bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3806: Cigarette smoke extract permits survival of TET2-/- and JAK2V617F hematopoietic cells
Authors: Ramanathan Gajalakshmi , Craver Brianna , Arechavala Rebecca J. , Herman David , Kleinman Michael T. , Fleischman Angela G. 
Published: September 2020

Abstract
Inactivating somatic mutations in the ten-eleven translocation 2 (TET2) gene is commonly found in clonal hematopoiesis (CH) and contributes to the pathogenesis of myeloid malignancies such as myeloproliferative neoplasms (MPN), myelodysplastic syndrome and acute myeloid leukemia. Incidentally, smoking behavior is associated with CH and we hypothesize that TET2 mutant hematopoietic cells are resistant to the deleterious effects of smoke exposure.
Cigarette smoke extract (CSE) was prepared from mainstream cigarette smoke using a standard protocol and volume/volume percentage (v/v %) dilutions used for in vitro exposures. Lineage negative bone marrow cells isolated from TET2 wildtype (TET2+/+) and TET2 knockout (TET-/-) mice were exposed to CSE and cultured in methylcellulose containing semi-solid medium to assess myeloid colony formation. Cells were also grown in liquid culture under long-term culture conditions to demonstrate the effect of CSE on cell proliferation. Peripheral blood mononuclear cells from JAK2V617F positive MPN patients were treated with CSE for colony forming assays.
TET2+/+ bone marrow cells showed decreased myeloid colony formation in the presence of CSE while TET2-/- cells were not significantly affected by CSE. Similarly in liquid cultures, TET2-/- lineage negative cells expanded significantly compared to their wildtype counterparts indicating resistance to the growth suppressive effects of CSE. The JAK2V617F mutation is the most common somatic mutation found in MPN patients and we observed that the suppressive effect of CSE on myeloid colony formation was more pronounced in normal subjects than JAK2V617F positive MPN subjects.
This suggests that CH associated somatic mutations such as in TET2 promote resistance of bone marrow progenitors to environmental stressors like cigarette smoke, possibly via anti-apoptotic mechanisms. Future studies include studying the effects of inhalation exposure of combustible and electronic cigarette smoke on TET2-/- and JAK2V617F hematopoiesis in competitive bone marrow transplanted mice.
Citation Format: Gajalakshmi Ramanathan, Brianna Craver, Rebecca J. Arechavala, David Herman, Michael T. Kleinman, Angela G. Fleischman. Cigarette smoke extract permits survival of TET2-/- and JAK2V617F hematopoietic cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3806.
Abstract 3807: Exposure to environmental chemicals leads to extensive branching and expression of cancer-promoting proteins in human mammary gland organoids
Authors: Hinrichs Johanna H. , Winkler Juliane , Liu Pengyuan , Ataii Nassim , Fisher Susan , Werb Zena 
Published: September 2020

Abstract
Bisphenol A (BPA) is a chemical compound used as a plasticizer in a variety of plastic goods. Environmental chemicals like BPA haven been detected in the blood of women and girls. Yet, they are thought to interact with the endocrine system, thereby having carcinogenic effects on the tissue. While BPA is removed from many commercial products because of its potential carcinogenic effect, surprisingly little is known about its replacement chemicals, such as bisphenol S (BPS) and bisphenol F (BPF). In this study, we aim to link the exposure to those environmental chemicals to changes in human mammary gland morphology and to the underlying alterations at the protein level.
We established organoid cultures from non-malignant primary human mammary gland tissue as advanced disease model. Human breast organoids were treated with physiological levels of various environmental chemicals, includingBPA, BPF, BPS, 17-β-estradiol (E2), or DMSO as vehicle control. Effects on organoid morphology were studied using brightfield and confocal microscopy. We analyzed relative protein abundances by quantitative mass spectrometry to define proteome changes between the different treatments.
The exposure to the environmental chemicals as well as E2 resulted in altered branching morphology while organoid size was not affected. It is already known that BPA has oncogenic potential. Surprisingly, in our study, BPA has the least severe effects when compared to the other bisphenols. BSF or BPS treatment lead to more extensive branching and changed morphology.On protein level, each environmental chemical resulted in distinct protein profiles which show proteins alterations independent of endocrine signaling. Moreover, the exposure to the different bisphenols resulted in the upregulation of proteins which are involved in carcinogenesis, tumor progression and metastasis in various cancer types including breast cancer and thus may support tumor development.
In this study, we identified BPS and BPF as potential carcinogens. Not only BPA, but also BPF and in particular BPS alter branching morphology of mammary organoids and influence various biological processes that are distinct from endocrine signaling. Our study highlights the desperate need of a thorough characterization of environmental chemicals to prevent the development of breast cancer caused by the exposure to potential harmful and carcinogenic substances.
Citation Format: Johanna H. Hinrichs, Juliane Winkler, Pengyuan Liu, Nassim Ataii, Susan Fisher, Zena Werb. Exposure to environmental chemicals leads to extensive branching and expression of cancer-promoting proteins in human mammary gland organoids [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3807.
Abstract 3834: Histone methyltransferase SET8 is regulated by miR-192/-215 and induces oncogene-induced senescence via p53-dependent DNA damage in human gastric carcinoma cells
Authors: Zhang Xiaojing , Peng Yin , Hu Fan , Gao Yuli , Zhe Jin , Meltzer Stephenj 
Published: September 2020

