bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 3938: The alkylphosphocholine erufosine induces ER and mitochondrial stress in OSCC cells
Authors: Ansari Shariq S. , Sharma Ashwini , Ali Doaa M. , Köng Rainer , Berger Martin R.
Published: August 2018

Endoplasmic reticulum (ER) plays an essential role in cell function and survival. Accumulation of unfolded or misfolded proteins in the lumen of the ER activates the unfolded protein response (UPR), resulting in ER-stress and subsequent apoptosis. The alkylphosphocholine erufosine is a known Akt-mTOR inhibitor in oral squamous cell carcinoma (OSCC). In the present study we decipher a new facet of erufosine's mechanism of action and evaluate its role to induce ER and mitochondrial stress leading to autophagy, apoptosis and ROS induction. The cellular toxicity of erufosine was determined by MTT assay after 24h, 48h and 72h exposure in two OSCC cell lines, HN-5 and SCC-61. Based on these results, HN-5 cells were exposed to erufosine at concentrations corresponding to IC25, IC50 and IC75 concentrations, and then their mRNA was isolated and analyzed by Illumina Chip array. Gene expression and gene set enrichment analyses were performed to identify core KEGG (Kyoto Encyclopedia of Genes and Genomes) pathways and GO (Gene Ontology) terms. A positive enrichment of ER stress upon erufosine exposure was observed, which was verified at protein level for the ER stress sensors and their downstream mediators. Upon erufosine exposure, calcium influx into the cytoplasm of the two OSCC cell lines was also observed. A differential regulation of genes and positive enrichment of processes pertaining to autophagy and apoptosis was also observed upon erufosine exposure. Autophagy was confirmed by increased acidic vacuoles and LC3-B levels. Apoptosis was confirmed by nuclear staining, annexin-V and immuno-blotting of caspases. The induction of mitochondrial stress upon erufosine exposure was predicted by gene enrichment analysis and shown by erufosine's effect on mitochondrial membrane potential, ATP and ROS production in OSCC cells. These data show that ER and mitochondrial targeting by erufosine represents a new facet of its mechanism of action as well as a promising new framework in the treatment of head and neck cancers. They furthermore support the future evaluation of erufosine as a therapeutic approach in cancer treatment alone, or in combination.

Citation Format: Shariq S. Ansari, Ashwini Sharma, Doaa M. Ali, Rainer Köng, Martin R. Berger. The alkylphosphocholine erufosine induces ER and mitochondrial stress in OSCC 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 3938.
Abstract 4030: Loss of E-cadherin expression is a critical step for peritoneal metastasis of gastric cancer with sub-serosal invasion
Authors: Togano Shingo , Yashiro Masakazu , Kuroda Kenji , Okuno Tomohisa , Miki Yuichiro , Hirakawa Kosei , Ohira Masaichi
Published: August 2018

Background: Peritoneal metastasis is the most frequent type of metastasis in gastric cancer. However, the actual mechanisms responsible for this metastasis are not understood clearly. The process of peritoneal metastasis requires some steps including the detachment of cancer cells from the primary tumor. E-cadherin, a calcium-dependent cell-cell adhesion molecule, is known to mediate intercellular adhesion. We examined the correlation between peritoneal metastasis of gastric cancer and reduced E-cadherin expression.

Methods: A total of 694 gastric cancer cases of which were resected at our department was enrolled in this study. The expression of E-cadherin of gastric cancer cells was evaluated by immunohistochemical staining. We analyzed the correlation between the E-cadherin expression and clinicopathological features. A case with less than 50% of E-cadherin expression was defined as E-cadherin negative.

Results: Among the 694 cases, 232 cases were E-cadherin negative. 171 cases developed peritoneal metastasis. Among the 171cases, 88cases were E-cadherin negative. E-cadherin-negative cancer was significantly (p<0.0001) associated with peritoneal metastasis. The 5-year survival rate of patients with E-cadherin-negative cancer (48.1%) was poor, in compared to that with E-cadherin-positive tumor (62.8%), while a multivariate analysis did not show significant difference. A multivariate analysis of sub-serosal invasion tumors (T3 stage tumor) indicated that only E-cadherin-negative was significantly associated with peritoneal metastasis (p=0.0005).

