bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5047: Exosomes derived from HIV-1-infected T cells activate ERK 1/2 through EGFR and TLR3 in cancer cells
Authors: Feng Zhimin , Chen Lechuang , Yue Hong , Wang Bingcheng , Jin Ge
Published: August 2018

Exosomes secreted from HIV-1-infected T cells play important roles in coordinating the immune responses and disease processes in HIV-infected people who are treated with antiretroviral therapy. We hypothesize that exosomes from HIV-infected T cells can promote growth and progression of non-AIDS-defining cancer cells. Activation of the MAPK ERK 1/2 is important for cancer cell survival, proliferation and invasion. We investigated the ability of exosomes from HIV-1-infected T cells to activate ERK 1/2 in head neck squamous cell carcinoma (HNSCC) and lung cancer cells for cancer cell proliferation and progression. Exosomes were isolated from Jurkat and J1.1, a latently HIV-1-infected Jurkat cells, using differential ultracentrifugation and quantified by acetylcholinesterase (AchE) activity. HNSCC cell line HSC3 and lung cancer cell lines NCI-H1299 and NCI-H1437 cells were treated with exosomes. To test the involvement of EGFR and TLR3 in ERK 1/2 activation by exosomes, EGFR neutralizing antibody Cetuximab or the TLR3/dsRNA inhibitor complex was added to cancer cells 30 min prior to adding exosomes. To test the entry of exosomes into cells, HEK293 cells were transiently transfected with EGFR-GFP, and seeded into glass bottom 35 mm dishes, time lapse Z-stacking imaging was performed and deconvolution was used to remove out of focus signals. By using Western blotting, we found that exosomes from J1.1 cells (HIV-1-infected), but not those from HIV-negative Jurkat cells, constantly induced ERK 1/2 phosphorylation in HNSCC and lung cancer cells. Cetuximab dramatically decreased ERK activation by exosomes from J1.1 cells, suggesting an EGFR-dependent ERK activation. While EGF induced a whole series of EGFR phosphorylation, exosomes from J1.1 cells failed to do so, suggesting that exosomes from J1.1 cells trigger ERK activation through a non-canonical EGFR pathway. Furthermore, ERK activation was significantly reduced in TLR3/dsRNA inhibitor pretreated cancer cells in response to exosomes from J1.1 cells, indicating exosomes triggered ERK activation is also TLR3 dependent. Time lapse imaging showed that exosomes quickly entered into cells within two minutes; however, Cetuximab greatly delayed exosome entry. To conclude, we found that exosomes from HIV-infected T cells can induce ERK activation in HNSCC and lung cancer cells through interaction with EGFR and TLR3.

Citation Format: Zhimin Feng, Lechuang Chen, Hong Yue, Bingcheng Wang, Ge Jin. Exosomes derived from HIV-1-infected T cells activate ERK 1/2 through EGFR and TLR3 in cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5047.
Abstract 5050: Molecular heterogeneity in HPV-dependent and -independent vulvar carcinogenesis: Chromosome 1 gain reflects the cancer risk in vulvar intraepithelial neoplasia
Authors: Bleeker Maaike CG , Swarts Dorian RA , Voorham Quirinus JM , Wilting Saskia M. , Beurden Marc van , Steenbergen Renske DM
Published: August 2018

Vulvar squamous cell carcinoma (VSCC) and precancerous high-grade vulvar intraepithelial neoplasia (VIN) can develop through human papillomavirus (HPV)-dependent and -independent pathways, indicating a heterogeneous disease. Only a minority of VIN progress, but current clinical and histological classifications are insufficient to predict the cancer risk. Here we analyzed copy number alterations (CNA) to assess the molecular heterogeneity of vulvar lesions in relation to HPV and cancer risk. HPV-status and CNA by means of whole-genome next-generation shallow-sequencing were assessed in 24 VSCC and 41 VIN lesions. The latter included VIN of women with associated VSCC (VINVSCC) and women who did not develop VSCC during >10 year follow-up (VINnoVSCC). HPV-testing resulted in 41 HPV-positive lesions (16 VINVSCC, 14 VINnoVSCC and 11 VSCC) and 24 HPV-negative lesions (11 VINVSCC and 13 VSCC). HPV-positive and -negative VSCC showed a partially overlapping pattern of recurrent CNA, including frequent gains of 3q and 8q. In contrast, amplification of 11q13/cyclinD1 was exclusively found in HPV-negative lesions. HPV-negative VINVSCC had less CNA than VSCC (P = 0.009), mainly 8q gains and 8p losses. In HPV-positive lesions, CNA frequencies were lower in VINnoVSCC than in VINVSCC (P = 0.022) and VSCC. Interestingly, a 1pq gain was detected in 81% of VINVSCC and only 29% of VINnoVSCC. In conclusion, HPV-dependent and -independent vulvar carcinogenesis is characterized by distinct CNA patterns at the VIN stage, while more comparable patterns are present at the cancer stage. Progression risk in VIN is reflected by the extent of CNA, in particularly chromosome 1 gain.

