bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4176: Abrogation of desmoglein 2-mediated c-met activation as a novel therapeutic strategy for reducing anaplastic thyroid cancer metastasis
Authors: Lee Dong G. , Min Jeong-Ki
Published: August 2018

Anaplastic thyroid cancer (ATC) is a fast growing and highly metastatic cancer with limited therapeutic options. Thus, targeted therapies need to be developed. This study aims to determine desmoglein 2 (Dsg2) expression in ATC and to elucidate its biological role and potential as a therapeutic target in ATC. Dsg2 expression was low or abnormal in ATC tissues. The ATC patients with reduced Dsg2 expression also had distant metastasis. Dsg2 depletion significantly increased cell migration and invasion, with relatively little effect on ATC cell proliferation in vitro, and increased distant metastasis in vivo. Dsg2 knockdown induced cell motility through the hepatocyte growth factor receptor (HGFR, c-Met)/Src/Rac1 signaling axis, with no alterations in the expression of EMT-related molecules. Further, specific targeting of c-Met significantly inhibited the motility of shDsg2-depleted ATC cells. Decreased membrane expression of Dsg2 increases the metastatic potential of ATC cells. Dsg2 plays an important role in migration and invasiveness in cell lines derived from ATC. Therapies targeting c-Met might be effective for ATC patients with decreased membrane expression of Dsg2. Thus, analysis of Dsg2 expression may provide a novel therapeutic strategy for ATC treatment.

Keywords Anaplastic Thyroid Cancer, Desmoglein 2, Metastasis, c-Met, Tivantinib

Citation Format: Dong G. Lee, Jeong-Ki Min. Abrogation of desmoglein 2-mediated c-met activation as a novel therapeutic strategy for reducing anaplastic thyroid cancer metastasis [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 4176.
Abstract 4181: Simultaneous inhibition of the Wnt/B-catenin and Ras pathways by KY1022 effectively suppresses tumor initiation and progression of colorectal cancer
Authors: Ro Eun Ji , Hwang Jeong-Ha , Cho Yong-Hee , Choi Kang-Yell
Published: August 2018

APC (80-90%) and K-Ras (40-50%) mutations frequently occur in human colorectal cancer (CRC) and these mutations cooperatively accelerate tumorigenesis including metastasis. In addition, both B-catenin and Ras levels are highly increased in CRC, especially in metastatic CRC (mCRC). Targeting these signaling pathways may therefore be an ideal strategy for treating mCRC; however, therapeutics targeting both of these pathways have not been developed because the mechanisms regulating these pathways are not well understood. Based on our recent identification of the B-catenin destruction complex-mediated Ras protein stability mechanism, we screened a small-molecule library to identify compounds that destabilize both B-catenin and Ras proteins via inhibition of the Wnt/B-catenin pathway. Among the effective small molecules destabilizing both B-catenin and Ras proteins via inhibition of the Wnt/β-catenin pathway, we characterized KY1022, a small molecule that most effectively inhibits EMT, an initial process of metastasis. As shown by in vitro and in vivo studies using APCMin/+/K-RasG12DLA2 mice, KY1022 effectively suppressed the development of mCRC at an early stage of tumorigenesis. Our results suggest that a small molecular approach degrading both B-catenin and Ras via inhibition of the Wnt/B-catenin signaling would be an ideal strategy for treatment of CRC including mCRC.

Citation Format: Eun Ji Ro, Jeong-Ha Hwang, Yong-Hee Cho, Kang-Yell Choi. Simultaneous inhibition of the Wnt/B-catenin and Ras pathways by KY1022 effectively suppresses tumor initiation and progression of colorectal 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 4181.
Abstract 4196: Aspirin inhibits metastasis in the intravascular phase through the blockade of COX-1-TXA2 pathway in platelets
Authors: Lucotti Serena , Cerutti Camilla , Soyer Magali , Gil-Bernabé Ana M. , Gomes Ana L. , Allen Philip D. , Smart Sean , Markelc Bostjan , Watson Karla , Armstrong Paul C. , Mitchell Jane A. , Warner Timothy D. , Ridley Anne J. , Muschel Ruth J.
Published: August 2018

Evidence from experimental studies and clinical trials has shown that aspirin reduces the incidence of distant metastases. Aspirin inhibits cyclooxygenase (COX)-1 and COX-2, triggering anti-thrombotic and anti-inflammatory effects, respectively. However, the mechanisms underlying the anti-metastatic effect of aspirin are still largely unknown.

By employing mouse models of experimental and spontaneous metastasis, we have shown that aspirin and a selective COX-1 inhibitor dramatically reduced metastatic lung nodules, an effect that was not replicated by selective inhibition of COX-2. Metastasis was impaired in COX-1 deficient mice as well, suggesting a pivotal role of microenvironmental COX-1 in the metastatic process. In particular, COX-1 activity was essential during the intravascular phase of pulmonary metastasis. Using a model of platelet depletion and re-infusion we identified platelet-derived thromboxane A2 (TXA2) as the main product of COX-1 responsible for its permissive effect on metastasis. The inhibition of the COX-1-TXA2 pathway in platelets decreased the aggregation of platelets on tumor cells and was associated with a reduction in endothelial activation, in tumor cell adhesion to the endothelium, in recruitment of metastasis-promoting monocytes/macrophages and in transendothelial migration. Thus, platelet-derived TXA2 orchestrates the generation of a favorable intravascular metastatic niche that promotes tumor cell seeding and metastasis.

