bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 2596: Toll like receptor 4 as a potential DNA repair modulator in U87MG-GBM cells
Authors: Moretti Isabele F. , Lerario Antonio , Oba-Shinjo Sueli M. , Marie Suely K. , Lima Marina Trombetta 
Published: September 2020

Abstract
Toll like receptor 4 (TLR4) has a dual role in cellular fate, depending on the stimuli and microenvironment context. Its activation may promotes cell survival and proliferative phenotype via MyD88 with NFkB activation, or leads to cell death via TRIF. Glioblastoma (GBM), grade IV astrocytoma, is the most common and aggressive primary malignancy in the brain. Surgical tumor cytoreduction followed by radiotherapy and adjunct chemotherapy with Temozolomide (TMZ) have been considered the standard of care for GBM. And, in spite of several combinatory rescue therapeutic trials, the median overall survival of GBM patients continue to be approximately 15 months. In this context, new therapeutic strategies are urgent, and immune modulation has demonstrated optimistic results in several tumor types. Therefore, the aim of this study was to analyze the role of TLR4 in human astrocytomas. First, we evaluated TLR4 expression in 140 human astrocytoma of different grades of malignancy, by qRT-PCR. TLR4 molecular signaling pathways were investigated through LPS stimuli in U87MG-GBM cell line. In astrocytoma samples, TLR4 was up-regulated when compared to brain non-neoplastic samples (p&lt;0.05, Kruskall Wallis and Dunn tests), with increased expression in Mesenchymal compare to Classical and Proneural subtypes. U87MG cells stimulation with LPS (10ug/ml) lead to NF-kB (p65) nuclear translocation after 12hr through western blot and immunofluorescence, which was unexpected as such translocation through the MyD88/TRAF6 canonical pathway occurs within 30min to 2hr after LPS stimulation. TRIF, IL1B, RIPK1/3 expressions were increased after LPS stimulation, suggesting an activation of inflammasome and ripoptosome pathways. Moreover, the combined LPS treatment with TMZ showed a higher apoptotic rate after 48hr when compared to TMZ alone. However, the transcriptomic analysis by RNASeq of U87MG cells after LPS stimulation showed an upregulation of DNA repair genes, particularly MBD4 and PLAUR. Both genes are upregulated in mesenchymal GBM subtype in the TCGA RNASeq dataset, and they present correlated expression with TLR4 (r=0.406, p=0.004 and r=0.338, p=0.019 by Spearman's test). Our results suggest that the anti-tumoral effect by TLR4 pathway activation by enhancement of tumor apoptosis might be more efficient with combined inhibition of DNA repair cascade. In fact, OSI-296 and BKM-120 have shown to decrease MBD4 expression, and increased overall survival was observed in a phase II clinical trial for solid tumor.
Citation Format: Isabele F. Moretti, Antonio Lerario, Sueli M. Oba-Shinjo, Suely K. Marie, Marina Trombetta Lima. Toll like receptor 4 as a potential DNA repair modulator in U87MG-GBM 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 2596.
Abstract 2601: Expression and clinicopathological association of RABs in pancreatic cancer
Authors: Anand Shashi , Zubair Haseeb , Khan Mohammad Aslam , Singh Seema , Singh Ajay P. 