Abstract
Purpose: This aim of study was to investigate the clinicopathologic significance and potential role of histone methyltransferase SET8 in the progression of gastric carcinoma (GC).
Experimental Design: SET8 was screened out by gene microarray in gastric cell lines with or without miR-192/-215 inhibitors or mimics. The effects of SET8 on cell proliferation, invasion, and metastasis were examined using EdU assays, invasion studies, subcutaneous tumor experiments, and tail-vein injection in nude mice. Immunohistochemical staining was performed to detect SET8 expression in GC tissues, and real-time PCR was performed to examine the correlations between miR-192/-215 and SET8. SA-β-Gal staining assays were performed to identify the occurrence of senescence. Senescence-associated proteins were examined by real-time PCR and Western blotting.
Results: SET8 regulated by miR-192/-215 displayed decreased expression in GC. Inhibition of miR-192/-215 resulted in weakened proliferation, migration, and invasion in GC. The 3'UTR of SET8 was targeted by miR-192/-215, and SET8 siRNA treatment successfully rescued the biological phenomenon. SET8 was found to trigger oncogene-induced senescence (OIS) in GC in vivo and in vitro, highlighting the functional mechanism of the miR-192/-215/SET8 axis in GC. During OIS, p53 and p21 were key genetic switch points. Taken together, these data show that the miR-192/-215/SET8/p53 circuit is a key regulator of GC promotion and is highly associated with proliferation, migration, and metastasis.
Conclusions: Our findings reveal that SET8 functions via the OIS-signaling pathway as a negative regulator of metastasis. This provides an experimental rationale for investigation of the OIS-signaling pathway as a potential therapeutic target in GC.
Supported by: National Nature Science Foundation of China (81772592) to Z.J.; National Nature Science Foundation of China (81871969) to X.Z.; National Natural Youth Science Foundation of China (31601028) to Y.P.; Guangdong Provincial Key Laboratory of Regional Immunity and Diseases (2019B030301009) to Z.J.
Citation Format: Xiaojing Zhang, Yin Peng, Fan Hu, Yuli Gao, Jin Zhe, Stephenj Meltzer. Histone methyltransferase SET8 is regulated by miR-192/-215 and induces oncogene-induced senescence via p53-dependent DNA damage in human gastric carcinoma cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3834.
Abstract 3842: Histone methyltransferase G9a promotes migration, invasion of non-small cell lung cancer through enhancing FAK and NF-kB signal pathways
Authors: Sun Ting , Zhang Keqiang , Pangeni Rajendra P. , Li Wendong , Raz Dan J. 
Published: September 2020

