bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 615: Roles of XRCC1 genetic polymorphism in head and neck cancer patients receiving radiation therapy in Taiwan
Authors: Huang Shiang-Fu , Chien Huei-Tzu , Liao Chun-Ta , Wang Hung-Ming , Wang Yuan-Hung
Published: August 2019

In Taiwan, oral cancer (including sub-sites in the oral cavity, oropharynx and hypopharynx) is the fourth most common cancer in men. The incidence of oral cancer still increased in the past decade. Oxidative stress generated from cigarette smoking, AQ chewing and alcohol drinking was shown to contribute to oral cancer development. The oxidative stress associated DNA lesions are repaired via the DNA base excision repair (BER) pathway. In constrast, radiation therapy was an important treatment modality either as primary or adjuvant in these patients. Ionizing radiation induced DNA damage was also repaired by BER related mechanisms. To investigate the role of BER genes XRCC1 in patients receiving radiation therapy in Taiwan, a comprehensive study on evaluation of polymorphisms of XRCC1 genes was carried out in 753 oral cancer male cases received radiation therapy as primary or adjuvant treatment. Patients with XRCC1 194Arg, XRCC1 280Arg, and XRCC1399Gln homozygous genotypes had worse disease-free survival (p = 0.015, p = 0.587 and p = 0.013, respectively). A linkage disequilibrium exists between XRCC1 194Arg and XRCC1 399Gln genotyping. We further investigated the influence of XRCC1 194Arg and XRCC1 399Gln haplotype in the radiation effects in oral cancer patients. XRCC1 194 Arg/Arg homozygous and its combination with XRCC1 399 Arg/Gln genotyping were selected. We found that the XRCC1 194Arg/Arg and XRCC1 399Gln/Gln haplotype had the worse survival (p = 0.022, ). Multivariate Cox regression adjusting tumor stage, lymph node metastasis and tumor differentiation confirmed that XRCC1 194Arg and XRCC1 399Gln haplotype was an independent prognostic factor for oral cancer receiving radiation therapy. These indicated that XRCC1 194Arg and XRCC1 399Gln genotypes had adverse influence in oral cancer patients receiving radiation therapy. XRCC1 194Arg and XRCC1 399Gln haplotype predicts poor prognosis in these patients.

Citation Format: Shiang-Fu Huang, Huei-Tzu Chien, Chun-Ta Liao, Hung-Ming Wang, Yuan-Hung Wang. Roles of XRCC1 genetic polymorphism in head and neck cancer patients receiving radiation therapy in Taiwan [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 615.
Abstract 62: HIC1 inhibits cells metastasis via epithelial-mesenchymal transition in esophageal squamous cell carcinoma
Authors: Sha Beibei , Chen Ping , Liu Xingge , Hu Xuanyu , Li Miaomiao , Li Pei
Published: August 2019

Invasion and metastasis are the main causes of death in patients with esophageal squamous cell carcinoma (ESCC). Epithelial-mesenchymal transition (EMT) has been reported to be closely related with tumor metastasis. Although HIC1 (hypermethylated in cancer 1) gene has been observed to be epigenetically modified in many cancers, its intrinsic role and mechanism in ESCC metastasis, especially in EMT, remains unclear. Our previous study showed that hypermethylation of the HIC1 promoter markedly elevated in metastatic ESCC tissues compared with primary and adjacent normal tissues and it was associated with poor patients’ survival. In the present study, we used immunohistochemistry (IHC) to detect the expression of HIC1 in ESCC tissues and adjacent normal tissues, and the results showed that HIC1 was down-expressed in ESCC tissues and overexpressed in adjacent normal tissues. Results from trans-well-migration and wound healing experiments showed that overexpression of HIC1 in ESCC cells (KYSE 30) can reduce cell migration and metastasis behavior. Moreover, Western blot showed that overexpression of HIC1 could down-regulate EphA2 expression, simultaneously induced the markers of EMT target changed, in which E-cadherin was observed up-regulated, Snail and Vimentin were down-regulated. In summary, our results suggests that HIC1 may regulate EphA2-SirT1-P53 signaling pathway to mitigate ESCC aggressiveness.

Note: This abstract was not presented at the meeting.

Citation Format: Beibei Sha, Ping Chen, Xingge Liu, Xuanyu Hu, Miaomiao Li, Pei Li. HIC1 inhibits cells metastasis via epithelial-mesenchymal transition in esophageal 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 62.
Abstract 626: Association of circulating vitamin D concentrations with colorectal cancer risk differs depending on inherited genotypes in combination with dietary lifestyle factors
Authors: Gibbs David C. , McCullough Marjorie L. , Song Mingyang , Um Caroline Y. , Wu Kana , Jenab Mazda , Giovannucci Edward , Bostick Roberd M. , Fedirko Veronika
Published: August 2019

