bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5191: Targeting CX3CR1 Impairs the reseeding and colonization of circulating tumor cells and decelerates metastatic progression
Authors: Qian Chen , Worrede-Mahdi Asurayya , Kaur Ramanpreet , Shen Fei , Salvino Joseph , Meucci Olimpia , Fatatis Alessandro
Published: August 2018

This study aims to understand the mechanisms and dynamics by which cancer cells, departing from existing metastases as circulating cancer cells (CTCs), further seed and colonize skeleton and soft tissues, expanding metastatic spreading and precipitating the clinical progression to terminal disease. We have previously shown that the chemokine receptor CX3CR1 is implicated in the metastatic seeding of breast cancer cells and that FX-68, a novel small-molecule antagonist for this receptor, effectively contains both number of lesions and total tumor burden in animal models of metastatic disease. Here we established that FX-68 impairs the reseeding of skeleton and soft tissue and that the CTCs unable to reseed and forced to remain in the blood eventually succumb to apoptotic death. The metastatic potential of CTCs was confirmed by reinoculation experiments as well as by forcefully mobilizing cancer cells from metastases back in the blood and assessing the number of additional lesions generated, in the presence and absence of FX-68. Furthermore, we found that prolonging the permanence of CTCs in the blood by targeting CX3CR1 with FX-68 improved exposure of cancer cells to the chemotherapy drugs doxorubicin or docetaxel significantly decreased tumor progression and extended overall survival. Finally, we found evidence that cancer cells seeding the skeleton despite the inhibition of CX3CR1 failed to develop into tumors, suggesting a role of CX3CR1 expression in the metastasis-initiating properties of breast cancer cells. In summary, our results strongly support developing CX3CR1 antagonists and promoting their clinical use, as this approach will provide novel and effective tools to contain the progression of metastatic disease in patients.

Citation Format: Chen Qian, Asurayya Worrede-Mahdi, Ramanpreet Kaur, Fei Shen, Joseph Salvino, Olimpia Meucci, Alessandro Fatatis. Targeting CX3CR1 Impairs the reseeding and colonization of circulating tumor cells and decelerates metastatic progression [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 5191.
Abstract 5209: Overexpression of TGF-β in Smad4 deficient head and neck squamous cell carcinomas promotes myeloid cell infiltration associated with metastatic progression
Authors: Yu Jingjing , Hernandez Ariel , Raben David , Wang Xiao-Jing
Published: August 2018

Purpose of study: Transforming growth factor-β (TGF-β) has been shown to promote tumor progression when overexpressed and could be an important biomarker for head and neck squamous cell carcinomas (HNSCC) prognosis and treatment. Overexpression of TGF-β in tumor epithelia and stroma is observed in Smad4 deficient HNSCCs and is known to upregulate inflammation and angiogenesis. Our preliminary studies in Smad4 deficient HNSCC mice have revealed that inhibition of the TGF-β receptor (TGF-βR), specifically of the intracellular kinase domain, did not affect tumor growth in an immune compromised background but significantly decreased the number of metastatic lesions. We aim to elucidate the specific mechanisms by which TGF-β signaling promotes this metastatic phenotype in Smad4 deficient HNSCCs. We hypothesized that Smad4 deficient SCCs overexpress TGF-β to create an inflammatory microenvironment and promote angiogenesis to drive tumor progression in vivo.

Methods: Nude mice were injected subcutaneously in the flank with SCC cells derived from keratinocyte-specific Smad4 knockout mice and treated with either a TGF-βR inhibitor (n=5) or a vehicle control (n=6) once primary tumors reached 1mm3. Using immunofluorescence (IF) or immunohistochemistry (IHC), sections of the primary tumor were probed for leukocyte marker CD45, macrophage markers F4/80, iNOS and Arg1, myeloid cell markers CD11b and Ly6G and endothelial cell marker CD31.

Results: Smad4 deficient SCCs treated with the TGF-βR inhibitor showed a significant decrease in CD45+ leukocytes, with a specific decrease in CD11b+Ly6G+ cells compared to the vehicle control group. However, there was no difference in the total number of F4/80+ macrophages between TGF-βR inhibitor treated and vehicle control groups. There was also no significant difference in angiogenesis between these two groups as measured by the number of blood vessels/mm2 and the percentage area of the tumor covered by blood vessels.

Conclusions: Smad4 deficient SCC tumors treated with the TGF-βR inhibitor showed a significant decrease in myeloid cell populations, suggesting that overexpressed TGF-β plays a role in mediating inflammation in the tumor stromal environment. Treatment of SCC tumors with the TGF-βR inhibitor, however, did not show an affect on angiogenesis compared to the vehicle control.

