bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2476: Discovery of an androgen-responsive long noncoding RNA that associates with progression of ERG-overexpressing prostate cancers
Authors: Kohvakka Annika , Shcherban Anastasia , Kivinummi Kati K. , Annala Matti , Urbanucci Alfonso , Nykter Matti , Visakorpi Tapio
Published: August 2018

Androgen receptor (AR) signaling pathway has an important role in the growth and development of normal prostate, but also in tumorigenesis and progression of prostate cancer (PC). Although the mechanisms of AR signaling have been widely studied and utilized for treatment in advanced PC, the exact role of AR in development of primary PC is unclear. Former studies have found that AR cistrome is reprogrammed during tumorigenesis to bind novel genomic loci by master regulators, including the ETS family transcription factor ERG. While many AR-induced target genes are known, the effect of AR signaling on regulation of long noncoding RNAs (lncRNAs) is poorly understood, especially in the context of PC progression. Previously, we discovered multiple novel PC-associated transcripts (PCATs) to be aberrantly expressed in PC. Here, we evaluated the expression of 39 Tampere PCATs (TPCATs) in 87 radical prostatectomy specimens using high-throughput real-time PCR, and studied their association with time to PSA progression after prostatectomy. Six TPCATs were significantly associated with time to PSA progression, and four of them also associated with extracapsular extension. In addition, we assessed the expression of TPCATs in the TCGA prostate adenocarcinoma cohort, and found many to be correlated with ERG expression. Moreover, publicly available AR ChIP-seq data from PC tumors indicated that several ERG-associated TPCATs had AR-binding sites on their promoters, some of which overlapped with ERG binding sites. Most notably, we found one progression-associated TPCAT that was regulated by AR in an androgen-sensitive manner according to AR siRNA knockdown and DHT stimulation experiments in vitro. The same TPCAT was also highly associated with overexpression of ERG, and further validated to be a highly PC-specific lncRNA that was abundantly expressed in primary PCs. Taken together, these findings give more insight into the role of AR cistrome in the regulation of lncRNAs in primary PC, and introduce a potential novel prognostic marker to be used in early detection of aggressive PC.

Citation Format: Annika Kohvakka, Anastasia Shcherban, Kati K. Kivinummi, Matti Annala, Alfonso Urbanucci, Matti Nykter, Tapio Visakorpi. Discovery of an androgen-responsive long noncoding RNA that associates with progression of ERG-overexpressing prostate cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2476.
Abstract 2479: LncRNA GAS8-AS1 directs epigenetic modulation of the tumor suppressor GAS8 via MLL1/WDR5 to suppress hepatocarcinogenesis
Authors: Yang Ming
Published: August 2018

Long noncoding RNAs (lncRNAs) are vital players in hepatocellular carcinoma (HCC). We previously identified a significantly mutated lncRNA GAS8-AS1 locating in intron 2 of GAS8. However, its involvement in HCC is still largely unknown. In this study, we thoroughly functional evaluated its critical role in regulating chromatin dynamics, gene expression, cancer development and sorafenib multi-kinase target therapy in HCC. We for the first time declare that GAS8-AS1 and its host gene GAS8 act as novel HCC tumor suppressors employing a series of in vitro and in vivo assays. Consistent with this notion, a significantly decreased GAS8-AS1 or GAS8 expression exists in HCC tissues and is associated with poor prognosis of HCC patients. Interestingly, lncRNA GAS8-AS1 could evidently promote GAS8 transcription. Although GAS8-AS1 does not impact promoter DNA methylation of GAS8, we successfully annotated two lncRNA interacting proteins, Mixed Lineage Leukemia 1 (MLL1), a histone 3 Lys4 (H3K4) methyltransferase, and its partner WD-40 Repeat Protein 5 (WDR5) based on bioinformatics methodology. Chromatin immunoprecipitation and RNA immunoprecipitation assays elucidate that lncRNA GAS8-AS1 is required to maintain the GAS8 promoter in an open chromatin state. In accord with this, knockdown of GAS8-AS1 results in reduced MLL1/WDR5 binding, decreased H3K4me3 levels, diminished RNA Pol II functions and gene silencing of GAS8. GAS8-AS1 may function as part of a surveillance mechanism that keep activation of GAS8 promoter and transcription, which thus prevents hepatocarcinogenesis. Our results highlight the prevalent involvement of regulatory lncRNAs in cancers and provide pathogenic insights into HCC development and treatment.

Citation Format: Ming Yang. LncRNA GAS8-AS1 directs epigenetic modulation of the tumor suppressor GAS8 via MLL1/WDR5 to suppress hepatocarcinogenesis [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 2479.
Abstract 2486: PTEN fusion gene promotes growth in cancer
Authors: Yu Yanping , Chang Silvia , Ren Baoguo , Michalopoulos George , Tseng George , Luo Jianhua
Published: August 2018

Deletion or mutation of PTEN is frequent in a variety of human cancers and is an underlying mechanism for cancer development. Here, we report a PTEN-involved fusion gene and the corresponding genome breakpoint in human cancers. Pten fusion is highly recurrent in cancers and their matched lymph nodes and is located nucleoli. Targeted knockout of this fusion gene in Du145 and MCF7 cells impeded tumor cell growths, retarded S phase entry, and reduced invasiveness, and was prone to the UV- induced apoptosis. Knockout of PTEN-fusion blocked xenografted-tumor growth in mice, while forced-expression of PTEN -fusion through hydrodynamic injection into mice led to hepatocellular carcinoma. These findings indicate that the genome rearrangement of PTEN-fusion results in an oncogenic fusion protein in addition to the loss of Pten as an underlying mechanism in driving cancer development.