Higher circulating 25-hydroxyvitamin-D [25(OH)D] concentrations - used clinically to assess vitamin D status - are inversely associated with colorectal cancer (CRC) risk. However, it remains unknown whether this association varies by common, inherited polymorphisms encoding common vitamin D-binding protein (DBP) isoforms linked to differences in vitamin D handling and metabolism. We pooled participant-level data from three prospective cohorts in the United States (Cancer Prevention Study-II [CPS-II], Nurses’ Health Study [NHS]) and Western Europe (European Prospective Investigation into Cancer and Nutrition [EPIC]), comprising 1,710 incident CRC cases diagnosed between 1990 and 2011, and 1,649 matched controls selected using incidence density sampling. There were 1,106, 246 and 358 CRC cases in EPIC, CPS-II and NHS, respectively. Pre-diagnostic 25(OH)D measurements were calibrated to the same assay and seasonally-adjusted. Study-specific relative risks (RRs) for 25(OH)D and CRC risk according to DBP2 isoform (determined by the GC rs4588 variant) were estimated using multivariable unconditional logistic regression and pooled using fixed effects models. We also stratified our models by dietary vitamin D intake to account for differences in fortification/supplementation in the US and Europe. Variation in 25(OH)D concentrations across studies reflected differences in vitamin D intake and supplement use. Among individuals with lower vitamin D intakes (≤ 330 IU/day), for each 25 nmol/L increase in 25(OH)D, CRC risk was 30% lower among those with the DBP2 isoform (RR = 0.70, 95% CI = 0.59-0.83), and 5% lower among those without DBP2 (RR = 0.95, 95% CI = 0.82-1.10) (Pheterogeneity by DBP isoform = 0.009). Among individuals with higher vitamin D intakes (>330 IU/day, CPS-II and NHS only), for each 25 nmol/L increase in 25(OH)D, CRC risk was 18% higher among those with DBP2 (RR = 1.18, 95% CI = 0.82-1.10), and 33% lower among those without the DBP2 isoform (RR = 0.67, 95% CI = 0.53-0.85) (Pheterogeneity by DBP isoform = 0.002). These findings suggest that the association of 25(OH)D with CRC risk differs depending on inheritance of the functional DBP2 isoform in combination with levels of dietary vitamin D intake.

Citation Format: David C. Gibbs, Marjorie L. McCullough, Mingyang Song, Caroline Y. Um, Kana Wu, Mazda Jenab, Edward Giovannucci, Roberd M. Bostick, Veronika Fedirko. Association of circulating vitamin D concentrations with colorectal cancer risk differs depending on inherited genotypes in combination with dietary lifestyle factors [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 626.
Abstract 64: Regulation and functional role of the cell-cell junction-associated RNAi machinery in colon cancer
Authors: Daulagala Amanda C. , Bridges Catherine , Rutledge Lauren , Nair-Menon Joyce , Yost Micheal , Kourtidis Antonis
Published: August 2019

Colon cancer is the third most common and the second deadliest type of cancer in the United States. A typical characteristic of colon tumors is the compromised integrity of the colonic epithelial monolayer. The Adherens Junction (AJ) is a cell-cell adhesion complex essential for the maintenance of the epithelial integrity, composed of E-cadherin and the catenin family of proteins. We have recently shown that the E-cadherin-p120 catenin partner PLEKHA7 is critical for epithelial monolayer integrity. We have also revealed that PLEKHA7 recruits the core components of the RNA interference (RNAi) machinery, such as Ago2, DROSHA and DGCR8, as well as miRNAs at the mature apical adherens junctions, to suppress expression of pro-tumorigenic markers, through miRNA-mediated silencing. However, we still haven’t examined the extend of the contribution of this mechanism to colon tumorigenesis and its modes of regulation. We hypothesize that the adherens junction-RNAi mechanism acts as an epithelial sensor that tethers monolayer integrity to cell behavior. To examine this, we generated 3D cultures of well-differentiated Caco2 colon epithelial cells on Matrigel to examine the distribution of RNAi proteins. Indeed, co-localization of PLEKHA7 with RNAi proteins occurs specifically at the apical surface of these spheroids, whereas PLEKHA7 depletion results in loss of junctional localization of RNAi proteins and in enlarged, multilayered structures. Corroborating these results, PLEKHA7 and RNAi components co-localize specifically at the apical surface of normal colon crypts, whereas extensive disruption or loss of this co-localization was observed in colon tumor patient samples. Fibrosis and extensive deposition of extracellular matrix (ECM) due to chronic inflammation is a precursor to colon cancer. ECM can affect adherens junctions integrity, through integrin-Src signaling. Indeed, we have found that Src activity disrupts localization of PLEKHA7 and RNAi proteins to the junctions. To investigate the potential role of ECM in the regulation of the junctional RNAi mechanism, we are now examining Caco2 cells grown on different ECM substrates, as well as under different conditions of mechanical strain, since fibrosis poses mechanical stress on cells. In summary, our data point towards a novel putative tumor suppressor mechanism, of which we are investigating its modes of regulation and its involvement in colon tumorigenesis.

Citation Format: Amanda C. Daulagala, Catherine Bridges, Lauren Rutledge, Joyce Nair-Menon, Micheal Yost, Antonis Kourtidis. Regulation and functional role of the cell-cell junction-associated RNAi machinery in colon 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 64.
Abstract 642: Increasing BMI is associated with tumor volume and uterine weight in endometrial cancer
Authors: Zhang Yingao , Desravines Nerlyne , Moore Dominic T. , Gehrig Paola A. , Clark Leslie H. , Bae-Jump Victoria L.
Published: August 2019