Taken together, our data suggest that COX-1 inhibition in platelets by aspirin is sufficient to exert an anti-metastatic effect and shed a new light on COX-1-TXA2 signalling in the context of tumor cell dissemination. From this perspective, TXA2 might present a more selective therapeutic target for the prevention of metastasis.

Citation Format: Serena Lucotti, Camilla Cerutti, Magali Soyer, Ana M. Gil-Bernabé, Ana L. Gomes, Philip D. Allen, Sean Smart, Bostjan Markelc, Karla Watson, Paul C. Armstrong, Jane A. Mitchell, Timothy D. Warner, Anne J. Ridley, Ruth J. Muschel. Aspirin inhibits metastasis in the intravascular phase through the blockade of COX-1-TXA2 pathway in platelets [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 4196.
Abstract 42: Triple negative breast cancer cell heterogeneity in zebrafish-host microenvironment
Authors: Snaar-Jagalska B.Ewa , Groenewoud Arwin , Astrologo Letizia , Chen Lanpeng , Julio Marianna Kruithof-de
Published: August 2018

Metastatic triple negative breast cancer (TNBC) can be considered as incurable endpoint in the progression of TNBC, prevention or treatment of metastasis therefore is of paramount importance. New insights into intra-tumoral heterogeneity have cast light on the existence of multiple functionally distinct cellular entities within one tumor, among others these cells can be divided based on their metastatic potential. The zebrafish provides us with the unique opportunity to study the first steps of metastatic initiation in great detail both microscopically and transcriptionally, on the side of both the host and cancer cells. Organotropic TNBC lines engrafted into this model recapitulate the tropism found in higher vertebrates. Using our zebrafish TNBC xenograft model we study these cellular sub-populations in vivo during metastatic initiation. These small metastatic lesions can be further analyzed ex vivo through the re-isolation of novel zebrafish specific organotropic cell lines. Re-engraftment of these greatly enhances the tumorigenic potential, and transcriptional analysis reveals increased EMT plasticity and an induction of stemness markers. Engraftment of these cells into murine tibia shows a significantly increased tumorigenic and osteolytic potential, further underscoring the critical role of this sub-population in cancer initiation. To determine the genetic drivers of this process we performed whole exome sequenced (NGS) of both, in vitro cultured cancer cells (maternal and re-isolated) and micro-dissected tissue samples derived from in situ zebrafish xenograft material. We are currently in the process of evaluating the candidate genes underpinning the enhanced tumorigenic potential in both the zebrafish and murine models used.

Citation Format: B.Ewa Snaar-Jagalska, Arwin Groenewoud, Letizia Astrologo, Lanpeng Chen, Marianna Kruithof-de Julio. Triple negative breast cancer cell heterogeneity in zebrafish-host 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 42.
Abstract 4204: Structure-activity relationship of diphenylamine derivatives to target epithelial to mesenchymal transition in triple negative breast cancer
Authors: Bhatt Akshita B. , Wright Thomas D. , Anna Katie , Gupta Mohit , Flaherty Patrick , Hoang Van , Burow Matthew , Cavanaugh Jane E.
Published: August 2018

Triple negative breast cancer (TNBC) is characterized by the loss of hormone receptors and high invasive potential. TNBC cells express high levels of the mesenchymal markers vimentin, fibronectin, N-cadherin, and cadherin11, which are involved in cell invasion and metastasis, and low levels of the epithelial markers E-cadherin, occludins and cytokeratins, which are involved in cellular contact and stability of the cell membrane. Mitogen activated protein kinases, including ERK5 and ERK1/2 are important pro-survival proteins and are known to be upregulated in most cancers. Of these pathways, the MEK5-ERK5 pathway is understudied in triple negative breast cancer, and there are few research tools available to selectively inhibit this pathway. The diphenylamine moiety was derived from the parent molecule Trametinib, a FDA approved MEK1/2 inhibitor for melanoma. Structural modifications were made on the lead molecule to gain selectivity towards MEK. SC-1-151, one of the compounds from this diphenylamine series was identified as a dual MEK1/2 (98.6% inhibition) and MEK5 (59% inhibition) inhibitor. Additionally, we have shown that SC-1-151 attenuates tumor growth in severe combined immunodeficient (SCID) xenograft mice and causes a mesenchymal to epithelial transition (MET) in MDA-MB-231 TNBC cells. The goal of this research is to determine the structural features of diphenylamines that are responsible for MET.

E-cadherin and cadherin-11 protein expression and cell morphology were examined to study MET after the treatment of MDA-MB-231 cells with different structural analogs of SC-1-151 for 5 days. Alkyl or N-Methyl piperazine substituents on the amide of ring 1 produced similar result as SC-1-151, and substituting the amide group with acid or ester also induced MET. In contrast, ortho-fluoro, para-iodo functional groups of the arene ring 2, when replaced with a meta-bromo substituent did not induce MET. Therefore, our data suggest the necessary functional groups for inducing MET include the hydrogen atom on the amine linkage, and the iodine atom on the arene ring 2. Future studies will be performed to determine the specific protein interactions of the promising compounds.

