bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2229: PD-L1 is expressed on CD11bhighCD14highHLA-DR+ myeloid cells in the tumor microenvironment of non-small cell lung cancer (NSCLC)
Authors: Sonn Chung-Hee , Oh Yunji , Kim Tae Eun , Ha Jung Sun , Kim Hong Kwan , Cho Jong Ho , Shin Sumin , Kim Jhingook
Published: July 2016

Understanding the immune system of cancer patients is essential for the development of optimal treatment. We analyzed tumor cells and infiltrating immune cells in tumor tissues of NSCLC to determine the type of cells which expressed immune checkpoint molecules and/or their ligands, and compared them to clinicopathological characteristics. To obtain a single cell suspension from the tumor tissues of lung cancer patients, each fresh tumor sample was chopped and treated with collagenase and DNase. The dissociated tumor tissue cells (DTCs) were stained with fluorochrome-labeled mAbs and the distribution of immune cell components and expression of surface markers in DTCs were determined by FACSuite analysis. The yield of DTCs was 5.87±4.73×10⁁7/gram. PD-L1, which is a ligand for immune checkpoint molecule PD-1, is expressed on CD11bhigh myeloid cells in the blood, regardless of HLA-DR expression. On the other hand, PD-L1 is expressed on CD11highCD14highSSClow myeloid cells in the tumor site. All PD-L1 expressing cells were positive for HLA-DR, a ligand for a T cell receptor of CD4 T cells. Furthermore, these cells also expressed both CD86 and CD124. Interestingly, most tumor cells expressed very low levels of the PD-L1 molecule. And we found that CD11highCD14+HLA-DR- myeloid cells did not express PD-L1. The proportion of CD11highCD14+HLA-DR-PD-L1- myeloid cells was higher in advanced stage patients than early stage patients. In conclusion, PD-L1 is expressed on CD11bhighCD14highHLA-DR+SSClow myeloid cells, but not CD11bhighCD14+HLA-DR- myeloid cells. We propose that the PD-L1+ myeloid cells are likely to play an important role in tumor immunity. Further functional studies will be necessary to determine how the interaction between the cells will affect immunity. code noagegendersmokingstagecell typePD-L1+HLA-DR+CD14highCD11bhigh cells (%)PD-L1-HLA-DR-CD14+CD11bhi cells (%)L-03257maleyesIAmucinous adenocarcinoma39.81.2L-05281maleyesIAadenocarcinoma19.44.1L-04849maleyesIIIAsquamous cell carcinoma4.09.0L-02471maleyesIIIAadenocarcinoma11.20.4L-03665maleyesIVadenocarcinoma11.45.3L-04273maleyesIVadenocarcinoma21.65.5L-04674malenoIAadenocarcinoma6.11.6L-02167femalenoIBlarge cell, adenocarcinoma14.72.3L-01976femalenoIBadenocarcinoma15.816.7L-27145femalenoIVadenocarcinoma11.217.2L-27245femalenoIVadenocarcinoma9.924.8

Citation Format: Chung-Hee Sonn, Yunji Oh, Tae Eun Kim, Jung Sun Ha, Hong Kwan Kim, Jong Ho Cho, Sumin Shin, Jhingook Kim. PD-L1 is expressed on CD11bhighCD14highHLA-DR+ myeloid cells in the tumor microenvironment of non-small cell lung cancer (NSCLC). [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 2229.
Abstract 2234: Exploring the oncolytic potential of Newcastle Disease Virus
Authors: Harper James A. , Travers Jon , Franks Ruth , Burke Shannon , Navarro Christel , Xing Cheng , Wang Weijia , Xu Qi , Wilkinson Robert W. , Jin Hong , Carroll Danielle
Published: July 2016

Oncolytic viruses are live, replication-competent viruses that infect and/or replicate selectively in tumour cells leading to the destruction of the infected cell. Cell death occurs as a natural consequence of the viral life cycle and newly produced virions can infect and kill neighbouring tumour cells leading to an amplified therapeutic effect. Oncolysis has the added benefit of potentially producing novel tumour antigens that can induce an immune-mediated therapeutic response.

Newcastle Disease Virus (NDV) is an avian paramyxovirus, which has proven safety and demonstrated efficacy against a variety of mammalian cancers in PhI clinical studies. Using reverse genetics, we generated a recombinant strain of NDV that overcomes environmental and regulatory concerns uncoupling oncolytic potency and avian pathogenicity. We have additionally inserted the GM-CSF gene in order to maximize anti-tumour immune response and provide an in situ, patient/tumour- specific vaccine, combined with potent oncolysis. We have evaluated the biological characteristics of recNDVGM-CSF in vivo and in vitro. In vitro recNDVGM-CSF selectively replicates in and kills a wide variety of tumour cell types. In vivo, using a fibrosarcoma model that supports viral replication, a single administration (intra-tumoural or systemic) was able to cure 80% of tumour bearing mice. Additionally, following IV administration recNDVGM-CSF localises to and replicates within the tumour leading to a localised GM-CSF production within the tumour. The inherent properties of NDV (self-propagation, tumour-selective replication, immune activation) coupled with the ability to genetically engineer NDV to express therapeutic transgenes may provide a multi-modal attack on the tumour, delivering greater benefit to patients.