Published: September 2020

Abstract
Rab proteins form the largest subfamily of small GTPases and play an indispensable role in the regulation of vesicle trafficking. They control a wide variety of endocytic, transcytic and exocytic transport pathways and thus play important roles in cellular growth and behavior. Altered expression and functions of Rab proteins have been reported in various malignancies and implicated in tumorigenic processes. Pancreatic cancer (PC) is a highly lethal malignancy and molecular mechanisms associated with its pathobiology are relatively less understood. Here we mined The Cancer Genome Atlas (TCGA) Database for differential expression and clinicopathological association of RAB genes in pancreatic cancer. Out of the 62 RAB genes analyzed, 5 (RAB3A, RAB26, RAB25, RAB21, and RAB22A) were found to have significant upregulation in cancer, while 5 (RAB6B, RAB8B, RABL2A, RABL2B and RAB32) exhibited significant downregulation. Racially disparate expression was also reported for some RABs. RAB3A and RAB26 had significantly higher and lower expression, respectively, in African American (AA) and Asian PC as compared to Caucasian Americans (CA) PC, whereas RAB25 was significantly higher in Asian PC than CA PC. No clear trend of altered expression was observed with increasing stage and grade, although stage- and grade-specific differences were recorded for some RABs. RAB21, RAB22A, and RAB8B showed significantly higher expression in PC from middle-aged patients (41-60 years) compared to older age groups, but no gender-associated differential expression was observed. PC from occasional drinkers had significantly higher expression of RAB21 compared to daily or weekly drinker, whereas RAB25 expression was significantly higher in social drinkers, compared to occasional ones. Expression of RABL2A was significantly reduced in PC of diabetic patients whereas RAB26 was significantly down in pancreatitis patients. Most importantly, a significant association of high expression of RAB21, RAB22A and RAB25, and low expression of RAB6B, RABL2A, and RABL2B was observed with poorer survival of PC patients. Together, our study is suggestive of potential diagnostic and prognostic significance of RABs in PC and warrants further investigations to define their functional and mechanistic significance.
Citation Format: Shashi Anand, Haseeb Zubair, Mohammad Aslam Khan, Seema Singh, Ajay P. Singh. Expression and clinicopathological association of RABs in pancreatic 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 2601.
Abstract 2611: Comparison of 3D culture conditions to develop an in vitro model for breast cancer cell emboli formation
Authors: Vitale Ariana , Sterneck Esta , Kuppusamy Balamurugan 
Published: September 2020

Abstract
Inflammatory breast cancer (IBC) is a rare type of breast cancer accounting for 2-4% of all breast cancer patients, but accounts for 7-10% of breast cancer related mortality. IBC is characterized by extensive skin invasion and the formation of tumor cell emboli within the breast and dermal lymphatic system. Characteristics of emboli include cell to cell adhesions mediated by E-cadherin, encoded by the gene CDH1. Studies from our laboratory show that CEBPD is expressed in IBC patient emboli and promotes emboli formation in vitro. Mechanistically we found that CEBPD promotes cell-cell adhesion within emboli at least in part by supporting the expression of E-cadherin adhesion complex proteins. In this study, we compared an in vitro model for intra-lymphatic emboli formation with conditions of tumor sphere formation to begin to address the role of fluid shear in emboli formation and emboli-specific molecular events, if any. We conducted experiments under slow rocking and non-rocking culture conditions with 4 different culture media. We found that the mechanical shear forces (achieved by low speed rocking) generated large, well-compacted emboli-like structures within 24 hours of suspension culture. In the absence of fluid shear, the cells assembled into smaller and more irregular aggregates. In agreement with this, our preliminary data suggest that E-cadherin expression is modestly higher under fluid shear conditions in addition to overall increase under 3D conditions. We are now validating the mechanisms for higher expression of emboli in 3D culture conditions and are designing experiments to address the functional relevance of the pathways for IBC emboli biology. Understanding the molecular mechanisms of emboli formation may provide valuable insights into the uniquely aggressive nature of IBC.
Citation Format: Ariana Vitale, Esta Sterneck, Balamurugan Kuppusamy. Comparison of 3D culture conditions to develop an in vitro model for breast cancer cell emboli formation [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 2611.