Citation Format: Yanping Yu, Silvia Chang, Baoguo Ren, George Michalopoulos, George Tseng, Jianhua Luo. PTEN fusion gene promotes growth in 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 2486.
Abstract 249: Genome-scale analysis of aberrant DNA methylation for emerging, high incidence of colorectal cancer in Taiwan
Authors: Chen KuoHsing , Lai Liang-Chuan , Chang Hsien-Fang , Chao Yu-Liang , Lin Been-Ren , Liang Jin-Tung , Cheng Ann-Lii , Chuang Eric Y. , Yeh Kun-Huei
Published: August 2018

Background:

The incidence of colorectal cancer (CRC) in Taiwan is increasing in decades, and the definite etiology remains elusive. This study aimed to explore the association of aberrant DNA methylations and the development of colorectal cancer in Taiwan.

Materials and methods: We prospectively collected 27 CRC patients' tumors, the adjacent normal colons (frozen specimens), and clinical characteristics, including age, sex, body mass index (BMI), stage at diagnosis and medical history of diabetes mellitus and hyperlipidemia. We conducted a genome-scale analysis of DNA methylation in these 27 CRC and 5 adjacent normal colon samples by using Illumina Methylation EPIC Beadchips. We used Wilcoxon rank-sum test (p < 0.05) and β value difference (≧ 0.25 or ≦ -0.25) to evaluate the methylation differences between CRC and adjacent normal colon samples.

Results: Overall, 7,672 differentially methylated genes (DMGs) were found between 27 CRCs and 5 adjacent normal colons. Principle component analysis demonstrated that a distinct cluster of adjacent normal colon and CRC tissue samples. Pathway analysis (β value difference ≧ 0.4 or ≦ -0.4) showed that ERK/MAPK signaling, GP6 signaling, CREB signaling in neurons, CDK5 and IGF-1 pathways were the top 5 pathways that were involved by DMGs. Subgroup analysis revealed most DMGs (N = 5,228) are shared by all 3 subgroups, however, patients who had metabolic syndrome had fewest subgroup-specific DMGs between tumors and adjacent normal colons, compared to patients who had no metabolic syndrome with normal weight or overweight (BMI < 25 or ≧ 25).

Conclusion: We demonstrated a genome-scale difference of aberrant DNA methylation in colorectal cancer in an East Asian population. These data will be validated further in a larger cohort.

Citation Format: KuoHsing Chen, Liang-Chuan Lai, Hsien-Fang Chang, Yu-Liang Chao, Been-Ren Lin, Jin-Tung Liang, Ann-Lii Cheng, Eric Y. Chuang, Kun-Huei Yeh. Genome-scale analysis of aberrant DNA methylation for emerging, high incidence of colorectal cancer in Taiwan [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 249.
Abstract 2490: Characterization of the role of DPPA3 in stemness regulation in hepatocellular carcinoma by DNA methylation
Authors: Yan Qian , Zhang Yu , Yu Dandan , Kam Ngar Woon , Guan Xin-Yuan
Published: August 2018

Hepatocellular carcinoma (HCC) is one of the most common cancers in the world with a very poor prognosis, which is closely associated with cancer stem cell (CSC). Global RNA-sequencing was used in different developmental stages of hepatocytes, including embryonic stem cell, endoderm, liver progenitor, and premature hepatocyte, as well as two pairs of nontumor and tumor tissues from HCC patients. We found that Developmental Pluripotency Associated 3 (DPPA3) was highest expressed in the stage of liver progenitor cells, and was down-regulated along with development. In HCC patients, DPPA3 was expressed in less than 1% tumor cells, while no staining was detected in normal counterparts. Functional assays revealed that DPPA3 overexpression increased HCC cells proliferation rate, the ability of foci formation and colony formation in soft agar, as well as migration and invasion. Furthermore, DPPA3 significantly increased HCC cells sphere formation frequency and sensitivity to chemotherapeutic agent including cisplatin, 5-Fu, and Sorafenib. In order to investigate the role of DPPA3 in CSC maintenance and stemness regulation in HCC, whole genome methylation sequencing was applied to compare methylation status between DPPA3- and empty vector-transfected cells. Results showed that DPPA3 reduced CG methylation level within functional regions in the genome. Of the 541 differentially methylated regions, the majority of them were involved in developmental process and cell differentiation. Overall, our findings give a novel insight into how methylation regulation controls the fate of cancer stem cells.

