bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1704: Predicting survival and improving treatment for uterine serous carcinoma patients using USC78, a genomic risk score
Authors: Tran Lynn K. , Myers Emily K. , Mysona David P. , Tran Paul M. , Lee Wonsok , Wallbillich John J. , Kleven Daniel , Ghamande Sharad , She Jin-Xiong
Published: August 2019

Uterine serous carcinoma (USC) is a rare but particularly invasive histological subtype of uterine cancer, the most common gynecologic malignancy in developed countries. Though previous studies with small sample sizes have been conducted to identify transcriptomic and proteomic biomarkers for USC, none have resulted in a clinical assay for patient risk stratification. I have discovered a panel of 78 genes highly upregulated in poor prognosis patients from The Cancer Genome Atlas (TCGA) USC dataset. To create a composite score for our biomarker panel, I applied a machine learning algorithm (elastic net regression) to the TCGA USC gene expression data set to generate a model which outputs a linear predictor score (USC78) based on individual patients’ expression of the 78 genes from our signature. I then quantified gene expression from formalin-fixed paraffin embedded (FFPE) tumor tissue in our cohort of Augusta University (AU) patients and calculated their USC78 score to demonstrate the ability of this model to separate good and poor survival prognosis in a single-center, retrospective validation study. In both TCGA and AU, higher USC78 scores are associated with worse overall survival. This score is also able to risk stratify serous ovarian carcinoma patients in the TCGA data set, suggesting that USC78 may be an important prognosis prediction tool in serous gynecological cancers. Additionally, network analysis of these genes reveals increased TGF-B signaling correlates with the poor-prognosis USC78-high tumor expression profile. TGF-B inhibition has demonstrated chemosensitization effects in primary USC tumor cell lines. I aim to use USC78 to better understand the biological mechanisms behind poor survival prognosis in USC patients and identify potential therapeutic targets for USC. One such target is TGF-B inhibition, and further knockdown, overexpression, and inhibitor studies will be conducted to test whether TGF-B inhibition can improve USC patient prognosis.

Citation Format: Lynn K. Tran, Emily K. Myers, David P. Mysona, Paul M. Tran, Wonsok Lee, John J. Wallbillich, Daniel Kleven, Sharad Ghamande, Jin-Xiong She. Predicting survival and improving treatment for uterine serous carcinoma patients using USC78, a genomic risk score [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1704.
Abstract 1710: Role of Nurr1 in cervical cancer pathogenic pathway
Authors: Wan Kok Ting , Leung Thomas Ho Yin , Siu Michelle Kwan Yee , Ngan Hextan Yuen Sheung
Published: August 2019

Aims: To study the oncogenic role of Nurr1 in cervical cancer and elucidate the underlying mechanism.

Background: Orphan nuclear receptor related-1 protein (Nurr1) belonging to the NR4A family is important for carcinogenesis in multiple biological processes such as cell proliferation and apoptosis. It has been reported that the expression of Nurr1 is significantly correlated with various types of cancer, such as prostate and breast cancer. However, the significance of Nurr1 in cervical cancer etiology remains poorly understood.

Methods: Primary cancer cells were established from tumor tissues resected from cervical cancer patients. Six pairs of attached and spheroid cells derived from primary cervical cancer tissues were established namely CP-1A/CP-1S, CP-2A/CP-2S, CP-3A/CP-3S, CP-4A/CP-4S, CP-5A/CP-5S and CP-6A/CP-6S. A microarray was performed to identify differentially expressed genes in two pairs of attached and sphere cells, followed by validation with qPCR. Stable clones of cervical cancer cell lines with Nurr1 overexpression and knock-down were generated. The role of the selected gene was then functionally characterized by XTT assay, migration assay, spheroid formation assay, apoptosis assay, in vitro and in vivo tumorigenicity assay in cervical cancer cell lines. Downstream proteins of Nurr1 were detected by western blot.

Results: Microarray analysis showed that Nurr1 was highly expressed in sphere cells as compared with attached cells. Relative expression of Nurr1 mRNA in sphere cells was approximately 200-fold more than that in attached cells in qPCR validation. Cancer cell lines, in which Nurr1 was transiently knocked down by siRNA demonstrated an attenuation in proliferation rate, migration, spheroid formation and colony formation. By flow cytometric analysis, more cells were detected in early apoptotic stage with Nurr1 siRNA transfection as compared with NTC siRNA transfected cells, suggesting the role of Nurr1 in preventing cervical cancer cells from apoptosis. Western blotting also demonstrated that there was an increased expression of Bim and p21 in Nurr1 silenced cells. This may indicate the involvement of Nurr1 in modulating the transcription of pro-apoptotic and tumor suppressor genes.

Conclusion: Nurr1 is a potential oncogenic target in cervical cancer pathogenic pathway. Suppression of Nurr1 expression attenuated cancer progression and promoted apoptosis.