Citation Format: Jingjing Yu, Ariel Hernandez, David Raben, Xiao-Jing Wang. Overexpression of TGF-β in Smad4 deficient head and neck squamous cell carcinomas promotes myeloid cell infiltration associated with metastatic progression [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 5209.
Abstract 5212: The inhibitory effect of iron control on breast cancer cells in tumor microenvironment
Authors: Goto Wataru , Kashiwagi Shinichiro , Takada Koji , Asano Yuka , Takashima Tsutomu , Noda Satoru , Onoda Naoyoshi , Hirakawa Kosei , Ohira Masaichi
Published: August 2018

Background: Iron is an essential element for both normal and cancer cells, and it is well known that iron depletion leads to G1-S arrest and apoptosis. Iron depletion is also known to reduce serum hemoglobin and oxygen supply to the tissue, which may induce angiogenesis. On the other hand, it is reported that hypoxia mediates epithelial-mesenchymal transition (EMT) and cancer cell metastasis with angiogenesis. In addition, it is thought that antitumor autoimmune response relates to suppression of EMT, and we investigated previously it with clinical samples. In this study, we examined the mechanism of iron control therapy in breast cancer based on hypoxia, EMT and immune microenvironment.

Material and Methods: Human breast cancer cell lines MDA-MB-231 (triple-negative breast cancer), MCF-7 (luminal type) and OCUB-1 (HER2-enriched type), and iron chelator Deferoxamine (DFO) were used in the present study. The effect of proliferation and migration of breast cancer cells by DFO was determined using MTT assay and wound healing assay. We also examined the influence of cell cycle by DFO using flow cytometry. Furthermore, we investigated the relationship between iron depletion of breast cancer cells and hypoxia, EMT and immune microenvironment using RT-PCR. We produced subcutaneous xenografts of mice, and examined the change in tumor sizes by administration of DFO.

Results: The iron-depleted condition suppressed breast cancer cell proliferation and migration. In cell cycle analysis, DFO increased the population of the G0-G1 phase. In addition, iron depletion induced hypoxia and angiogenesis. Also, N-cadherin and PD-L1 expression of breast cancer cells were increased by iron-depleted condition. Subcutaneous tumors in the DFO administration group were suppressed tumor growth in vivo.

Conclusion: Iron control by DFO showed the effect of tumor suppression, and suggested the possibility of novel treatment for patients with breast cancer. In addition, this mechanism related to hypoxia, EMT and immune microenvironment.

Citation Format: Wataru Goto, Shinichiro Kashiwagi, Koji Takada, Yuka Asano, Tsutomu Takashima, Satoru Noda, Naoyoshi Onoda, Kosei Hirakawa, Masaichi Ohira. The inhibitory effect of iron control on breast cancer cells in tumor 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 5212.
Abstract 5215: Vitamin D receptor regulates intestinal barriers in colon cancer
Authors: Zhang Yong-guo , Lu Rong , Bakke Danika , Xia Yinglin , Sun Jun
Published: August 2018

Background Vitamin D and Vitamin D receptor (VDR) play important roles in the development of colon cancer. Tight junction structures are essential in intestinal barrier integrity, inflammation, and cancer. The disruption of tight junctions is a common manifestation of various diseases including cancers. Claudin-5 is a tight junction protein that is expressed in epithelia and endothelia and form paracellular barriers and pores that determine permeability. It is downregulated in colon cancer. Although VDR and Claudin-5 are all involved in colon cancer, it remains unclear whether they are closely related or function independently. Methods In the current study, we investigate the novel role of VDR in regulating Claudin-5, using whole body VDR knockout mice (VDR-/-) and intestinal epithelial VDR knockout mice (VDRΔIEC). In vitro, VDR-/- embryonic fibroblasts (MEF) cells, cultured human intestinal epithelial cells, organoids, and human colon cancer samples were used to determine the molecular mechanism. Results In intestinal samples of colon cancer patients, VDR expression is low, whereas the expression of Claudin-5 is also decreased. In the colon of VDR-/- mice and VDRΔIEC mice, deletion of VDR led to lower claudin-5 at the protein and mRNA levels. This observation was also confirmed by immunostaining of Claudin-5. In VDR-/- MEF cells, both Western blot and real time PCR revealed that VDR deletion led to lower protein and mRNA levels of claudin-5 in cells without any treatment. Vitamin D3 is known to increase VDR expression and activate VDR signaling. Protein and mRNA levels of Claudin-5 were significantly elevated in mice, mouse organoids, and human intestinal epithelial SKCO15 cells treated with 1, 25 vitamin D3. In contrast, knockdown of Claudin-5 using siRNA did not change the expression level of VDR protein in human colonic epithelial cells. Our data suggest that the Claudin-5 gene is a downstream target of the VDR signaling pathway. Conclusion Taken together, we show that VDR is important for the maintenance of physiological level of Claudin-5. This study also reveals a complex role for vitamin D/VDR regulation of intestinal barrier functions in colon cancer.