Citation Format: Qian Yan, Yu Zhang, Dandan Yu, Ngar Woon Kam, Xin-Yuan Guan. Characterization of the role of DPPA3 in stemness regulation in hepatocellular carcinoma by DNA methylation [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 2490.
Abstract 2491: Nuclear exporter protein XPO1 a novel prognostic and therapeutic target in gastric cancer
Authors: Muqbil Irfana , Mahdi Zaid , Choudhary Rahman , Baloglu Erkan , Senapedis William , Landesman Yosef , Shacham Sharon , Kauffman Michael , Kim Steve , Beydoun Rafic , Berri Richard N. , Shields Anthony , Mohammad Ramzi M. , Azmi Asfar S.
Published: August 2018

The high mortality rate associated with Gastric Cancer (GC) indicates the urgent need for actionable therapeutic targets. The nuclear exporter protein exportin 1 (XPO1/CRM1) is the exclusive exporter of many tumor suppressor proteins (TPSs) and growth regulators. XPO1 is often over-expressed in different malignancies leading to aberrant cytoplasmic localization of TSPs and subsequent inactivation. A detailed analysis on the correlation of XPO1 with inflammation-metaplasia-dysplasia-carcinoma sequence progression was performed using immunohistochemistry in 70 GC cases: (a) 10 cases from normal gastric mucosa, (b) 10 cases of stomach with intestinal metaplasia with and without inflammation, (c) 10 cases of mucosa with low-grade dysplasia (d) 10 cases of mucosa with high-grade dysplasia, (e) 10 cases of gastric adenocarcinoma and (f) 20 cases of metastatic gastric carcinoma. A correlation between XPO1 expression, the pathological and clinical features of the disease as well as survival were analyzed. Gastric cancer cell lines were exposed to the Selective Inhibitors of Nuclear Export (SINE) compounds (selinexor, KPT-8602, KPT-185, or the natural agent, leptomycin B (LMB), then analyzed using cytotoxicity and molecular assays. In addition, the activity of selinexor was evaluated in a sub-cutaneous xenograft of gastric cancer cell line NCI-N87. XPO1 served as a prognostic marker for poor outcome as positive staining of XPO1 in GC correlated with aggressive behavior of the disease. Targeting XPO1 using SINE compounds or LMB resulted in inhibition of GC cellular growth (IC50<200 nM), induction of apoptosis and suppression of colony formation (p<0.01). Molecular analysis revealed nuclear retention of several important TSPs, induction of pro-apoptotic proteins and suppression of pro-survival factor Bcl-2. Selinexor given orally at doses of 15 mg/kg twice a week for three weeks caused statistically significant reduction of NCI-N87 tumors in mice (p<0.05). Efficacy studies of SINE compounds in patient derived models of GC are ongoing. Our findings strongly demonstrate the potential of XPO1 to serve not only as a prognostic marker but also as a therapeutic marker in GC that warrants further clinical investigations.

Citation Format: Irfana Muqbil, Zaid Mahdi, Rahman Choudhary, Erkan Baloglu, William Senapedis, Yosef Landesman, Sharon Shacham, Michael Kauffman, Steve Kim, Rafic Beydoun, Richard N. Berri, Anthony Shields, Ramzi M. Mohammad, Asfar S. Azmi. Nuclear exporter protein XPO1 a novel prognostic and therapeutic target in gastric 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 2491.
Abstract 2496: YY1 regulates the germinal center reaction by inhibiting apoptosis
Authors: Trabucco Sally E. , Zhang Hong
Published: August 2018

The germinal center (GC) reaction produces high-affinity antibodies for a robust adaptive immune response. If dysregulated, the same processes cause GC B cells to become susceptible to lymphomagenesis. It is important to understand how the GC reaction is regulated. In this study, we show that transcription factor YY1 is required to maintain a robust GC reaction in mice. Selective ablation of YY1 significantly decreased in the frequency and number of GC B cells during the GC reaction. This decrease of GC B cells was accompanied by increased apoptosis in these cells. Further, we found that loss of YY1 disrupted the balance between dark zones and light zones, leading to a preferential decrease in dark zone cells. Collectively, these results indicate that YY1 plays an important role in regulating the balance between dark zone and light zone cells in GCs and between survival and death of GC B cells.

Citation Format: Sally E. Trabucco, Hong Zhang. YY1 regulates the germinal center reaction by inhibiting apoptosis [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 2496.
Abstract 250: Multiple lines of evidence make ZDHHC3 a possible marker for breast cancer in African American women
Authors: Kinney Nicholas A. , Varghese Robin T. , Anandakrishnan Ramu , Garner Harold R.
Published: August 2018

Rates of breast cancer mortality continue to be divided among race and ethnicity without a definitive explanation. Between the years 2005-2009 the racial disparity in breast cancer mortality caused about 9 extra death per 100 breast cancer cases in African American women compared to Caucasian women. In addition, young African Americans tend to have more aggressive forms of breast cancer. The source of this health disparity likely stems from a complex combination of mammography rate and frequency; segregation; socioeconomic status; and genetics. We contribute to what is known about genetics by investigating microsatellites - a type of repetitive DNA - and possible links to the breast cancer mortality gap. Microsatellites are understudied compared to single nucleotide polymorphisms and have the capacity to affect gene expression. We screen 33,854 microsatellites in germline DNA of African American women with and without breast cancer: four are statistically significant. We preform further analysis of genes harboring these microsatellites; the results reveal that ZDHHC3 is particularly interesting. Its location (3p21) is already linked to early invasive breast cancer. Kaplan-Meier analysis shows that ZDHHC3 alterations may be associated with poor breast cancer survival. Data from cBioPortal suggests that ZDHHC3 mRNA expression is lower in African Americans compared to Caucasians. These independent lines of evidence make ZDHHC3 a candidate for further investigation.