Conclusion: The loss of E-cadherin is a critical step in the promotion of cancer cells to a more malignant phenotype. E-cadherin is a useful prognostic factor for peritoneal metastasis in gastric carcinoma with sub-serosal tumor.

Citation Format: Shingo Togano, Masakazu Yashiro, Kenji Kuroda, Tomohisa Okuno, Yuichiro Miki, Kosei Hirakawa, Masaichi Ohira. Loss of E-cadherin expression is a critical step for peritoneal metastasis of gastric cancer with sub-serosal invasion [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 4030.
Abstract 4086: Efficient primary culture model of patient-derived epithelial cells from colorectal cancer using a ROCK inhibitor and feeder cells
Authors: Hong Hyekyung , Yun Nakhyeon , Kim Taewon , Lee Yeosong , Song Sujeong , Bae Jaemoon , Lee Wooyong , Cho Yongbeom
Published: August 2018

In vitro culture for patient-derived cells offers many advances in the development of novel therapies for colorectal cancer. Although various culture systems have been developed, experimental systems in which the long-term expansion of patient-derived epithelial cells still remained difficult. Here, we demonstrated that epithelial cells isolated from colorectal cancer patient-derived xenograft can be efficiently immortalized in conditioned medium from irradiated feeder cells containing a Rho kinase inhibitor Y-27632. Patient-derived tumor cells were rapidly proliferated to reach confluence in approximately 6 days. Short tandem repeat (STR) analysis of tumor tissue and cultured cells demonstrate that they have 13 identical STR loci and Amelogenin, Penta E, Penta D, D2S1338 and D19S433. The epithelial character was confirmed by staining for EpCAM, cytokeratin 20 and vimentin as a mesenchymal marker. Cells were continued to proliferate and formed well-defined tumor spheroids when transferred to 3D culture. To study the mechanism for promoting unrestricted cell proliferation, we investigated the expression of several cellular genes that fall into two designated pathways, induction of telomerase and cytoskeletal remodeling/p16 pathway. Moreover, patient-derived tumor cells were utilized for high-throughput screening of FDA approved drugs allowing detection of gene-drug associations. Collectively, we demonstrate that the long-term expansion of patient-derived tumor epithelial cells and these cells could be a useful tool in order to investigate a personalized treatment approach.

Citation Format: Hyekyung Hong, Nakhyeon Yun, Taewon Kim, Yeosong Lee, Sujeong Song, Jaemoon Bae, Wooyong Lee, Yongbeom Cho. Efficient primary culture model of patient-derived epithelial cells from colorectal cancer using a ROCK inhibitor and feeder 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 4086.
Abstract 4191: Effect of a novel orally bioavailable CXCR4 inhibitor, AMD070, on the metastasis of oral cancer cells
Authors: Kuribayashi Nobuyuki , Uchida Daisuke , Kinouchi Makoto , Kuribayashi Kyoko , Hasegawa Tomonori , Yagisawa Shuma , Kawamata Hitoshi
Published: August 2018

We have previously demonstrated that the stromal cell-derived factor (SDF)-1/CXCR4 system is involved in the metastasis of head and neck cancer. Additionally, it has been shown that the blockade of CXCR4 by subcutaneous daily injection with AMD3100, a CXCR4 antagonist, may be effective in preventing metastasis in CXCR4-related head and neck cancer. Recent investigations have suggested that AMD070, a novel orally bioavailable inhibitor of CXCR4, may be minimally invasive compared with AMD3100. In the present study, we examined the effect of AMD070 on metastasis induced by the SDF-1/CXCR4 axis in B88-SDF-1 oral cancer cells, which express high levels of SDF-1 and CXCR4. Although treatment with AMD070 did not affect the anchorage-dependent growth of B88-SDF-1 cells, it significantly suppressed the anchorage-independent growth. Moreover, the SDF-1/CXCR4-dependent migration and invasion of B88-SDF-1 cells was significantly inhibited following treatment with AMD070. Subsequently, we performed an experimental therapy using AMD070 to prevent the distant metastasis of B88-SDF-1 cells in vivo. Daily oral administration of AMD070 significantly inhibited the lung metastasis of B88-SDF-1 cells in nude mice. Furthermore, AMD070 treatment did not induce body weight loss despite the tumor bearing-condition of the mice. In addition, we could not detect the apparent macroscopic organ abnormalities associated with the daily oral administration of AMD070. These results indicated that AMD070 could be useful as a novel orally bioavailable inhibitor of oral cancer metastasis.