Obesity and diabetes is associated with increased risk and worse outcomes in endometrial cancer (EC). Our previous work has found that obesity leads to more aggressive tumor behavior and a doubling of tumor size in EC mouse models. Thus, we explored the association between endometrial tumor volume (TV) and uterine weight (UW) as a function of BMI in EC patients. We performed a retrospective review of EC patients diagnosed and treated from 2005-2010 at a single tertiary care center. Tumor histology, dimensions, stage, grade, and UW were extracted from surgical pathology reports. Patient age, BMI, and diabetic status were also collected. TV was determined using the formula TV=l*d*w*pi/6. Independent variables include obesity classification, BMI, and diabetic status. Univariable and multivariable regression models were used to assess strength of association. A total of 564 EC patients were evaluated, with either endometrioid (n=522) or serous (n=42) histologies (Table 1). Within the endometrioid subgroup, associations were noted for both increased TV and UW with obesity (TV p=0.02; UW p<0.001) and BMI (TV p=0.007; UW p<0.001). Within the serous subgroup, increased TV was associated with obesity and BMI (p<0.001; p=0.007), but UW was only associated with obesity (p=0.005) and not BMI (p=0.07). Controlling for stage and grade, multivariable models demonstrated similar patterns with obesity status and BMI within both subgroups. Multivariable analyses also demonstrated that having diabetes (n=160) was associated with both higher TV and UW in the endometrioid subgroup (TV p=0.004; UW p=0.02), but not in the serous subgroup (TV p=0.71; UW p=0.12). Obesity and increasing BMI appear to be associated with both increased TV and UW in EC patients, consistent with our previously described findings in murine models. Diabetes was also associated with higher TV and UW, but was only significant in patients with endometrioid ECs. Our results may partially account for the worse outcomes seen in obese EC patients.

Table 1EC HistologyMedianIQREndometrioid (n=522)Uterine Weight (g)Non-obese118.183.0-157.5Obese140.0103.8-200.0Tumor Volume (cm3)Non-obese3.6950.927-9.664Obese4.3591.419-14.216Serous (n=42)Uterine Weight (g)Non-obese116.667.4-166.6Obese189.8127.6-252.6Tumor Volume (cm3)Non-obese3.2490.601-9.382Obese23.8247.356-59.494

Citation Format: Yingao Zhang, Nerlyne Desravines, Dominic T. Moore, Paola A. Gehrig, Leslie H. Clark, Victoria L. Bae-Jump. Increasing BMI is associated with tumor volume and uterine weight in endometrial 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 642.
Abstract 647: Immunoprevention of triple negative breast cancer by a TOP2A multi-peptide vaccine
Authors: Lee Sang Beom , Pan Jing , Jiang Yueqiang , Palen Katie , Johnson Bryon , Fernando Romaine Ingrid , Sei Shizuko , Lubet Ronald , You Ming , Wang Yian
Published: August 2019

Triple negative breast cancer (TNBC) comprises approximately 20% of all breast cancers, and it has a poor prognosis due to aggressive growth characteristics and lack of effective targeted therapies. Emerging data suggest vaccination as a promising approach to prevent TNBC as well as other cancer types, but appropriate vaccine targets need to be identified. In the present study, we employed in silico analyses to select genes that are uniquely expressed in TNBC. Topoisomerase 2 alpha (Top2A) was found to be highly expressed in human TNBC. Top2A is a key enzyme in DNA replication and has been identified as a target of anti-cancer therapy. Overexpression of Top2A was confirmed in human breast cancer tissue microarrays and in mouse TNBCs derived from C3(1)/TAg mice. Top2A-specific MHC class II epitopes with optimal binding affinity were identified using the “combined scoring system”, based on their potential to elicit a Th1 immune response. Furthermore, all peptides were selected to have 100% amino acid sequence homology between humans and mice for potential clinical translation. Four Top2A peptides with strong immunogenic responses were selected by immunization of tumor naïve mice followed by IFN-γ-based ELISPOT assays. These peptides were combined to form a multi-peptide Top2A vaccine. The anti-tumor efficacy of the Top2A vaccine was tested in a preventive setting using a syngeneic TNBC model in C3(1)/TAg-REAR mice. Vaccinated animals demonstrated a significant reduction in tumor growth when compared to the adjuvant control with tumor size decreasing from 278.1 mm3 in adjuvant control to 150.4 mm3 in vaccinated group (p < 0.05). Taken together, our data clearly demonstrate that the Top2A multi-peptide vaccine is highly immunogenic and provides immunopreventive efficacy against TNBC in mice.

Citation Format: Sang Beom Lee, Jing Pan, Yueqiang Jiang, Katie Palen, Bryon Johnson, Romaine Ingrid Fernando, Shizuko Sei, Ronald Lubet, Ming You, Yian Wang. Immunoprevention of triple negative breast cancer by a TOP2A multi-peptide vaccine [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 647.
Abstract 663: Diagnostic screen for lung cancer using high throughput metabolomics screen
Authors: Plumb Robert S. , Gethings Lee , Peck Andrew
Published: August 2019

Lung cancer is one of the most common cancers in the world. It is a leading cause of cancer death in men and women in the United States, cigarette smoking being leading cause of lung cancer. There are two main types of lung cancer small cell and non-small cell lung cancer (NSCLC). The most common type being NSCLC which accounts for 80-85% of all reported cases. The ability to rapidly detect / diagnose lung cancer at an early stage is critical to successful treatment selection and patient survival.

The use of metabolic profiling in biomedical applications metabolic profiling is being deployed as a method for finding novel, mechanistic, biomarkers of disease with obvious potential for improving diagnosis, and patient stratification. Here we present a rapid, simple and reliable high throughput targeted LC/MS single platform, for the quantification/monitoring of small molecule metabolites, lipids and peptides. The methodology employs a single set of LC/MS conditions which facilitate TCA cycle, bile acids, biogenic amine, free fatty acids, acyl carnitines, lipids and 100 protein panel without need for user intervention.