Citation Format: Nicholas A. Kinney, Robin T. Varghese, Ramu Anandakrishnan, Harold R. Garner. Multiple lines of evidence make ZDHHC3 a possible marker for breast cancer in African American women [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 250.
Abstract 2502: Role of TMEM97/σ2 receptor in breast cancer cell proliferation and responses to chemotherapy
Authors: Zhang Yuanqin , Wang Jiuhui , Nie Daotai
Published: August 2018

Introduction:

Breast cancer is the most prevalent cancer for women both in the developed and the developing countries. In 2017, about 40,610 women are expected to die from breast cancer in the U.S. The sigma receptors (σRs) are transmembrane receptors with two subtypes, termed sigma-1 receptor and sigma-2 receptor. Sigma-2 receptor has been found overexpressed in numerous human cancer tissues, and compound targeting it are now in clinical trials for breast cancer diagnosis. But it's biological role in breast cancer is unknown because the encoded gene has not been clearly identified. Recently, it has been demonstrated that TMEM97, not progesterone Receptor Membrane Component 1 (PGRMC1), is a bona fide sigma-2 receptor. The identification of sigma-2 (σ2) receptor will make it possible to define its biological functions in breast cancer and develop modulator of σ2 receptor as potential cancer therapeutics.

Methods:

The expression of TMEM97/σ2 receptor in multiple cancer cells was evaluated by RT-qPCR. Regulation of P53 on TMEM97/σ2 receptor was determined by Western Blot after 48 hours Doxorubicin treatment of MCF7 cell and HCT116 wild type and P53-/- cells. PB28, an agonist of σ2 receptor and an antagonist of σ1 receptor, was used (0, 1μM, 10μM and 20μM for 4 days) to test its effects on MCF7 cell growth and colony formation. TMEM97/σ2 receptor was cloned and sequenced. We overexpressed TMEM97/σ2 receptor in MCF7 cells to characterize its role in cell proliferation, survival, and colony formation, with or without treatment of its agonist PB28.

Results and conclusion:

TMEM97/σ2 receptor is expressed in various cancer cell lines including lung, breast and prostate cancer, albeit at different levels. TMEM97 expression was reduced in MCF7 cells when p53 expression was stimulated by doxorubicin. PB28 reduced the number of viable cells and also reduced colony formation of MCF7 cells in a dose dependent manner. Meanwhile overexpression of TMEM97/σ2 receptor retarded cell growth in MCF7 cells and sensitized them to PB28 as evidenced by reduced IC50 when compared with control cells. The data suggest that TMEM97/σ2 receptor is expressed in breast cancer cells, with its expression suppressed by p53, and activation of this receptor led to reduced tumor cell proliferation.

Citation Format: Yuanqin Zhang, Jiuhui Wang, Daotai Nie. Role of TMEM97/σ2 receptor in breast cancer cell proliferation and responses to chemotherapy [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 2502.
Abstract 2505: Baicalein reverses chronic stress-induced ovarian cancer promotion through GABA receptors
Authors: Li Ke , Kang Yu , Yu Yinhua
Published: August 2018

Studies suggest that chronic stress accelerate the progression of ovarian cancer, and the promotion effects can be abrogated by β-blockers. However, β-blockers should be assessed and tailored to each cancer patient as they will exert toxic effects in overdose. Baicalein, a widely used Chinese herbal medicine, identified as a new type of anticancer drug with an obvious function of anti-anxiety in animal experiments and whose mechanism may be mediated by GABA receptors. The focus of the current study was to determine whether baicalein could block the effects of chronic stress on tumor growth and investigate the underlying mechanism that relates to GABA receptors. Two kinds of ovarian cancer cell lines (OVCAR-8 and CAOV-3) were selected to be investigated in the inhibitory effect of baicalein on cancer cells. Cell CCK-8 assay, wound healing assay, transwell assay and flow cytometry were used for in vitro experiments. Chronic stress was induced by a restrained stress model in nude mice carrying xenografts in the presence and absence of treatment with baicalein. Xenograft sizes and weight were measured after 4 weeks. Norepinephrine was measured by HPLC and GABA was measured by immunoassays in blood and tumor tissues. Expression of GABA receptors in the xenografts was analyzed by real-time quantitative PCR, western blotting and immunohistochemistry (IHC). Our results showed that ovarian cancer cell lines exhibited significant promotion in the presence of NE concentrations of 10Μm and baicalein concentrations of 10μM could reverse its effects by Cell CCK-8 assay and wound healing assay. Daily restraint stress resulted in significantly increased tumor growth in tumor model of ovarian cancer in nude mice. And this increase was completely blocked with daily baicalein treatment. Chronic stress attenuated expression of GABA receptors in vivo and in vitro; conversely, baicalein increased expression of GABA receptors. Overall, our data suggest that baicalein stimulated GABA receptors activation to reduce tumor growth in ovarian cancer. Considering the stimulatory effects of chronic stress on cancer growth, our study implicates baicalein as a potential medicine for blocking the deleterious effects of chronic stress.