Citation Format: Akshita B. Bhatt, Thomas D. Wright, Katie Anna, Mohit Gupta, Patrick Flaherty, Van Hoang, Matthew Burow, Jane E. Cavanaugh. Structure-activity relationship of diphenylamine derivatives to target epithelial to mesenchymal transition in triple negative 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 4204.
Abstract 4220: Heterogeneous distribution of prognostic protein markers in glioblastoma
Authors: Stetson Lindsay C. , Ostrom Quinn T. , Liao Peter , Sloan Andrew E. , Chance Mark R. , Barnholtz-Sloan Jill S.
Published: August 2018

Background: Glioblastoma (GBM) is the most common and lethal primary malignant brain tumor in adults. The heterogeneity of the disease leads to significant variability in response to standard therapy (surgery plus concurrent radiation and temozolomide). An accurate and reliable predictor of patient prognosis represents an unmet need to improve the care of GBM patients. While protein markers are an effective readout of cellular function, proteomics has rarely been utilized in GBM prognostic marker discovery. Experimental Procedures: GBM patients were prospectively recruited (Ohio Brain Tumor Study) and proteomics discovery using liquid chromatography mass spectrometry analysis (LC MS/MS) was performed in a discovery set of 27 patients including 13 short-term survivors (< 9 months, STS) and 14 long-term survivors (>= 18 months, LTS). Statistically significant proteins were evaluated in two independent datasets, including in18 samples micro-dissected from multiple tumor areas of 6 GBM patients. Results: Proteomics discovery identified 11,941 peptides in 2,495 unique proteins, with 172 proteins exhibiting significant dysregulation when comparing STS and LTS. Proteins involved in glycolysis/TCA cycle were up-regulated in STS compared to LTS by examination of individual targets as well as upon application of a novel protein and peptide pathway enrichment analysis. Validation of these dysregulated proteins and other protein markers from the literature in our first validation set (18 samples micro-dissected from n=6 patients) demonstrated that they were very unevenly distributed throughout individual patients' tumors. Several proteins overcame the heterogeneous nature of the tumors and were both prognostic markers differentially expressed between LTS and STS, as well as potential drug targets owing to their even distribution throughout the tumor. Protein abundance of significant marker proteins were verified in a second independent validation set using Western immunoblots. Conclusion: The current study verified the importance of metabolism in GBM pathology and demonstrated the heterogeneous nature of GBMs at the protein level.

Citation Format: Lindsay C. Stetson, Quinn T. Ostrom, Peter Liao, Andrew E. Sloan, Mark R. Chance, Jill S. Barnholtz-Sloan. Heterogeneous distribution of prognostic protein markers in glioblastoma [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 4220.
Abstract 4223: Personal history of non-melanoma skin cancer and the risk of death from invasive cutaneous melanoma in men
Authors: Song Fengju , Han Jiali
Published: August 2018

BACKGROUND: Previous studies have found an increased risk of invasive cutaneous melanoma (CM) among those with a history of non-melanoma skin cancer (NMSC). OBJECTIVE: The aim of this study is to evaluate the risk of CM death after NMSC. METHODS: The study was based on the Health Professionals Follow-up Study (HPFS). Cox proportional hazards model was used to examine the hazard ratio (HR) of CM death associated with personal history of NMSC, among the entire study population (primary analysis) and invasive CM cases (secondary analysis) respectively. RESULTS: We documented a total of 908 invasive CM cases over a total of 0.7 million person-years of follow-up. Among all participants, the risk of developing either lethal or non-lethal invasive CM increased for those with NMSC history. The risk of melanoma death based on NMSC history was non-significantly increased, with HR (95% CI) of 1.53 (0.95 - 2.46). In the case-only analysis, those with NMSC history had a significantly lower risk of melanoma death compared with those with no such history, HR=0.60 (0.35-0.94). LIMITATIONS: Because HPFS consists exclusively of male health professionals, the results of this study may not be extended to the entire population. CONCLUSION: Personal history of NMSC is associated with a decreased risk of melanoma-specific death among male patients with invasive CM. KEYWORDS: invasive cutaneous melanoma; non-melanoma skin cancer; mortality; cohort study

Citation Format: Fengju Song, Jiali Han. Personal history of non-melanoma skin cancer and the risk of death from invasive cutaneous melanoma in men [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 4223.
Abstract 4226: Association between NSAID, statins, and bisphosphonates and prostate cancer survival during androgen deprivation therapy
Authors: Raittinen Paavo V. , Talala Kirsi , Taari Kimmo , Tammela Teuvo L. , Ilmonen Pauliina , Auvinen Anssi , Murtola Teemu J.
Published: August 2018

We study the association between non-steroidal anti-inflammatory drugs (NSAIDs), Statins, and Bisphosphonates (BPs), and prostate cancer (PCa) survival during androgen deprivation therapy (ADT). STAMPEDE trial has demonstrated better PCa-specific survival in men using combination of celecoxib (CEL) and zoledronic acid (ZA) during ADT compared to ADT alone. The mechanism is unclear. ZA inhibits mevalonate pathway (MevP) previously linked with cancer growth. We evaluated PCa survival among men on ADT and simultaneously using BPs including ZA or statins, another drug group inhibiting MevP, and NSAIDs including CEL. We hypothesized that combined use of a MevP inhibitor and NSAID would be associated with improved PCa survival.