Citation Format: Maaike CG Bleeker, Dorian RA Swarts, Quirinus JM Voorham, Saskia M. Wilting, Marc van Beurden, Renske DM Steenbergen. Molecular heterogeneity in HPV-dependent and -independent vulvar carcinogenesis: Chromosome 1 gain reflects the cancer risk in vulvar intraepithelial neoplasia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5050.
Abstract 5056: HPV, HIV and male gender as major risk factors for anal neoplastic transformation in African Americans
Authors: Brim Hassan , Afsari Ali , Atefi Nazli , Retland Nicole , Abbas Muneer , Naab Tammey , Shokrani Babak , Laiyemo Adeyinka O. , Lee Edward L. , Nouraie Seyed M. , Ashktorab Hassan
Published: August 2018

Background: Human Papilloma Virus (HPV) is the most common sexually transmitted infectious agent. The incidence HPV-related anal cancers increased among HIV infected people. Aim: To assess risk factors of anal lesions among African Americans in an inner city hospital. Methods: We reviewed medical records of 370 African-Americans with anal lesions at Howard University Hospital from Jan. 2007 to Dec. 2015. Demographic, clinical and pathological data including; HPV, HIV, HCV (hepatitis C virus), diabetes mellitus (DM), hypertension (HTN) and body mass index (BMI) were collected. Statistical analysis was performed using Chi-square and Student's t-test. Results: 276 (75%) patients were male, with median age of 44 years and BMI of 25.8 kg/m2. The frequency of condyloma, high grade dysplasia, squamous cell carcinoma (SCC) and adenocarcinoma was 191 (52%), 26 (7%), 31 (8%) and 8 (2%), respectively. The frequency of HPV, HIV, and HCV was 231 (68%), 147 (43%) and 42 (12%), respectively. HPV and HIV were risk factors for condyloma and dysplasia (P<0.05). Forty two percent of patients had diabetes (DM) and hypertension (HTN). Conclusion: Our results shows that majority of patients with condyloma are male and young with HPV and HIV infection. A clinical utility of HPV as risk factor may be use for prediction of anal lesions.

Citation Format: Hassan Brim, Ali Afsari, Nazli Atefi, Nicole Retland, Muneer Abbas, Tammey Naab, Babak Shokrani, Adeyinka O. Laiyemo, Edward L. Lee, Seyed M. Nouraie, Hassan Ashktorab. HPV, HIV and male gender as major risk factors for anal neoplastic transformation in African Americans [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5056.
Abstract 5059: Comprehensive molecular profiling of HPV-induced transformation over time
Authors: Babion Iris , Miok Viktorian , Jaspers Annelieke , Wieringen Wessel N. van , Steenbergen Renske D. , Wilting Saskia M.
Published: August 2018

Although persistent infection with high-risk human papillomavirus (HPV) is generally acknowledged as necessary cause for cervical cancer, additional molecular changes are required for the development of cancer. Those changes include chromosomal aberrations that result in deregulated expression of coding and non-coding RNAs. In this study we performed a comprehensive and longitudinal molecular characterization of HPV-transformed keratinocyte cell lines, to identify the sequential order and relevance of these molecular alterations for HPV-induced transformation. Genome-wide chromosomal, mRNA and miRNA expression profiles were generated from 4 HPV-transformed keratinocyte cell lines at 8 different passages representing progressive stages of transformation. Unsupervised hierarchical clustering of DNA copy number, mRNA and miRNA profiles revealed that in all 4 cell lines most pronounced changes were associated with the acquisition of anchorage independence, a hallmark of transformation. Approximately one third of differentially expressed mRNAs and miRNAs was directly attributable to DNA copy number alterations. Focal adhesion, TGF-β signalling and mTOR signalling pathways were enriched among these genes. In addition, longitudinal analysis identified more than 600 potential miRNA-mRNA interactions that were negatively correlated during passaging. Four interactions were also predicted by 3 independent prediction algorithms, and have in part already been functionally validated, thereby demonstrating the validity of our approach. In conclusion, integrated longitudinal analysis of our HPV-induced transformation model enabled us to pinpoint relevant interconnected molecular changes and affected signalling pathways. Increased understanding of the interplay between different molecular alterations and affected pathways will be of importance for the clinical management of patients diagnosed with HPV-induced (pre)cancerous lesions.

Citation Format: Iris Babion, Viktorian Miok, Annelieke Jaspers, Wessel N. van Wieringen, Renske D. Steenbergen, Saskia M. Wilting. Comprehensive molecular profiling of HPV-induced transformation over time [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5059.
Abstract 5073: Involvement of TGFa-EGFR-Akt axis on enhanced proinflammatory chemokines in triple-negative breast cancer cells
Authors: Ignacio Rosa Mistica Coles , Gibbs Carla , Lee Eunsook , Son Deok-Soo
Published: August 2018

Triple-negative breast cancer (TNBC) is aggressive, leading to poorer outcomes. Chemokines have chemoattractive potential for cancer metastasis. We investigated the signature of chemokine network between TNBC and non-TNBC cells followed by underlining mechanisms on enhanced proinflammatory chemokines in TNBC. Analysis from microarray dataset revealed that basal-like BC subtype representing TNBC expressed dominantly proinflammatory chemokines, such as CXCL1 and 8, compared to non-TNBC. Chemokine PCR array confirmed the dominant proinflammatory chemokines in TNBC cells. As a driving factor for proinflammatory chemokines in TNBC cells, we checked expression profiles for epidermal growth factor receptor (EGFR) family and its downstream signaling. TNBC cells showed higher expression levels of EGFR and phosphorylated Akt compared to non-TNBC cells. In addition, EGF enhanced the proinflammatory chemokines in TNBC cells. Knockdown of Akt reduced the CXCL2 promoter activity, while overexpression of Akt enhanced the activity. MKK2206, an Akt inhibitor, abrogated the CXCL2 promoter activity, but targeting Erk using inhibitor and knockdown did not reduce the activity. As a driving factor for EGFR-mediated Akt activation in TNBC cells, we found transforming growth factor alpha (TGFα) among ligands of EGFR family. MK2206 decreased the TGFα promoter activity, while overexpression of Akt significantly increased the activity in TNBC cells. MK2206 abrogated TGFα protein released by TNBC cells. MK2206 downregulated CXCL2 mRNA, while TGFα upregulated the CXCL2 expression. Taken together, higher expression of proinflammatory chemokines in TNBC involves TGFα-EGFR-Akt axis, probably contributing to the inflammatory burden followed by promoted cancer progression and higher mortality in TNBC.