Citation Format: Nobuyuki Kuribayashi, Daisuke Uchida, Makoto Kinouchi, Kyoko Kuribayashi, Tomonori Hasegawa, Shuma Yagisawa, Hitoshi Kawamata. Effect of a novel orally bioavailable CXCR4 inhibitor, AMD070, on the metastasis of oral 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 4191.
Abstract 4203: Alpha 9 nicotinic acetylcholine receptor promotes tumor progression in triple negative breast cancer
Authors: Kuo Yung-Che , Chu Jan-Show , Lee Kha-Liang , Wang Hsiao-Feng , Drew Victor James , Zhuang Wei-Zhan , Chen Chi-Long , Ho Yuan-Soon , Huang Yen-Hua
Published: August 2018

Purpose: To assess the potential therapeutic target on the alpha 9 nicotinic acetylcholine receptor (α9-nAChR) and the insulin-like growth factor (IGF)-I receptor (IGF-IR) for preventing distant metastasis in TNBC patients. Experimental Design: Expression levels of stemness-related genes, α9-nAChR and IGF-IR in clinical tissues (n = 67) were analyzed by RT-qPCR and the Pearson correlation analysis. The α9-nAChR, OCT4, IGF-1R, and p-IGF-1R protein in tissues were detected by immunohistochemical staining (n = 200). The effects of either α9-nAChR activation or IGF-1R transactivation on stemness expression in TNBC were examined using Western blot, ALDH activity assay with flow cytometry, shRNA silencing, wound closure/migration/invasion assay, as well as microCT scanning and IVIS assay for animal models. Results: A high positive correlation between the gene expression levels of OCT4/NANOG/CD24/CD44 and α9-nAChR/IGF-IR in human breast cancer tissues was observed. IGF-1R and OCT4 highly expressed in tumor part compared than that of in normal part. Moreover, IGF-1R positively correlated with OCT4 in tumor tissues. We also show that nicotine induced α9-nAChR activation trans-activates the expression of IGF-1R and pluripotent protein OCT4, which mediated stemness-related properties in TNBC cell lines. Our results reveal that knockdown α9nAChR and IGF-1R diminish the pernicious effects of nicotine, inhibit stemness activities, and suppress metastasis both in vitro and in vivo. Conclusions: A crosstalk between α9-nAChR and IGF-IR signal mediates the stemness-related properties and metastasis therefore convene poor prognosis in TNBCs.

Citation Format: Yung-Che Kuo, Jan-Show Chu, Kha-Liang Lee, Hsiao-Feng Wang, Victor James Drew, Wei-Zhan Zhuang, Chi-Long Chen, Yuan-Soon Ho, Yen-Hua Huang. Alpha 9 nicotinic acetylcholine receptor promotes tumor progression in triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4203.
Abstract 4408: Development and validation of a serum miRNA biomarker panel for the detection of early stage lung cancer
Authors: Ying Lisha , Zou Ruiyang , Du Lingbin , Shi Lei , Zhang Nan , Jin Jiaoyue , Wu Junzhou , Zhang Fanrong , Chen Kaiyan , Yu Herbert , Mao Weimin , Su Dan
Published: August 2018