Our study cohort includes 4,428 men from the Finnish Randomized Study of PCa Screening (FinRSPC) initiating ADT in 1995-2015. Cox proportional hazards model with adjustment for age, FinRSPC study arm, tumor clinical characteristics and co-morbidities (obtained from national registries) was used to calculate HRs and 95% CI for PCa death. Medication use was analyzed as time-dependent variable.

Compared to non-users, the risk of PCa death was increased in users of NSAIDs or acetaminophen, and lowered in statin users. Use of BPs or coxibs alone were not associated with the risk. Coxibs and statins together were associated with lowered risk to a similar degree as statins alone. No statistically significant risk differences were observed for other combinations.

Statin users with high-risk prostate cancer undergoing ADT have lowered risk of PCa death. NSAID users have increased risk of PCa death, which becomes statistically insignificant when used with statins. Statin and BP use together shows no statistically significant evidence of negating the effects of statin. No clear additive benefit was observed for statins and coxibs together over statins alone. Our findings do not support additive benefits of MevP inhibitor and NSAIDs.

Statistical significance codes: *** : p = 0.001, ** : p = 0.01, * : p = 0.1DrugHR95 % CISignificanceStatin0.780.680.90***Acetylsalisylic acid0.900.761.07Coxib1.070.941.22NSAID1.171.041.31**Acetaminophen1.661.511.82***Bisphosphonate0.790.381.64Bisphosphonate and NSAID0.890.551.42Statin and NSAID1.070.901.27Statin and Bisphosphonate1.140.861.50Coxib and Bisphosphonate0.850.421.74Coxib and Statin0.800.621.02*EAU tumor Risk Group2.662.353.00***

Citation Format: Paavo V. Raittinen, Kirsi Talala, Kimmo Taari, Teuvo L. Tammela, Pauliina Ilmonen, Anssi Auvinen, Teemu J. Murtola. Association between NSAID, statins, and bisphosphonates and prostate cancer survival during androgen deprivation therapy [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 4226.
Abstract 4243: Association of oral health with lung cancer risk: Results from the Southern Community Cohort Study
Authors: Yoon Hyung-Suk , Wen Wanqing , Long Jirong , Zheng Wei , Blot William J. , Cai Qiuyin
Published: August 2018

Increasing evidence suggests that oral health may be associated with cancer and other chronic diseases. Oral bacterial infections, which induce inflammation of the gums and tooth loss, can progress to systemic inflammation. In addition, poor oral health is often the result of smoking and/or limited access to regular dental care. Previous studies have suggested that poor oral health may increase the risk of lung cancer among individuals of European and Asian descent, but the evidence remains elusive. In this nested case-control study, we investigated whether oral health status was associated with lung cancer risk among African Americans and European Americans and whether the association varied by race and smoking habits. A total of 403 incident lung cancer cases and 1,612 matched controls from the Southern Community Cohort Study were included in the current study. The controls were matched to cases on age, sex, race, and recruitment site. Multivariate conditional logistic regression models were fitted to evaluate the risk of lung cancer linked to tooth loss, tooth decay, and history of periodontal disease (gingivitis or periodontitis). Stratified analyses were conducted by race, smoking status, and pack-years, and the interactions of these variables with oral health were further evaluated. Tooth loss was significantly associated with an increased risk of lung cancer. Compared to those without any loss of teeth, the odds ratio (OR) of loss of >10 teeth was 1.64 (95% confidence interval [CI]: 1.00-2.69) after adjusting for confounders. Tooth decay was also significantly associated with increased lung cancer risk. Those with ≥ 6 decaying teeth had an OR of 1.65 (95% CI: 1.18-2.31), compared to those with no tooth decay. An increased lung cancer risk was significantly associated with a history of periodontal disease (OR: 1.44, 95% CI: 1.09-1.19). While there was no evidence of interaction by race, the increase in risk associated with tooth loss, tooth decay, and periodontal disease was observed only among smokers who had smoked more than the race-specific median pack-year. Results from our study indicate that poor oral health, as measured by tooth loss, tooth decay, and periodontal disease is associated with an increased risk of lung cancer and that this association is more evident among heavy smokers.

Citation Format: Hyung-Suk Yoon, Wanqing Wen, Jirong Long, Wei Zheng, William J. Blot, Qiuyin Cai. Association of oral health with lung cancer risk: Results from the Southern Community Cohort 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 4243.
Abstract 4251: Neighborhood data system to enable health disparities research in Chicago
Authors: Press David J. , Gomez Scarlett Lin , Aschebrook-Kilfoy Briseis , Ahsan Habibul
Published: August 2018

Background: Chicago has become the focus of national concern following findings of striking disparities in cancer incidence and mortality, with worse outcomes for Non-Hispanic Black persons. Chicago also has communities with very high crime rates, racial/ethnic segregation, and resource-poor communities. Social stressors, including crime, social isolation, low socioeconomic status (SES), interpersonal and institutional discrimination, and residence in resource-poor settings intersect at individual- and neighborhood- levels across the lifespan and are associated with poor health outcomes. However, the extent to which these social determinants separately and together contribute to cancer health disparities is unclear.