Citation Format: Ke Li, Yu Kang, Yinhua Yu. Baicalein reverses chronic stress-induced ovarian cancer promotion through GABA receptors [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 2505.
Abstract 251: Luteolin-mediated increase in miR-26a inhibits prostate cancer cell growth and induces cell cycle arrest targeting EZH2
Authors: Kanwal Rajnee , Moreton Stephen , Franco Daniel , Gupta Sanjay
Published: August 2018

Luteolin (3',4',5,7-tetrahydroxyflavone), a flavonoid found in several vegetables and fruits, has been reported to possess anticancer and apoptosis inducing properties. However, the molecular targets of luteolin action are presently unknown. The enhancer of zeste homolog 2 (EZH2) is a catalytic subunit of polycomb repressive complex 2 (PRC2), which catalyzes the trimethylation of histone H3 on Lysine 27 (H3K27) and is involved in chromatin remodeling and gene silencing. EZH2-mediated epigenetic gene silencing plays an important role in several human cancers including prostate neoplasm. Emerging studies demonstrate that miR-26a, a non-coding microRNA, regulates EZH2 expression and is frequently lost during cancer progression. Here we investigated the potential involvement of luteolin-mediated miR-26a upregulation and EZH2 suppression leading to cell cycle arrest and apoptosis in cancer cells. Human prostate cancer DU145 and PC-3 cells, which possess high constitutive EZH2 expression, were treated with 5-20 µM luteolin at various times significantly inhibited EZH2 and H3K27 trimethylation in dose and time dependent manner. Luteolin treatment further resulted in increased expression of miR-26a in both cell lines, as exhibited by EZH2 inhibitor GSK126. Mechanistic investigations revealed that miR-26a overexpression suppressed cell cycle regulatory molecules such as cyclin D and E, cyclin dependent kinases CDK4 and CDK6, and CDK inhibitors p14/ARF, p16/INK4A and p21/WAF1 in EZH2-dependent manner. Interestingly, similar observations were noted in prostate cancer cells with a marked decrease in cyclins and CDKs along with concomitant increase in CDK inhibitors after luteolin treatment resulting in G0/G1 phase cell cycle arrest. In plasma specimens, obtained from healthy individuals, high-risk subjects and prostate cancer patients exhibit a progressive decrease in miR-26a expression. Taken together, our results indicate that miR-26a functions as a growth suppressive miRNA lost in prostate cancer, whereas luteolin is a promising agent which demonstrate anti-proliferative effects mediated by upregulation of miR-26 repressing EZH2 in prostate cancer cells.

Citation Format: Rajnee Kanwal, Stephen Moreton, Daniel Franco, Sanjay Gupta. Luteolin-mediated increase in miR-26a inhibits prostate cancer cell growth and induces cell cycle arrest targeting EZH2 [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 251.
Abstract 2518: Folate hydrolase-1 is a novel target for J591-brachytherapy in Merkel cell carcinoma
Authors: Meier Barbara , Ramirez-Fort Marigdalia K. , Lachance Kristina , Church Candice D. , Mahase Sean S. , Jenrette Joseph M. , Lange Christopher S. , French Lars E. , Nghiem Paul , Bander Neil H.
Published: August 2018

Folate hydrolase-1 (FOLH1) is a type II transmembrane protein. Oncologically, FOLH1 is upregulated throughout prostate cancer cells; it is also luminally expressed by the neovasculature of most solid tumors but not by normal vessels. J591, a monoclonal antibody (AB), is specific to, and is effectively endocytosed after extracellular binding to, FOLH1. J591 is presently being developed in clinical trials as a vehicle for AB-based brachytherapy in FOLH1+ cancers. Merkel cell carcinoma (MCC) is a rare, neuroendocrine tumor; metastatic (m) MCC is associated with poor survival. We characterized FOLH1 expression in MCC to determine its target potential for J591-brachytherapy. Paraffin sections from primary (p) and mMCC were deparaffinized and rehydrated. Samples were stained with 3E6 (DAKO), a mouse IgG1 monoclonal anti-human FOLH1. Mouse IgG1 (10 ug/mL in 1% bovine serum albumin) was used as an isotype-matched negative control. Anti-CD31 (IgG1) was used as a positive control. Kaplan-Meier survival curves were calculated based on patient outcome data and FOLH1 expression. 81 MCC tumors were evaluated. 67% (54/81) of all cases with 77% (24/31) of pMCC and 60% (30/50) of mMCC tumors demonstrated FOLH1+ neovessels. No cellular staining of tumor cells was identified. 34 patients with FOLH1 +/- MCC were demographically homogeneous in terms of sex, age, immunosuppression status, prior therapies, stage at diagnosis, and local or distant recurrences. No significant differences were detected based on FOLH1 status, in regards to MCC specific survival (P=0.905), or overall survival (P=0.687), as measured from time of diagnosis. FOLH1 is expressed in the majority of pMCC and mMCC cases and is nonprognostic. Our findings support further investigation of targeted therapy with J591-brachytherapy for the management of MCC.

Citation Format: Barbara Meier, Marigdalia K. Ramirez-Fort, Kristina Lachance, Candice D. Church, Sean S. Mahase, Joseph M. Jenrette, Christopher S. Lange, Lars E. French, Paul Nghiem, Neil H. Bander. Folate hydrolase-1 is a novel target for J591-brachytherapy in Merkel cell carcinoma [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 2518.
Abstract 2521: Patient GBM cell originated secretome analysis identifies cytokine A, as a potent therapeutic target
Authors: Han Suji , Shin Kayoung , Lee Kyoungmin , Kim Sungsoo , Kang Hyunju , Lee Jin-Ku , Nam Hyun , Joo Kyeung Min , Nam Do-Hyun
Published: August 2018

GBM is the most aggressive human primary brain tumor and leads to poor clinical outcomes. According to previous studies, exogenous factors secreted from primary brain tumor cells can change the growth pattern of normal cells. We analyzed the ingredients of cell culture media from two patient glioblastoma cells which can grow without growth factors. As a result, we selected a significant factor for tumor cell growth which is called cytokine A, the Heparin-binding growth factor. Because of the cell viability maintenance ability of cytokine A, we tried to investigate detailed mechanism of cytokine A and applied the result to the combination therapy. Cytokine A is critical for tumorigenesis of GBM by modulating DNA damage signaling and cell cycle through ROS. For that reason, cytokine A is effective therapeutic target and can be used in combination therapy.