Lung cancer is the most prevalent cancer and leading cause of cancer death worldwide. Low-dose spiral computed tomography (LDCT) scan is being recommended as a screening test for smokers in the U.S. However, the poor specificity (73% specificity at 94% sensitivity) of LDCT has raised significant concern for its high chance of false positive (96% of LDCT positives are benign nodules). This study aims to develop a blood (serum) miRNA based molecular diagnostic test for the detection of early stage lung cancer. In the discovery phase, more than 400 miRNAs were profiled with MiRXES's qPCR based high throughput assay platform through a highly defined Chinese male smoker case-control cohort (n=424) where the cases were collected from Zhejiang Cancer Hospital and the controls were collected from the LDCT screening program in Zhejiang province, China. 29 miRNA biomarkers with p-value (FDR) < 0.01 and more than one z-score (standardized score) difference were identified and validated in another Chinese case-control cohort (n=432) collected from similar sources and a Caucasian case-control cohort (n=218) collected from EU and US. 18 out of the 29 miRNA biomarkers were validated in both cohorts (p-value < 0.01 and z-score >0.4). With multiple time of two-fold cross-validation, 5 miRNAs were found to be minimally required to form the biomarker panel for the accurate prediction of early stage lung cancer and the panel gives 0.984 (95% CI 0.973-0.991) AUC for the discovery cohort, 0.936 (95% CI 0.912-0.957) AUC for the Chinese validation cohort and 0.970 (95% CI 0.939-0.986) AUC for the Caucasian validation cohort. The 5-miRNA biomarker panel were then further validated in three additional Asian cohorts: a Chinese cohort collected from the similar sources (0.973, 95% CI 0.950-0.986, AUC), a Chinese cohort collected from independent sources (0.916, 95% CI 0.852-0.949, AUC) and a Singaporean cohort (Chinese, Malay and Indian population) (0.911, 95% CI 0.822-0.963, AUC).

Citation Format: Lisha Ying, Ruiyang Zou, Lingbin Du, Lei Shi, Nan Zhang, Jiaoyue Jin, Junzhou Wu, Fanrong Zhang, Kaiyan Chen, Herbert Yu, Weimin Mao, Dan Su. Development and validation of a serum miRNA biomarker panel for the detection of early stage lung 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 4408.
Abstract 45: Snai2 is a new target to mediate glucocorticoid signaling on breast cancer cell migration
Authors: Ling Jun , Singhal Adit , Lopez-Dee Zenaida P. , Porreca Brittany , Sprague Trinity
Published: August 2018

Steroid hormone receptors such as estrogen and progesterone receptors are well studied in breast cancer pathology; they are also used as drug targets for breast cancer therapy in the clinic. In contrast, glucocorticoid (GC) as a ubiquitous stress activated steroid hormone is less investigated in breast cancer. However, GC is frequently used as a co-treatment for breast cancer chemotherapy that generates some controversial effects to even promote cancer progression or recurrence in certain subtypes of breast cancer. To address this clinical issue, we focused on investigating the specificity of GC signaling on breast cancer cell behaviors in this study. When breast cancer cells were treated by dexamethasone (Dex), they were more responsive to cell migration (measured by electric cell-substrate impedance sensing, ECIS) than the effects on cell proliferation and apoptosis, wherein MDA-231 (triple negative breast cancer) cell was more sensitive to Dex than MCF7 cell (luminal A subtype). Further gene expression analysis by qRT-PCR microarray revealed that the glucocorticoid receptor (GR) responsive gene patterns are different between these two subtypes of breast cancer cells. One of remarked changes was that Snai2 (a zinc finger transcriptional factor) was highly activated in both cell lines. Western blotting analysis confirmed that MDA-231 cells have much higher basal level of Snai2 but with less fold increase upon Dex treatment than MCF-7. GR response elements (GREs) were also identified in the promoter region of Snai2, thus confirming Snai2 as a new target gene of GR in breast cancer. When Snai2 was knocked out by CRISPR, it was found that the basal migration rate was decreased in both cell lines. More importantly, their response rates to Dex treatment were also decreased as compared to the wild type cells. These decreases in cell migration are similar to those treated by GR antagonist RU486, thus confirming that Snai2 is a mediator to regulate the effects of GC/GR signaling on breast cancer cell migration. In summary, this study identified Snai2 as a new target gene of GR to regulate cell migration and potentially metastasis in response to GC signaling. The differential regulation of different subtypes of breast cancer cells by GC signaling was also confirmed. (+ equal contribution; * Corresponding author).