Citation Format: Rosa Mistica Coles Ignacio, Carla Gibbs, Eunsook Lee, Deok-Soo Son. Involvement of TGFa-EGFR-Akt axis on enhanced proinflammatory chemokines in triple-negative breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5073.
Abstract 51: Role of hematopoietic cell kinase (HCK) in breast cancer dormancy
Authors: Khiste Sachin , Eton Elliot , Hodgetts Kevin , Lusic Hrvoje , Gopinath Kodaganur , Krishnamurthy S , Sengupta Shiladitya , Goldman Aaron
Published: August 2018

Background: Triple negative breast cancer (TNBC) is an aggressive basal-like malignancy, which recurs more frequently than any other subtype of breast cancer. We recently discovered that TNBC can overcome drug pressure by switching to a hybrid basal-like (CD44Hi) and epithelial-like (CD24Hi) cell state, which is accompanied by a transient period of quiescence, or dormancy. These dormant, ‘hybrid' cells highly express the phosphorylated SRC family kinase, hematopoietic cell kinase (HCK). Methods: Here, we used a panel of novel HCK inhibitors to study the effect of signaling-disruption in drug tolerance and tumor cell dormancy. Using flow cytometry and immunofluorescent microscopy we examined how HCK inhibition perturbs common features associated with dormancy such as proliferation and cell cycle status, intracellular levels of reactive oxygen species, and state of glucose metabolism. Finally, we validated the results using an ex-vivo human tumor model that predicts clinical response and recreates the native tumor microenvirnoment, CANscriptTM. Results: We determined that targeted inhibition of HCK down-regulates the features of tumor cell dormancy and drug tolerance, eliminating cells with a hybrid mesenchymal-epithelial phenotype, diminishing polyploidy and hyper-G2 endoreduplication, and suppressing features of a dysregulated glycolytic state. Conclusion: Expression of HCK drives cancer cells to a quiescent cancer cells subpopulation, resulting in tolerance to chemotherapy. Targeted disruption of HCK overcomes these features of dormancy. HCK inhibitors could emerge as a novel treatment for chemotherapy-induced breast cancer dormancy.

Citation Format: Sachin Khiste, Elliot Eton, Kevin Hodgetts, Hrvoje Lusic, Kodaganur Gopinath, S Krishnamurthy, Shiladitya Sengupta, Aaron Goldman. Role of hematopoietic cell kinase (HCK) in breast cancer dormancy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 51.
Abstract 5111: Gene editing in the NSG mouse strain and its genetic derivatives
Authors: Grass David S. , Buaas F William , Wiles Michael V. , Low Benjamin E. , Kutny Peter
Published: August 2018

Mouse models continue to be powerful tools in preclinical oncology and immuno-oncology studies. The development of the inbred highly immunocompromised mouse strain NSG (NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ) has dramatically improved the ability to work with human tumors in an organismal context using xenograft approaches. Furthermore, technical innovations have expanded the utility of this personalized tumor platform via the reconstitution of parts of the human immune system. An example of this is the NSG-SGM3 inbred mouse strain, in which transgenes expressing human IL3, GMCSF and SCF have been stably integrated into the NSG background. This NSG “derivative” strain can effectively support the stable engraftment of human myeloid lineages and the regulatory T cell populations. Direct genetic modification of the NSG genome in mouse embryos was a key technical capability to attain this NSG “derivative” strain rapidly and avoid extensive breeding. Modification of NSG and derivative strains represent an opportunity to optimize the engraftment of other human lineages. The ability to apply modern genome modification technology to different inbred mouse strains is highly desirable but must overcome two major technical hurdles - 1) Reproductive biology constraints that prevent live born progeny after reagent delivery methods such as microinjection or electroporation and 2) Cell intrinsic DNA-repair processes that are a prerequisite enabling gene editing tools. Given the SCID mutation (Prkdcscid) in the NSG background compromises some DNA repair pathways, it was not clear whether nuclease assisted recombination technologies would be a robust tool to modify NSG (and derivatives) genomes at the zygote stage. Here, we demonstrate that CRISPR-Cas9 is an effective tool for introducing genome modifications in NSG and present a specific example using the Hprt locus. Furthermore, we will present general experience with genomic modifications using NSG, and its derivatives, to emphasize direct modification of these genomes is technically feasible and is enabling the rapid sequential genetic tailoring in this high value cancer model strain.