Citation Format: James A. Harper, Jon Travers, Ruth Franks, Shannon Burke, Christel Navarro, Cheng Xing, Weijia Wang, Qi Xu, Robert W. Wilkinson, Hong Jin, Danielle Carroll. Exploring the oncolytic potential of Newcastle Disease Virus. [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 2234.
Abstract 2236: Long lasting CR status of stage IV gall bladder cancer and colon cancer after combined treatment including autologous formalin-fixed tumor vaccine (AFTV)
Authors: Kuranishi Fumito , Sumi Yusuke , Uemae yoij , Ohno Tadao
Published: July 2016

(INTRODUCTION) The prognosis of advanced (stage IV) cancer is very poor. However, we have already reported two cases of eradication of advanced breast cancer with bone metastasis (AACR 2013 and AACR 2015) after combined treatments including AFTV. We here additionally report successful cases of gall bladder cancer (stage IV, liver metastasis) and colon cancer (stage IV, lung metastasis).

(CASE 1) A 61-year-old female with stage IV gall bladder cancer (T3N1M1: liver metastasis) was operated in May 2011 including partial resection of liver. She was treated with, as the first-line adjuvant therapy, AFTV followed with conventional chemotherapy. This patient is still alive without any recurrence confirmed by CT etc.

(CASE 2) A 60 year-old male with stage IV (N3, H0, P1, M1: lung, lt. adrenal gland, abdominal wall) was operated in August 2009. Then conventional chemotherapy, 2 courses of AFTV and radiation were applied sequentially. No recurrence was observed for more than 5 years.

(DISCUSSION) We diagnosed both patients staying at long lasting complete remission. The prognoses of stage IV gall bladder cancer and abdominal wall-metastasized colon cancer are very poor despite of conventional treatments. Adverse effects of AFTV were less than grade 2, therefore combination therapies including AFTV are worth to consider, although larger scale clinical trial is essential.

Citation Format: Fumito Kuranishi, Yusuke Sumi, yoij Uemae, Tadao Ohno. Long lasting CR status of stage IV gall bladder cancer and colon cancer after combined treatment including autologous formalin-fixed tumor vaccine (AFTV). [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 2236.
Abstract 2249: Detection of EGFR mutations in circulating cell-free DNA of non-small cell lung cancer patients by next-generation sequencing
Authors: Suzawa Ken , Tomida Shuta , Matsubara Takahiro , Ohashi Kadoaki , Maki Yuho , Yamamoto Hiromasa , Morita Mizuki , Soh Junichi , Asano Hiroaki , Tsukuda Kazunori , Kiura Katuyuki , Miyoshi Shinichiro , Toyooka Shinichi
Published: July 2016

[Background] Mutational analysis is essential for recent personalized therapeutic approaches of non-small cell lung cancer (NSCLC) patients. Circulating cell-free DNA (cfDNA) from plasma is an emerging topic as a tool for liquid biopsy and possibly enable repetitive genetic diagnosis or monitoring of cancer status. However, detection of cfDNA from the circulation still remains challenging because of its highly fragmented and low concentration nature. In this study, we analyzed cfDNA samples by targeted deep sequencing at high coverage.

[Materials and Methods] Twenty matched DNA samples were collected from metastatic NSCLC patients with EGFR mutation, which was detected by a clinical EGFR mutant enriched PCR assay using PNA-LNA PCR clamp method. These samples were extracted from tissue samples of fresh tumor tissue except one formalin fixed paraffin embedded (FFPE) sample (tDNA), and plasma samples (cfDNA). Mutation hotspots were enriched by multiplex PCR using the GeneRead DNAseq Targeted Panels V2 Human Tumor Actionable Mutations panel (QIAGEN, Hilden, Germany), followed by library construction, and sequenced on the MiSeq sequencer. Resulting data was analyzed using CLC Genomics Workbench and Genomics Server and mapped to the full human genome.

[Results and discussion] The mean read depth in target region was 59,000 reads. Matched EGFR driver mutations between cfDNA and tDNA were detected with an overall concordance of 25% (5 of 20 samples). The concordance rate of exon21 L858R point mutation, exon19 deletions and exon20 T790M point mutation were 32.5% (3 of 8 samples), 16.7% (2 of 12 samples), and 66.7% (2 of 3 samples), respectively. The mutant allele frequencies in cfDNA ranged from 0.1% to 44.5% (median, 0.3%). While further improvement is mandatory, these results suggest that targeted deep sequencing of cfDNA at high coverage may have a potential utility as a non-invasive mutational analysis. In addition, there seemed to be a tendency of better detection rate for point mutations (45.5%) than deletions (16.7%), while the difference didn't reach statistical significance (p = 0.19), which were corresponding to previous reports.