Plasma from pilot cohort of lung cancer a health control samples were evaluated using this new methodology. The methodology showed excellent reproducibility and accuracy. The peptide analysis showed that 10 peptides were shown to be markers of lung cancer, these are listed in Table 1. Acylcarnitines were quantified over a range of 5 - 625ng/mL data showed that the C14:2 tetradecadienoyl carnitine and C16:1 palmitoleoyl carnitine were elevated in lung cancer samples whereas the C8:1 octenoyl carnitine level was reduced in the lung cancer samples . Although deoxycholic acid and chendeoxycholic acid appeared to be reduced in the lung cancer samples this was determined to be not statistically relevant. A total of 29 amino acids were measured during the analysis. Using t-test with p-value FDR cutoff adjusted to 0.01 the data clearly showed that sarcosine was highly over-expressed in lung cancer samples.

Table 1P02749Apolipoprotein HP19652Alpha-1-acid glycoprotein 2P68871Hemoglobin subunit betaP02671Fibrogen alpha chainP02656Apolipoprotein C-IIIP02750Leucine-rich alpha 2-glycoprotein (LRG)P00738HaptoglobinP02748Complement component C9P02763Alpha-1-acid glycoprotein 1P01011Alpha-1-antichymotrypsin

Note: This abstract was not presented at the meeting.

Citation Format: Robert S. Plumb, Lee Gethings, Andrew Peck. Diagnostic screen for lung cancer using high throughput metabolomics screen [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 663.
Abstract 673: Impact of PD-L1 feedback modulation on the sensitivity of anti-PD-1/PD-L1
Authors: Zhu Jian , Zhu Rebecca L.
Published: August 2019

The long-term benefits of anti-PD-1/PD-L1 are limited in most cancer patients. Resistance and insensitivity are major challenges of this therapeutic class. Feedback regulation of PD-L1 could serve as a mechanism of drug resistant, especially negative feedback loops where inhibition of PD-L1 may lead to up-regulation of its own expression and function. In this study, computational models were established to investigate PD-L1 feedback modulations to facilitate discovery of biomarkers for drug sensitivity and to inform design of drug combination studies. As of October 2018, over 10,000 publications on cancer and immunosuppressive factors (e.g. PD-1, PD-L1, CTLA4, IDO1, TIM-3, LAG3, TIGIT) were found in PubMed. About 2500 abstracts of these articles were downloaded and systematically analyzed through natural language processing and text mining. Information on gene/drug, pathway/disease along with results from cell/animal/clinical studies were extracted, standardized and integrated into a directed graph-based database. Algorithms were designed to construct networks and identify PD-L1 feedback loops (FBLs). Fisher's Exact tests were performed to evaluate the significance of factors involved in (+) and (-)FBLs. The computationally generated PD-L1 feedback network comprises 102 nodes, 216 interactions with 1080 possible FBLs (502 positive vs 578 negative). In particular, up-regulation of IL10, pEGFR, SRC, HIF1A, ISX, NF-kappaB, pSMAD2/SMAD2, IFN-gamma were strongly associated with up-regulation of PD-L1 upon inhibition of PD-1/PD-L1 (p<0.0001, (-)FBLs). Interestingly, TIM-3, a potential IO target and commonly up-regulated by anti-PD-1/PD-L1, is involved in both (+) and (-)FBLs. Inhibition of TIM-3 (e.g. anti-TIM-3) could decrease IL-10 in Tregs and enhance efficacy of anti-PD-1/PD-L1, but meanwhile, anti-TIM-3 could activate NF-kappaB, IRF7 and STAT1, leading to up-regulation of PD-L1 expression and potential resistance to anti-PD-1/PD-L1. In conclusion, the literature-based network analysis proved to be effective to identify potential regulatory pathways driving resistance to immunotherapies, to prioritize the selection of biomarkers, and to assess risk of drug combinations. Particularly, through the analysis of feedback regulations of PD-L1, novel therapeutic approaches to boost efficacy of anti-PD-1/PD-L1 could be postulated. Targeting genes involved in both (+) and (-)FBLs of PD-L1, such as TIM-3, needs to be carefully evaluated.

Citation Format: Jian Zhu, Rebecca L. Zhu. Impact of PD-L1 feedback modulation on the sensitivity of anti-PD-1/PD-L1 [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 673.
Abstract 676: Leveraging TCGA gene expression data to build predictive models for cancer drug response
Authors: Clayton Evan
Published: August 2019

Personalized oncology promises to increase the success rate of cancer drug therapy by using molecular tumor profiles to determine the optimal therapeutic for an individual. 5-Fluorouracil and Gemcitabine are two Food and Drug Administration approved chemotherapeutics commonly prescribed to cancer patients. Here, we build machine learning models using gene expression data from patients’ primary tumors to predict their response, positive or negative, to these drugs. We compared several clustering and classification methods for predicting response. Of those tested, Clara was found to be the best clustering algorithm while random forest was the best classification method, with prediction accuracy up to 86%. We determined that models trained across several cancers outperform single cancer models. We also found that genes most informative for predicting drug response were enriched in well-known cancer signaling pathways, highlighting their potential significance in chemotherapy prognosis. Ultimately, such predictive models may aid oncologists with making critical treatment decisions.