Abstract 264: Long noncoding RNA LNC01534 regulates cancer stem cells in colorectal cancer
Authors: Ohtsuka Masahisa , Xin Wu , Ichihara Momoko , Hirose Haruka , Takahashi Hidekazu , Matsuda Chu , Akamatsu Hiroki , Mizushima Tsunekazu , Doki Yuichiro , Yamamoto Hirofumi 
Published: September 2020

Abstract
Introduction The clinical importance of cancer stem cells (CSCs) has been widely reported, and clinical studies targeting CSCs are in progress. Additionally, accumulating evidence suggests that long noncoding RNAs (lncRNAs) have essential roles in cancer initiation and progression, while deregulated lncRNA expression is found in a variety of cancer types. In this study, we focused on lncRNAs regulating CSCs in colorectal cancer (CRC).
Objectives To identify the lncRNAs associated with CSCs and to evaluate their clinical relevance and function.
Materials and Methods We separated CSCs and other cells in CRC using the ornithine decarboxylase (ODC) degron system to visualize CSCs based on the correlation between low proteasome activity and CSC properties. We performed comprehensive gene analysis comparing CSCs and other cells to identify a candidate lncRNA that regulates the stemness of CRC cells. Finally, we analyzed the clinical relevance and function of the lncRNA.
Results The lncRNA LNC01534 showed high expression in CSCs compared with the level in other CRC cells. Using gene expression and clinical data from the Cancer Genome Atlas Project for CRC patients, we showed that LNC01534 was associated with overall survival in CRC. LNC01534 was also associated with overall survival and disease-free survival in 187 separate CRC samples. Functional studies revealed that the loss of LNC01534 function reduced proliferation and arrested the cell cycle in CRC cells. Furthermore, genes that underwent expression changes upon the knockdown of LNC01534 were evaluated by RNA-seq. This showed that the knockdown of LNC01534 affected the expression of genes related to endoplasmic reticulum (ER) stress and autophagy. Specifically, the expression levels of HSPA5, LC3-I, p62, and Beclin-1–genes related to ER stress and autophagy–increased upon suppressing LNC01534.
Conclusions We demonstrated the clinical and biological relevance of LNC01534 in association with CSCs, and discovered the effect of LNC01534 on ER stress and autophagy to maintain stemness in CRC.
Citation Format: Masahisa Ohtsuka, Wu Xin, Momoko Ichihara, Haruka Hirose, Hidekazu Takahashi, Chu Matsuda, Hiroki Akamatsu, Tsunekazu Mizushima, Yuichiro Doki, Hirofumi Yamamoto. Long noncoding RNA LNC01534 regulates cancer stem cells in colorectal 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 264.
Abstract 2688: Pan-cancer analysis of clonality and the timing of systemic spread in paired primary tumors and metastases
Authors: Hu Zheng , Li Zan , Ma Zhicheng , Curtis Christina 
Published: September 2020

Abstract
Metastasis is the primary cause of cancer-related deaths, but the natural history, clonal evolution and patterns of systemic spread are poorly understood. We analyzed exome sequencing data from 458 paired primary tumors (P) or metastasis (M) samples from 136 breast, colorectal and lung cancer patients, including both untreated (n=98) and treated (n=101) metastases. We find that treated metastases often harbored private driver gene mutations whereas untreated metastases did not, suggesting the acquisition and selection of additional mutations is not required for untreated metastases and treatment promotes clonal evolution. Additionally, the ratio of nonsynonymous versus synonymous mutations (dN/dS) showed decreased selection pressures in the metastasis relative to primary tumors. Polyclonal seeding was common in lymph node metastases (n=19/35, 54%; mostly untreated) and untreated distant metastases (n=20/70, 29%), but less frequent in treated metastases (n=9/90, 10%). Mathematical modeling shows that the low number of metastasis-private clonal mutations is consistent with early metastatic seeding, which commonly occurred several years prior to diagnosis in breast (2.4 years, range 0-3.3), lung (3.6 years, range 2.8-3.7) and colorectal (4.1 years, range 3.1-4.6) cancers. Thus, this pan-cancer analysis reveals early systemic spread in three common cancer types. Further, these data suggest that the natural course of metastasis is selectively relaxed relative to early tumor development and that metastasis-private mutations are not drivers of cancer spread but are instead associated with drug resistance.