Citation Format: David S. Grass, F William Buaas, Michael V. Wiles, Benjamin E. Low, Peter Kutny. Gene editing in the NSG mouse strain and its genetic derivatives [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5111.
Abstract 5119: In vivo characterization of the Duffy Antigen Receptor for Chemokines (DARC) in the breast cancer microenvironment
Authors: Martini Rachel , Manley Nancy , Davis Melissa
Published: August 2018

Chemokines are small inflammatory molecules that, in combination with their receptors, recruit lymphocytes to influence immune and inflammatory responses. This work specifically follows an atypical chemokine receptor, known as the Duffy Antigen Receptor for Chemokines (DARC). DARC is a non-signaling receptor able to bind both the CC and CXC chemokine classes, and mainly functions to regulate homeostatic levels of chemokines in circulation, and aid in chemokine transcytosis from sites of inflammation to mediate immune response. DARC serves as a portal of entry for the malaria-causing parasite Plasmodium vivax. In Sub-Saharan Africa, where malaria is endemic, a regulatory variant arose removing DARC expression from red blood cells (RBCs). This mutation, known as the Duffy-Null allele, was highly beneficial in this population, as removal of DARC from RBCs conferred immunity to Duffy-Null individuals. The Duffy-Null allele quickly rose to fixation in this population, and today recent decedents of Sub-Saharan Africans carry this allele. In the context of breast cancer (BCa) we are particularly interested in how Duffy-Null status impacts the tumor microenvironment. In clinic, we observe that pre-menopausal women of African descent have higher incidence rates of BCa, and that regardless of age, these women are disproportionately diagnosed with the most aggressive molecular subtype of BCa, triple-negative BCa. Knowing the important role that DARC plays in immune response, and that the Duffy-Null allele is prevalent in this population of women, we aim to investigate how Duffy-Null status impacts the tumor microenvironment in vivo through establishment of mouse models that depict Duffy-Null status in the BCa environment. To do this, we are using the C3(1)-TAg BCa transgenic mouse, alongside a DARC-KO mouse to characterize cancer progression and aggressiveness in BCa transgenic mice with varying global DARC expression. At 6 and 12 week time points, mice were sacrificed and the following information was obtained: total number of tumors, tumor location, tumor size and weight, and hematoxylin and eosin staining of tumor sections. Removal of DARC could lead to an excess of chemokines in the tumor microenvironment, promoting processes of tumor progression and metastasis, leading to the more aggressing tumor phenotypes we observe in women of African descent in clinic.

Citation Format: Rachel Martini, Nancy Manley, Melissa Davis. In vivo characterization of the Duffy Antigen Receptor for Chemokines (DARC) in the breast cancer microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5119.
Abstract 5121: Obesity and prostate cancer progression: Role of the chemokine CXCL12
Authors: Ahn Songyeon , Saha Achinto , Kolonin Mikhail G. , DiGiovanni John
Published: August 2018

Prostate cancer (PCa) is the third leading cause of cancer in men in the United States. Many epidemiologic studies suggest that obesity increases PCa progression and mortality. However, the mechanistic relationship between PCa progression and obesity is very complex and not fully understood. In previous studies, we have shown that obesity enhances PCa growth and progression in the HiMyc mouse model of PCa by changing the surrounding tumor microenvironment, especially in white adipose tissue (WAT). WAT consists of many cells including mature adipocytes, adipose stromal cells (ASCs) and inflammatory cells, which secrete chemokines, cytokines and proinflammatory molecules. ASCs are an important component of the stromal vascular fraction (SVF) from WAT that secretes such factors. In this study, we evaluated the role of ASCs and ASC-derived CXCL12 in driving PCa progression in obesity. We observed upregulation of mRNA expression of CXCL12 in the SVF of the periprostatic WAT from obese HiMyc mice. In HMVP2 cells, a PCa cell line derived from HiMyc mice expressing high levels of CXCR4 and CXCR7, CXCL12 treatment increased activation of MAPKs (JNK and ERK), AKT, STAT3 and NFkB. In further studies, HMVP2 cells were treated with the CXCR4 antagonist, AMD3100, alone or together with CXCL12 and the CXCL12-induced signaling was abolished in AMD3100-treated cells. Similar results were observed in CXCR4 knockdown cells and CXCR7 knockdown cells. In addition, knockdown of CXCR4 and CXCR7 significantly decreased migration and invasion of the HMVP2 cells. Lastly, a tumor study was performed by injecting HMVP2 cells into the flank of FVB/N mice, which were fed either a control diet or obese diet. AMD3100 significantly inhibited the growth of HMVP2 tumors in both obese and control diet groups. AMD3100 also inhibited proliferation and epithelial-mesenchymal transition (EMT) in both obese and control diet groups. In conclusion, this study provides evidence that the CXCL12/CXCR4/CXCR7 signaling pathway is important for obesity-driven PCa progression in HiMyc mice. Furthermore, this pathway may represent a novel therapeutic target for inhibiting obesity-driven PCa growth and progression.