Citation Format: Evan Clayton. Leveraging TCGA gene expression data to build predictive models for cancer drug response [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 676.
Abstract 68: Comparative transcriptome analysis of primary tumor and matched tumor thrombus reveals that coagulation cascades activation contributes to the venous metastases of osteosarcoma
Authors: Ji Tao , Wang Yifei , Guo Yi , Li Dasen , Liang Haijie , Ren Tingting , Guo Wei
Published: August 2019

Comparative transcriptome analysis of primary tumor and matched tumor thrombus reveals that coagulation cascades activation contributes to the venous metastases of osteosarcoma

Osteosarcoma is the most frequent malignant primary tumor of bone in children and adolescents with an approximately 5-year survival about 50-60%. Lung metastasis remains as the most challenging condition during treatment and follow-up, which has been regarded as the main reason for bottle neck of prognosis. Tumor thrombus is occasionally diagnosed in patients with malignant tumor. It usually exists during tumor angiogenesis associated with proliferation and metastasis. The presentation shows aggressive features and advanced stage of the disease. Tumor thrombus is associated with highly metastasis potential. Based on the evolution theory of tumor metastasis, tumor thrombus can be the intermediate phase between local invasion and distal metastasis. In order to investigate the underlying mechanism and identify candidate metastasis driver genes, we collected a set of high quality paired tumor tissue, both the primary site and corresponding tumor thrombus that were treated by thrombectomy. In this study, we present a large-scale transcriptome analysis, by RNA sequencing, of 4 patients diagnosed with large pelvic osteosarcoma with tumor thrombus. The dysregulated genes between primary site and thrombus suggested that the complement and coagulation cascades, focal adhesion and extracellular matrix receptor was correlated with the formation of tumor thrombus. Tissue factor was identified as a major marker in coagulation pathway activation. We also identified NOTCH2NL-NBPF1 fusion genes in all four patients. All of these findings broaden our knowledge of osteosarcoma thrombus and may also contribute to the novel mechanism of metastasis in osteosarcoma.

Citation Format: Tao Ji, Yifei Wang, Yi Guo, Dasen Li, Haijie Liang, Tingting Ren, Wei Guo. Comparative transcriptome analysis of primary tumor and matched tumor thrombus reveals that coagulation cascades activation contributes to the venous metastases of osteosarcoma [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 68.
Abstract 681: Methylation biomarkers of cellular response to demethylating drug
Authors: Wang Xuewei , Kocher Jean-Pierre , Sun Zhifu
Published: August 2019

DNA methylation plays an import role in both tumorigenesis and therapy resistance. Demethylating drug regimens have shown efficacy in hematological malignancies, and are actively studied for other cancer types. High variability in patients’ responses to these drugs has been observed and highlights the demand to identify predictive biomarkers for these demethylating drugs. In particular, decitabine (latest FDA-approved demethylating drug) demonstrated as the most potent agent and has the best response on hematological cell lines. Two public datasets were collected to identify methylation biomarkers of decitabine. The first one is from Genomics of Drug Sensitivity in Cancer, with methylation data from Illumina 450K array and drug response from cell lines. The other one is a publicly available patient dataset (methylations from RRBS sequencing and decitabine response from clinic). 64900 CpG sites common to the two methylation platforms were used to train machine learning model (Elastic Net), based on the methylation data and decitabine response (AUC) from 108 hematological cell lines in GDSC. 73 CpGs were identified as predictive biomarkers by Elastic Net. The trained model was then used to make predictions on 39 patients (20 responders and 19 non-responders) with their methylation profiles of the same set of CpGs. The predicted response of responder group is indeed better than non-responder group (p-value = 0.0501), indicating the validity of identified methylation biomarkers. The top one CpG identified by Elastic Net resides in the promoter region of gene C20orf85, and is significantly hypermethylated in non-responder group (p-value = 0.0458), suggesting its potential involvement in the resistance to decitabine.

Citation Format: Xuewei Wang, Jean-Pierre Kocher, Zhifu Sun. Methylation biomarkers of cellular response to demethylating drug [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 681.
Abstract 697: Degradation of cancer mitochondria by the mitochondria-eating protein induces iron-dependent cancer cell death
Authors: Nakamura Yasuyuki , Ikari Naoki , Arakawa Hirofumi
Published: August 2019

Parkin/Pink1-mediated mitophagy plays a critical role in mitochondrial quality control, in which the unhealthy mitochondria are sequestered by autophagosomes, and degraded by fusion between autophagosomes and lysosomes. Here, we report non-canonical mitophagy as a new function of tumor suppressor p53. The mitochondria-eating protein (Mieap) was originally identified as a p53-target gene. Mieap induced large vacuole-like structures (Mieap-induced vacuoles: MIVs) in cancer cells. MIVs were generated from around the mitochondria, and directly ate and degraded cancer mitochondria without autophagosomes. Therefore, we defined the function as the Mieap-regulated non-canonical mitophagy. UVRAG was found to be a Mieap-interacting protein that mediates the MIVs formation. Interestingly, degradation of cancer mitochondria by the MIVs induced cell death via iron-dependent production of reactive oxygen species. Introduction of exogenous Mieap strikingly suppressed in vivo tumor growth of Mieap-deficient colorectal cancer cells. Deficiency of endogenous Mieap dramatically promoted cancer development, malignancy, and aggressiveness in colorectal and gastric cancer model mice. The Mieap-induced cell death was found to occur in in vivo hypoxic tumor microenvironment of colorectal cancer tissues. These results suggest that degradation of cancer mitochondria by Mieap-regulated non-canonical mitophagy plays a critical role in p53 tumor suppression via iron-dependent cancer cell death.