Citation Format: Suji Han, Kayoung Shin, Kyoungmin Lee, Sungsoo Kim, Hyunju Kang, Jin-Ku Lee, Hyun Nam, Kyeung Min Joo, Do-Hyun Nam. Patient GBM cell originated secretome analysis identifies cytokine A, as a potent therapeutic target [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 2521.
Abstract 2522: Luminal epithelial cell expression of amphiregulin is required for mammary gland development and facilitates mutant p53-driven tumorigenesis
Authors: Lofgren Kristopher A. , Meier David R. , Girtman Megan A. , Peterson Esther A. , Jenkins E. Charles , Kenny Paraic A.
Published: August 2018

Amphiregulin (AREG) is an estrogen-responsive ligand of the epidermal growth factor receptor. Mice with germline deletion of AREG are defective in pubertal mammary expansion. Using shRNA, we have previously reported that the estrogen-responsive growth of MCF7 xenograft tumors in immunocompromised mice is also AREG-dependent. Here we use a series of genetically engineered mouse AREG models (lacZ-knock-in, conditional knockout and germline knockout) to address two outstanding questions: (1) Among the numerous AREG-expressing cell types in the mammary gland and stroma, does the requirement for AREG in mammary gland development reside specifically in the luminal epithelial cells? (2) Is AREG required for mammary tumorigenesis in immunologically-intact mice? An initial characterization of mammary gland development using an AREG-lacZ knock-in allele identified AREG promoter activity in the luminal epithelial but not the myoepithelial compartment. We generated AREGfl/fl mice and achieved luminal epithelial cell specific AREG deletion using MMTV-Cre. Mice lacking AREG in the luminal epithelial compartment (but otherwise proficient in AREG expression) phenocopied the deficit in outgrowth seen in the AREG germline deletion, indicating that luminal epithelial AREG expression is essential for mammary gland development. To determine the extent to which AREG deletion affects mammary tumor latency and multiplicity in immune-intact animals, we used a Cre-regulated model of p53 mutation in mammary tissue (MMTV-Cre x p53LSL-R172H). We used this p53-dependent model to generate at-risk cohorts with AREG deficient (AREG-/-) and AREG proficient (AREG+/+ & AREG+/-) genotypes. Although the cohorts are still maturing, the median age of mammary tumors observed in AREG proficient mice is 258 days (median tumor multiplicity is 2 per animal), while only a single tumor has been detected in the AREG deficient cohort (at 415 days). Together, these data point to two temporally distinct roles for amphiregulin–an essential function in mammary ductal expansion during puberty and a subversion of this developmental role during tumorigenesis later in life.

Citation Format: Kristopher A. Lofgren, David R. Meier, Megan A. Girtman, Esther A. Peterson, E. Charles Jenkins, Paraic A. Kenny. Luminal epithelial cell expression of amphiregulin is required for mammary gland development and facilitates mutant p53-driven tumorigenesis [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 2522.
Abstract 2526: Modulation of metastatic factors in prostate cancer cells by DAX-1 (NR0B1), an androgen-induced orphan nuclear receptor
Authors: Valencia Roxxana Valeria Beltran , Tzagarakis-Foster Christina
Published: August 2018

DAX-1, a member of the nuclear hormone receptor superfamily, has a pivotal role in adrenal and gonadal development. However, recent studies have shown that DAX-1 acts as a transcriptional repressor influencing the progression of many types of cancers. We hypothesize that DAX-1 has key roles in regulating metastatic genes during prostate cancer progression. We found that introducing DAX-1 into prostate cancer cells lacking endogenous expression of DAX-1 leads to a decrease in metastatic rates. Consequently, we analyzed DAX-1 transcriptional regulation over different markers associated with metastasis. Since the androgen receptor is a key regulator of prostate cancer progression, we used CRISPR to knockout DAX-1 expression in hormone-dependent prostate cancer cells, allowing us to understand the role DAX-1 has as a negative regulator of metastatic genes after androgen receptor activation. We have shown that treatment of hormone-dependent prostate cancer cells with a non-aromatizable androgen leads to upregulation of DAX-1 expression. Here we use the same mechanism to induce DAX-1 expression and analyze the effects on metastatic markers. Results have shown that DAX-1 is sufficient to decrease the expression of metastatic markers and inhibit metastatic rates in prostate cancer cells. These results further explain some of the intrinsic mechanisms involved in prostate cancer metastasis and could reveal new therapeutic routes in the treatment of metastatic prostate cancer.