Citation Format: Yong-guo Zhang, Rong Lu, Danika Bakke, Yinglin Xia, Jun Sun. Vitamin D receptor regulates intestinal barriers in colon 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 5215.
Abstract 5219: Effect of extracellular vesicles from gastric cancer cells on the morphologic and genetic changes of peritoneal mesothelial cells
Authors: Okuno Tomohisa , Yashiro Masakazu , Togano Shingo , Kuroda Kenji , Miki Yuichiro , Kasashima Hiroaki , Hirakawa Kosei , Ohira Masaichi
Published: August 2018

Background: Gastric cancer is a major cause of cancer death, particularly on the basis of peritoneal metastasis. Human scirrhous gastric carcinoma (SGC), diffusely infiltrating carcinoma, or Borrman type 4 also known as linitis plastica-type carcinoma is characterized by rapid cancer cell infiltration and proliferation accompanied by extensive stromal fibrosis. SGC account for about 10% of all gastric carcinomas, and carry an extremely worse prognosis than other types of gastric carcinoma, reflecting their frequent peritoneal metastasis. We previously reported that SGC cells (“seed”) might change the peritoneum to favorable environment, so-called pre-metastatic niche (“soil”), (Cancer 77:1668-75, 1996). Recently, it has been clarified that extracellular vesicles (EVs) play an important role for cell-cell interactions. In this study, we investigated the effect of EVs from SGC cells on the pre-metastatic niche formation of the peritoneum.

Methods: A SGC cell line, OCUM-2MD3, OCUM-12, KATO-III, gastric cancer cell line, MKN74, MKN45, and peritoneal mesothelial cells (PM cells) were used. PKH26-labeled EVs from OCUM-2MD3 cells were intravenously injected into nude mice, and the distribution of EVs was examined by a fluorescence microscope. The morphology of PM cells was investigated in the addiction of EVs from various gastric cancer cell lines using a phase-contrast microscope. And gene expression was examined by RT-PCR in the presence or absence of EVs from OCUM-2MD3. Clinical significance of EVs markers CD9 and CD63 was immunohistochemically evaluated using 63 human gastric cancer specimens.

Results: PKH26-labeled EVs were frequently found in the peritoneum, stomach and liver, but rare in lung and brain of mice. Peritoneal mesothelial cells uptake EVs of gastric cancer cells and change morphology from cobblestone shape to spindle shape in vitro. mRNA expression level of MMPs and integrins in PM cells was increased following the addiction of EVs, in compared with the control. The high expression of CD9 and CD63 was significantly correlated with distant metastasis.

Conclusion: EVs from gastric cancer cells might induce a pre-metastatic niche at peritoneum by the morphologic and genetic changes of mesothelial cells.

Citation Format: Tomohisa Okuno, Masakazu Yashiro, Shingo Togano, Kenji Kuroda, Yuichiro Miki, Hiroaki Kasashima, Kosei Hirakawa, Masaichi Ohira. Effect of extracellular vesicles from gastric cancer cells on the morphologic and genetic changes of peritoneal mesothelial 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 5219.
Abstract 5234: Transcriptome profiling of single prostate cancer cells following androgen deprivation
Authors: Clark Ashlee K. , Lawrence Mitchell , Nim Hieu , Lister Natalie , Frydengerg Mark , Risbridger Gail , Taylor Renea
Published: August 2018

Androgen withdrawal is the standard of care for men with metastatic prostate cancer. While all patients initially respond, resistance is inevitable and lethal castration-resistant prostate cancer ensues. Using patient-derived xenografts (PDX) of localized tumors, we have identified a subpopulation of castration-tolerant prostate cancer cells that survive following castration. Identifying the unique biologic characteristics of these cells is essential in determining their role in tumor progression and potential to be targeted by therapeutic agents. To study the genomic features of castrate-tolerant cells, we enriched for prostate cancer cells from PDXs using FACS and subjected them to single-cell isolation and RNA seq. We efficiently captured and sequenced >50 cells from pre- and post-castration PDXs using the Fluidigm C1 platform. Sequencing of isolated single cells was performed using the Illumina HiSeq in rapid mode with 50 bp fragment sequencing chemistry (3 million reads/cell). Multidimensional scaling showed that the response to castration is not uniform in all human cells. A unique gene set was identified in intact versus castrate-tolerant cells; we identified distinct changes in energy metabolism, including suppression of ATP production to aid cell survival. We also detected consistent upregulation of the retinoic acid signaling pathway, involving upregulation of CRABP2 and RARRES3 expression in castrate-tolerant cells. This is the first study to report on gene expression in single human prostate cells and revealed novel endocrine-related changes prior to and following androgen deprivation. Our data suggest that further and/or alternative hormone suppression may be effective in targeting castration-tolerant prostate cancer cells.

Citation Format: Ashlee K. Clark, Mitchell Lawrence, Hieu Nim, Natalie Lister, Mark Frydengerg, Gail Risbridger, Renea Taylor. Transcriptome profiling of single prostate cancer cells following androgen deprivation [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 5234.
Abstract 5242: NFĸB pathway activation is a key determinant of tamoxifen tolerance and recurrence in breast cancer
Authors: Kastrati Irida , Brovkovych Svitlana D. , Stender Joshua D. , Alarid Elaine T. , Frasor Jonna
Published: August 2018