Citation Format: Ken Suzawa, Shuta Tomida, Takahiro Matsubara, Kadoaki Ohashi, Yuho Maki, Hiromasa Yamamoto, Mizuki Morita, Junichi Soh, Hiroaki Asano, Kazunori Tsukuda, Katuyuki Kiura, Shinichiro Miyoshi, Shinichi Toyooka. Detection of EGFR mutations in circulating cell-free DNA of non-small cell lung cancer patients by next-generation sequencing. [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 2249.
Abstract 2255: Evaluation of the diagnostic value of serum anti-cyclin B1 autoantibody as a biomarker in lung cancer
Authors: Xing Mengtao , Li Pei , Shi Jianxiang , Dai Liping , Li Jitian , Zhang Jianying
Published: July 2016

To detect the diagnostic value of cancer-associated autoantibodies in lung cancer, a panel of 9 tumor-associated antigens (TAAs) were used in this study. The TAAs were composed of p62, p16, Koc, p53, Cyclin B1, Cyclin E, Survivin, HCC1, and Rol-A full-length recombinant proteins. Enzyme-linked immunosorbent assay and immunoblotting were used to detect antibodies in 50 lung cancer sera with clinicopathological characteristics and 42 sera from normal individuals. Following, antibodies to Cyclin B1, survivin, p53 and HCC1 were selected and confirmed in another 62 sera from patients with lung cancer but without clinical date and evaluate the diagnostic value of these TAAs. The results showed that the positive rate of autoantibodies against nine selected TAAs (cyclin B1, survivin, HCC1,p53, KOC,p16,p62, RalA and cyclin E) reached 36%, 36%, 26%, 12%, 8%, 6%, 6%, 2%, 0% in 50 lung cancer sera and 4.8%, 2.4%, 0%, 0%, 2.4%, 4.8%, 2.4%, 2.4%, 4.8% in 42 normal sera, respectively. The AUC values for distinguishing lung cancers from normal sera were 0.767, 0.400, 0.567, 0.395, 0.230, 0.517, 0.635, 0.194, 0.411, respectively. These data suggest that cyclin B1, survivin, HCC1 and p53 has higher clinical diagnostic quality and value than other TAAs in lung cancer. In the confirmation group with 62 lung cancer sear, autoantibodies against these four TAAs were also significantly higher in patients with lung cancer (AUC values of 0.764,0.653,0.623 and 0.622, p<0.05). Evaluation of the diagnostic value of these four TAAs in 50 sera from lung cancer patients with clinical characteristics, and results revealed that serum anti-cyclin B1, anti-survivin and anti-p53 autoantibodies significantly increased in patients with stage I but only serum anti-cyclin B1 increased in stage II and III lung cancer (p <0.05, respectively), ROC curve analysis has shown that anti-cyclin B1 and anti-p53 autoantibodies can discriminate stage I lung cancer from normal controls with AUC value of 0.757,0.697, respectively (p<0.01), and anti-survivin and anti-HCC1 autoantibodies failed to discriminate stage I lung cancer from normal controls. Our results suggest that these four TAAs has higher clinical diagnostic quality and value than other TAAs in lung cancer and serum cyclin-B1 may have the potential to serve as novel non-invasive diagnostic biomarkers in patients with lung cancer, especially in early stage lung cancer.

Citation Format: Mengtao Xing, Pei Li, Jianxiang Shi, Liping Dai, Jitian Li, Jianying Zhang. Evaluation of the diagnostic value of serum anti-cyclin B1 autoantibody as a biomarker in lung 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 2255.
Abstract 2259: Comprehensive evaluation of promising biomarkers for lung cancer risk prediction
Authors: Johansson Mattias A. , Taguchi Ayumu , Muller David , Hanash Samir , Brennan Paul
Published: July 2016

Introductory sentence indicating the purposes of the study:

Lung cancer will remain the number one cancer killer worldwide for the foreseeable future (Globocan 2012). Although low-dose CT screening has been shown to reduce mortality, there is an urgent need to improve risk prediction models to identify those subjects at high risk and most likely to benefit from screening in order its improve the screening efficacy (PMID: 23697514, 25372087).

Brief description of pertinent experimental procedures:

We have assembled a compendium of promising blood-based risk biomarkers for lung cancer that are currently being validated in order to identify a panel of validated risk markers for use in lung cancer risk assessment. Using pre-diagnostic serum samples from CARET cohort participants (current or former heavy smokers) we have recently validated pro-surfactant protein B (Pro-SFTPB) and Diacetyl spermine (DAS) as risk markers for non-small cell lung cancer (PMID: 26282655). Validation of additional markers from the compendium using Pro-SFTPB and DAS as anchor markers with further refinement of the risk prediction models is currently being performed in two large independent cohort studies (EPIC and NSHDS). Biospecimens from these cohorts include pre-diagnostic plasma from 550 former and current smoking lung cancer cases that were diagnosed within 5 years of blood draw, along with 1,100 matched controls.

Summary of the new, unpublished data and conclusion:

Preliminary data indicate that blood-based biomarkers of lung cancer risk have a strong potential to improve on questionnaire-based risk prediction models for lung cancer (PMID: 26282655). During the meeting we will present detailed results of the individual risk biomarkers and their associations with lung cancer risk up to 2 years prior to diagnosis, as well as an assessment of the extent to which they can improve traditional risk prediction models.

Citation Format: Mattias A. Johansson, Ayumu Taguchi, David Muller, Samir Hanash, Paul Brennan. Comprehensive evaluation of promising biomarkers for lung cancer risk prediction. [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 2259.
Abstract 2261: PKR and Jagged1 associated with lymph node metastasis in lung cancer
Authors: Pataer Apar , Hao Chuncheng , Correa Arlene M. , Weissferdt Annikka , Behrens Carmen , Wistuba Ignacio I. , Swisher Stephen G.
Published: July 2016

In the current study, we analyzed the association between lymph node metastasis and over 50 biomarkers in 319 NSCLC patients. We observed that tumor location and the protein expressions of Jagged1 and RNA-dependent protein kinase (PKR) have significantly association with the lymph node metastasis. We further investigated the mechanism of interaction between Jagged1 and PKR, and demonstrated that induction of PKR promotes Jagged1 protein degradation through proteasome system in lung cancer cells. We also found that combination of Jagged1 with PKR improved prognostic value in all stages NSCLC patients. However, combination of Jagged1 with PKR did not improve lymph node metastatic value. Taken together, our data suggest that PKR and Jagged1 are independent markers for predicting lymph node metastasis, and PKR may reduce lymph node metastasis partly by promote Jagged1 proteins degradation in NSCLC patients.