Purpose: To elucidate health disparities attributable to neighborhood social stressors in the Chicago setting, a pilot Chicago Neighborhood Data System (ChNDS) is currently in production, using the Chicago Police Department crimes database, data from the 2010 Census short form, 2008-2012 American Community Survey (ACS), University of Chicago MAPSCorps project, and Environmental Systems Research Institute (ESRI) geodatabases.

Approach: We use principal components analysis (PCA) to generate composite indices of neighborhood-level factors at the small area level (census tract or block group): racial/ethnic composition; SES; immigration/ acculturation; other sociodemographic characteristics; crime; and community assets including density/proximity to types of businesses, retail food, and recreational facilities.

Future directions: Data record linkages at the small area-level will be performed between the pilot ChNDS and observational and population-based datasets to examine the impact of neighborhood-level characteristics on research study participation, disease occurrence, healthcare utilization, and disease outcomes in Chicago.

Citation Format: David J. Press, Scarlett Lin Gomez, Briseis Aschebrook-Kilfoy, Habibul Ahsan. Neighborhood data system to enable health disparities research in Chicago [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 4251.
Abstract 4260: Dynamic modeling of macrophage plasticity in bone metastatic prostate cancer
Authors: Baratchart Etienne A. , Lo Chen Hao , Lynch Conor , Basanta David
Published: August 2018

Tumor-associated macrophages, particularly those with an anti-inflammatory phenotype, have long been implicated in the progression of primary solid malignancies, including prostate cancer. Metastatic prostate cancer typically manifests in the bone where it induces painful osteogenic lesions that are incurable. Bone is naturally rich in myeloid-derived macrophages whose temporal polarization into pro- (M1) and anti-inflammatory (M2) phenotypes is critical for regulating the bone repair program mediated by bone-resorbing osteoclasts and bone-building osteoblasts. However, the dynamics of macrophage polarization in the context of bone metastatic prostate cancer are underexplored and difficult to address with traditional biologic approaches. To address the role of macrophage polarization in the context of bone metastatic prostate cancer, we first investigated the macrophage polarization temporal dynamics in normal bones and analyzed macrophage plasticity in vivo subsequent to intratibial injury. Bone marrows were isolated at several time points and profiled by flow cytometry for pro- and anti-inflammatory monocyte macrophage content. Contralateral tibias were analyzed for bone volume, osteoblast and osteoclast numbers. We then designed a mathematical model describing the different cell population dynamics in the bones. Generation of the model required testing a number of assumptions regarding macrophage polarization behavior. For example, it is unknown whether M1 resolves at the initial stages of bone injury repair through death or by repolarizing into M2. For each aspect, competing mechanistic assumptions were proposed and simulated by sets of ODEs. The best-fitting assumptions for each aspect were integrated into a single comprehensive ODE model to fully describe the dynamics of the bone resident cell populations during bone remodeling. This experimentally validated model is now being used to address how bone resident cells respond to metastatic prostate cancer cells. In conclusion, we have generated an ODE model that describes macrophage polarization over time during bone repair and how pro- and anti-inflammatory macrophages interact with bone stromal cells. The mathematical model predictions are in agreement with our biologic experiments in vivo and will allow us to interrogate how macrophage polarization impacts the behavior of metastatic prostate cancer cells in the bone microenvironment.

Citation Format: Etienne A. Baratchart, Chen Hao Lo, Conor Lynch, David Basanta. Dynamic modeling of macrophage plasticity in bone metastatic prostate 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 4260.
Abstract 4290: Mad1 promotes tumor progression through destabilization of p53
Authors: Wan Jun , Weaver Beth A.
Published: August 2018

Mitotic arrest deficient 1 (Mad1) plays a well-characterized role in the mitotic checkpoint. However, interphase roles of Mad1 that do not impact mitotic checkpoint function are still largely uncharacterized. We have found that upregulation of Mad1, which is common in human breast cancer, results in decreased protein stability of the tumor suppressor p53 in multiple cell types. To gain mechanistic insight into this process, we first determined whether increasing expression of Mad1 alters its localization. Upregulated Mad1 localizes to puncta within interphase nuclei in both breast cancer tissue and cultured cells. We found that upregulated Mad1 localizes to ProMyelocytic Leukemia Nuclear Bodies (PML NBs), which have been implicated in the stabilization of p53. Immunoprecipitation results indicate that Mad1 and PML interact and that the C-terminal domain (CTD) of Mad1 and the N-terminal domain of PML are required for this interaction. Within the CTD of Mad1, the SUMO-Interacting Motif (SIM) is critical for the localization of Mad1 to PML NBs. MDM2 is an E3 ubiquitin ligase that targets p53 for degradation. In response to DNA damage, PML sequesters Mdm2 to the nucleolus, which stabilizes p53. In cells with elevated levels of Mad1, Mad1 interrupts the interaction between PML and MDM2. Mad1, rather than MDM2, is sequestered to the nucleolus after DNA damage, and Mad1 displaces MDM2 from PML. The displaced MDM2 is no longer sequestered in the nucleolus, freeing it to ubiquitinate p53, resulting in p53 destabilization. Upregulation of Mad1 promotes the growth of orthotopic mammary tumors. Mammary tumors with upregulated Mad1 show decreased levels of p53 and its downstream effector p21. Our results show that, in addition to causing a low rate of chromosome missegregation, Mad1 upregulation has an unexpected interphase role in promoting tumor formation and progression by destabilizing p53. Thus, Mad1 upregulation promotes tumors in through at least two distinct pathways.