Citation Format: Yasuyuki Nakamura, Naoki Ikari, Hirofumi Arakawa. Degradation of cancer mitochondria by the mitochondria-eating protein induces iron-dependent cancer cell death [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 697.
Abstract 698: Enhancing apoptosis to improve the therapy of EGFR-mutant lung cancer
Authors: Tanaka Kosuke , Han Song , Ganesan Yogesh T. , Hsieh James J. , Cheng Emily H.
Published: August 2019

EGFR-TKIs have shown clinical efficacy in EGFR-mutant lung cancer patients, but most of the cases eventually develop resistance to the drugs, including the third-generation EGFR-TKI. Here, we develop a cell death mechanism-based combination strategy to enhance the therapeutic effect of EGFR-TKIs as well as to overcome the acquired resistance to EGFT-TKI in EGFR-mutant lung cancer. Through high-throughput screening of a custom pathway inhibitor library (-1000 drugs), we discovered Aurora kinase inhibitors that synergized with a third-generation EGFR-TKI (AZD9291) to induce apoptosis in EGFR-mutant lung cancer cells. Aurora kinase inhibitors enhanced AZD9291-mediated induction of BIM and PUMA, two important sentinels interconnecting the EGFR signal transduction pathway and the mitochondrial apoptotic machinery BAX and BAK. Combined inhibition of Aurora kinases and EGFR prevented the emergence of EGFR-TKI resistant clones in clonogenic assays. Interestingly, AZD9291 resistant lung cancer cells exhibited greater sensitivity to Aurora kinase inhibitors compared with parental cells. In conclusion, we found that the combination of AZD9291 and Aurora kinase inhibitors synergistically induced BIM and PUMA-mediated apoptosis, which may render a promising strategy to improve the therapy of EGFR-mutant lung cancer.

Citation Format: Kosuke Tanaka, Song Han, Yogesh T. Ganesan, James J. Hsieh, Emily H. Cheng. Enhancing apoptosis to improve the therapy of EGFR-mutant lung 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 698.
Abstract 699: KMT2A regulates cervical cancer cell growth andapoptosisthrough cooperation with VDAC1
Authors: Zhang Changlin , Li Yizhuo , Long Qian , Liu Tianze , Qin Ge , Shi Dingbo , Chen Miao , Zhang Kefang , Shi Shuihan , Deng Wuguo
Published: August 2019

Cervical cancer is an aggressive cutaneous malignancy, illuminating the exact mechanisms of tumorigenesis and discovering novel therapeutic targets are urgently needed. Lysine methyltransferase 2A (KMT2A) is a transcriptional co-activator regulating gene expression during early development and hematopoiesis, but the role of KMT2A in cervical cancer remains largely unknown. Here, we showed that KMT2A regulated cervical cancer cell growth and apoptosis through cooperation with VDAC1 (voltage dependent anion channel 1). Knockdown of KMT2A significantly inhibited cell viability and cell migration and induced apoptosis in cervical cancer cells. Overexpression of VADC1 reversed the KMT2A knockdown-mediated suppression of cell viability and cell migration and induction of apoptosis. The results from a xenograft mouse model confirmed that KMT2A regulated cervical cancer growth via the VDAC1 signaling. Moreover, analyses of clinical samples demonstrated that the expression of KMT2A and VDAC1 were positively correlated in cervical cancer tissues, especially in cervical intraepithelial neoplasia (CIN) samples. Collectively, our results indicated that KMT2A promoted cervical cancer growth by cooperating with the VDAC1, suggesting that the KMT2A/VDAC1 signaling pathway may be a potential therapeutic target for cervical cancer.

Citation Format: Changlin Zhang, Yizhuo Li, Qian Long, Tianze Liu, Ge Qin, Dingbo Shi, Miao Chen, Kefang Zhang, Shuihan Shi, Wuguo Deng. KMT2A regulates cervical cancer cell growth andapoptosisthrough cooperation with VDAC1 [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 699.
Abstract 7: Discovery of OICR-10268: A potent and selective BCL6 inhibitor
Authors: Watson Iain D. , Isaac Methvin , Wilson Brian , Chau Anh , Morin Justin , Subramanian Pandiaraju , Mamai Ahmed , Joseph Babu , Prakesch Michael , Uehling David , Abibi Ayome , Marcellus Richard , Strathdee Craig , Subramaniam Ratheesh , Theriault Brigitte , Winston Jeffrey , Chan Manuel , Griffin Carly , Cheung Herman , Kiyota Taira , Undzys Elijus , Aman Ahmed , Poda Gennady , Kuntz Doug , Pomroy Neil C. , Privé Gil G. , Al-awar Rima
Published: August 2019

The transcription factor B cell lymphoma 6 (BCL6) is required for the generation of an effective humoral immune response through the development and maintenance of germinal centers (GCs). The inhibition of the protein−protein interaction between BCL6 and its corepressors has been implicated as a therapeutic target for diffuse large B-cell lymphoma (DLBCL), a type of non-Hodgkin’s lymphoma (NHL). Using structure-based drug design, we initiated a program to identify novel BCL6 inhibitors. We identified a high micromolar virtual screening hit which was then optimized for potent biophysical binding and anti-proliferative cellular activity resulting in the identification of OICR-10268, a potent and selective Bcl6 inhibitor.