Citation Format: Apar Pataer, Chuncheng Hao, Arlene M. Correa, Annikka Weissferdt, Carmen Behrens, Ignacio I. Wistuba, Stephen G. Swisher. PKR and Jagged1 associated with lymph node metastasis in lung 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 2261.
Abstract 2264: Tumor fragmentation is a poor prognostic histologic biomarker for lung squamous cell carcinoma
Authors: Casanova Ruben , Soltermann Alex
Published: July 2016

Introduction: Lung squamous cell carcinoma (SCC) is the second most frequent histologic type of non-small cell lung carcinoma (NSCLC). Next to pTNM staging, grading is also important for histomorphologic classification of tumors into relevant prognostic groups. SCC forms cohesive tumor sheets of various sizes, ranging from large pushing boundaries to small detached clusters. The presence of small clusters such as single cells, tumor buds 15 cells) could also contain prognostic information and hence be pertinent for tumor grading. Herein we propose a computer-based morphometric analysis for the global assessment of lung SCC fragmentation.

Methods: Surgically resected lung SCC tumors (discovery tissue microarray cohort n = 208, 2 cores per patient; validation whole section cohort n = 99, 2 blocks per patient) were immuno-histochemically stained with pan-cytokeratin. Color-based segmentation (tumor epithelia brown, stroma blue-grey) allowed to separate tumor from stroma and to compute the number of tumor fragments. Associations with disease-free (DFS) and overall survival (OS) were assessed by univariate Cox regressions.

Results: Tumor fragments were defined as tumor clusters >3000μm2 (circa >15 cells) entirely surrounded by stroma. Criteria for high fragmentation were: > 5 fragments summed over 2 TMA cores (range: 2-18, median = 5) and > 700 fragments summed over 2 whole sections (range: 94-3988, median = 570). Respectively, 44% and 42% of the patients had highly fragmenting tumors in the discovery (D) and validation cohort (V), correlating with blood vessel infiltration (cc = 0.181, p<0.001 (D); cc = 0.220, p = 0.029 (V)). High fragmentation was also associated with decreased OS (HR = 1.77, p = 0.001) and DFS (HR = 1.83, p = 0.027) in (D). This was confirmed in (V): OS (HR = 2.09, p = 0.004), DFS (HR = 2.60, p = 0.005).

Conclusions: We used an automated morphometric approach for unbiased scoring of tumor fragmentation on digitalized histological sections. This versatile method could potentially be applied to virtually any solid human carcinoma. Our analysis revealed that tumor fragmentation is a poor prognosticator for lung squamous cell carcinoma, e.g. due to increased likelihood of vessel infiltration.

Citation Format: Ruben Casanova, Alex Soltermann. Tumor fragmentation is a poor prognostic histologic biomarker for lung squamous cell carcinoma. [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 2264.
Abstract 2265: An integrated analysis of gene mutations and gene sets for predicting paclitaxel response in lung adenocarcinoma
Authors: Huang Chia-Yu , Chiu Yu-Chiao , Lu Tzu-Pin , Lai Liang-Chuan , Tsai Mong-Hsun , Hsiao Tzu-Hung , Chuang Eric Y.
Published: July 2016

Lung cancer is the leading cause of cancer death worldwide. A prevalent histological subtype of lung cancer is adenocarcinoma. Chemotherapies and targeted therapies have been developed to treat such malignancy. However, due to the heterogeneity of cancer genomes, drug responses vary considerably among patients and the average survival rate remains quite unsatisfactory. Therefore, integrated biomarkers for predicting drug responses are greatly needed. Addressing this, in the present study we aimed to develop a prediction model based on an integrated analysis of gene mutations and gene sets. Briefly, the two-tailed Student's t-test was performed to identify the gene mutations and gene sets of which activities were associated with drug sensitivity, and classification trees were derived from these genomic features. We applied the analysis to genomic datasets and drug sensitivity data from the Cancer Cell Line Encyclopedia (CCLE) and gene sets defined in the Molecular Signatures Database (MSigDB), and constructed a prediction model for response to paclitaxel, a widely used drug for cancers, in lung adenocarcinoma. Taking KRAS mutation as an example, we identified 20 and 15 drug response-associated gene sets in KRAS-mutant and KRAS-wild type cell lines, respectively. The two lists of gene sets were mutually exclusive, suggesting the need of building individual prediction models for groups of cancer subtypes. We then built a classification tree for each of the two groups and tested their prediction performance by leave one out cross-validation tests; ∼64% and ∼81% accuracy was achieved for KRAS-mutant and KRAS-wild type cell lines, respectively. Gene sets of “SIG_CHEMOTAXIS” and “PID_ERB_GENOMIC_PATHWAY” served as crucial nodes for the trees of KRAS-mutant and KRAS-wild type cells, respectively. In conclusion, we developed a novel method that integrates gene mutations and gene sets for predicting drug responses and demonstrated its high performance in lung adenocarcinoma. Our model is widely applicable to identify potent biomarkers for anticancer drugs in cancers and accelerate the realization of precision medicine.

