suggesting that part of the individual immunoprofile is stabile over time.
In conclusion, preliminary results suggests that Mimotope Variance Analysis generates individual-specific immunological profiles. This profile most likely reflect prior exposure environmental pathogens. Further, we aim to decode the differences in the immunological profiles between cancer patients and cancer-free subjects.
Citation Format: Espen Enerly, Mari Nygård, Madleen Orumaa, Arno Pihlak, Susan Pihelgas, Hilde Langseth, Toomas Neuman, Kaia Palm. Mimotope variance analysis: A novel immunoprofiling method to monitor progression of cervical cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4039.
Abstract 4048: Stem cell properties of normal human keratinocytes influence transformation responses to HPV16
Authors: Woappi Yvon , Creek Kim E. , Pirisi Lucia 
Published: July 2016

Abstract
Human papillomavirus (HPV) infection is the primary cause of cervical cancer. While HPV infection is very common, cervical cancer is a rare outcome. Why some women develop cervical cancer and others do not is not well understood, but likely involves host cell-specific factors and a woman's ability to get rid of HPV-infected cells. We have previously reported marked variability of epidermal growth factor receptor (EGFR) expression in normal human epidermal tissue from different individuals and demonstrated that increased immortalization by HPV16 E6 and E7 oncogenes correlated with lower basal levels of EGFR. We now report a significantly greater transformation response in stem cell-like keratinocytes after transfection with the full length HPV16 genome. Primary normal human keratinocyte strains (NHKc) from 59 individuals were cultured in 3-D anchorage-free suspension to assess spheroid forming ability and to determine resistance to anchorage-independent cell death. Only about 40% of NHKc strains survived and formed spheroids in 3-D culture and grew when plated back in monolayers as spheroid-derived NHKc (SD-NHKc). Fluorescence-activated cell sorting analysis found that SD-NHKc were enriched in cells expressing known keratinocyte stem cell markers: low levels of EGFR and high levels of integrin α6, while spheroid-non forming strains (SNF-NHKc) had dramatically reduced numbers of EGFRlo/ITGα6hi cells. Isolation and cultivation of EGFRlo/ITGα6hi keratinocytes isolated from primary monolayer NHKc cultures revealed this cell population to behave like SD-NHKc with respect to anchorage-independent survival, proliferative potential, cell morphology, and clonogenicity. Upon transfection with HPV16 DNA, most NHKc strains acquired anchorage-independence and exhibited increased expression of EGFR. Importantly, EGFRlo/ITGα6hi keratinocytes and SD-NHKc displayed exuberant growth responses after transfection with HPV16 DNA and significantly higher transformation efficiency by HPV16. In contrast, SNF-NHKc grew more sluggishly and were relatively resistant to HPV16-mediated immortalization. Our study indicates that stem cell-like keratinocytes are preferentially transformed by HPV16 DNA and that stemness is a phenotypic determinant of susceptibility to HPV16-mediated transformation in primary human keratinocytes.
Citation Format: Yvon Woappi, Kim E. Creek, Lucia Pirisi. Stem cell properties of normal human keratinocytes influence transformation responses to HPV16. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4048.
Abstract 4049: Conserved region 2 of Human Papilloma Virus 18 protein E7 is required for E7 binding to centromere protein C
Authors: Tokuda Aoi , Takahashi Shoko , Yoshimoto Masafumi , Yaginuma Yuji 
Published: July 2016

Abstract
The molecular genetics of cervical cancer have not been thoroughly elucidated, despite the fact that cervical cancer is responsible for the deaths of over 250,000 women annually worldwide. Infection with human papillomaviruses (HPVs) is linked to the pathogenesis of a variety of human diseases, including genital warts, laryngeal papillomas, and most notably, cervical cancer. Recently HPV infection was reported to be associated with head and neck cancers.High-risk human papillomaviruses (HPVs) are etiologically linked to human cervical and oral cancers. HPV infection leads to aneuploidy, a numerical chromosomal aberration caused by dysregulation of chromosomal segregation. We found that high-risk HPV18 and HPV58 E7 proteins bind to centromere protein C (CENP-C), a component of the kinetochore that is essential for proper chromosomal segregation. Low-risk HPV4, HPV6, and HPV11 E7s do not bind to CENP-C. The PxDLLCxE sequence in conserved region 2 (CR2) of HPV18 E7 is required for E7 binding to CENP-C. Our results indicate that differences in the ability of high- and low-risk HPV E7s to bind to CENP-C reflect the different oncogenic potentials of high- and low-risk type HPVs.We demonstrated that the E7 proteins from low-risk HPVs do not bind to CENP-C, and that the PxDLLCxE sequence in conserved region 2 of the HPV18 E7 protein is required for E7 binding to CENP-C. CENP-C is a component of the inner kinetochore, and as such, it plays an essential role in proper chromosome segregation, mitotic checkpoint function, and kinetochore assembly. Therefore, we hypothesize that differences in the capability of various E7s from low- and high-risk type HPVs to bind to CENP-C reflect the different oncogenic potential of high- and low-risk type HPVs.
Citation Format: Aoi Tokuda, Shoko Takahashi, Masafumi Yoshimoto, Yuji Yaginuma. Conserved region 2 of Human Papilloma Virus 18 protein E7 is required for E7 binding to centromere protein C. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 4049.