Nearly 75% of breast tumors express estrogen receptor (ER) and will be treated with endocrine therapies, such as tamoxifen or aromatase inhibitors. Despite their proven success, it is estimated that up to 50% of tumors fail on endocrine therapy and recur as aggressive therapy-resistant tumors. Therefore, preventing recurrence remains a major clinical problem. One hypothesis is that recurrences are driven by cells that survive therapeutic intervention. This poorly understood population is referred to as minimal residual disease that consists of drug-tolerant cells capable of surviving by entering a state of negligible growth. In trying to model tamoxifen tolerance and minimal residual disease in vitro, we have utilized the clonogenic assay where ER+ breast cancer cells' clonal growth and survival in the presence of tamoxifen is examined. Tamoxifen-tolerant cells show activation of the pro-inflammatory NFĸB pathway measured by an NFĸB-reporter, elevated expression of bona-fide target genes, and enhanced nuclear localization of p65 and p50, two major transcription factors of the NFĸB pathway. Tamoxifen does not directly activate the NFĸB pathway, but rather cells with high NFĸB activity proliferate independently of tamoxifen despite retaining ER expression. Mechanistically, NFĸB-dependent ER phosphorylation at S305 residue may contribute to failure to respond to tamoxifen. More importantly blocking NFĸB with multiple NFĸB inhibitors is sufficient to eradicate this population of tamoxifen-tolerant cells, suggesting NFĸB is required and can be exploited for therapy. We find that dimethyl fumarate, an FDA-approved anti-inflammatory and immune-modulatory drug, which we have established as an effective NFĸB inhibitor in breast cancer cells, prevents recurrence modeled both in vitro and in vivo. Altogether this work shows a novel role for the NFĸB pathway in promoting a population of cells that are tamoxifen tolerant and capable of seeding recurrent breast cancer disease. Furthermore, therapeutic targeting by an NFĸB inhibitor such as dimethyl fumarate can be used to eradicate residual disease and prevent breast cancer recurrence.

Citation Format: Irida Kastrati, Svitlana D. Brovkovych, Joshua D. Stender, Elaine T. Alarid, Jonna Frasor. NFĸB pathway activation is a key determinant of tamoxifen tolerance and recurrence in breast 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 5242.
Abstract 5261: Indices of diet quality and risk of lung cancer incidence and mortality in the Women's Health Initiative Observational Study
Authors: Myneni Ajay A. , Giovino Gary A. , Millen Amy E. , LaMonte Michael J. , Wactawski-Wende Jean , Neuhouser Marian L. , Zhao Jiwei , Shikany James M. , Mu Lina
Published: August 2018

Background: Prospective evidence on the association between diet patterns and lung cancer risk is limited, particularly in older women. We investigated whether commonly used diet quality indices - Healthy Eating Index 2010 (HEI), Alternate Healthy Eating Index 2010 (AHEI), alternate Mediterranean Diet (aMED) and Dietary Approaches to Stop Hypertension (DASH) - were associated with lung cancer incidence and mortality in the Women's Health Initiative Observational Study (WHI OS).

Methods: Self-reported diet intake (food frequency questionnaires) and relevant covariate information was obtained from 86,090 race-ethnically diverse postmenopausal women aged 50-79 years at baseline (1993-1998). During a median follow-up of approximately 17 years, 1,491 lung cancer cases and 1,393 lung cancer deaths were documented. We used cox proportional hazards regression models to estimate hazards risks and 95% confidence intervals [HR (95% CI)] after controlling for age, race/ethnicity, education, body mass index, smoking, physical activity and energy intake.

Results and conclusions: Diet quality indices were not associated with lung cancer incidence overall but all four dietary indices showed a protective association (highest versus lowest quintile) against squamous cell carcinoma cases (12.8% of total lung cancer cases) [HEI: 0.56 (0.33-0.96), AHEI: 0.42 (0.24-0.76), aMED: 0.65 (0.39-1.08), DASH: 0.56 (0.32-0.97)]. Diet quality indices were not associated with lung cancer mortality overall or when stratified by histological type, except for HEI with lung cancer mortality overall [fourth versus lowest quintile: 0.82 (0.69-0.99)]. Smoking status did not modify the association between diet quality indices and lung cancer incidence or mortality. Further studies in other populations may help elucidate the relationship between diet patterns and lung cancer, especially by histological type.

Citation Format: Ajay A. Myneni, Gary A. Giovino, Amy E. Millen, Michael J. LaMonte, Jean Wactawski-Wende, Marian L. Neuhouser, Jiwei Zhao, James M. Shikany, Lina Mu. Indices of diet quality and risk of lung cancer incidence and mortality in the Women's Health Initiative Observational Study [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 5261.
Abstract 5279: Prevalence and correlates of perceived harmfulness and addictiveness to traditional and alternative tobacco products among US adults
Authors: Mohammed Kahee A. , Schoen Martin W. , Osazuwa-Peters Nosayaba , Al-Taee Ahmad M. , Khneizer Gebran W. , Arnold Lauren D. , Hinyard Leslie , Burroughs Thomas E.
Published: August 2018

Purpose: Beliefs about addictiveness and harmfulness of tobacco products are associated with intentions to use them. This study aimed to (1) evaluate perceived harmfulness and addictiveness patterns towards multiple tobacco products and (2) examine sociodemographic variables that influence those perceptions.