Citation Format: Chia-Yu Huang, Yu-Chiao Chiu, Tzu-Pin Lu, Liang-Chuan Lai, Mong-Hsun Tsai, Tzu-Hung Hsiao, Eric Y. Chuang. An integrated analysis of gene mutations and gene sets for predicting paclitaxel response 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 2265.
Abstract 227: Leptin induces a fibroblast and endothelial cell crosstalk in tumor stroma
Authors: Lanier Viola
Published: July 2016

Background: The tissue microenvironment plays a major role in tumor development and recurrence. It is known that leptin secreted by cancer and adipose cells can modify the tumor stroma, which is composed of immune, fibroblast, and endothelial cells (EC). Leptin affects both normal and cancer associated fibroblasts. We have previously shown that leptin can modify EC directly via an increase of angiogenic features; which was linked to VEGFR signaling and Notch expression independent of VEGF. It is hypothesized that leptin induces a communication between fibroblast and endothelial cells via induction of Notch signaling, which may be related to exosome production.

Methods: Non-expressing Ob-R fibroblasts, and fibroblasts transfected with Ob-R plasmid, were used to investigate leptin's actions on Notch activation. Fibroblasts were challenged with leptin (0.6, 1.2, 6.2 nM) and an inhibitor of Notch signaling activation (DAPT). Fibroblast culture supernatants were added to and human umbilical vein endothelial cells (HUVEC) cultures. Notch proteins and RNA expression and molecular markers of activated fibroblasts and derived exosomes were determined via Western blot, RT-PCR, and multiplex analyses, respectively. EC angiogenic features were also determined.

Results: Our findings suggest that leptin induces fibroblast activation, which triggers the expression of Notch and the development of angiogenic features by EC.

Conclusions: A novel mechanism is proposed by which leptin promotes tumor stroma development via Notch activation and a crosstalk between fibroblasts and EC. Cancer therapies targeting leptin signaling could be a new strategy for interfering with cellular processes that promotes changes of tumor stroma towards a more aggressive tumor environment.

Citation Format: Viola Lanier. Leptin induces a fibroblast and endothelial cell crosstalk in tumor stroma. [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 227.
Abstract 228: Tumor suppressor functions of BNIP3 and mitophagy in pancreatic ductal adenocarcinoma
Authors: Chourasia Aparajita H. , Springer Maya Z. , Macleod Kay F.
Published: July 2016

The role of mitochondrial dysfunction in cancer is an emerging research field fuelled by the growing realization that mitochondrial dynamics are subject to oncogenic control, with mitochondrial biogenesis and mitophagy in particular modulated directly or indirectly by key oncogenes (c-Myc, B-Raf, K-ras) and tumor suppressors (p53, pRB). Work from our lab has focused on understanding how defects in mitophagy influence tumorigenesis with a particular focus on how BNIP3 loss during tumor progression influences disease outcome. BNIP3 functions as a dimer at the outer mitochondrial membrane to target mitochondria for degradation at the autophagolysosome through its interaction with processed LC3 at nascent phagophores. BNIP3 is a transcriptional target of HIF-1, E2F and FoxO3A transcription factors and is induced by activated Ras and suppressed by the RB and p53 tumor suppressors. Various studies have shown BNIP3 expression to be induced at early stages of human cancers and then lost as cancers progress to malignancy. In particular, epigenetic silencing of BNIP3 in human pancreas cancer has been correlated with reduced median survival. However, the mechanisms by which loss of BNIP3 may be contributing to cancer progression are not well delineated and a better understanding of how BNIP3-dependent mitophagy modulates pancreatic cancer in particular could provide new insight to the development of this deadly disease. We set out to examine how loss of BNIP3 affects the progression of pancreatic ductal adenocarcinoma using human cell lines and genetically engineered mouse models. We show that loss of BNip3 markedly accelerates tumor initiation and progression to invasive PDAC in the Pdx1-Cre;LSL-KRasG12D mouse model with metastases to the liver and lungs evident in BNip3 null mice but not in age-matched wild-type mice. Supporting a role for BNIP3 in suppressing pancreatic tumorigenesis, we also showed that re-expression of exogenous BNIP3 in Miapaca-2 cells (in which endogenous BNIP3 is silenced) resulted in reduced growth while knockdown of BNIP3 in CFPAC1 cells (that express BNIP3) promotes cell growth. Work described here examines the underlying mechanisms explaining these tumor suppressor functions of BNIP3 in pancreatic ductal adenocarcinoma.

Citation Format: Aparajita H. Chourasia, Maya Z. Springer, Kay F. Macleod. Tumor suppressor functions of BNIP3 and mitophagy in pancreatic ductal 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 228.
Abstract 2284: Synergistic anti-tumor effect of the combination of amrubicin and erlotinib in non-small cell lung cancer with wild type EGFR
Authors: Otani Sakiko , Sasaki Jiichiro , Masuda Kuniko , Okamoto Kazuya , Namubu Akiko , Matsumoto Toshihide , Tanaka Masumi , Ishihara Mikiko , Nakahara Yoshiro , Fukui Tomoya , Igawa Satoshi , Masuda Noriyuki
Published: July 2016

Background: Standard second-line chemotherapy against advanced non-small-cell lung cancer (NSCLC) is a single agent such as docetaxel, pemetrexed, or erlotinib (Erl). We recently reported that the combination of amrubicin (AMR) and Erl was clinically safe and effective for previously treated NSCLC patients. The purpose of this research is to confirm the experimental effectiveness of the combination and to investigate mechanism of the action.