Citation Format: Iain D. Watson, Methvin Isaac, Brian Wilson, Anh Chau, Justin Morin, Pandiaraju Subramanian, Ahmed Mamai, Babu Joseph, Michael Prakesch, David Uehling, Ayome Abibi, Richard Marcellus, Craig Strathdee, Ratheesh Subramaniam, Brigitte Theriault, Jeffrey Winston, Manuel Chan, Carly Griffin, Herman Cheung, Taira Kiyota, Elijus Undzys, Ahmed Aman, Gennady Poda, Doug Kuntz, Neil C. Pomroy, Gil G. Privé, Rima Al-awar. Discovery of OICR-10268: A potent and selective BCL6 inhibitor [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 7.
Abstract 70: Oxidative stress and cell death in multidrug-resistant cancer
Authors: Aryal Baikuntha P. , Gillet Jean-Pierre , Rao V Ashutosh
Published: August 2019

Reactive oxygen species (ROS) are constantly generated in mitochondria as a consequence of normal cellular metabolic reactions and play vital roles in cell signaling and homeostasis. Prolong production of elevated levels of ROS may lead to oxidative damage, genetic instability, and genetic mutation causing cancer initiation and progression. One proposed therapeutic strategy is to alter oxidative stress from a ROS-induced tumor promoting event to a ROS-induced apoptotic signaling to kill cancer cells. Most ROS inducing therapeutic agents initially induce ROS to kill cancer cells by apoptosis; however, prolonged exposure of such drugs may increase ROS scavenging enzymes resulting in a reduced ROS levels and an adaptive microenvironment that makes cancer cells resistant to the same ROS-inducing therapeutic agents. Chronic exposure of ROS may also upregulate the multidrug resistance associated protein expression in cancer cells. We hypothesize that induction of oxidative stress may sensitize drug resistant cancer cells leading to cell death. We used two pairs of cell lines, a mitoxantrone sensitive and mitoxantrone resistant MCF7 human breast adenocarcinoma (MCF7/MCF7MX) and a H460 non-small lung cancer cell line (H460/H460MX) for our experiments to study the impact of oxidative stress on drug resistance. We employed western blot, ELISA assays, and FACS analysis to determine the total and specific protein oxidative carbonylation, protein aggregation, and cell death by apoptosis or necrosis. At basal level, both mitoxantrone resistant MCF7MX and H460MX cells showed slightly lower level of total protein carbonylation compared to their mitoxantrone sensitive counterparts. Chemical induction of oxidative stress by external sources increased the total protein carbonylation and cell death in both mitoxantrone sensitive and resistance cancer cells. We also observed an increase in protein aggregation with increasing oxidative stress. Our results indicate that oxidative stress-induced protein oxidation by carbonylation and aggregation of oxidized protein which may sensitize drug resistant cancer cells leading to cell death.

Citation Format: Baikuntha P. Aryal, Jean-Pierre Gillet, V Ashutosh Rao. Oxidative stress and cell death in multidrug-resistant 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 70.
Abstract 705: Tumor intrinsic PD-L1 is mediated by BRAFV600E and can regulate drug-induced apoptosis in human colon cancer
Authors: Feng Daofu , Pal Krishnendu , Mukhopadhyay Debabrata , Liu Xin , Dong Haidong , Huang Shengbing , Sinicrope Frank A.
Published: August 2019

Resistance to chemotherapy remains a major obstacle to improving outcomes of cancer treatment. Recent data suggest intrinsic functions of immune checkpoint proteins, including PD-L1, that may be distinct from their immunoregulatory roles. We found that colorectal cancer (CRC) cells with mutant BRAF(V600E) or KRASalleles show increased PD-L1 transcripts and expression of cell surface PD-L1 protein compared to those with wild-type copies. Furthermore, ectopic expression of BRAFV600E or mutant KRAS induced PD-L1 whose upregulation was attenuated by inhibition of RAS-RAF-MEK-ERK signaling. Combined knockdown of MEK/ERK effectors c-JUN and YAP markedly reduced PD-L1. Knockout of PD-L1 in RKO cells resulted in a reduction in chemotherapy-induced DNA double strand breaks (pH2AX) and caspase-3 cleavage compared to parental cells. Results were confirmed in PD-L1 knockout MC38 murine CRC cells where re-expression of PD-L1, but not its deletion mutants, was shown to induced pH2AX and apoptosis. In cells with knockout of PD-L1, reductions in p-AKT and the BH3-only proteins BIM and BIK were observed that could be restored by re-expression of PD-L1. Treatment of cells with an anti-PD-L1 antibody reduced p-AKT, BIM and BIK, and also attenuated chemotherapy-induced apoptosis. Re-expression of BIM in PD-L1 knockout cells restored apoptosis. Murine tumor xenografts generated from cells with knockout of PD-L1 displayed resistance to oxaliplatin-induced tumor regression compared to tumors derived from parental cells. Using TCGA datasets, human CRCs with high vs low PD-L1 mRNA levels were significantly associated with better survival. In summary, we identified a non-immune function of tumor cell-intrinsic PD-L1 that upregulates BH3-only proteins BIM and BIK to promote apoptosis, thereby suggesting its potential as a predictive biomarker.