Methods: Weighted multivariate ordinal logistic regression analyses were performed on 5,474 US adults aged ≥18 years who participated in the 2015 and 2017 cycles of the Health Information National Trends Survey-FDA. The two primary outcomes were perceived harmfulness and addictiveness of traditional (cigarette, cigar, and pipe filled with tobacco) vs alternative tobacco (e-cigarette, hookah, and “roll your own” cigarettes). All models were adjusted for age, gender, race/ethnicity, education, household income, census region, and smoking status.

Results: Overall, 7.9% of adults in the U.S. perceived alternative tobacco as “not harmful at all” compared to 2.4% for the traditional products. Similarly, 16.2% of adults perceived alternative tobacco as “not addictive at all” compared to 13.6% for the traditional products. Beliefs about the harmfulness and addictiveness of traditional and alternative tobacco products differed by sociodemographic characteristics and smoking status: Those who believed traditional and alternative tobacco products are more harmful were more likely to be female, older (compared with those aged 18-34), Black (compared with White), former or never smokers (compared with current smokers), and adults living in West (compared with Northeast). Similarly, those who believed traditional and alternative tobacco products are more addictive were more likely to be female, older, Black, and adults living in West. However, no association between smoking status and perceived addictiveness was noted.

Conclusion: Adults in U.S. perceive tobacco harmfulness and addictiveness differently based on their sociodemographic characteristics and smoking status. These findings are potential areas for targeted behavioral interventions to increase individuals' perceived risk of harm and addiction to all forms of tobacco products in the United States.

Citation Format: Kahee A. Mohammed, Martin W. Schoen, Nosayaba Osazuwa-Peters, Ahmad M. Al-Taee, Gebran W. Khneizer, Lauren D. Arnold, Leslie Hinyard, Thomas E. Burroughs. Prevalence and correlates of perceived harmfulness and addictiveness to traditional and alternative tobacco products among US adults [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 5279.
Abstract 5285: Evaluation of a postpartum human papillomavirus vaccination program
Authors: Berenson Abbey B. , Hirth Jacqueline M. , Kuo Yong-Fang , Rupp Richard E.
Published: August 2018

Evaluation of a postpartum human papillomavirus vaccination program

Berenson AB, Hirth JM, Kuo YF, Rupp RE

Purpose: To evaluate the effectiveness of a large vaccination program which offers the human papillomavirus (HPV) vaccine to women during their postpartum hospitalization.

Methods: Based on the success of a pilot which offered the HPV vaccine to a select group of women, a postpartum HPV vaccination program was expanded in 2016 to include all eligible mothers who gave birth in a large Texas hospital serving low income women. Women from 23 different counties were included. With funding from the Cancer Prevention Research Institute of Texas (CPRIT), patient navigators were hired to identify and personally counsel all eligible women who met catch-up recommendation guidelines prior to their hospital discharge. Women were informed about the need and availability of the HPV vaccine during their hospitalization and offered it free of charge. Follow-up doses are coordinated with postpartum visits as well as well-baby visits in collaboration with the Department of Pediatrics. With Institutional Review Board approval, records of women who were approached during the first 11 months of the expanded program were reviewed to determine the number who chose to participate.

Results: Baseline HPV vaccination rates were low. A total of 1,782 postpartum women were counseled about the HPV vaccine of which 168 (9.4%) were already vaccinated and 5 others were ineligible. Of the remaining 1,609 women, 71.2% received a dose of the HPV vaccine during their hospitalization. The refusal rate among eligible women was 24%. Another 4.3% (n-76) agreed to get vaccinated, but were released from the hospital before the injection was given. Of these, 48 (63.2%) received the dose at a later clinic visit.

Conclusions: Offering the HPV vaccine as part of postpartum care can increase catch-up vaccination of young women who did not have the opportunity to be vaccinated at a younger age.

Citation Format: Abbey B. Berenson, Jacqueline M. Hirth, Yong-Fang Kuo, Richard E. Rupp. Evaluation of a postpartum human papillomavirus vaccination program [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 5285.
Abstract 5292: Visceral adiposity-stimulated genotoxicity and malignant transformation of epithelial cells
Authors: Chakraborty Debrup , Jdanov Vladislav , Benham Vanessa , Bullard Blair , Bernard Jamie J.
Published: August 2018