Citation Format: Kok Ting Wan, Thomas Ho Yin Leung, Michelle Kwan Yee Siu, Hextan Yuen Sheung Ngan. Role of Nurr1 in cervical cancer pathogenic pathway [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1710.
Abstract 172: BCL11A regulation of extracellular matrix genes may be necessary for invasion of triple-negative breast cancer
Authors: Ingles Natasha N. , Seachrist Darcie , Keri Ruth
Published: August 2019

Triple-negative breast cancer (TNBC) represents 15% of all breast cancers, yet conveys the poorest patient prognoses due to the lack of targeted therapies and the high incidence of metastatic recurrence. This prompt recurrence is thought to be due, in part, to the high proportion of cancer stem cells (CSC) within TNBC tumors. Therefore, understanding what regulates CSCs and metastasis will allow us to significantly reduce cancer-related deaths. One of the transcription factors that is upregulated in TNBC is BCL11A, which is critical for CSC biology. We have found that BCL11A is necessary for TNBC invasion and metastasis. However, the genes that BCL11A targets to control invasion and metastasis are unknown. To identify the BCL11A-regulated transcriptome, we performed RNA-seq analysis of MDA-MD-231 TNBC cells transiently transfected with BCL11A-targeted siRNA or a non-silencing control. Gene set enrichment analysis of the differentially expressed genes revealed enrichment of genes involved in extracellular matrix and matrisome genes. Notably, BCL11A silencing reduced expression of several matrix-metalloproteases, with MMP1 being the most significantly differentially expressed gene. Thus, we hypothesize that BCL11A promotes TNBC invasiveness and metastasis through regulation of MMP expression. We confirmed downregulation of MMP1 and MMP9 mRNA and protein in response to BCL11A silencing by both qRT-PCR and western blot in independent MDA-MB-231 cells, as well as in the HCC1143 TNBC cell line. MMP activity was also reduced when assessed by fluorokine assay and gelatin zymography, for MMP1 and MMP9, respectively. Furthermore, both BCL11A and MMP expression is increased in brain-seeking clones of MDA-MB-231 cells and MMP1 and 9 expression is also reduced with BCL11A silencing. Together, these data suggest that BCL11A regulates TNBC invasion by reducing the expression of MMP1 and MMP9. Current studies are directly assessing the functional impact of MMP regulation by BCL11A on invasion and metastatic progression of TNBC.

Citation Format: Natasha N. Ingles, Darcie Seachrist, Ruth Keri. BCL11A regulation of extracellular matrix genes may be necessary for invasion of triple-negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 172.
Abstract 1730: DACH1 gene deletion extends portraits of human prostate cancer
Authors: Jiao Xuanmao , Sante Gabriele Di , Li Zhiping , Rocco Agnese Di , Wang Min , Ertel Adam , McCue Peter A. , South Andrew P. , Cordon-Cardo Carlos , Stokes Matthew P. , Marra Marco , Jones Steven J. , Kossenkov Andrew , Pestell Richard G.
Published: August 2019

Purpose of the study: This study was conducted to define the role of Dachshund in prostate cancer, through assessing human prostate cancer samples and through genetic deletion in the mouse. Prostate cancer (PCa), the second leading cause of death in American men, is a genetically heterogeneous disease, likely representing distinct genetic drivers, with terminal events caused primarily by metastasis. Substratification of PCa into genetic subtypes, forms the basis of rational therapy for PCa. A better molecular understanding of the disease is necessary in order to develop novel targeted therapies of metastatic PCa. Known genetic drivers to tumor initiation include PTEN and NKX3.1 deletions, rearrangements of the TMPRSS2 gene to the oncogenic ETS transcription factor, ERG, and genetic predisposing factors include germline DNA-repair gene mutations. The DACH1 gene, initially cloned as an inhibitor of Elipse, the hyperactive epidermal growth factor (EGFR) in Drosophila, was found to be reduced in abundance in several malignancies including breast and prostate cancer.

Results: In order to determine whether the DACH1 gene is deleted or mutated in prostate cancer we interrogated the genomic sequencing analysis of over 490 patients from 5 population cohorts. Homozygous deletion of DACH1 was identified in 18% (N=61), 11% (N=136), 10% (N=492), 7% (N=103) and 3% (N=150) of prostate cancer in 5 distinct cohorts. The prevalence of DACH1 gene deep deletions was higher in the metastasis than in the primary tumors. The Transgenic Adenocarcinoma Mouse Prostate (TRAMP) transgenic, Dach1fl/fl, and Probasin-Cre, ROSA26mT/mG transgenic mice were used to generate a prostate epithelial cell specific Dach1 gene knockout mouse (Probasin-Cre-Dach1fl/fl ROSA26mT/mG-TRAMP) lines. Prostate specific deletion of the murine Dach1 gene enhanced progression of prostatic intraepithelial neoplasia (PIN), associated with increased prostate epithelial cell proliferation, epithelial mesenchymal transition (EMT), DNA damage and inflammation.

Conclusions: DACH1 gene deletion may define a distinct subclass of prostate cancer that may benefit from PARP inhibitors, and platinum-based chemotherapy.