Citation Format: Songyeon Ahn, Achinto Saha, Mikhail G. Kolonin, John DiGiovanni. Obesity and prostate cancer progression: Role of the chemokine CXCL12 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5121.
Abstract 5123: Changes of oral microbiome profiles associated with oral squamous cell carcinoma
Authors: Hashimoto Kengo , Nomoto Shuji , Hirabayashi Sho , Ueda Sei , Oh-iwa Ichiro , Nagao Toru , Shimozato Kazuo
Published: August 2018

[Background] Oral squamous cell carcinoma (OSCC) is the most frequently occurring cancer among the head and neck SCCs worldwide. Despite significant efforts in OSCC research and advanced surgical treatments, radiotherapy, and chemotherapy, the 5-year overall survival rate has remained < 50% for the last few decades. OSCC prognosis remains poor due to late diagnosis. Therefore, clinically effective biomarkers for the early detection of OSCC can greatly improve the survival rate and prognosis. Approximately 15% of oral cancers cannot be explained by major risk factors, such as tobacco and alcohol use. A growing body of evidence supports an association between specific microorganisms, including those in the oral cavity, and various types of cancers. Because the role of bacteria in OSCC development has not been completely described, we investigated the changes in abundance of oral microbiota associated with OSCC. [AIMS] To identify the association between the oral microbiome and OSCC for the early detection and prevention. [Materials and methods] Unstimulated saliva samples were collected from 4 OSCC-free, 6 leukoplakia, and 6 OSCC subjects. Total bacterial genomic DNA from saliva was isolated, and 16S rRNA gene survey was performed by next-generation sequencing of the V4 region. The sequences were analyzed against reference sequences from the Human Oral Microbiome Database, and the relative abundance distribution for phylogenetic groups in 3 different libraries was compared. [Results] In total, 448 bacterial species from combined (OSCC-free, leukoplakia, and OSCC) libraries were detected, which were classified into 133 genera, 69 families, 41 orders, 26 classes, and 12 phyla. The frequency of phylum Bacteroidetes was more notably abundant in OSCC subjects compared with leukoplakia and OSCC-free subjects. In contrast, the abundances of genera Streptococcus and Rothia were significantly lower in OSCC subjects compared with leukoplakia and OSCC-free subjects using the Mann-Whitney U test. [Conclusion] This study revealed some OSCC-associated shifts in oral microbial communities. Salivary screening is suitable for the primary screening procedure because of its noninvasiveness and easy accessibility. Therefore, these changes in the saliva microbiome may have potential application as a useful diagnostic tool for the early detection of OSCC and malignant transformation of pre-cancerous regions.

Citation Format: Kengo Hashimoto, Shuji Nomoto, Sho Hirabayashi, Sei Ueda, Ichiro Oh-iwa, Toru Nagao, Kazuo Shimozato. Changes of oral microbiome profiles associated with oral squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5123.
Abstract 5125: The relationship between microbiome Fusobacterium nucleatum and autophagy in esophageal cancer
Authors: Liu Yang , Baba Yoshifumi , Nakagawa Shigeki , Sawayama Hiroshi , Ishimoto Takatsugu , Iwatsuki Masaaki , Yamashita Yoichi , Miyamoto Yuji , Yoshida Naoya , Baba Hideo
Published: August 2018

Background: Research into the microbiome is a rapidly advancing field in human cancers. The gut microbiome has recently been shown to play an important role in health, as well as in diseases such as obesity, inflammatory bowel disease, diabetes, non-alcoholic fatty liver disease, and several types of cancers. Fusobacterium nucleatum is a component of the human microbiome that primarily inhabits the oral cavity. It causes periodontal disease and has also been implicated in the development of human cancers. We have recently reported that Fusobacterium nucleatum in esophageal cancer tissues was associated with shorter survival, suggesting a potential role as a prognostic biomarker. In addition, It has been revealed that Fusobacterium nucleatum promotes chemoresistance to colorectal cancer by modulating autophagy. Nonetheless, the relationship between Fusobacterium nucleatum and autophagy in esophageal cancers have been still unknown. We constructed this study to examine the relationship between Fusobacterium nucleatum and autophagy.

Methods: Using 284 esophageal cancers, we detected ATG-7 and ULK -1 expression by IHC staining in tumors.

Results: 21% cases showed ATG-7 positive and 25% cases showed ULK-1 positive. ATG-7 and ULK-1 expression in Fusobacterium nucleatum-positive tumors were significantly higher compared with that of Fusobacterium nucleatum-negative tumors (P=0.026 and P=0.0077). In addition, ATG-7 expression was associated with cancer-specific survival rate in esophageal cancer patients (P=0.0496). There was no relationship of ULK-1 expression and prognosis.

Conclusion: Fusobacterium nucleatum was related with ATG-7 and ULK-1. Our findings may shed lights on understanding the mechanism by which Fusobacterium nucleatum affects chemoresistance in esophageal cancer.

Citation Format: Yang Liu, Yoshifumi Baba, Shigeki Nakagawa, Hiroshi Sawayama, Takatsugu Ishimoto, Masaaki Iwatsuki, Yoichi Yamashita, Yuji Miyamoto, Naoya Yoshida, Hideo Baba. The relationship between microbiome Fusobacterium nucleatum and autophagy in esophageal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5125.
Abstract 5142: Elevated glycolysis in pre-malignant ovarian cancer
Authors: Lokshin Anna E. , Azher Mounia Alaoui El , Velikokhatnaya Liudmila , Prosser Denise
Published: August 2018