Methods: We perform to evaluate the NSCLC cell lines proliferation inhibition by using XTT assay and to make two-dimension isobolograms for assessment of the combination effect. Flowcytemetry analysis (FACS) and Western blot analysis (WB) is also done in vitro. In vivo growth inhibition assay using subcutaneous model on SCID mice is performed. In addition, we measure intracellular concentration of amrubicinol(AMR-OH) which is converted as active metabolite from AMR in both cells treated with AMR alone and the combination.

Results: The combination of AMR and Erl had significant synergistic effect on EGFR wild type NSCLC cell line A549, while additive effect on EGFR mutant cell line PC-9. Sub G0-G1 population on FACS was significantly increased in A549 cells treated with combination. The combination activated apoptosis related proteins in WB. The strong growth inhibition of subcutaneous tumors on SCID mice was observed in the combination cohort. Intracellular concentration of AMR-OH was significantly increased in the combination than in AMR monotherapy.

Conclusion: The combination of AMR and Erl is synergistically induced apoptosis and growth inhibition on A549 cells via increasing intracellular concentration of AMR-OH. This combination may be a promising therapeutic option against advanced NSCLC with wild type EGFR.

Citation Format: Sakiko Otani, Jiichiro Sasaki, Kuniko Masuda, Kazuya Okamoto, Akiko Namubu, Toshihide Matsumoto, Masumi Tanaka, Mikiko Ishihara, Yoshiro Nakahara, Tomoya Fukui, Satoshi Igawa, Noriyuki Masuda. Synergistic anti-tumor effect of the combination of amrubicin and erlotinib in non-small cell lung cancer with wild type EGFR. [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 2284.
Abstract 2311: Adenovirus-transduced allogeneic dendritic cells for cancer immunotherapy
Authors: Essand Magnus , Fotaki Grammatiki , Jin Chuan , Ramachandran Mohanraj , Ma Jing , Karlsson-Parra Alex , Yu Di
Published: July 2016

Immunotherapy is currently being established as standard treatment for cancer. Immune checkpoint blockade antibodies can cure a large proportion of patients with refractory melanoma and lung cancer, and adoptive transfer of patient-derived CD19 chimeric antigen receptor (CAR) T cells results in complete remission in a majority of patient with refractory leukemia. Cancer vaccines based on patient-derived dendritic cells (DC) modified ex vivo to present tumor antigens are promising for cancer treatment especially when combined with immune checkpoint blockade antibodies. Ex vivo-modified DCs are however poor in migrating to secondary lymphoid organs after re-administration. In reality, it is more likely endogenous DC that endocytos the injected DC, mature and then migrate to lymph nodes where they present peptide fragments of tumor antigen to T cells together with required co-stimulation.

Herein, we present a new strategy utilizing adenovirus (Ad)-transduced allogeneic DCs (alloDCs) as immunostimulatory adjuvant to induce both innate and adaptive immune response. Ad/alloDC can be prepared, tested and made ready for release in advance and quickly be prepared for injection in cancer patients upon request, thereby minimizing GMP-processing and handling of cells. Besides encoding tumor antigens, adenovirus also provides stimulatory danger signals for DC activation. We show that Ad/alloDCs secret a panel of proinflammatory cytokines and chemokines including IL-12 and CXCL10 at high levels in a sustainable fashion. We also show that Ad/alloDC recruit and activate neutrophils and NKs to the site of injection. Ad/alloDC can also recruit and interact with allo-reactive T cells to create a proinflammatory milieu, which further recruit and activate endogenous DCs. We verified that endogenous DCs activated in situ by an Ad/alloDC injection migrate to draining lymph node, where they present the engulfed tumor antigen to T cells. We show that these migratory endogenous DCs facilitate T cell activation and proliferation. Moreover, when injected intratumorally, Ad/alloDCs modulate the immunosuppressive environment by reducing the ratio of monocytic to granulocytic myeloid-derived suppressor cells (MDSC). To evaluate the therapeutic efficacy, we use B16 melanoma tumor-bearing C57Bl/6 mice injected with adenovirus-transduced DCs from Balb/c origin to obtain an allogeneic reaction. We show that Ad/alloDC treatment could significantly delay tumor growth and prolong survival of tumor-bearing mice.