Citation Format: Xuanmao Jiao, Gabriele Di Sante, Zhiping Li, Agnese Di Rocco, Min Wang, Adam Ertel, Peter A. McCue, Andrew P. South, Carlos Cordon-Cardo, Matthew P. Stokes, Marco Marra, Steven J. Jones, Andrew Kossenkov, Richard G. Pestell. DACH1 gene deletion extends portraits of human prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1730.
Abstract 1737: Association of inherited variants with chromosomal breaks in cancer
Authors: He Meng Xiao , Shahamatdar Sahar , Taylor-Weiner Amaro , Zack Travis , Ramachandran Sohini , Allen Eliezer Van
Published: August 2019

Background: Cancer cells frequently harbor genomic rearrangements and somatic copy number alterations that result from chromosome breaks, which vary in frequency across the genome. The distribution of breakpoint locations depends on features of the local genomic context, such as presence of certain retrotransposons. Moreover, inherited variants have been associated with specific chromosomal alterations in myeloproliferative neoplasms and in blood from cancer-free individuals. Here, we developed and applied computational methodology to broadly identify associations between inherited variants and the location of somatic breakpoints across cancers.

Methods: We analyzed TCGA Affymetrix Genome-Wide Human SNP Array 6.0 data from matched tumor-normal pairs (n = 5,923 samples remaining after quality control and selecting for individuals of predominantly European ancestry). Germline genotypes were called from normal samples using Birdseed v2. Allele-specific copy number analysis using both normal and tumor samples for each individual was performed using ASCAT v2.5.1. Breakpoints were inferred as the intervals between ASCAT-called somatic chromosomal segments.

We assigned individuals binary phenotypes corresponding to whether a somatic breakpoint was detected in non-overlapping 5 megabase bins of the genome. We then performed an association analysis between each phenotype and variants located on the same chromosome as the corresponding bin, adjusting for sex and ancestry via logistic regression with plink2. Analyses were performed separately for each disease type followed by a meta-analysis of association summary statistics with METAL.

Results and Conclusions: We identify several candidate associations between inherited variants and the presence of somatic breakpoints in specific regions of the genome (using a p-value cutoff of 5e-8 for each phenotype). Our results suggest that heritable genetic variation may be associated with risk of development of localized chromosome breaks in somatic cells. Ongoing work is aimed at validating the putative associations (for example using data from the PanCancer Analysis of Whole Genomes study) and identifying potential mechanisms associated with putative risk alleles.

Citation Format: Meng Xiao He, Sahar Shahamatdar, Amaro Taylor-Weiner, Travis Zack, Sohini Ramachandran, Eliezer Van Allen. Association of inherited variants with chromosomal breaks in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1737.
Abstract 1739: Diminished SKP1 or CUL1 expression induces chromosome instability in high-grade serous ovarian cancer precursor cells
Authors: Lepage Chloe C. , Nachtigal Mark W. , McManus Kirk J.
Published: August 2019

High-grade serous ovarian cancer (HGSOC) is the most common ovarian cancer subtype as well as the most lethal. High mortality rates associated with HGSOC are in part due to a lack of reliable early detection methods and effective therapies. Importantly, chromosome instability (CIN; an increased rate of chromosome gains or losses), is causally implicated in cancer development, progression, and resistance to treatment, and is just beginning to be evaluated in an HGSOC context. Overexpression of Cyclin E1 protein induces CIN and genomic amplification contributes to HGSOC pathogenesis in 20% of HGSOC patients. Cyclin E1 protein levels are normally regulated in a cell cycle-dependent manner by the SCF (SKP1-CUL1-FBOX) complex, an E3 ubiquitin ligase that targets substrates for proteolytic degradation. In cases where genomic amplification is absent, we hypothesize that loss of SCF complex function stemming from diminished expression of individual SCF complex components underlies increases in Cyclin E1 levels and induces CIN. The current study focuses on the SCF complex components SKP1 and CUL1 and characterizes their role in early HGSOC pathogenesis within a fallopian tube secretory epithelial cell model (FT; a cell of origin for HGSOC). Using siRNA-based approaches, we show that diminished SKP1 and CUL1 expression results in increased Cyclin E1 protein levels in FT cells. Using quantitative imaging microscopy techniques, we further identify statistically significant changes in CIN-associated phenotypes within the silenced populations, including changes in nuclear areas, increases in micronucleus formation (i.e. small DNA-containing bodies outside of the primary nucleus), and increases in numerical chromosome abnormalities identified in mitotic chromosome spreads. Thus, the current study explores the early origins of HGSOC and identifies SKP1 and CUL1 as two promising new molecular players that may contribute to CIN to drive FT cell transformation, drug resistance and disease recurrence.

Citation Format: Chloe C. Lepage, Mark W. Nachtigal, Kirk J. McManus. Diminished SKP1 or CUL1 expression induces chromosome instability in high-grade serous ovarian cancer precursor cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1739.
Abstract 1741: Identification of microsatellite instability in hematologic malignant cell lines
Authors: Bhattacharya Priyanjali , Chikan Naveed Anjum , Sharma Arati , Desai Dhimant , Annageldiyev Charyguly , Patel Pinaki , Patel Trupti N.
Published: August 2019