Citation Format: Jun Wan, Beth A. Weaver. Mad1 promotes tumor progression through destabilization of p53 [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 4290.
Abstract 4300: Cell cycle pathway gene regulation in glioblastoma multiforme (GBM) and GBM derived stem cells: Implicating Pentraxin 3 upregulation
Authors: Wesley Umadevi V. , Clark Paul , Jaeger Jacob , Kuo John , Dempsey Robert
Published: August 2018

Introduction: Glioblastoma Multiforme (GBM) is the most aggressive type of brain cancer and progresses at a rapid rate. The major obstacle in the treatment of GBM is selective survival of a subpopulation of GBM derived stem cells (GSC) which are highly proliferative and resistant to drug induced apoptosis. The proteins involved in the regulation of the cell cycle impact survival potential of GBM and GSCs. Pentraxin 3 (PTX3), an angiogenic and survival factor plays a key role in cell proliferation, angiogenesis and invasion. However, little is known about the association of PTX3 and cell cycle regulation in GBM derived stem cells. Mehods: GBM patient tumor lines, and GSC derived from the same GBM lines were used. These cell lines were exposed to hypoxia and re-oxygenation. Cell cycle profiling and analysis were carried out using flow cytometry and FlowJo Single Cell Data Analysis Software. Xeno-transplanted mouse brain sections were used for PTX3 expression analysis by immunohistochemistry and immunofluorescence staining. Targeted proteomic approach was used to determine PTX3 levels in GBM and GSC. Alterations in the cell cycle regulatory gene expression in GSC and GBM tumor line in presence and absence of PTX3 was identified by transcriptome array analysis. Results: Our targeted proteomic approach using antibody array identified substantially higher levels of PTX3 in GSCs. GBM tumor lines showed higher percent of cells in G1 phase, where as GSCs showed increased number of cells in G2 and S-phase, both under normoxia and hypoxia. GSCs showed increased levels of GADD45A, known to protect GBM cells from apoptosis. Increased PTX3 expression correlated with increased cell cycle progression. Furthermore, exogenous PTX3 increased CyclinD1, a protein required for progression through the G1 phase of the cell cycle. Conclusion: GSC cells show cell cycle progression, presumably with an increased growth advantage. PTX3 expression is associated with GBM malignancy, and may contribute to GBM cell cycle progression and invasion. Overall, these results indicate that PTX3 and the cell cycle pathway are strongly associated with malignant GBM.

Citation Format: Umadevi V. Wesley, Paul Clark, Jacob Jaeger, John Kuo, Robert Dempsey. Cell cycle pathway gene regulation in glioblastoma multiforme (GBM) and GBM derived stem cells: Implicating Pentraxin 3 upregulation [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 4300.
Abstract 4303: Modulation of protein interaction states through the cell cycle
Authors: Dai Lingyun , Zhao Tianyun , Bisteau Xavier , Sun Wendi , Prabhu Nayana , Lim Yan Ting , Sobota Radoslaw , Kaldis Philipp , Nordlund Pär
Published: August 2018

Global profiling of protein expression through the cell cycle has revealed subsets of periodically expressed proteins. However, expression levels alone only give a partial view of processes determining cellular events. The cell cycle progression events are to a large extent controlled by the dynamic biochemical interactions of proteins with physiological ligands such as other proteins, metabolites, lipids, nucleic acids or low molecular weight effectors. Using a proteome-wide implementation of the Cellular Thermal Shift Assay (CETSA) to study specific cell cycle phases in K562 cell, we uncover modulations of interaction states for more than 750 proteins along the cell cycle. Notably, many protein complexes are modulated in specific cell cycle phases, reflecting their roles in processes such as DNA replication, chromatin remodeling, transcription, translation, and nuclear membrane decomposition. Surprisingly, only small differences in interaction states were seen between G1 and G2 phases, suggesting similar hardwiring of biochemical processes in these two phases. The present work reveals novel molecular details of the cell cycle. CETSA, therefore emerge as a novel approach to discover cellular modulations during cancer development and therapy.

Citation Format: Lingyun Dai, Tianyun Zhao, Xavier Bisteau, Wendi Sun, Nayana Prabhu, Yan Ting Lim, Radoslaw Sobota, Philipp Kaldis, Pär Nordlund. Modulation of protein interaction states through the cell cycle [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 4303.
Abstract 4314: Chromatin state alterations during melanoma progression
Authors: Rai Kunal , Terranova Christopher , Maitituoheti Mayinuer , Tang Ming , Akdemir Kadir , Orouji Elias , Chin Lynda , Fiziev Petko , Dhamdhere Mayura , Samant Neha S. , Shah Amikhsha , Sharma Sneha , Ernst Jason
Published: August 2018