Citation Format: Magnus Essand, Grammatiki Fotaki, Chuan Jin, Mohanraj Ramachandran, Jing Ma, Alex Karlsson-Parra, Di Yu. Adenovirus-transduced allogeneic dendritic cells for cancer immunotherapy. [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 2311.
Abstract 2340: PD-1 blockade suppresses gastric cancer development by promoting antitumor immunity in mice
Authors: Kim Woosook , Hayakawa Yoku , Fox James G. , Wang Timothy C.
Published: July 2016

Since advanced gastric cancer is difficult to cure with chemo and/or radiation therapy, new therapeutic approaches are clearly needed. Targeting immune checkpoints such as CTLA-4 and PD-1/PD-L1 has shown promising antitumor immunity in many malignancies by modulating T cell activity. PD-L1 and to a lesser extent of PD-L2 are expressed in human gastric cancer and expression of PD-L1 is associated with poor prognosis. Therefore, developing immunotherapies is an attractive strategy for the treatment of gastric cancer. We evaluated the efficacy of anti-PD-1 therapy in a mouse model of gastric cancer. Gastrin-deficient (GAS-KO) mice have been shown to develop large antral tumors with the combination of N-methyl-N-nitrosourea (MNU) and Helicobacter felis (H. felis) infection. Tumors developed in GAS-KO mice exhibited increased expression of PD-L1 and higher mutation rates than those in wild-type mice. To study the effect of anti-PD-1 antibody in tumor initiation or an early stage of progression, GAS-KO mice were induced with H.felis/MNU to develop tumors, one week later (10 weeks post-MNU) treated with anti-PD-1 antibody or isotype control (10 mg/kg IP qw) and sacrificed at 30 weeks post-MNU. We confirmed that GAS-KO mice treated with isotype control had moderate tumor burdens. Of interest, however, less and much smaller tumors were observed in mice treated with anti-PD-1. To compare immune cell populations in spleen tissue and peripheral blood from each group of mice, flow cytometric analysis was performed. Anti-PD-1-treated mice showed an increase in T cells including cytotoxic CD8+ T cells and a decrease in regulatory T cells and myeloid-derived suppressor cell (MDSC) populations compared with controls. In accordance with this, CD3-positive T lymphocytes were more frequently found in gastric tumors of anti-PD-1-treated mice than controls. Furthermore, qRT-PCR analysis revealed differential cytokine gene expression profiles including Il6, Il1b, and Il23 following anti-PD-1 treatment. Together, these data implicate that PD-1 blockade promotes immune infiltrates and therefore potentiates antitumor immune responses, suggesting targeting PD-1 as a potential new anticancer strategy in gastric cancer. Understanding anti-PD-1 therapy at the cellular and molecular level could provide new strategies for developing highly efficacious combination therapies.

Citation Format: Woosook Kim, Yoku Hayakawa, James G. Fox, Timothy C. Wang. PD-1 blockade suppresses gastric cancer development by promoting antitumor immunity in mice. [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 2340.
Abstract 2357: Therapeutic potential of the natural human IgG1k antibody pritumumab
Authors: Babic Ivan , Mukthavaram Rajesh , Nomura Natsuko , Pingle Sandeep , Glassy Mark C. , Kesari Santosh
Published: July 2016

Pritumumab is a natural human IgG1 kappa antibody derived from a regional draining lymph node of a patient with cervical carcinoma. The recognized antigen is an altered form of vimentin, called ecto-domain vimentin (EDV), that is expressed on the cell surface of epithelial tumor cells. A recombinant version of the mAb was made using the GPEx® system in CHO cells. In a series of comparable studies the CHO mAb was compared with the original hybridoma. Binding specificity with both flow cytometry and immunohistochemical analysis, Western blot analysis, and Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) activity confirms the comparability of the two versions. A concentration-dependent study of pritumumab-mediated ADCC suggests an EC50 of 39.6ng/ml. Further studies include establishing a xenograft model with both SCID and athymic nude mice in which pritumumab was effective in preventing tumor growth in nude mice but not in SCID mice. Analysis of a blood brain barrier model suggests pritumumab shows minimal distribution in normal brain tissues and significant binding in tumor areas of brain tissues indicating the mAb crosses the tumor brain barrier. Overall, these data together suggest pritumumab is suitable for development as an anti-tumor therapeutic.

Citation Format: Ivan Babic, Rajesh Mukthavaram, Natsuko Nomura, Sandeep Pingle, Mark C. Glassy, Santosh Kesari. Therapeutic potential of the natural human IgG1k antibody pritumumab. [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 2357.
Abstract 2363: Combinational therapy with specific immunization plus checkpoint inhibition results in enhanced tumor regression and survival benefit
Authors: Gabitzsch Elizabeth , Rice Adrian , Latchman Yvette , Balint Joseph P. , Jones Frank
Published: July 2016

Effective immunotherapeutic treatment of various cancers may require combinational approaches to induce specific immunity to the tumor as well as inhibition of immune check-points. We have developed a viral gene delivery platform to immunize against numerous tumor associated antigens (TAA) such as carcinoembryonic antigen (CEA), HER2/neu and HPV 16. We investigated the therapeutic benefit of the immunotherapeutics alone and in combination with check-point inhibitors such as programmed death-ligand 1 (PD-1) blockade and LAG3 blockade in murine tumor models. As single agents, immunization with the viral delivery platform encoding the TAA induced target-specific cell-mediated immunity and anti-tumor responses in the respective model. When the immunotherapies were combined with immune checkpoint blockade, an increased level of anti-tumor activity against was observed in addition to an improvement in survival. Tumor microenvironment analysis immunized mice revealed an increase in CD8+ tumor-infiltrating lymphocytes (TILs). In addition, we observed induction of suppressive mechanisms such as programmed death-ligand 1 (PD-L1) expression on tumor cells and an increase in PD-1+ TILs. When immunotherapy was combined with anti-PD-1 antibody, we observed CD8+ TILs at the same increased level but found that a smaller fraction of these were PD-1+. Furthermore, we observed a reduction in PD-L1 expression on tumor cells, providing a mechanism by which combination therapy favors tumor clearance and a rationale for pairing antigen-specific vaccines with checkpoint inhibitors in future clinical trials.