Defective DNA repair machinery which is a consequence of chromosomal or molecular alterations is associated with variety of diseases including solid tumors. Cytogenetic analysis is an important tool in diagnosis and treatment of hematologic malignancies. Our aim is to understand the role of microsatellite instability in initiation and progression of blood cancers. Here we hypothesize that leukemogenic cells may carry defective repair mechanisms that contribute to other genomic instabilities. In order to explore the possible role of microsatellite instability (MSI) in leukemia and lymphoma, eight different DNA repair genes involved in mismatch repair (MMR) and direct reversal of damage (DRD) were analyzed in K562 (CML), Molt-4 (T-ALL) and Daudi (Burkitt’s lymphoma) cell lines. Fragment analysis was carried out using novel primers considering repeat motifs enriched with (A)n, (CA)n and (AT)n for instability. Almost all the cell lines showed moderate to high level of microsatellite instability and exhibited a shift in selected repeat regions indicating possible loss (66.7%) (MSH2, MSH4, MSH6, MLH1, MLH3, PMS1) or gain (25%) (MSH3, MGMT) of nucleotide repeats as compared to normal DNA. The K562, Philadelphia negative CML cell line appeared to be microsatellite stable (MSS) for genes MSH2 and MLH1. Treatment induced microsatellite instability is known in leukemic patient cells as also seen in Molt-4 cells. Our present and previous observations suggest that MSI is a non-random event in leukemia/lymphoma which needs confirmatory future studies in newly diagnosed untreated patient samples. Considering the fact that both MSI and MSS represents two distinct molecular pathways of carcinogenesis, assessment of MSI, if added as distinct diagnostic criteria, can possibly refine early detection and help in identifying novel targets in management of hematologic malignancies.

Citation Format: Priyanjali Bhattacharya, Naveed Anjum Chikan, Arati Sharma, Dhimant Desai, Charyguly Annageldiyev, Pinaki Patel, Trupti N. Patel. Identification of microsatellite instability in hematologic malignant cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1741.
Abstract 1754: Monoubiquitinated γ-H2AX - a more specific biomarker of DNA double-strand breaks
Authors: Luczak Michal W. , Zhitkovich Anatoly
Published: August 2019

DNA double-strand brakes (DSBs) are a highly toxic form of DNA damage originating from many endogenous and exogenous sources. Cells employ specialized mechanisms for rapid detection and repair of these lesions. Histone H2AX phosphorylated at Ser139 (known as γ-H2AX) acts as a central platform for recruitment of other components of DSB repair. γ-H2AX assessment is currently the most widely used approach for quantitation of DSBs. However, γ-H2AX can be also induced by certain nongenotoxic stressors and during apoptosis, which can lead to false positive observations, especially in prolonged exposures. γ-H2AX undergoes RNF168-mediated K13/K15 monoubiquitination which is necessary for the recruitment of the downstream DSB repair factors. This ubiquitination event is rarely analyzed despite its functional importance. We examined formation and detection of γ-H2AX and its mono- (ub1) and diubiquitinated (ub2) forms in several human cell lines in response to mechanistically distinct inducers of DSBs and false-positive DSBs stressors. We found that γ-H2AX-ub forms were poorly detected using some common blotting procedures and antibodies. Under optimized conditions, γ-H2AX-ub1 was the predominant form accounting up to 80-90% of total γ-H2AX in primary human cells and its formation was strictly associated with the presence of DSBs. γ-H2AX and γ-H2AX-ub1 showed similar dose dependence and disappearance kinetics whereas H2AX-ub2 form was a more stable and dose-independent product. Unlike γ-H2AX, its ubiquitinated forms were not observed in the absence of DSBs (heat-shock or during apoptotic DNA cleavage). Apoptotic cells showed cleavage of the γ-H2AX-targeting E3 ubiquitin ligase RNF168. Our findings demonstrate that the ub1 form is a major fraction in the overall formation of γ-H2AX and its quantitation offers advantage in specificity for nonapoptotic DSBs.

Citation Format: Michal W. Luczak, Anatoly Zhitkovich. Monoubiquitinated γ-H2AX - a more specific biomarker of DNA double-strand breaks [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1754.
Abstract 1771: Role of Tsc2-mTORC1 signaling in homeostasis of intestinal epithelium and its relation to inflammation and cancer
Authors: Kotani Takenori , Setiawan Jajar , Murata Yoji , Saito Yasuyuki , Matozaki Takashi
Published: August 2019

Intestinal tumorigenesis driven by mutations in adenomatous polyposis coli gene requires the mammalian target of rapamycin complex 1 (mTORC1) activity, whereas the physiological roles of mTORC1 in the homeostasis of intestinal epithelial cells (IECs) remain virtually unknown. We here generated mice, in which tuberous sclerosis complex 2 (Tsc2), a negative regulator of mTORC1, was specifically ablated in IECs (Tsc2 CKO mice). Tsc2 CKO mice manifested the enhanced proliferative activity of IECs in intestinal crypts as well as the promoted migration of these cells along the crypt-villus axis. In vitro study showed that conditional ablation of Tsc2 in IECs promoted development of intestinal organoids, while mTORC1 inhibitor, rapamycin, diminished this phenotype. Tsc2 CKO mice also manifested the increased apoptotic rate of IECs as well as the increased ectopic Paneth cells, which are one of the major differentiated IECs. In addition, such conditional ablation of Tsc2 resulted in a suppressed expression of Wnt target genes in the colonic epithelium. Finally, we found that Tsc2 CKO mice manifested the increased susceptibility for dextran sulfate sodium (DSS)-induced colitis. Although mTORC1 and Wnt signaling pathways are thought to cooperated to promote oncogenesis in the intestinal epithelium, our results thus suggest that the mTORC1 signaling unexpectedly counter-regulates the Wnt signaling with the maintenance of intestinal homeostasis.