Citation Format: Daofu Feng, Krishnendu Pal, Debabrata Mukhopadhyay, Xin Liu, Haidong Dong, Shengbing Huang, Frank A. Sinicrope. Tumor intrinsic PD-L1 is mediated by BRAFV600E and can regulate drug-induced apoptosis in human colon 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 705.
Abstract 709: Cordycepin induces apoptosis of human ovarian cancer cells by inhibiting CCL5-mediated Akt/NF-κB signaling pathway
Authors: Jang Ik-Soon
Published: August 2019

The chemokine, CCL5, is a key mediator for the recruitment of immune cells into tumors and tissues. Akt/NF-κB signaling is significantly activated by CCL5. However, the role of NF-κB inactivation in apoptosis induced by negative regulation of CCL5 remains unclear. Here, we analyzed the effect of cordycepin on NF-κB activity in SKOV-3 cells and found that cordycepin-mediated inhibition of NF-κB signaling induced apoptosis in SKOV-3 cells via the serial activation of caspases. In addition, immune-blotting analysis showed that CCL5 is highly expressed in SKOV-3 cells. In addition to activating caspases, we show that, cordycepin prevents TNF-α-induced increase in CCL5, Akt, NF-κB, and c-FLIPLactivation and that CCL5 siRNA could inhibit Akt/NF-κB signaling. Moreover, cordycepin negatively regulated the TNF-α-mediated IκB/NF-κB pathway and c-FLIPL activation to promote JNK phosphorylation, resulting in caspase-3 activation and apoptosis. Also, we show that c-FLIPL is rapidly lost in NF-κB activation-deficient. siRNA mediated c-FLIP inhibition increased JNK. SP600125, a selective JNK inhibitor, downregulated p-JNK expression in cordycepin-treated SKOV-3 cells, leading to suppression of cordycepin-induced apoptosis. Thus, these results indicate that cordycepin inhibits CCL5-mediated Akt/NF-κB signaling, which upregulates caspase-3 activation in SKOV-3 cells, supporting the potential of cordycepin as a therapeutic agent for ovarian cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Ik-Soon Jang. Cordycepin induces apoptosis of human ovarian cancer cells by inhibiting CCL5-mediated Akt/NF-κB signaling 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 709.
Abstract 710: Real-time apoptosis and necrosis detection in 3D spheroid cellmodels
Authors: Niles Andrew L. , Kupcho Kevin R. , Riss Terry L. , Lazar Dan F. , Cali James J.
Published: August 2019

The structural and biological complexity of cancer spheroids make them uniquely suited to address or approximate the pharmacodynamics of new chemical entities directed at chemotherapeutic reduction of tumor masses. Unfortunately, this same complexity represents a substantial technical challenge for measuring apoptosis and necrosis using conventional methods in real time. To address this challenge, we explored the use of a multiplexed bioluminescent and fluorescent, real-time assay reagent which utilizes annexin V fusion proteins containing binary elements of a complementing luciferase, a time-released luciferase substrate and an excludable DNA membrane integrity dye. The assay reagent was added to HCT-116 and HepG2 spheroids dosed with serial dilutions of panobinostat, bortezomib, or paclitaxel to examine the magnitude, potency and kinetics of the responses. Luminescent and fluorescent data were continuously collected throughout the 48h exposure using a conventional multimode plate reader equipped with atmospheric control. After the exposure period, the treated spheroids were imaged by fluorescence microscopy to assess necrotic burden as well as analyzed for remaining cellular viability using ATP content compared to untreated control. Consistent with their known modes of cytotoxic action, bortezomib produced the first measurable phoshatidylserine (PS) exposure (~8h), followed by panobinostat (~12h) and paclitaxel (18-22h). The apoptotic potencies and magnitudes of response matured thereafter as a function of time until 48h. Secondary necrosis resulting from completion of the apoptotic program lagged PS exposure in both time and magnitude. Imaging results for necrosis and ATP determinations provided orthogonal and inversely complementary verification of data values obtained with the apoptosis assay reagent. Taken together, this workflow may help define the in vitro pharmacological disposition of new chemical entities with respect to their capability to penetrate and induce apoptosis in spheroids and/or other more complex multicellular models.

Citation Format: Andrew L. Niles, Kevin R. Kupcho, Terry L. Riss, Dan F. Lazar, James J. Cali. Real-time apoptosis and necrosis detection in 3D spheroid cellmodels [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 710.
Abstract 714: Cannabidiol-induced apoptosis is mediated by activation of Noxa in human colorectal cancer cells
Authors: Yun Hye Kyeong , Jeong Soyeon , Jeong Yoon A , Jo Min Jee , Kim Jung lim , Park Seong Hye , Kim Bo Ram , Na Yoo Jin , Kim Dae Young , Kim Bu Gyeom , Lee Dae Hee , Oh Sang Cheul
Published: August 2019

Cannabidiol (CBD), one of the compounds present in the marijuana plant, has anti-tumor properties, but its mechanism is not well known. The authors aimed to evaluate the apoptotic action of CBD in colorectal cancer cells (CRC), and focused on its effects on the novel pro-apoptotic Noxa-reactive oxygen species (ROS) signaling pathway. CBD experiments were performed using the colorectal cancer cell lines HCT116 and DLD-1. CBD induced an apoptosis by regulating many pro- and anti-apoptotic proteins, of which Noxa showed a significantly higher expression. To understand the relationship between Noxa and CBD-induced apoptosis, authors downregulated Noxa levels using siRNA and confirmed that the expression of apoptosis markers was decreased. After ROS production was blocked, the level of Noxa also decreased, suggesting that ROS is involved in the regulation of Noxa. The Noxa and ROS are well-known pro-apoptotic signaling agents. As a result, CBD was found to induce apoptosis in a Noxa-and-ROS-dependent manner. The effects of CBD treatment were re-demonstrated in vivo, thus confirming its role as a novel, reliable anticancer drug

Citation Format: Hye Kyeong Yun, Soyeon Jeong, Yoon A Jeong, Min Jee Jo, Jung lim Kim, Seong Hye Park, Bo Ram Kim, Yoo Jin Na, Dae Young Kim, Bu Gyeom Kim, Dae Hee Lee, Sang Cheul Oh. Cannabidiol-induced apoptosis is mediated by activation of Noxa in human colorectal 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 714.