Obesity is a global epidemic with a predicted rate of 42% in the U.S.A by 2050. Epidemiologic studies show that obesity is a risk factor for developing cancer; however, the molecular mechanism has not been fully elucidated. Our published data demonstrate that fibroblast growth factor-2 (FGF2) released from fat cells (adipocytes) in the visceral adipose tissue (VAT) induces transformation/tumorigenicity in the skin and mammary epithelial cells. Specifically, FGF2 released from VAT stimulates epithelial cell growth in soft agar by inducing the proto-oncogene c-Myc. Growth in soft agar is a measure of transformation/tumorigenicity; neither transformation nor c-Myc induction in epithelial cells was reversible. c-Myc overexpression can initiate a process of genetic instability linked to tumor initiation. Our discovery of this novel direct path of VAT-stimulated tumorigenesis adds mechanistic insight to our earlier discovery that VAT secretions promote UVR-induced nonmelamoma skin cancer. The objective of our current study was to determine the mechanism by which FGF2 stimulates malignant transformation. We hypothesized that FGF2 from VAT induces c-Myc and subsequent genomic instability in epithelial cells, leading to increased carcinogenesis. To test hypothesis we generated a filtered conditioned-medium from the human VAT treated MCF-10A (mammary epithelial) and JB6 P+ (skin epithelial) cells and measured several downstream mediators of FGF2 and activation of FGFR-1 (FGF2 receptor). Following VAT treatment, epithelial cells demonstrated induced c-Myc protein expression along with ROS accumulation, elevated γ-H2AX foci, and increased micronucleus (MN) formation. We found that inhibition of c-Myc attenuated VAT-induced neoplastic transformation of MCF-10A and JB6 P+ cells, while constitutive activation of the c-Myc-induced spontaneous neoplastic transformation of JB6 P+ cells. Collectively, our data suggested FGF2 released from VAT interacts with FGFR-1 and activates c-Myc. The role of c-Myc in the formation of MN and DNA damage is under investigation. Determining the impact of excess VAT on cancer will lead to strategies to help prevent adiposity-associated cancers and identify individuals at risk for disease or individuals who may be susceptible to compounded genotoxicity due to DNA-damaging environmental exposures.

Citation Format: Debrup Chakraborty, Vladislav Jdanov, Vanessa Benham, Blair Bullard, Jamie J. Bernard. Visceral adiposity-stimulated genotoxicity and malignant transformation of epithelial 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 5292.
Abstract 5307: Inferring parsimonious migration histories for metastatic cancers
Authors: El-Kebir Mohammed , Satas Gryte , Raphael Benjamin J.
Published: August 2018

Recent studies have attempted to infer the pattern of cellular migrations between a primary tumor and distant metastases using phylogenetic trees constructed from somatic mutations. Several of these studies have challenged the conventional view of monoclonal seeding of a metastasis, and reported complex patterns of migration between primary tumors and metastases, including polyclonal seeding and reseeding.

Most current analyses that attempt to infer migration patterns from somatic mutations rely on two key assumptions, sample homogeneity and mutation-migration concordance, that do not typically hold for cancer sequencing datasets. Consequently, standard phylogenetic analysis of mutations may result in incorrect or misleading patterns of metastasis.

We introduce a computational model to evaluate migration patterns in a rigorous manner. We use this model as a basis for Metastatic And Clonal History INtegrative Analysis (MACHINA), an algorithm that jointly infers parsimonious clone trees and migration histories of metastatic cancers from DNA sequencing data. We show that MACHINA accurately recovers clone trees and migration histories on simulated data. We apply MACHINA to sequencing data from metastatic ovarian, breast, prostate and skin cancer samples. In several cases, we find simpler migration histories than previously reported, and these alter conclusions regarding metastasis-to-metastasis spread, the anatomical site of the primary tumor, or the occurrence of polyclonal seeding and reseeding in individual patients. MACHINA enables researchers to rigorously assess the validity of different migration patterns in individual patients with metastatic disease and evaluate the prevalence of different migration patterns across large cohorts of patients and tumor types.

Citation Format: Mohammed El-Kebir, Gryte Satas, Benjamin J. Raphael. Inferring parsimonious migration histories for metastatic cancers [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 5307.
Abstract 5338: Comprehensive pan-cancer genomic analysis reveals link between 3'UTR DNA methylation and increased gene expression
Authors: McGuire Michael H.
Published: August 2018

Investigations into the function of non-promoter DNA methylation have yielded new insights into epigenetic regulation of gene expression. Previous studies have highlighted the importance of distinguishing between DNA methylation in discrete functional regions (e.g. promoter, gene body, enhancer); however, the relationship between DNA methylation outside of the promoter region and gene expression in cancer requires further exploration. Through integrated analysis of gene expression and DNA methylation profiles (>3,000 samples across multiple cancer types and corresponding normal tissues), we uncovered an enrichment of DNA methylation sites within the 3'UTR in which DNA methylation is strongly positively correlated with gene expression. In 5 of 10 tumor types examined, DNA methylation of the 3'UTR is associated with patient survival in a significant number of genes. Filtering for genes in which 3'UTR DNA methylation, relative to gene body DNA methylation, is more strongly correlated with gene expression yields a list of genes enriched for functions involving T cell activation. The immune checkpoint gene HAVCR2 (TIM-3) shows a substantial increase in 3'UTR DNA methylation, but not adjacent exonic/intronic, or promoter DNA methylation, upon T cell stimulation and upregulation of gene expression. Furthermore, this increase in HAVCR2 gene expression can be abrogated by treatment with demethylating agents. These findings implicate the 3'UTR as a functionally relevant DNA methylation site, particularly in regard to T cell activity. Additionally, they reveal a novel mechanism by which HAVCR2 is upregulated in T cells, providing a new means of targeting immune checkpoint blockade. Finally, these data offer a selective marker for choosing patients that could respond to the combination therapy of PD-1/PD-L1 inhibition and demethylating agents.