The extent and nature of epigenomic changes associated with melanoma progression is poorly understood. To determine chromatin state patterns of human metastatic melanomas, we defined epigenome of melanoma by profiling 6 histone modification marks (H3K4me, H3K27Ac, H3K4me3, H3K79me2, H3K27me3 and H3K9me3) in 48 metastatic melanoma tumors, cell lines and short-term cultures. Computation of chromatin states identified specific chromatin state aberrations in human tumors and their relationship with specific genotypes. Specifically, we defined patterns of enhancer elements, broad H3K4me3 domains and bivalent chromatin states enriched in specific genotypes of BRAF, NRAS, and WT groups of melanomas. Importantly, we identified switch from specific bivalent chromatin states in melanocytes to active states leading to upregulation of EMT genes in NRAS mutant melanomas. Similarly, we noted gains of broad H3K4me3 domains in particular classes of oncogenes in specific gentoypes as well as during transitions from melanocytes to melanomas. Finally, we identified clusters of enhancers that are specifically enriched in subset of melanoma samples defining intertumor epigenetic heterogeneity in melanoma tumors. In addition, to investigate chromatin state changes associated with early stages of melanoma progression, we systematically profiled 35 epigenetic modifications in a cell phenotypic model of non-tumorigenic and tumorigenic states (and verified in human nevi and melanoma samples). Specific chromatin state transitions featuring loss of histone acetylations and H3K4me2/3 were observed on specific promoter and enhancer elements targeting cancer-regulatory genes in important melanoma-driving cell signaling pathways. Intriguingly, only a small fraction of chromatin state transitions correlated with expected changes in gene expression patterns. Restoration of acetylation levels on deacetylated loci by histone deacetylase (HDAC) inhibitors selectively blocked excessive proliferation in tumorigenic cells and human melanoma cells, suggesting functional roles of observed chromatin state transitions in driving hyperproliferative phenotype. Overall, our study reveals principles of epigenetic reprogramming in melanoma progression stages and how they may establish transcriptional programs that impart aggressive characteristics to melanomas.

Citation Format: Kunal Rai, Christopher Terranova, Mayinuer Maitituoheti, Ming Tang, Kadir Akdemir, Elias Orouji, Lynda Chin, Petko Fiziev, Mayura Dhamdhere, Neha S. Samant, Amikhsha Shah, Sneha Sharma, Jason Ernst. Chromatin state alterations during melanoma 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 4314.
Abstract 4318: Identifying drivers of SMARCA4/BRG1-deficient SCCOHT tumorigenesis by integrative multi-omic analysis
Authors: Orlando Krystal A. , Raab Jesse R. , Lang Jessica D. , Hendricks William P. , Wang Yemin , Huntsman David G. , Trent Jeffrey M. , Parker Joel S. , Weissman Bernard E.
Published: August 2018

Over 94% of small cell carcinomas of the ovary, hypercalcemic type (SCCOHT), a rare and aggressive form of ovarian cancer, have mutations and concomitant protein loss in SMARCA4 (BRG1), one of the two mutually exclusive ATPases of the SWI/SNF chromatin remodeling complex. SCCOHT tumors rarely have secondary mutations, making them an excellent model for understanding the role BRG1 and SWI/SNF complexes play in tumor suppression. We hypothesize that BRG1 loss drives SCCOHT tumorigenesis by altering chromatin accessibility and gene expression. We have previously shown that BRG1 re-expression in SCCOHT cell lines suppresses cell growth and induces an elongated, neuronal-like morphology. To identify the top genes and transcription factors driving SCCOHT tumorigenesis, we performed ATAC-seq and RNA-seq in a SCCOHT cell line +/- BRG1 re-expression. BRG1 re-expression increased overall chromatin accessibly, shown by an increase in the number of ATAC-seq peaks. ATAC peaks gained following BRG1 re-expression were enriched in transcription factor binding motifs from FOS/JUN/AP-1, TEAD, and SOX family members. RNA-seq analysis demonstrated that BRG1 re-expression upregulated more genes than those downregulated, consistent with the increase in ATAC-seq peaks. Preliminary pathway analysis identified gene enrichments in epithelial-mesenchymal transition pathway and extracellular matrix remodeling following BRG1 re-expression, consistent with the observed morphology change. Cell types enrichment analysis (xCell) identified a shift from a mesenchymal stem cell-like gene signature in the control SCCOHT cells to an epithelial cell-like gene signature following BRG1 re-expression, further suggesting a possible mesenchymal-epithelial transition in SCCOHT cells after BRG1 re-expression. Future studies include integration of the ATAC/RNA-seq data to further identify correlations between gene expression changes and enriched transcription factor binding motifs, integrative ChIP-seq analysis for BRG1 following re-expression, and proteomic analyses. These studies will uncover the key genes, proteins, and transcription factors affected by BRG1 loss in other adult cancers, provide insight into BRG1's role in SCCOHT tumorigenesis, and potentially yield novel therapeutic targets.

Citation Format: Krystal A. Orlando, Jesse R. Raab, Jessica D. Lang, William P. Hendricks, Yemin Wang, David G. Huntsman, Jeffrey M. Trent, Joel S. Parker, Bernard E. Weissman. Identifying drivers of SMARCA4/BRG1-deficient SCCOHT tumorigenesis by integrative multi-omic analysis [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 4318.
Abstract 4321: Up-regulation of METTL3 promoted m6A modification and epigenetic silencing of SOCS2 in human liver cancer
Authors: Chen Mengnuo , Wei Lai , Law Cheuk-Ting , Tsang Felice H. , Shen Jialing , Cheng Carol L. , Tsang Long-Hin , Ho Daniel W. , Chiu David K. , Lee Joyce M. , Wong Carmen C. , Ng Irene O. , Wong Chun-Ming
Published: August 2018