Abstract
Overexpression of euchromatic histone methyltransferase 2 (EHMT2 or G9a) is associated with carcinogenesis and poor prognosis in a number of cancer types. In this study, we investigated the effect of G9a on invasion of non-small cell lung cancer (NSCLC) cells and the mechanism underlying cancer cell invasion. Knockdown of G9a was shown to drastically suppress in vitro cellular proliferation, migration, and invasion of A549 and H1299 NSCLC cells, while overexpression of G9a significantly enhanced the proliferation and invasion of these NSCLC cells. Additionally, knockdown of G9a led to significant decrease in expression of focal adhesion kinase (FAK) gene and activation of FAK pathway. UNC0638, a selective inhibitor of G9a, significantly suppressed both the proliferation, migration and invasion of these NSCLC cells, and concurrently led to a dramatic drop of global demethylation of H3K9 (H3K9me2) and suppressed the activation of FAK pathway. In addition, Defactinib, a potent FAK inhibitor, partially abolished the enhanced invasion by overexpression of G9a in these NSCLC cells. Furthermore, G9a was proved to regulate nuclear NF-κB transcriptional activity in NSCLC cells, and NF-κB inhibitor partially abolished the activated FAK signaling pathway by overexpressed G9a, which indicates that G9a-enhanced invasion and activation of FAK pathway may be mediated by NF-κB transcriptional activity. Notably, immunohistochemistry analysis further revealed that activation of FAK pathway was associated with overexpressed G9a in about 150 cases of NSCLC tissues. Altogether, these data have strongly demonstrated that G9a may promote invasion of NSCLC cells by upregulating FAK protein and pathway via enhancing NF-κB transcriptional activity, indicating a therapeutic implication of targeting these two pathways in NSCLC with overexpressed G9a.
Citation Format: Ting Sun, Keqiang Zhang, Rajendra P. Pangeni, Wendong Li, Dan J. Raz. Histone methyltransferase G9a promotes migration, invasion of non-small cell lung cancer through enhancing FAK and NF-kB signal pathways [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3842.
Abstract 3854: Prevalence and prognostic role of FoxP3regulatory T lymphocytes in cancer: A tissue microarray study on &gt;20,000 cancers
Authors: Mandelkow Tim , Bady Elena , Blessin Niclas , Hube-Magg Claudia , Simon Ronald , Sauter Guido , Fraune Christoph , Lennartz Maximilian , Möller Katharin , Weidemann Sören , Luebke Andreas , Höflmayer Doris 
Published: September 2020

Abstract
Regulatory FoxP3+ lymphocytes function as suppressors of T-cell activity. The clinical impact of high FoxP3+ cell density in cancers is not fully understood, as some studies have linked high FoxP3+ cell density to good prognosis and others to poor prognosis in tumor cohorts with associated clinical data. While some data suggest that these variable data are due to biological differences between tumor entities, it is also possible that methodological differences have caused these discrepancies. This study was undertaken to analyze the density of FoxP3+ cells in various different cancer types by employing standardized methods. Tissue microarrays and large sections made from &gt;20,000 prostate, breast, colorectal, ovarian, pancreatic, bladder and stomach cancers were analyzed together with various normal and inflamed tissues by conventional brightfield FoxP3 immunohistochemistry. Samples were also analyzed by fluorescent multiplex immunohistochemistry to assess the fraction of Ki67+ FoxP3+ cells. Our results indeed suggested a variable role of FoxP3+ cells in different tumor types. High FoxP3+ density was linked to high Gleason grade (p=0.0003) and early biochemical recurrence (p&lt;0.0001) in 16923 prostate cancers, but to low tumor stage (p=0.027) and prolonged survival (p=0.0029) in 1341 breast cancers, and to low tumor stage (p&lt;0.0001) in 744 colorectal cancers. No significant associations were found to tumor phenotype in 549 ovarian, 574 pancreatic, 549 bladder and 346 stomach cancers. Multiplex fluorescence IHC analysis of FoxP3 and Ki67 revealed comparable fractions of proliferating FoxP3+ cells in healthy tissues (average 12.3%, range 5.8-18.5%) and inflammatory conditions (average 7.6%, range 2.6-17.2%). Interestingly, the rate of Ki67+FoxP3+ cells was markedly higher in 36 bladder cancers (average 14.2%, range 0-49.3%) suggesting active expansion of FOXP3+ cells in cancer.
Our data demonstrate an inverse prognostic impact of the FoxP3+ cell density in prostate and breast cancers. The increased proliferation rate of immune-regulatory FoxP3+ cells in some bladder cancer is interesting in the light of the variable response of these tumors to immune checkpoint inhibitors.
Citation Format: Tim Mandelkow, Elena Bady, Niclas Blessin, Claudia Hube-Magg, Ronald Simon, Guido Sauter, Christoph Fraune, Maximilian Lennartz, Katharin Möller, Sören Weidemann, Andreas Luebke, Doris Höflmayer. Prevalence and prognostic role of FoxP3regulatory T lymphocytes in cancer: A tissue microarray study on &gt;20,000 cancers [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3854.
Abstract 3870: Patterns of CD112R expression in normal lymphatic tissues, inflammation and cancer
Authors: Blessin Niclas Christian , Mandelkow Tim , Bady Elena , Hube-Magg Claudia , Sauter Guido , Simon Ronald , Fraune Christoph , Lennartz Maximilian , Weidemann Sören A. , Möller Katharina , Höflmayer Doris 
Published: September 2020