Citation Format: Takenori Kotani, Jajar Setiawan, Yoji Murata, Yasuyuki Saito, Takashi Matozaki. Role of Tsc2-mTORC1 signaling in homeostasis of intestinal epithelium and its relation to inflammation and cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1771.
Abstract 1778: 5’-nucleotidases are involved in the biology of human lung cancer cell lines
Authors: Raza Muhammad Zawwad , Cadassou Octavia , Cros-Perrial Emeline , Puisieux Alain , Dumontet Charles , Jordheim Lars Petter
Published: August 2019

Cytosolic-5’-nucleotidase II (cN-II) and ecto-5′-nucleotidase (CD73) are enzymes involved in the nucleotide metabolism by dephosphorylating intracellular and extracellular purine nucleotide monophosphates, respectively. Both enzymes have been shown to be involved in cancer by modifying anticancer drug activity, cancer cell biology and immune modulation. To increase the knowledge of cN-II and CD73 roles in cancer cell biology, we developed lung cancer cell models (NCI-H292) with a complete knockout of either or both enzymes using the CRISPR-Cas9 technique. These cell models are used to study cell proliferation and migration, cancer drug sensitivity, cellular behaviour and apoptosis using CFSE staining, IncuCyte relative confluence and wound healing assay, MTT assay, xCELLigence RTCA and caspase-3/7 apoptotic assay. SCID CB17 mice are also used to study the in vivo tumor growth using these cell models. Our results show that there is no significant difference in proliferation between different cell models exposed or not to different concentrations of either adenosine or AMP under normoxic and hypoxic conditions. On the other hand, using xCELLigence RTCA technique, we have observed that CD73-deficient cells have higher cell index as compared to CD73 expressing cells under basal conditions during the first 72 hours of culture. However, adenosine globally decreases the cell index of all NCI-H292 cells, unlike AMP, which influences the cell index of cN-II-deficient cells but not CD73-deficient cells. Under hypoxic condition, cN-II-deficient cells showed higher cell index than the other phenotypes in untreated conditions, whereas cells lacking CD73 showed higher sensitivity to adenosine or AMP. As compared to other phenotypes, CD73 deficient cells are also more sensitive towards vincristine but less sensitive towards mitomycin and fludarabine. These cells are also more prone to adapt to hypoxic conditions and exhibit slower migration rate than their corresponding control cells. However, the effect of adenosine and AMP on migration rate is higher on cN-II-deficient cells. Mice experiments showed no difference in in vivo tumor growth using these cell models. Overall, we show that both CD73 and cN-II are important players in the cell biology of lung cancer cells and in their response to purines. Upcoming experiments should help us understand the molecular mechanisms underlying these observed differences.

Citation Format: Muhammad Zawwad Raza, Octavia Cadassou, Emeline Cros-Perrial, Alain Puisieux, Charles Dumontet, Lars Petter Jordheim. 5’-nucleotidases are involved in the biology of human lung cancer cell lines [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1778.
Abstract 1781: Transcriptional activation of CBFβ by CDK11p110 is necessary to promote osteosarcoma cell proliferation
Authors: Feng Yong , Hornicek Francis , Duan Zhenfeng
Published: August 2019

Aberrant expression and activation of cyclin-dependent protein kinase (CDK) is a hallmark of cancer. CDK11 is a protein kinase in the CDK family and plays a crucial role in cancer cell growth and proliferation. However, the molecular mechanisms of CDK11 in osteosarcoma and CDK11 transcriptional regulated genes are largely unknown. In this study, we performed global transcriptional analysis using gene array technology to investigate the transcriptional role of CDK11 in osteosarcoma. The promoter luciferase assay, chromatin immunoprecipitation assay, and Gel Shift assay were used to identify direct transcriptional target of CDK11. Clinical significance relevance and function of CBFβ were further accessed in osteosarcoma tissue microarray and in osteosarcoma cell lines. We identified a transcriptional role of protein-DNA interaction for CDK11p110, but not CDK11p58, in the regulation of core-binding factor subunit beta (CBFβ) expression in osteosarcoma cells. The CBFβ promoter luciferase assay, chromatin immunoprecipitation assay, and Gel Shift assay confirmed that CBFβ is a direct transcriptional target of CDK11. High expression of CBFβ is associated with poor outcome in osteosarcoma patients. Expression of CBFβ contributes to the proliferation and metastatic behavior of osteosarcoma cells. These data establish CBFβas a mediator of CDK11p110dependent oncogenesis and suggest that targeting the CDK11-CBFβ pathway may be a promising therapeutic strategy for osteosarcoma treatment.

Citation Format: Yong Feng, Francis Hornicek, Zhenfeng Duan. Transcriptional activation of CBFβ by CDK11p110 is necessary to promote osteosarcoma cell proliferation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1781.
Abstract 1787: Selective activation of a PI3K catalytic isoform by G protein-coupled receptors in glioblastoma
Authors: Winn Abigail R. , Sheng Zhi
Published: August 2019

Objective: To determine how PI3K catalytic isoforms become dysregulated in glioblastoma.