Ovarian cancer is the fifth leading cause of cancer death in women and the most lethal gynecologic malignancy. High-grade serous ovarian carcinoma (HGSC) originates mainly from the fallopian tube epithelium, and is characterized by TP53 mutation identified in all HGSCs as well as the fallopian tubal precursor of serous tubal intraepithelial carcinomas (STICs). The loss of a functional TP53 is an early event in the malignant transformation of fallopian tube epithelium. We have established 4 primary cell cultures of normal fallopian tube epithelial (FTE) cells, and further generated immortalized FTE cells using SV40 large plus small T antigens (FTE-TAg) to disrupt TP53 function. This model mimics the primary lesions of ovarian cancer and is suitable to identify biomarkers and molecular targets of ovarian cancer. Therefore, we compared the secretomes of primary FTE cells with that of their parental isogenic pairs FTE-TAg cells using mass spectrometry analysis. We identified 813 proteins, 228 of which were differentially expressed. Ingenuity Pathway Analysis (IPA) core analysis revealed glycolysis as a major canonical pathway. Overexpression of key enzymes of glycolytic pathway was confirmed by IHC in human STIC lesions. Molecular mechanisms of glycolysis regulation in premalignant lesion of ovarian cancer were determined. Role of glycolysis in the establishment of premalignant ovarian cancer microenvironment (PME) was characterized. Prevention approaches targeting glycolysis were tested in transgenic animal model of ovarian cancer.

Citation Format: Anna E. Lokshin, Mounia Alaoui El Azher, Liudmila Velikokhatnaya, Denise Prosser. Elevated glycolysis in pre-malignant ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5142.
Abstract 5144: Cooperative effects of mushroom polysaccharides and herbal saponins on tumor growth and gut microenvironment in ApcMin/+mice
Authors: Li Xiao-ang , Huang Guoxin Huang , Khan Imran , Xia Wen-rui , Leong Wai kit , Hsiao W.L. Wendy
Published: August 2018

Prebiotics are considered as non-digestible food ingredients that stimulate the growth of beneficial gut bacteria, thus contribute to the health of the host. Recent evidences have suggested that prebiotics treatment may become one of the key therapeutic strategies for many diseases, especially for cancer. In our previous studies, both Gynostemma pentaphyllum saponin (GpS) and Ganoderma lucidum polysaccharides (GlP) demonstrated significant prebiotic effects in mouse models. In addition, GpS alone exerted significant anti-tumor effect by restoring the dysbiosis of the gut microbiota and microenvironment of the ApcMin/+mice. Thus, we hypothesized that supplement with GlP might enhance the anti-cancer effect of GpS through the modulation of the gut microbiota. To test the hypothesis, we treated ApcMin/+mice with four different treatments, i.e. GlP (750mg/kg), GpS (300mg/kg), GlP (750mg/kg) + GpS (300mg/kg) and the drinking water control. The results showed that GlP+GpS was more effective in suppressing the polyp growth compared to the GpS or GlP groups. GlP+GpS also improved the health of gut barriers by inducing Paneth cells and goblet cells, and reinstating the E-cadherin and N-cadherin expressions in a greater extend compared to the other groups. All treatments facilitated the polarization of M1 macrophage to M2 macrophage, and increased the anti-inflammatory cytokines (IL-4 and IL-10). Interestingly, expressions the SCFA receptors, GPR 43, 49 and 119 were markedly elevated in all treatment groups, with the most prominent expression observed in the GlP+GpS group. 16S rRNA sequencing data were analyzed by using alpha diversity analysis and weighted UniFrac distance analysis. Differential alterations of the composition of gut microbiota were observed in all treatment groups. Certain bacteria (Anoerodtipe, Bacteroides, Clostridium, Citribacter, Subdoligranulum, C.

leptum) associated colitis and colorectal carcinoma were significant decreased, while some SCFA-producing bacteria (Blautia sp., Eubacterium saphenum, Clostridium aminobutyricum) were enhanced in the treated groups. We concluded that the anti-tumor effects of GpS and GlP might be in part through the modulation of the gut microbiota and the improvement of epithelial microenvironment, and the stronger effects were produced by the GpS+ GlP combined treatment.

[This study was supported by Macau FDCT 015/2014/A1 and 103/2016/A3]

Citation Format: Xiao-ang Li, Guoxin Huang Huang, Imran Khan, Wen-rui Xia, Wai kit Leong, W.L. Wendy Hsiao. Cooperative effects of mushroom polysaccharides and herbal saponins on tumor growth and gut microenvironment in ApcMin/+mice [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5144.
Abstract 5149: CMTM6 is a potential metastasis regulator of EWS cells
Authors: Nishiyama Yuko , Asano Naofumi , Fujiwara Yuko , Kondou Tadashi , Tsuchiya Naoto
Published: August 2018

Ewing's sarcoma (EWS) is a highly aggressive and metastatic malignant tumor of bone and soft tissues, whose incidence is highest in children and young adulthood. Control of metastasis based on the molecular targeting contributes to improve patient prognosis. To this end, we screened expression of microRNAs in clinical samples to identify target molecules, which regulate malignant properties of EWS cells. Microarray analysis indicated that miR-451a is significantly downregulated in the cases with poor prognosis. Although miR-451a did not affect cell proliferation, it strongly repressed migration of EWS cells. Gene expression analysis revealed that miR-451a targeted CMTM6, a chemokine-like membrane protein with unknown function. Depletion of CMTM6 by siRNAs strongly inhibited migration of EWS cells. Our data suggest that at least the expression of miR-451a and CMTM6 is involved in metastasis onset.