Citation Format: Elizabeth Gabitzsch, Adrian Rice, Yvette Latchman, Joseph P. Balint, Frank Jones. Combinational therapy with specific immunization plus checkpoint inhibition results in enhanced tumor regression and survival benefit. [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 2363.
Abstract 2365: Checkpoint inhibitors and a multivalent melanoma vaccine as a novel combinatorial therapy
Authors: Maniyar Rachana R. , Tuli Neha , Suriano Robert , Geliebter Jan , Wallack Marc , Tiwari Raj K.
Published: July 2016

Over 1 million cases of skin cancer are diagnosed in the US every year, of which 75% deaths result due to melanoma, one of the deadliest forms of skin cancer, arising from melanocytes. We have isolated five patient derived melanoma cell lines (MEL-V, 3MM, GLM2, Mel2, KFM) and characterized the expression of various melanoma associated antigens (MAAs). The presence of a multitude repertoire of common MAAs (Gp100, MART-1, MAGE-A1, NY-ESO-1, Tyrosinase, TRP-1 (Gp75), TRP-2, Melanotransferrin, CD71, CD146) enabled us to use composite cell membrane preparations of these cells as a multivalent vaccine. To this end, we designed a vaccinia virus vaccine that was composed of membrane lysate preparations of the five primary cells and recombinant IL-2. When tested in a pilot study, delayed type hypersensitivity and a multivalent immune response were noted. In an effort to correlate genetic lesions with MAA expression we observed that in BRAF mutant melanoma cells, treatment with a BRAFv600E inhibitor, PLX4032, enhanced the expression of MAAs making the melanoma cells more visible to the vaccination driven immune response. We also observed that melanoma cells constitutively expressed CTLA-4 as determined by Immunofluorescence and Western Blots. This expression was found on the membranes as well as in the cytoplasm in comparatively high levels on wild type as well as the BRAFV600E mutant melanoma cell lines. Further our results suggest that CTLA-4 expression on melanoma cells promotes the immunosuppressive tumor microenvironment enabling tumor evasion. Given that CTLA-4 monoclonal antibodies such as Ipilimumab are approved for clinical treatment, this study opens up a new avenue into a plausible combinatorial therapy for our vaccinia virus based vaccine with Ipilimumab for BRAF mutant as well as wild type melanoma cells.

Citation Format: Rachana R. Maniyar, Neha Tuli, Robert Suriano, Jan Geliebter, Marc Wallack, Raj K. Tiwari. Checkpoint inhibitors and a multivalent melanoma vaccine as a novel combinatorial therapy. [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 2365.
Abstract 2369: Molecular profile of a GM-CSF overexpressing breast cancer whole-cell vaccine with systemic anti-tumor activity
Authors: Lacher Markus Daniel , Wiseman Charles L. , Wagner Joseph
Published: July 2016

BACKGROUND AND PURPOSE OF STUDY: The allogeneic whole-cell cancer vaccine BriaVax(TM) (formerly SV-BR-1-GM) is an ER/PR negative, HER2/neu positive breast cancer cell line (SV-BR-1) we engineered to stably overexpress GM-CSF. BriaVax, rendered proliferation incompetent by irradiation, has thus far been applied to 4 advanced stage cancer patients (3 subjects with breast cancer, 1 subject with ovarian cancer). One breast cancer subject responded to BriaVax with complete remission of a measurable lung lesion and near complete remission of multiple breast lesions after only 3 inoculations. Nevertheless, she relapsed 3 months after completing the protocol, with brain metastases as well as multiple breast lesions. We obtained FDA permission to resume vaccinations, and, upon doing so, all metastatic sites responded with a prompt tumor regression after only 3 inoculations. To prospectively identify patients with a high likelihood of benefiting from BriaVax therapy we began a program to identify molecular factors of diagnostic potential. Here, we describe a gene expression signature that might both be informative about BriaVax’ mechanism of action and helpful for developing diagnostic or monitoring biomarkers.

METHODS: To prospectively identify patients with tumors responsive to BriaVax we began a molecular analysis of both the BriaVax cell line and cells obtained from the special clinical responder's blood. BriaVax gene expression profiles were obtained through Illumina BeadArray and NanoString nCounter technologies and compared to gene expression data sets publically available through the Gene Expression Omnibus (GEO; National Center for Biotechnology Information) portal.

RESULTS: BriaVax expresses a gene signature consistent with a mechanism of action involving not only the activation of cytotoxic T cells but also the induction of a humoral response. In addition, BriaVax expresses known cancer antigens. Notably, blood-derived cells of the special clinical responder expressed genes complementing BriaVax’ gene expression signature, thus possibly explaining the unusually prompt and robust clinical response to BriaVax.

CONCLUSIONS: Our findings suggest that BriaVax exerts its therapeutic effects via multiple modes. We identified both candidate immunogens overexpressed in BriaVax compared to normal breast cells and unraveled a potential mechanism of action explaining the encouraging clinical response observed.

Citation Format: Markus Daniel Lacher, Charles L. Wiseman, Joseph Wagner. Molecular profile of a GM-CSF overexpressing breast cancer whole-cell vaccine with systemic anti-tumor activity. [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 2369.