Background: Recurrence in glioblastoma (GBM) is common and even more resistant to treatment than initial GBM. The Sheng Lab has previously identified that PIK3CB expression levels, but not other PI3K subtypes, are higher in recurrent GBM and are associated with GBM recurrence in newly diagnosed individuals. PI3KCB encodes for PI3K catalytic subunit p110β, and PI3K/AKT pathway activation is a well-known promoter of cell survival. Understanding mechanisms of PIK3CB activation in GBM may identify new therapeutic targets and improve GBM survival. Current knowledge of PI3K isoform activity in GBM is insufficient and contradictory. Given previous studies, it was hypothesized that G-Coupled Protein Receptor (GPCR) promoters preferentially activate PI3K/AKT in PIK3CBhigh GBM cells. Further, that knockdown of PIK3CB, but not other PI3K isoforms, inhibits PI3K/AKT reactivation in PIK3CBhigh cell lines.

Design/Methods: U87MG cells, an established p110βhigh GBM cell line, were starved overnight in serum free media, and then treated with growth factors, promoters or inhibitors of several previously hypothesized pathways. Cell lysates were collected and PI3K/AKT pathway activity was measured with immunoblotting. The ratio of pAKT/AKT/ACTB quantified PI3K/AKT pathway activity.

Results: Overnight serum starvation depleted pAKT, and incubation with 1% FBS restored pAKT signaling in a time dependent manner. Neither EGF nor FGF-2 activated PI3K/AKT signaling. Gefitinib, an EGFR inhibitor, failed to inhibit reactivation when cells were co-treated with 1% FBS. However, GPCR stimulation by Lysophosphatidic acid (LPA) increased pAKT/AKT ratios in a dose dependent manner. More significantly, Gallein, a GPCR inhibitor, suppressed PI3K/AKT signaling when cells were co-treated with 1% FBS. Selective PI3K inhibitors showed little PI3K/AKT reactivation at multiple doses.

Conclusions: These results suggest that GPCR signaling is responsible for PI3K/AKT pathway activation in p110βhigh cells.

Citation Format: Abigail R. Winn, Zhi Sheng. Selective activation of a PI3K catalytic isoform by G protein-coupled receptors in glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1787.
Abstract 1789: The importance of the phosphatase PRL family sub-cellular localization during the cellular cycle
Authors: O'Leary Anna K. , Blackburn Jessica
Published: August 2019

Acute lymphoblastic leukemia (ALL) is one of the most prevalent cancers diagnosed in adolescents. The survival rate for ALL is good during the initial diagnosis but is much less optimistic for patients who have relapsed ALL, due to limited treatment options. Additionally, the high dose cytotoxic chemotherapies given to ALL patients have long term adverse consequences. New and less toxic targeted therapies are needed for the treatment of ALL. We previously performed a large scale transplantation screen in zebrafish to identify genes associated with high relapse potential in ALL, and a top hit from this screen was the protein tyrosine phosphatase 4A3, PTP4A3 or PRL3. PRL3 overexpression in zebrafish enhances ALL progression and penetrance, and small molecule targeting of PRL3 in human cells rapidly induces apoptosis. The mechanisms through which PRL3 drive ALL progression and survival are unknown. PRL3 has a predicted nuclear localization site as well as a predicted prenylation motif, causing its localization to the plasma membrane. My preliminary data suggest that PRL3 shifts its localization between the plasma membrane and nucleus during the cell cycle. I am currently examining the localization of PRL3 and other PRL family members during different phases of the cell cycle and identifying signaling pathways that they may be involved with at these distinct sites to promote ALL progression, using CLIP-tag and SNAP-tag technologies that will allow me to image directly PRL localization in vitro. Biotin ligase approaches (Turbo-ID) will be used to determine the substrates of PRL activity and PRL interacting partners at these distinct locations. My project will provide scientific data that supports our hypothesis that PRL3 can transition between the membrane and nucleus, and plays a role in cell cycle. Identification of substrates of PRL3 that promote ALL progression may provide novel drug targets for this disease.

Citation Format: Anna K. O'Leary, Jessica Blackburn. The importance of the phosphatase PRL family sub-cellular localization during the cellular cycle [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1789.
Abstract 179: M6A demethylation enzyme FTO upregulates LDHA and promotes glycolysis and invasion and migration in oral squamous cell carcinoma
Authors: Hou Jinsong
Published: August 2019

Objective: To investigate the role of lactate dehydrogenase A (LDHA) in the glycolysis and invasion and metastasis of oral squamous cell carcinoma (OSCC), and to explore the molecular mechanism of fat mass and obesity-associated protein (FTO) regulating LDHA on the glycolysis and invasion and metastasis of OSCC.

Methods: Immunohistochemistry and RT-qPCR were performed to detect the expression of LDHA protein and gene in OSCC and adjacent non-cancerous tissues, and their correlation with clinical prognosis and pathological parameters were analyzed. Lentivirus-mediated shRNA silencing of LDHA and lactate dehydrogenase inhibitor (oxamate) were used to establish stable OSCC Cell lines HSC3 and SCC15, Cell Counting Kit-8 and colony formation assay were applied to evaluate cell proliferation, scratch test and Transwell assay were performed to detect cell migration and invasion, Glucose utilization assay and Lactate production assay were carried out to assess cell uptake of glucose and lactate production respectively. The molecular mechanism of FTO regulating LDHA on glycolysis, invasion and metastasis of oral squamous cell carcinoma was further explored, OSCC cell lines HSC3 and SCC15 transfected with siRNAs to silence FTO expression, EpiQuik™ m6A RNA Methylation Quantification Kit was applied to assess the m6A% in total RNA, the effect of FTO on LDHA expression was detected by Western blot and RT-qPCR. The biological function of FTO on oral squamous cell carcinoma cells was demonstrated by cell experiments in vitro.