Citation Format: Michael H. McGuire. Comprehensive pan-cancer genomic analysis reveals link between 3'UTR DNA methylation and increased gene expression [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 5338.
Abstract 5350: Single cell RNA-seq reveals transcriptionally distinct subpopulations within patient-derived pancreatic cancer organoids
Authors: Ray Mohana , Nandi Rajib K. , Calvo Isabel Romero , Kirby Kori , Weber Christopher , Kindler Hedy L. , Buschmann Mary , Roggin Kevin , White Kevin P.
Published: August 2018

Advances in 3D cell culture over the last decade have enabled the establishment of organoid models from a multitude of organs in varying stages of development and disease. Organoids recapitulate the tissue-specific cell types and exhibit similar morphology and spatial organization as the organ it is derived from. However, to the best of our knowledge, tumor organoids have never been profiled at a single cell level.

In this study, we applied Drop-seq, a high throughput single cell RNA-seq platform, to characterize the different cell types present in pancreatic cancer organoids. We sequenced the transcriptomes of 5494 single cells from organoids derived from a patient undergoing a Whipple procedure for pancreatic ductal adenocarcinoma (PDAC). We then developed a novel clustering approach based on a class of probabilistic generative models called topic models, leading to the identification of two subpopulations of cells. One subgroup expressed marker genes known to play important roles in PDAC biology, such as MUC1, CLDN4 and GAS5. Interestingly, this subgroup also bore markers for cancer stem cells (HES1 and SQSTM1) that have been found by multiple studies to mediate chemoresistance in PDAC. The other subpopulation differentially expressed AGR2 and TMBIM6, which are associated with endoplasmic reticulum (ER) stress and correlates with invasiveness of PDAC cell lines. Functional studies are underway to determine the exact role of these subgroups, but our preliminary findings are suggestive of a subpopulation of cells that maybe involved in resistance to chemotherapy. This is consistent with the clinical progression of this patient's cancer, suggesting that rapid, inexpensive single cell transcriptomics might provide more precise molecular information to personalized medicine programs than traditional RNA-seq alone.

Citation Format: Mohana Ray, Rajib K. Nandi, Isabel Romero Calvo, Kori Kirby, Christopher Weber, Hedy L. Kindler, Mary Buschmann, Kevin Roggin, Kevin P. White. Single cell RNA-seq reveals transcriptionally distinct subpopulations within patient-derived pancreatic cancer organoids [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 5350.
Abstract 5357: Mutational and transcriptomic profiling identify distinct angiogenic and inflammatory subtypes of angiosarcoma
Authors: Kim Jong Hyuk , Megquier Kate , Sarver Aaron L. , Thomas Rachael , Wang Chao , Elvers Ingegerd , Karlsson Elinor , Breen Matthew , Lindblad-Toh Kerstin , Modiano Jaime F.
Published: August 2018

Angiosarcoma is an aggressive, albeit rare cancer in humans. The cause of the vast majority of sporadic angiosarcomas is unknown, mortality is high, and no therapeutic targets have been identified to improve outcomes. Hemangiosarcoma (HSA) is a common cancer of dogs, and it shares histopathologic features with human angiosarcoma. In our previous work, canine HSAs were classified into angiogenic, inflammatory, and adipogenic subtypes based on transcriptional profiles. However, the genetic and molecular events that regulate transcriptional subtypes in angiosarcoma are not currently understood. Our goal was to use a comparative genomics approach to apply knowledge from appropriately powered canine studies to inform our research into human sarcomas. In this study, we identified recurrent mutations in RNASeq data from 93 HSAs and 16 nonmalignant controls, based on mutations first identified in exomes from 42 paired tumor and normal samples. In addition to identifying recurrent somatic mutations we also identified translocation fusions, allowing elucidation of oncogenic mechanisms for vascular endothelial growth factor receptors (VEGFR), phosphoinositide-3 kinase (PIK3) signaling pathways, and the p53 DNA damage repair pathway in canine HSA. Significantly, mutational signatures were associated with distinct molecular subtypes of canine hemangiosarcomas, and both the angiogenic and the inflammatory subtypes were apparent in RNASeq data from human angiosarcomas (n=14), suggesting that comparable etiologic mechanisms are operative in the canine and human disease. Our ongoing work seeks to understand how the molecular mechanisms give rise to molecular subtypes of angiosarcoma by defining the association between driver mutations, signaling pathway alterations and transcriptional patterns, which should allow us to identify rational therapeutic targets.

Citation Format: Jong Hyuk Kim, Kate Megquier, Aaron L. Sarver, Rachael Thomas, Chao Wang, Ingegerd Elvers, Elinor Karlsson, Matthew Breen, Kerstin Lindblad-Toh, Jaime F. Modiano. Mutational and transcriptomic profiling identify distinct angiogenic and inflammatory subtypes of angiosarcoma [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 5357.
Abstract 5364: Somatic mutations in African American and non-Hispanic White young-onset colorectal cancers
Authors: Rozek Laura S. , Qin Tingting , Koeppe Erika , Ulintz Peter , Greenson Joel K. , Cote Michele L. , Stoffel Elean M.
Published: August 2018