Epigenetic alteration is a common trait of human cancers. Chemical modifications on DNA and core histone proteins are the major mechanisms for epigenetic regulation. Recently, emerging evidence suggested that reversible chemical modifications on RNA also play a critical role in epigenetic control of gene expression. N6-methyladenosine (m6A) is the most abundant modification found in mammalian mRNA. m6A is involved in regulating mRNA stability, splicing, and translation. However, the implications of m6A modification in human carcinogenesis remain poorly understood. In this study, we investigated the expression of m6A methyltransferases and demethylases in human hepatocellular carcinoma (HCC). We found the major m6A methyltransferase METTL3 was remarkably up-regulated in HCC. High METTL3 expression was associated with poor overall and disease-free survival in HCC patients. Consistent with the METTL3 overexpression, we found that mRNA m6A level was significantly elevated in human HCC. To investigate the roles of METTL3 in human HCC, we employed lentiviral-based shRNA and CRISPR/Cas9 systems to inactivation METTL3 in HCC cell lines. We showed that knockdown and knockout of METTL3 drastically suppressed HCC proliferation and migration in vitro and abolished HCC tumorigenicity and lung metastasis in vivo. In contrast, overexpression of METTL3 by CRISPR/dCas9 SAM system promoted HCC growth. Using RNA-seq and m6A-Seq, we identified tumor suppressor gene SOCS2 as a novel target of METTL3. We detected m6A modification in SOCS2 mRNA and the m6A modification was diminished upon METTL3 knockdown. m6A modification promoted SOCS2 mRNA degradation. We demonstrated that knockdown of METTL3, mutation of m6A modification sites, and treatment of demethylating agent augmented SOCS2 mRNA expression in HCC cells. In addition, knockdown of YTHDF2, an m6A reader protein, also rescued SOCS2 mRNA expression in HCC cells. The above findings together demonstrated that up-regulation of METTL3 lead to m6A modification of SOCS2, which in turn promotes SOCS2 mRNA degradation through YTHDF2 dependent mechanism. Our findings provided a proof-of-concept model to demonstrate the importance of aberrant m6A modification in epigenetic silencing of tumor suppressor genes.

Citation Format: Mengnuo Chen, Lai Wei, Cheuk-Ting Law, Felice H. Tsang, Jialing Shen, Carol L. Cheng, Long-Hin Tsang, Daniel W. Ho, David K. Chiu, Joyce M. Lee, Carmen C. Wong, Irene O. Ng, Chun-Ming Wong. Up-regulation of METTL3 promoted m6A modification and epigenetic silencing of SOCS2 in human liver 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 4321.
Abstract 4322: Atrx inactivation drives motility and dysregulates differentiation in glioma cells of origin through global epigenomic remodeling
Authors: Danussi Carla , Bose Promita , Silberman Pedro , Arnam John S. Van , Vitucci Mark , Tang Oliver , Heguy Adriana , Chan Timothy A. , Sulman Erik P. , Lang Frederick , Creighton Chad J. , Deneen Benjamin , Miller C Ryan , Picketts David J. , Kannan Kasthuri , Huse Jason T.
Published: August 2018

Comprehensive genomic profiling in cancer continues to reveal frequent alterations in epigenetic regulators, firmly implicating chromatin biology in the oncogenic process. For instance, genetic inactivation of the SWI/SNF chromatin regulator ATRX (α-thalassemia mental retardation X-linked) represents a defining molecular alteration in both adult and pediatric malignant glioma, and occurs frequently in other cancers as well. ATRX deficiency has been linked to a wide spectrum of physiological dysfunction, including aberrant gene regulation, abnormal telomere maintenance, genomic instability, and aneuploidy. However, the precise oncogenic mechanism(s) induced by ATRX deficiency remain unclear, particularly those involving epigenomic dysregulation. To model these events in putative glioma cells of origin, we inactivated Atrx in primary murine neuroepithelial progenitors (mNPCs). Atrx loss, especially when coupled with Tp53 inactivation, promoted mNPC motility while also modulating differentiation state and potential, effectively recapitulating characteristic disease phenotypes and molecular features. Moreover, these phenotypes correlated with altered gene expression profiles in functionally relevant molecular networks (e.g. cell differentiation and migration). Integrating these transcriptional changes with shifts in chromatin accessibility occurring with Atrx deficiency, along with genome-wide Atrx distribution as determined by ChIP-seq, revealed highly significant spatial correlations between differentially expressed genes, regions of altered chromatin compaction, and genomic sites normally occupied by Atrx. Finally, target genes mediating specific Atrx-deficient phenotypes in vitro exhibited similarly selective misexpression in ATRX-mutant human glioma tissues and cell lines. These findings demonstrate that, in appropriate cellular and molecular contexts, ATRX deficiency and its epigenomic sequelae are sufficient to induce disease-defining oncogenic phenotypes.

Citation Format: Carla Danussi, Promita Bose, Pedro Silberman, John S. Van Arnam, Mark Vitucci, Oliver Tang, Adriana Heguy, Timothy A. Chan, Erik P. Sulman, Frederick Lang, Chad J. Creighton, Benjamin Deneen, C Ryan Miller, David J. Picketts, Kasthuri Kannan, Jason T. Huse. Atrx inactivation drives motility and dysregulates differentiation in glioma cells of origin through global epigenomic remodeling [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 4322.