Abstract
CD112R is an inhibitory immune checkpoint receptor and a putative target for novel immune therapies, but little is known about its molecular epidemiology in healthy and diseased tissues. To study the prevalence and expression level of CD112R+ immune cells, we analyzed more than 200 samples of normal lymphatic, inflamed and cancerous tissues in a microenvironment tissue microarray format (4mm tissue spot diameter) and large sections using fluorescent multiplex immunohistochemistry.
CD112R expression was detected at variable intensity levels in 47% of CD8+ cytotoxic lymphocytes, 49% of CD4+ T helper cells, 30% of FOXP3+ regulatory T helper cells and in 25% of CD56+ natural killer cells, but no expression was seen in CD11c+ dendritic cells and CD68+ macrophages. All analyzed compartments across normal and diseased tissues showed a small subset (CD8: 9±18%, CD4: 5±15%, FOXP3: 2±5%) of immune cells with supramaximal CD112R expression. The highest fraction of cells with supramaximal CD112R expression was found in the subset of CD8+ cytotoxic T cells in the Peyer's patches of ileum (62%), the intergranuloma area of lymph node sarcoidosis (27%) and in ovarian cancer (37%). In cancerous tissues, the density and the fraction cytotoxic T cells with supramaximal CD112R expression was highly variable and ranged from 5% in bladder cancer to 3% in lung cancer and 36% in ovarian cancer. A high variability of the number of cells with supramaximal CD112R expression was also seen within every tumor entity.
In summary, our analysis shows that CD112R expression is abundant in various subsets of immune cells but identifies a small fraction of cells with exceedingly high CD112R levels. The widespread occurrence of CD112R+ cytotoxic T cells in the cancer microenvironment may suggest considerable opportunities for checkpoint inhibitors targeting CD112R.
Citation Format: Niclas Christian Blessin, Tim Mandelkow, Elena Bady, Claudia Hube-Magg, Guido Sauter, Ronald Simon, Christoph Fraune, Maximilian Lennartz, Sören A. Weidemann, Katharina Möller, Doris Höflmayer. Patterns of CD112R expression in normal lymphatic tissues, inflammation and cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3870.
Abstract 3935: Runt related transcription factor-1 (runx1) plays a central role in vessel co-option of colorectal cancer liver metastases
Authors: Rada Miran , Kapelanski-Lamoureux Audrey , Petrillo Stephanie , Tabariès Sébastien , Siegel Peter , Lazaris Anthoula , Metrakos Peter 
Published: September 2020

Abstract
Colorectal cancer liver metastasis (CRCLM) has two major morphological growth pattern subtypes including desmoplastic (DHGP) and replacement (RHGP). The DHGP tumors depend on angiogenesis for their growth. Conversely, RHGP tumors are non-angiogenic and obtain their blood supply through vessel co-option, which the cancer cells hijacking pre-existing blood vessels of the surrounding liver tissue. Therefore, anti-angiogenic therapies have shown modest efficacy in CRCLM patients with RHGP lesions. In this study, we found overexpression of Runt Related Transcription Factor-1 (RUNX1) in the cancer cells of RHGP lesions. Upregulation of RUNX1 is positively correlated with cancer cell motility and epithelial mesenchymal transition (EMT) in vitro. Our results also showed that RUNX1 regulated by Transforming Growth Factor Beta-1 (TGFβ1) through TGFβ Receptor II (TGFβRII). Significantly, RUNX1 knockdown induced conversion RHGP lesions to DHGP in vivo. Collectively, this study suggests RUNX1 as a potential target therapy to overcome resistance to anti-angiogenic therapy in CRCLM.
Citation Format: Miran Rada, Audrey Kapelanski-Lamoureux, Stephanie Petrillo, Sébastien Tabariès, Peter Siegel, Anthoula Lazaris, Peter Metrakos. Runt related transcription factor-1 (runx1) plays a central role in vessel co-option of colorectal cancer liver metastases [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3935.
Abstract 396: The bile acid receptor FXR suppress colitis-induced colon cancer
Authors: Fu Ting , Evans Ronald 
Published: September 2020