Citation Format: Roxxana Valeria Beltran Valencia, Christina Tzagarakis-Foster. Modulation of metastatic factors in prostate cancer cells by DAX-1 (NR0B1), an androgen-induced orphan nuclear receptor [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 2526.
Abstract 2529: GENT 2.0, a newly updated database: Gene expression across normal and tumor tissues
Authors: Park Seung-Jin
Published: August 2018

We previously developed GENT1.0 (Gene Expression Database of Normal and Tumor tissues), a web-accessible database that provides gene expression patterns across diverse human cancer and normal tissues. GENT shows gene expression patterns in various tissues easily and has more than 40,000 samples for 25 normal-cancer paired tissues. Here, we made updated database called GENT2.0 based on 60,000 microarray data for 36 normal-cancer paired tissues with various functions. GENT2.0 1) has collected over 60,000 amounts of data for 36 normal cancer paired tissues; 2) can analyze histologic or molecular subtypes or tumor stages for 19 tissues; 3) provides all sorts of plots in svg (Scalable Vector Graphics) format with a high-resolution figure; 4) has twice the amount of data compared to GENT1.0, and the web page loading speed is almost the same as GENT1.0; and 5) data will be accessible on the web. We hope GENT2.0 will be a useful resource for cancer researchers in many stages from target discovery to target validation.

Citation Format: Seung-Jin Park. GENT 2.0, a newly updated database: Gene expression across normal and tumor tissues [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 2529.
Abstract 2543: Concurrent chemotherapy and temozolomide-resistant CAR-T immunotherapy enhances glioblastoma clearance in experimental animals
Authors: Sengupta Sadhak , Gergi Mansour , Guha Prajna , Sengupta Sudarshana , Junghans Richard P. , Sampath Prakash
Published: August 2018

Dismal long-term progression-free survival of glioblastoma (GBM) patients, who are concurrently treated with radiation and chemotherapy, demands development of alternative strategies for management of this disease. Recent clinical success of chimeric antigen receptors expressing T cells (CAR-T) in GBM patients, interest in tumor-directed immunotherapy has been reinvigorated. However as per standard care, immunotherapy must be concurrent with or following the standard care protocol for glioma therapy, which includes Temozolomide (TMZ) chemotherapy. Concurrent chemotherapy with TMZ reduces the efficacy of adoptive T cell immunotherapy by inducing severe lymphopenia, while post-chemotherapy treatment minimizes the window of opportunity for effective immunotherapy, because of inherent tumor heterogeneity resulting in an altered antigenic profile in recurrent GBMs. To address this, we have created a TMZ-resistant CAR-T cell directed against IL13Rα2+ GBM tumors by introducing a mutated O6-methylguanine-DNA methyltransferase (MGMT) moiety in the existing IL13CAR backbone. This specific mutation in MGMT (P140KMGMT) blocks the interaction of TMZ to MGMT and therefore protects the rapidly proliferating anti-GBM CAR-T cells from TMZ-induced lymphopenia. Activated P140KMGMT-IL13CAR-T cells were protected from TMZ-induced cytotoxicity in vitro without any loss of anti-tumor effects of these modified CAR-T cells. Experimental xenograft animals treated with P140KMGMT-IL13CAR-T cells and concurrent TMZ therapy showed 2-fold increase in median survival over groups that were treated with TMZ alone, and 2.8-fold increase in overall survival over groups that were treated with TMZ-sensitive second generation IL13CAR-T cells and concurrent TMZ. These encouraging results point us strongly in the direction of developing this CAR-T technology towards a promising future of concurrent chemotherapy and immunotherapy of GBM.

Citation Format: Sadhak Sengupta, Mansour Gergi, Prajna Guha, Sudarshana Sengupta, Richard P. Junghans, Prakash Sampath. Concurrent chemotherapy and temozolomide-resistant CAR-T immunotherapy enhances glioblastoma clearance in experimental animals [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 2543.
Abstract 2548: An image-based method to detect and quantify T cell mediated cytotoxicity of 2D and 3D target cell models
Authors: Larson Brad , Kambach Diane , Luty Wini , Souza Glauco
Published: August 2018

T cell-mediated cytotoxicity plays an important role in a suite of new methods being developed with the goal of incorporating a patient's immune system to combat cancer. The purpose of this study was to develop a sensitive, image-based in vitro method to evaluate and optimize adoptive T cell immunotherapies. Experiments were performed using MDA-MB-231 and fibroblast target cells co-cultured in 2D and 3D models, along with T cell to target cell ratios of 20:1, 10:1 and 5:1. T cell activation procedures were also tested where T cells were activated in the presence of anti-CD3 and anti-CD28 antibodies plus an IL-2 superkine only, or in a directed fashion with antibodies, superkine, and varying concentrations of target breast cancer cells. Experimental data show that directing the activation of T cells to recognize surface antigens on specific target cancer cells enhances the killing ability of the cells when compared to results from nondirected cells. In addition, increased T cell to target cell ratios induce higher levels of target cell cytotoxicity. Finally, it was evident that assay performance using 2D cultured target cells could be reliably carried out through a 72-hour incubation period. However, it exhibited limited utility beyond this point due to increasing numbers of uninduced necrotic cells within wells, negating the effect of test conditions. This phenomenon was unseen from 3D cultured target cells, indicating their ability to maintain a higher level of viability when cultured in this manner. Necrotic induction can then be reliably attributed to cell treatments using multiday incubations. The combination confirms the ability of the image-based method to deliver dependable results to assess T cell-mediated cytotoxicity using 2D and 3D target cell models.