Citation Format: Yuko Nishiyama, Naofumi Asano, Yuko Fujiwara, Tadashi Kondou, Naoto Tsuchiya. CMTM6 is a potential metastasis regulator of EWS cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5149.
Abstract 5155: The modulation of actin dynamics via atypical protein kinase-C activated cofilin regulates migration of colorectal cancer cells
Authors: Islam S M Anisul , Khalid Khandker Mohammad , Bommareddy Raja Reddy , Patel Rekha , Acevedo-Duncan Mildred
Published: August 2018

Colorectal cancer (CRC) is the second most common cancer in both men and women in the United States. The exact mechanism of CRC cells migration is poorly understood. Actin polymerization is thought to be an initial step in cancer cells motility cycle which drives the formation of cell protrusions that defines the direction of migration and initiate cell crawling. Cofilin is a significant actin regulating molecule that can regulate the migration of cancer cells by the formation of lamellipodia and filopodia. In this study, the effect of atypical Protein Kinase C (aPKC) on cofilin in CRC was studied by using two inhibitors of aPKC: 1) ICA-I (5-amino-1-(2,3-dihydroxy-4-hydroxymethyl) cyclopentyl)-1H-imidazole-4-carboxamide) is a specific inhibitor of PKC-ι and 2) ζ-Stat (8-hydroxy-1, 3, 6-naphthalenetrisulfonic acid) is a specific inhibitor of PKC-ζ. The cell lines tested were CCD18CO normal colon fibroblast and LOVO & RKO metastatic CRC cells. The inhibition of aPKC did not bring any significant toxicity in CCD18CO colon fibroblasts cells. However, our data showed that the inhibition of aPKC blocks the migration of CRC cells migration by increasing the level of cofilin phosphorylation at serine-3. Additionally, the aPKCs inhibition brought a significant reduction of CRC cells proliferation along with the reduction of survival markers. Our findings suggest that the PKC-ι and/or PKC-ζ may be responsible for the migration of CRC cells as well as carcinogenesis. These results suggest the possibility of utilizing aPKCs as the potential therapeutic targets for the CRC cells metastasis.

Citation Format: S M Anisul Islam, Khandker Mohammad Khalid, Raja Reddy Bommareddy, Rekha Patel, Mildred Acevedo-Duncan. The modulation of actin dynamics via atypical protein kinase-C activated cofilin regulates migration of colorectal cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5155.
Abstract 5167: Rab27a promotes melanoma cell invasion and metastasis via the regulation of exosome secretion
Authors: GUO DAJIANG , Lui Goldie Y. , Sharp Danae M. , Lai Siew Li , Tikoo Shweta , Kwan Rain , Geldermalsen Michelle van , Holst Jeff , Haass Nikolas K. , Weninger Wolfgang , Beaumont Kimberley A.
Published: August 2018

Introduction: The Rab GTPase family has been increasingly related to carcinogenesis and cancer biology during the past decade. In melanoma, Rab27a has been identified as a tumor dependency gene. Rab27a is thought to regulate exosome secretion in several cell types, however the exact role of Rab27a in melanoma biology and the underlying mechanisms are not well investigated.

Experimental Procedures: To investigate the influence of Rab27a on melanoma biology and exosome secretion, Rab27a shRNA knockdown (KD) or CRISPR knockout (KO) was performed in human and murine melanoma cell lines. The effect of Rab27a loss on melanoma cell invasion and motility was analyzed by in vitro 3D spheroid invasion and live cell imaging. Rab27a function on melanoma metastasis was investigated using a mouse melanoma spontaneous metastasis model. Exosomes secreted by melanoma cell lines with/without Rab27a KD/KO were purified and characterized by electron microscopy, NanoSight analysis, BCA assay, western blotting and mass spectrometry.

Results: Our results indicate that Rab27a loss in Rab27a high metastatic melanoma cell lines reduced 3D spheroid invasion and cell motility and Rab27a loss also reduced spontaneous melanoma metastasis in vivo. Rab27a KD invasion phenotype can be partially rescued by addition of exosomes from Rab27a replete cell conditioned media, but not exosomes derived from Rab27a KD cells. Loss of Rab27a does not alter the number of exosomes secreted from melanoma cells, but does alter exosome protein composition and morphology. Mass spectrometry analysis has shown that Rab27a replete cell secreted exosomes are more involved in regulating cell invasion and motility than the exosomes from Rab27a KD cells.

Conclusion: In summary, Rab27a promotes the invasion and metastasis of a subset of melanoma cells via the regulation of pro-invasive exosomes, which indicates Rab27a as a potential therapeutic target for preventing melanoma progression.

Citation Format: DAJIANG GUO, Goldie Y. Lui, Danae M. Sharp, Siew Li Lai, Shweta Tikoo, Rain Kwan, Michelle van Geldermalsen, Jeff Holst, Nikolas K. Haass, Wolfgang Weninger, Kimberley A. Beaumont. Rab27a promotes melanoma cell invasion and metastasis via the regulation of exosome secretion [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5167.
Abstract 517: LncRNA expression at the different site of high-grades serous carcinoma
Authors: Filippov-Levy Natalie , Smith Yoav , Trope Claes Gøran , Davidson Ben , Reich Reuven
Published: August 2018

Background: High-grades serous carcinoma (HGSC) is the most aggressive subtype of ovary cancer. The disease appears at three anatomic sites: ovary tumor, solid metastasis and effusions. Not much is known about the regulatory pathways involved in cancer progression. Long non coding RNA (lncRNA) is non protein coding RNA with size >200bp. Exosomes are 30-100nm endosomal-derived vesicles containing mRNA, miRNA, lncRNA, transcription factors, proteins and lipids. They are secreted from various cells and their substance and amount alter between healthy and diseases state.