Results: Compared with adjacent non-cancerous tissues, LDHA was highly expressed in OSCC (P<0.001), which was closely related to the differentiation, clinical stage and T stage of OSCC (P<0.05). Silencing LDHA, FTO or treating oxamate in HSC3 and SCC15 cells reduced glucose consumption, lactate production and cell proliferation, invasion and migration (P<0.05). In addition, after FTO silencing, the m6A level of total RNA increased (P<0.05), while the expression level of LDHA protein and mRNA decreased (P<0.01).

Conclusion: FTO may promote the expression of LDHA protein and mRNA in a m6A-dependent manner and enhance the ability of glycolysis, proliferation, invasion and

metastasis of OSCC.

Note: This abstract was not presented at the meeting.

Citation Format: Jinsong Hou. M6A demethylation enzyme FTO upregulates LDHA and promotes glycolysis and invasion and migration in oral squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 179.
Abstract 1800: miR-376b and miR-4668 predict therapeutic response to sunitinib in metastatic renal cell carcinoma
Authors: Kovacova Julia , Juracek Jaroslav , Poprach Alexandr , Buchler Tomas , Fiala Ondrej , Radova Lenka , Svoboda Marek , Kopkova Alena , Vecera Marek , Slaby Ondrej
Published: August 2019

Introduction: Metastatic renal cell carcinoma (mRCC) is routinely treated with sunitinib, a tyrosine kinase inhibitor (TKI) of VEGF signalisation. Although disease eventually progresses in most mRCC patients, length of progression-free survival (PFS) is ranging from few weeks up to more than 24 months. Patients with initial resistance to sunitinib could be redirected to other therapeutical options, as there are several other TKIs available for use in second and third line. However, without a reliable biomarker no result can be predicted. MicroRNAs (miRNAs) belong to class of short non-coding RNAs and could serve as biomarkers of therapy response due to their unique character and presence in tissues and body fluids. Their biomarker potential has been discussed concerning many diseases including mRCC, but current knowledge is very weak, has several discrepancies and is acquired on relatively small cohorts.

Material and method: Candidate microRNAs have been chosen based on global expression profiling using Affymetrix GeneChip 4.0 in 47 samples of FFPE mRCC tissue of patients treated with sunitinib (good response n=25, PFS longer than 17 months; poor response n=22, PFS shorter than 9 months). Validation was performed using qRT-PCR TaqMan assays on an independent cohort of 136 FFPE samples from mRCC patients treated with sunitinib with variable length of PFS. Local ethical committees at all involved centres approved the study protocol. The study was performed according to the World Medical Association Declaration of Helsinki and all patients signed an informed consent.

Results and discussion: Of all tested miRNAs, expression of miR-4668-5p and miR-376b was the most significantly deregulated in non-responding patients with high statistical significance (p>0,005) and combined AUC higher than 0,8. Although other independent validations are necessary, microRNAs presented here seem to be very promising as tools for therapy personalization.

Conclusion: We have successfully validated miR-4668-5p and miR-376b to have predictive potential in estimation of therapeutical response to sunitinib. Further functional analyses could shed more light on their involvement in development of therapy resistance. This work was supported by Ministry of Health of the Czech Republic, grant nr. NV18-03-00554. All rights reserved.

Citation Format: Julia Kovacova, Jaroslav Juracek, Alexandr Poprach, Tomas Buchler, Ondrej Fiala, Lenka Radova, Marek Svoboda, Alena Kopkova, Marek Vecera, Ondrej Slaby. miR-376b and miR-4668 predict therapeutic response to sunitinib in metastatic renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1800.
Abstract 1803: Openarray profiling of the systemic change of microRNAs in rhenium-188 liposome treated human head and neck cancer cells
Authors: Lin Bing-Ze , Chang Chun-Yuan , Chang Chih-Hsien , Lee Yi-Jang
Published: August 2019

Purpose188Re-liposome has been used for evaluating the theranostic efficacy in various human cancers. However, whether the therapeutic efficacy of 188Re-liposome directly related to microRNAs regulation and specific signaling pathways are largely unknown. In this project, we investigated the 188Re-liposome induced systemic microRNA expressive responses in human head and neck cancer (HNSCC) cells.

Methods

The FaDu-3R cells were implanted into the nude mice at the position of buccal cavity. After two weeks of implantation, the tumors were palpable and the bioluminescent imaging indicated the location of this orthotopic tumor model. The PEGylated 188Re-liposome was intravenously injected single and dual dose respectively. We harvested all of the tumors after 188Re-liposome injection one month ago, and used Taqman Openarray MicroRNA (Thermo, USA) to detect the expression of miRNAs in different groups. The dataset we observed was further analyzed by using Ingenuity Pathway Analysis (IPA) software to establish the molecular signaling pathway of 188Re-liposome radiopharmaceutical.