Abstract
Colorectal cancer (CRC) is the third most common cancer and a major cause of cancer related death. Colitis-associated colorectal carcinoma (CAC) is a type of CRC that develops in patients with inflammatory bowel disease (IBD), and is associated with chronic, idiopathic intestinal inflammation. While the underlying cause(s) are poorly understood, environmental factors including high fat diets and aberrant intestinal immune responses contribute to disease progression. Bile acid (BA) overproduction is implicated in diarrhea associated with IBD and CAC. As early sensors of dietary status, BAs are critical mediators of gut inflammation and immunity, in part through their ability to affect the activity of the Farnesoid X Receptor (FXR). FXR is considered the master regulator of BA homeostasis, also a key modulator of the pro- and anti-inflammatory responses in the intestinal epithelium, whose function severely compromised in both CAC patients and in AOM/DSS mouse model. Here, we show that inflammation-induced epithelial abnormalities altered the BAs profile, given the role of the crypt-villus architecture in establishing a BA gradient necessary for homeostasis. In addition, AOM/DSS treatment enhanced immune cell infiltration, as well as induced the formation of massive tumors in the colon. Given that intestinally-restricted FXR agonist FexD can reduced tumorigenesis in APCmin/+ mice, we also treated AOM/DSS mice with FexD. Notably, FexD activated FXR signaling pathway and restored BA homeostasis. Further, it reduced fecal bleeding, intestinal permeability, and decreased serum levels of the late carcinogenesis markers CEA and CA19. Treatment of FexD also resulted in a smaller spleen and reduced pro-inflammatory cytokine levels such as IL17 and IL6. The present study highlights the role of FXR in inflammation induced colon cancer, opening new therapeutic avenues for the treatment of CAC.
Citation Format: Ting Fu, Ronald Evans. The bile acid receptor FXR suppress colitis-induced colon cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 396.
Abstract 397: Myeloma derived mitochondrial DNA activates TLR9 driven pro-tumoral expansion
Authors: Jibril Aisha , Mistry Jayna J. , Kumar Prakrit , Moore Jamie A. , Hellmich Charlotte , Gomez Cesar , Bowles Kristian , Rushworth Stuart A. 
Published: September 2020

Abstract
Multiple myeloma (MM) is reported to cause 85,000 global deaths per year which is estimated to double by 2040. MM is an incurable malignancy of antibody (Ig) secreting differentiated B cells (plasma cells) characterized by the accumulation and localisation of tumor cells in the bone marrow microenvironment (BMM). Mitochondrial genome (mtDNA) contains CpG DNA repeats and CpG DNA has been shown to activate and induce proliferation of memory B cells to secrete Ig. Here we determine if malignant plasma cells maintain their proliferative advantage by releasing mtDNA into the tumour microenvironment which acts as feedback mechanism to induce further proliferation and Ig production by the malignant plasma cell.
Human MM cell lines U266 and MM1S were engrafted into NSG immunocompromised mice. Blood sample from control and engrafted mice were collected at various times and plasma was isolated and spun at 15,000g to remove cell debris and large extracellular vesicles. Human and mouse mtDNA was determine by real-time PCR. Results show that human mtDNA can be detected in the plasma from mice engrafted with U266 and MM1S at 2 weeks and 3 weeks post injection of cells. Next, CpG oligonucleotides were used to determine if MM cells can be activated by mtDNA CpG repeats. Primary MM, U226 and MM1S cells were all treated with CpG and CpG control for 4 and 24 hours. Real-time PCR was then used to detect the pro-survival signature showing that IL-1beta, IL-6 and IL-8 were all upregulated by CpG. We then analysed primary MM and MM cell lines for TLR9 expression by flow cytometry and showed that all samples tested had high levels of cytosolic TLR9 expression. Using TLR9 antagonist (ODN TTAGGG (A151)) we show that primary MM cells and MM cell lines have reduced IL-6 and IL-8 expression and decreased cell proliferation. Finally, U266 and MM1S transduced with TLR9 knockdown lentivirus had reduced engraftment, as measured by bioluminescence, in NSG mice.
Here we show that MM release mtDNA in vivo and this can induce a tumour directed proliferative response through the engagement of TLR9. This process is necessary for optimum tumor growth and forms part of the malignant phenotype of MM.
Citation Format: Aisha Jibril, Jayna J. Mistry, Prakrit Kumar, Jamie A. Moore, Charlotte Hellmich, Cesar Gomez, Kristian Bowles, Stuart A. Rushworth. Myeloma derived mitochondrial DNA activates TLR9 driven pro-tumoral expansion [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 397.