The proportion of colorectal cancers (CRCs) diagnosed at age <50 years is two-fold higher among African-Americans (AAs) compared to non-Hispanic Whites (NHW). Additionally, despite the rising incidence of young-onset CRC, little is known about the molecular characteristics of these as only 23 young-onset tumors were profiled in the TCGA cohort. Studies of CRC diagnosed in older individuals (age >50) demonstrate that CRC tumors in AAs are more often right-sided and MMR proficient, with higher prevalence of KRAS mutations. Risk of death from CRC among AAs with MMR proficient tumors is 73% higher than for NHW, after adjusting for treatment and known prognostic factors. We analyzed archival FFPE CRC tumors diagnosed in individuals age <50 (NHW enrolled in the UM Cancer Genetics Registry N=38 and African American subjects ascertained through the Wayne State University EpiCore N=13) for somatic mutations. Mutation profiling of tumor DNA from 51 CRCs diagnosed age <50 was performed using NGS sequencing with multigene panels in 2 sets (Qiagen GeneRead Cancer panel of 124 genes [N=36 tumors], Qiagen Qiaseq Comprehensive Cancer Panel of 275 genes [15 tumors]). We observed differences in CRC somatic mutations by race and by age. There was marked heterogeneity among tumors with regard to numbers of somatic mutations, with an average of 659 (range 39-10,267) variants detected per tumor, with 19/51 (37%) tumors classified as hypermutated. Among the most frequently mutated genes in young CRC tumors were APC (98% of tumors), PIK3CA (92% of tumors), NOTCH2 (86% of tumors), PTCH1 (82%), BRCA2 (67%), BRCA1 (84%), HNF1A (100%), RAD50 (29%), POLE (27%), MSH6 (39%), and FGFR3 (39%). These mutation profiles differ markedly from those of TCGA older-onset, nonhypermutated tumors. We identified differences in somatic mutations by race, including mutations in PTEN (NHW: 21% vs. AA: 8%), TGFBR2 (NHW: 32% vs. AA: 15%), FBXW7 (NHW: 47% vs. AA: 8%), and KRAS (NHW: 45% vs. AA: 31%). The relatively high prevalence of hypermutated tumors in this racially diverse cohort highlights the critical gaps in knowledge about pathways involved in pathogenesis of young-onset CRCs and how genomic information could guide precision therapies.

Citation Format: Laura S. Rozek, Tingting Qin, Erika Koeppe, Peter Ulintz, Joel K. Greenson, Michele L. Cote, Elean M. Stoffel. Somatic mutations in African American and non-Hispanic White young-onset colorectal cancers [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 5364.
Abstract 5365: Prospective patient stratification into robust cancer-cell intrinsic subtypes from colorectal cancer biopsies
Authors: Alderdice Matthew , Richman Susan D. , Gollins Simon , Stewart Peter , Hurt Chris , Adams Richard , McCorry Amy M. , Roddy Aideen , Vimalachandran Dale , Isella Claudio , Medico Enzo , Maughan Tim , McArt Darrgh G. , Lawler Mark , Dunne Philip D.
Published: August 2018

Colorectal cancer (CRC) biopsies underpin accurate diagnosis, but are also relevant for patient stratification in molecularly-guided clinical trials. The consensus molecular subtypes (CMS) and colorectal cancer intrinsic subtypes (CRIS) transcriptional signatures have potential clinical utility for improving prognostic/predictive patient assignment. However, their ability to provide robust classification, in pre-treatment biopsies from multiple regions or at different time points remains untested.

In this study, we undertook a comprehensive assessment of the robustness of CRC transcriptional signatures, including CRIS and CMS, using a range of tumour sampling methodologies currently employed in clinical and translational research. These include analyses using (i) laser-capture microdissected CRC tissue, (ii) eight publically available rectal cancer biopsy data sets (n=543), (iii) serial biopsies (from AXEBeam trial, NCT00828672; n=10), (iv) multi-regional biopsies from colon tumours (n=29 biopsies, n=7 tumours) and (v) pre-treatment biopsies from the phase II rectal cancer trial COPERNCIUS (NCT01263171; n=44). Compared to previous results obtained using CRC resection material, we now demonstrate that CMS classification in biopsy tissue is significantly less capable of reliably classifying patient subtype (43% unknown in biopsy versus 13% unknown in resections, p=0.0001). In contrast, there was no significant difference in classification rate between biopsies and resections when using the CRIS classifier. Additionally, we demonstrated that CRIS provides significantly better spatially- and temporally- robust classification of molecular subtypes in CRC primary tumour tissue compared to CMS (p= 0.003 and p=0.02, respectively).

These findings may have potential to inform ongoing biopsy-based patient stratification in CRC, enabling robust and stable assignment of patients into clinically-informative arms of prospective multi-arm, multi-stage clinical trials.

Citation Format: Matthew Alderdice, Susan D. Richman, Simon Gollins, Peter Stewart, Chris Hurt, Richard Adams, Amy M. McCorry, Aideen Roddy, Dale Vimalachandran, Claudio Isella, Enzo Medico, Tim Maughan, Darrgh G. McArt, Mark Lawler, Philip D. Dunne. Prospective patient stratification into robust cancer-cell intrinsic subtypes from colorectal cancer biopsies [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 5365.