Citation Format: Brad Larson, Diane Kambach, Wini Luty, Glauco Souza. An image-based method to detect and quantify T cell mediated cytotoxicity of 2D and 3D target cell models [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 2548.
Abstract 2551: Allogeneic CRISPR engineered anti CD70 CAR T cells demonstrate potent preclinical activity against both solid and hematological cancer cells
Authors: Padalia Zinkal , Porras Ashley , Dequeant Mary Lee , Sagert Jason , Nguyen Thao , John Matthias , Allen Melanie , Dar Henia , Henderson Daniel , Police Seshidar , Mu Dakai , Maeng Kelly , Huang Elaine , Spencer Sarah , Lorson Nickolaus , Gonzales Paul , Holmquist Chelsea , Hirschfeld Gregg , Terrett Jonathan A. , Kalaitzidis Demetrios
Published: August 2018

Autologous CAR T therapeutics have recently been approved for use in B-cell malignancies. While responses have been impressive using CD19 directed CAR T cells there has been a lack of comparable success for CAR T cells directed at solid tumor antigens. In an effort to address the need for effective and durable off-the shelf therapies for both hematologic and solid tumors we have developed allogeneic CAR T cells targeting the CD70 antigen. CD70 is expressed in both hematologic malignancies as well as in solid cancers such as renal cell carcinoma (RCC), while its expression in normal tissues is restricted to a subset of lymphoid cell types. CAR-T cells expressing a CD70 targeting CAR were generated by CRISPR Cas9 genome editing. T cells from healthy donors were edited to express a CD70 CAR by knocking this construct into the concurrently knocked out TCR alpha constant region (TRAC). Loss of the TCR reduces the risk of graft versus host disease enabling an allogeneic therapeutic. CD70 CAR T cells displayed potent cell killing function in vitro against CD70 expressing lymphoid and renal cancer derived cell lines across a broad range of antigen expression levels. CD70 CAR T cells also secreted IFNg, released granzyme B and proliferated in a CD70 specific manner. Furthermore, the CD70 targeting CAR T cells were able to eliminate established ccRCC tumor xenografts in mice

Citation Format: Zinkal Padalia, Ashley Porras, Mary Lee Dequeant, Jason Sagert, Thao Nguyen, Matthias John, Melanie Allen, Henia Dar, Daniel Henderson, Seshidar Police, Dakai Mu, Kelly Maeng, Elaine Huang, Sarah Spencer, Nickolaus Lorson, Paul Gonzales, Chelsea Holmquist, Gregg Hirschfeld, Jonathan A. Terrett, Demetrios Kalaitzidis. Allogeneic CRISPR engineered anti CD70 CAR T cells demonstrate potent preclinical activity against both solid and hematological cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 2551.
Abstract 2561: Fully human immunoglobulin heavy chain only-derived CD33 CAR for the treatment of acute myeloid leukemia
Authors: Schneider Dina , Xiong Ying , Chen Weizao , Zhu Zhongyu , Wu Darong , Hwang Jennifer , Dimitrov Dimiter S. , Dropulic Boro , Orentas Rimas J.
Published: August 2018

CD33 antigen is a promising target expressed on non-solid cancers, including acute myeloid leukemia (AML). Present investigative approaches to treatment of CD33-positive AML include antibody drug conjugates (My96, Mylotarg®) and CART cells incorporating CD33-targeting domains derived from a humanized scFv. Here, we designed a chimeric antigen receptor utilizing targeting domain derived from a fully human CD33 fragment variable heavy chain sequence, termed CAR33VH, and examined its in vitro and in vivo potency against AML.

Primary human CD4+ CD8+ T cells derived from three healthy donors were transduced with lentiviral constructs (LV) encoding CAR33VH, or control CAR construct based on scFv My96 (My96CAR). Flow cytometric analysis revealed expression of CAR33VH at 32%-45%, and expression of My96CAR at 77% - 86%. When challenged with CD33+ AML tumor lines HL60 and MOLM-14 in vitro, both constructs demonstrated efficient target killing. CD33- tumor lines K562 and Reh were not sensitive to CAR killing, underscoring CAR specificity to CD33 antigen. Pro-inflammatory cytokines IFN gamma, TNF alpha and IL-2 in culture supernatants of CART cells incubated with CD33+ HL-60 and MOLM-14 tumors, but not with CD33- K562 cells overnight, were induced, as measured by ELISA. Long-term co-incubation assay of CART cells with HL-60 leukemia at E:T ratios 1:5 to 1: 0.04, suggested similar killing potency and persistence of CAR33VH to the positive control scFv-based CAR. In in vivo AML model, NSG mice engrafted with MOLM-14 cells stably expressing firefly luciferase, both CAR33VH, and My96CAR control were equally efficient in tumor elimination.

In conclusion, CAR33VH, comprised of a fully human heavy-chain variable fragment only antigen binding domain, was efficient in tumor killing in vitro and in vivo, and may be used clinically for treatment of CD33+ hematologic malignancies. To our knowledge, this is one of the first instances demonstrating the feasibility of employing heavy chain only binder sequence in CART design.

Citation Format: Dina Schneider, Ying Xiong, Weizao Chen, Zhongyu Zhu, Darong Wu, Jennifer Hwang, Dimiter S. Dimitrov, Boro Dropulic, Rimas J. Orentas. Fully human immunoglobulin heavy chain only-derived CD33 CAR for the treatment of acute myeloid leukemia [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 2561.