Citation Format: Zheng Hu, Zan Li, Zhicheng Ma, Christina Curtis. Pan-cancer analysis of clonality and the timing of systemic spread in paired primary tumors and 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 2688.
Abstract 2732: NLRP12 suppresses colorectal cancer progression and invasion via downregulation of the Wnt/b-catenin pathway
Authors: Khan Shahanshah , Zaki Hasan 
Published: September 2020

Abstract
Metastatic CRC is associated with increased mortality, underscoring the importance of understanding the molecular mechanism of CRC invasion and metastasis. We recently demonstrated that mice deficient in NLRP12, an innate immune sensor, are susceptible to CRC with increased incidence of invasive adenocarcinoma. Here, we investigated the molecular mechanism of NLRP12-mediated suppression of cancer progression and invasion using mouse models and in vitro biochemical studies. Nlrp12−/− mice treated with azoxymethane plus dextran sodium sulfate developed higher tumor burden and invasive adenocarcinoma compared to WT mice. RNA-seq analysis showed significantly higher levels of protooncogenes, matrix metalloproteinases (Mmp 3, 7,8, 10, 12), and epithelial to mesenchymal transition (EMT) markers (Vim, Fn1, Foxc2, Zeb2, Ezh2) in Nlrp12−/− tumors. To understand the underlying mechanism of increased expression of cancer promoting genes in Nlrp12−/− tumors, we analyzed activation of various cell signaling pathways. To our surprise, there was no major difference in the activation NF-κB, ERK, JNK, and STAT3, which have previously been shown to be downregulated by NLRP12, between WT and Nlrp12−/- tumors. However, we observed strikingly higher levels of β-catenin and expression of Wnt target genes (Ctnnb1, Axin2, Yap1, Tcf3, Lgr5, etc.) in Nlrp12−/− tumors. Consistently, deletion of NLRP12 in APCmin/+ mice led to higher polyp development and activation of the Wnt/β-catenin pathway. Notably, hyperactivation of the Wnt/β-catenin pathway is a hallmark of CRC and about 80% of CRC patients carry mutations in the Wnt/β-catenin pathway. To further explore the link between NLRP12 and the Wnt/β-catenin pathway, we overexpressed NLRP12 in HEK293T cells and CRC cell line MC38. NLRP12-overexpressed cells showed reduced activation of β-catenin and expression of Wnt target genes following stimulation with Wnt3a. An opposite result was observed when NLRP12 was knocked out from these cell lines. Consistently, Nlrp12-KO MC38 cells exhibited increased proliferation, colony formation, and migration. These data for the first time demonstrate that NLRP12 is an intrinsic negative regulator of the Wnt/β-catenin pathway. Overall, these findings reveal a novel CRC suppression pathway involving NLRP12 and suggest that the NLRP12/β-catenin signaling axis could be therapeutically targeted for the treatment of CRC invasion and metastasis.
Citation Format: Shahanshah Khan, Hasan Zaki. NLRP12 suppresses colorectal cancer progression and invasion via downregulation of the Wnt/b-catenin pathway [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 2732.
Abstract 2736: AKT1E17K activates focal adhesion kinase and promotes melanoma brain metastasis
Authors: Kircher David A. , Trombetti Kirby A. , Silvis Mark R. , Parkman Gennie L. , Fischer Grant M. , Angel Stephanie N. , Stehn Christopher M. , Strain Sean C. , Grossmann Allie H. , Duffy Keith L. , McMahon Martin , Davies Michael A. , Mendoza Michelle C. , VanBrocklin Matthew W. , Holmen Sheri L. 