Results

Quantification of the photon signals showed that the tumor growth of the dual 188Re-liposome treated mice was slower than that of control ones. The survival rate of 188Re-liposome treated animals were also better than control ones. The microRNA Openarray showed that, compared to untreated control, 17 miRNA were significantly changed in expressive levels over 5 fold after 188Re-liposome treatment. Subsequently, we used IPA to determine the role of these miRNA, and found these miRNA with oncogenic and tumor suppressive properties were down-regulated and up-regulated, respectively. Among them, we focused on miR-152-5p that is closed related to cancer occurrence and development, and showed that knockdown of miR-152-5p could decreased the effects of 188Re-liposome on suppressing the growth of HNSCC xenograft tumors.

Conclusion

HNSCC tumor model treated with dual dose of 188Re-liposome exhibited significant response to this radiopharmaceutical. We also found that 17 miRNA could be gsignificantly re-regulated, and miR-152-5p is one example to be involved in mediating the efficacy of 188Re-liposome. Targeting on specific miRNA may enhance the application value of 188Re-liposome.in cancer treatment.

Citation Format: Bing-Ze Lin, Chun-Yuan Chang, Chih-Hsien Chang, Yi-Jang Lee. Openarray profiling of the systemic change of microRNAs in rhenium-188 liposome treated human head and neck cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1803.
Abstract 1813: Depletion of PCAT1 in head and neck cancer cells limits tumor growth by regulating c-Myc, AKT1 and MAPK signalling pathways
Authors: Sur Subhayan , Steele Robert , Ray Ratna B.
Published: August 2019

Head and neck squamous cell carcinoma (HNSCC) represents one of the most common malignancies worldwide with a high mortality rate mainly due to lack of early detection markers, frequent metastasis and aggressive phenotype. Recently, long non-coding RNAs (lncRNAs) have been shown to have important regulatory roles in human cancers. Up-regulation of prostate cancer-associated transcript 1 (PCAT1) in TCGA dataset of HNSCC was noted. In this study, we aimed to evaluate role of the PCAT1 in HNSCC. During validation, we observed up-regulation of PCAT1 in HNSCC patient samples and cell lines. siRNA mediated knock-down of PCAT1 significantly reduced cell proliferation, sphere formation and wound healing in HNSCC cell lines (JHU29 and Cal27). Mechanistic study revealed a significant down-regulation of c-Myc and AKT1 in both RNA and protein levels upon knockdown of PCAT1 in HNSCC cell lines. In-silico analysis suggested a significant positive correlation between PCAT1 and c-Myc or PCAT1 and AKT1 in HNSCC cell lines. Subsequently, we observed modulation of MAPK signalling by activation of p38 and apoptosis signal-regulating kinase 1 (ASK1) upon knockdown of PCAT1, and induction of apoptosis as evident of caspase 9 activation and PARP cleavage. Introduction of PCAT1 into HNSCC xenograft tumors displayed a reduction in tumor volume. Together our data demonstrated an important role of the PCAT1 in HNSCC progression and has potential as a therapeutic candidate.

Citation Format: Subhayan Sur, Robert Steele, Ratna B. Ray. Depletion of PCAT1 in head and neck cancer cells limits tumor growth by regulating c-Myc, AKT1 and MAPK signalling pathways [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1813.
Abstract 1816: Tumor suppressor LINC02487 inhibits oral cancer cell migration and invasion through directly binding protein USP17
Authors: Feng Lu , Qiu Feng , Zhang Jianjun , Sun Minglei , Chen Wantao , Qiu Weiliu
Published: August 2019

Objective: The aim of this study is to explore the functions and mechanisms of long non-coding RNA LINC02487 in oral squamous cell carcinoma(OSCC).

Materials and methods: Relative expression level of LINC02487 in OSCC cell lines and samples were examined by qRT-PCR. Intracellular localization was done using RNA fluorescence in situ hybridization. LINC02487 was cloned into pCMV-puro vector, and transfected into OSCC cell lines HN6 and HN30 by lentivirus. Cell functions as proliferation, apoptosis, cell cycle, migration and invasion were examined subsequently. RNA binding protein was explored by ChIRP-MS. And protein expression was determined by western-blotting assay.

Results: Previous data showed a downregulation of LINC02487 in OSCCs. Here we confirmed the lower expression of LINC02487 in 6 OSCC cell lines comparing with immortalized normal oral epithelial cell lines, and in 50 OSCC samples comparing with paired adjacent normal tissue. LINC02487 was located in cytoplasm, aggregated around nuclei membrane. Overexpression of LINC02487 significantly suppressed cell migration and invasion, increased the ratio of cells in G1 phase, with a minor influence on cell proliferation and apoptosis. Furtherly, LINC02487 was found directly binding to USP17, a deubiquitinating enzyme, and regulating cell migration and invasion through USP17 expression.

Conclusion: Our study confirmed that long non-coding RNA LINC02487 was downregulated in paired OSCC samples and cell lines. It acts as a tumor suppressor through its directly binding protein USP17.

Citation Format: Lu Feng, Feng Qiu, Jianjun Zhang, Minglei Sun, Wantao Chen, Weiliu Qiu. Tumor suppressor LINC02487 inhibits oral cancer cell migration and invasion through directly binding protein USP17 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1816.