Objective: Identification of lncRNA effecting cancer progression and patient survival.

Methods: Custom made lncRNA microarray (composed off˜17,000 lncRNAs and˜22,000 mRNAs) followed by validation series of 180 specimens from three anatomic sites, including 77 effusions, 40 ovarian HGSC specimens, 21 solid metastases and exosomes from 42 effusion supernatants. 10 selected lncRNA were subjected to clinicopathologic parameters and survival association analysis.

Results: out of 10 lncRNA chosen for validation, 5 lncRNA- ESRG, Link-A, MEG3, GATS and PVT1 - were differently expressed between the anatomic sites and exosomes (all p<0.001). In univariate survival analysis, higher ESRG levels were significantly related to longer overall survival (OS) in the entire cohort (p=0.023), Higher Link-A levels in post-chemotherapy specimens was significantly associated with longer OS (p=0.015). MEG3 levels were significantly higher in pre-chemotherapy effusions tapped at diagnosis (n=40) compared to post-chemotherapy specimens (p=37; p=0.017). Higher GATS expression was seen in specimens from patients who were sub-optimally debulked (p=0.046). Moreover, H19 which did not change significantly between the anatomic sites, was higher in effusions from patients whose tumors showed primary resistance to chemotherapy (PFS≤6 months).

Conclusion: lncRNAs are differently express at different anatomic site and exosomes in HGSC and can provide new therapeutic targets. Furthermore, their level in effusion fluids can serve as a prognostic marker.

Citation Format: Natalie Filippov-Levy, Yoav Smith, Claes Gøran Trope, Ben Davidson, Reuven Reich. LncRNA expression at the different site of high-grades serous carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 517.
Abstract 5203: Bitter melon extract modulates immune signaling in prevention of tobacco associated carcinogen induced oral carcinogenesis
Authors: Sur Subhayan , Steele Robert , Ray Ratna B.
Published: August 2018

Oral squamous cell carcinoma (OSCC) is a complex malignancy and tobacco habit is one of the main etiological factors. Our previous study showed potential chemopreventive effect of bitter melon (Momordica charantia) extract (BME) in different pre-clinical cancer models like head and neck, breast, prostate. The aim of this study is to understand the molecular mechanism of BME in prevention of carcinogen induced in-vivo oral carcinogenesis. For this purpose, we induced oral cancer in immunocompetent mice by using tobacco associated carcinogen 4-nitroquinoline 1-oxide (4-NQO), and treated with BME through their drinking water. We observed that BME feeding significantly reduced the incidence of 4-NQO induced oral cancer. Histological analysis showed moderate dysplasia to invasive squamous cell carcinoma in 4NQO treated mice tongue, whereas no remarkable changes were seen in the BME-fed mice. RNA-seq data showed differential expression of several genes during carcinogenesis and during prevention of the cancer. We previously showed that BME treatment decreases the infiltrating regulatory T (Treg) cells by inhibiting FoxP3+ populations in the tumors and in spleens, suggesting an immunomodulatory role of BME in prevention. Here, we observed elevated expression of pro-inflammatory genes s100a9, IL23a, IL-1β and immune check point gene PDCD1/PD1 of the pathway, during oral cancer development. Interestingly, BME treatment significantly reduced the expression of these molecules. Subsequently, we observed the enhancement of MMP9 during carcinogenesis, which was reduced in BME-fed mouse tongue tissues. Taken together, our observation strongly demonstrated a chemopreventive effect of BME on carcinogen induced oral cancer.

Citation Format: Subhayan Sur, Robert Steele, Ratna B. Ray. Bitter melon extract modulates immune signaling in prevention of tobacco associated carcinogen induced oral carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5203.
Abstract 5206: The biological significance of CCL 16 regulated by thyroid hormone in liver cancer cells
Authors: Wu Cheng-Heng , Lin Kwang-Huei
Published: August 2018

Thyroid hormones(THs) are multi-function mediators that modulate various physiological processes. Thyroid hormone receptors(TRs) are type II nuclear receptors, TRs binds to thyroid hormone response elements (TREs) of target promoter regions modulates target gene expression at the transcriptional level upon THs binding. Previous literatures illustrate an association between aberrant TR regulation and human hepatocellular carcinoma (HCC). We employed cDNA microarrays and SILAC secretome analysis to identify the T3-regulated secreted proteins after T3 treatment in TR-expressing hepatoma cell lines, Chemokine (C-C motif) ligand 16 (CCL16) expression is positively regulated by T3 treatment. The patients with higher expression of CCL16 have poor recurrence survival rate. Ectopic expression of CCL16 in HCC cell lines increased tumor growth rate in vivo but not in vitro. CCL16 induced chemotaxis of monocytes and macrophages via interacting with its receptors CCR1 and CCR5. Expression of proangiogenic genes in macrophages were increased while co-cultured with HCC cell lines. The motility of HUVECs were increased after treated with conditional medium of macrophages co-cultured with HCC cell lines. Our study revealed that CCL16 recruited macrophage and promoted tumor progression.

Citation Format: Cheng-Heng Wu, Kwang-Huei Lin. The biological significance of CCL 16 regulated by thyroid hormone in liver cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5206.