Published: September 2020

Abstract
Hyper-activation of the PI3K/AKT signaling pathway occurs in most metastatic melanomas and increased PI3K/AKT pathway activity correlates with disease progression. The serine/threonine kinase, AKT, represents a major signaling hub within the pathway and consists of three highly conserved paralogs that have both distinct and overlapping functions. Activating mutations of AKT1 and AKT3 occur in human melanoma but their role in melanoma formation and metastasis remains unclear. Using an established melanoma mouse model, we evaluated the ability of constitutively active E17K, E40K, or Q79K mutants of each AKT paralog to promote tumor progression and metastasis in the context of BRAFV600E expression and loss of Cdkn2a and Pten. Expression of AKT1E17K promoted highly aggressive melanomas that metastasized to the lungs and brain. This metastatic phenotype was not significantly observed in the case of other mutant AKT-positive tumors, suggesting that the AKT paralogs have distinct, non-overlapping roles in the development of melanoma metastases. AKT1E17K-positive tumors showed AKT1E17K-dependent up-regulation of multiple focal adhesion (FA) factors, which are key components of focal adhesions and established stimulators of cell motility, as well as phosphorylation of focal adhesion kinase (FAK). Ectopic expression of AKT1E17K in non-metastatic melanoma cells increased cell invasion, a phenotype abrogated by pharmacological inhibition of AKT or FAK. These findings strongly suggest that one mechanism by which AKT1 promotes melanoma metastasis is through regulation and activation of proteins involved in focal adhesions. This has important implications for the development of therapeutic strategies aimed at preventing or treating disseminated disease.
Citation Format: David A. Kircher, Kirby A. Trombetti, Mark R. Silvis, Gennie L. Parkman, Grant M. Fischer, Stephanie N. Angel, Christopher M. Stehn, Sean C. Strain, Allie H. Grossmann, Keith L. Duffy, Martin McMahon, Michael A. Davies, Michelle C. Mendoza, Matthew W. VanBrocklin, Sheri L. Holmen. AKT1E17K activates focal adhesion kinase and promotes melanoma brain metastasis [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 2736.
Abstract 2738: Role of KLF8 for drug-resistant metastasis of breast cancer
Authors: Hao Jie , Lu Heng , Mukherjee Debarati , Sun Aiqin , Yu Lin , Zhao Jihe 
Published: September 2020

Abstract
Metastasis and drug resistance are intertwined processes that are responsible for the vast majority of patient deaths from breast cancer. The underlying mechanisms still need to be investigated extensively. We previously demonstrated that KLF8 activates CXCR4 transcription in metastatic breast cancer. Here, we report a novel signaling mechanism that turns a localized metastasis into multiple organ metastases in response to chemotherapy. We show that KLF8 expression in breast cancer cells can be over-induced by chemotherapeutic drugs. Data from large-cohorts of patients indicates that post-chemotherapy there is a close correlation between the aberrant high levels of KLF8 and CXCR4, and this correlation is well associated with drug resistance, metastasis, and poor prognosis. To mimic this upregulation, we overexpressed KLF8 or CXCR4 in a human breast cancer cell line known to metastasize to the lungs only after i.v. injection in the nude mice. Surprisingly, we found that these KLF8 or CXCR4 overexpressing cells metastasized all over the body after orthotopically implanted. Tube formation assay, Ki67 staining, and western blotting revealed that KLF8 or CXCR4 overexpression promoted angiogenic vascular growth involving increased expression and secretion of VEGF protein. We also performed the matrigel-on-top 3-D culture to mimic the colonization step of metastasis and found that KLF8 or CXCR4 overexpression strongly enhances the formation of filopodium-like protrusions and proliferation via CXCR4 stimulation. Taken together, these results suggest that the chemo-induction of KLF8 upregulation is critical for drug resistance and systemic metastasis through enhanced tumor angiogenesis and colonization via CXCR4 over-activation. This discovery identifies the KLF8-CXCR4 signaling axis as a potentially new therapeutic target for breast cancer.
Citation Format: Jie Hao, Heng Lu, Debarati Mukherjee, Aiqin Sun, Lin Yu, Jihe Zhao. Role of KLF8 for drug-resistant metastasis of breast 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 2738.