Citation Format: Jun Ling, Adit Singhal, Zenaida P. Lopez-Dee, Brittany Porreca, Trinity Sprague. Snai2 is a new target to mediate glucocorticoid signaling on breast cancer cell migration [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 45.
Abstract 4500: BET proteins regulate hyaluronan synthases in pancreatic ductal adenocarcinoma
Authors: Kumar Krishan , Ebine Kazumi , Pham Thao , Shang Meng , Munshi Hidayatullah G.
Published: August 2018

Background: Pancreatic ductal adenocarcinoma (PDAC) is associated with an extensive desmoplasia which is composed of multiple extracellular matrix components, including collagen, fibronectin, laminin, and hyaluronan (HA). Inhibitors targeting bromodomain and extra-terminal (BETi) proteins have emerged as potential therapeutic agents and are in clinical trials for a range of malignancies. We have previously shown that BETi attenuate fibrosis in vivo by decreasing collagen expression; however, the effect of BETi on HA has not been previously evaluated. Increased HA, which is synthesized by HA synthases (HAS), is associated with poor outcome in PDAC. Targeting HA has been postulated to provide a potential therapeutic opportunity to overcome PDAC chemo-resistance. Preclinical data have demonstrated that enzymatic degradation of HA with PEGPH20 remodels the tumor microenvironment, decreases the interstitial pressure and enhance the drug entry in tumors. Several ongoing clinical trials are evaluating PEGPH20 in combination with chemotherapies. We now report that BET inhibitors markedly decrease expression of HAS2 and HAS3, which are overexpressed in PDAC.

Purpose: The aim of this study is to investigate the role of BET proteins in HA synthesis.

Methods: mRNA expression levels HAS2 and HAS3 were determined in a panel of 7 PDAC cell lines by quantitative real-time RT-PCR. Protein expression was determined by western blotting.

Results: We found that treatment with BET inhibitors decrease the expression of HAS2 and HAS3 in all cell lines. Furthermore, siRNA mediated knockdown of individual BET proteins demonstrated that HAS2 is primarily regulated by BRD2, while HAS3 is primarily regulated by BRD4.

Conclusion: Our results demonstrate that BET proteins play a role in HA synthesis by regulating expression of HAS2 and HAS3 enzymes.

Citation Format: Krishan Kumar, Kazumi Ebine, Thao Pham, Meng Shang, Hidayatullah G. Munshi. BET proteins regulate hyaluronan synthases in pancreatic ductal adenocarcinoma [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 4500.
Abstract 4553: A novel bispecific antibody for EGFR-directed blockade of the PD-1/PD-L1 immune checkpoint
Authors: Koopmans Iris , Samplonius Douwe F. , Ginkel Robert J. van , Wierstra Peter J. , Heskamp Sandra , Bremer Edwin , Helfrich Wijnand
Published: August 2018

PD-L1-blocking antibodies produce significant clinical benefit in selected cancer patients by reactivating functionally-impaired anticancer T cells. However, their efficacy is potentially reduced by ‘on-target/off-tumor' binding to PD-L1 widely expressed on normal cells. This lack of tumor selectivity potentially induces a generalized activation of all antigen-experienced T cells as is evidenced by frequent autoimmune-related adverse events during and after treatment. To address these issues, we constructed a tetravalent bispecific antibody (bsAb), designated PD-L1xEGFR, to direct PD-L1-blockade to EGFR-expressing cancer cells and thus reactivate anticancer T cells more selectively. The IC50 of PD-L1xEGFR for binding to EGFR+ cancer cells was ~140 fold lower compared to an analogous PD-L1-blocking bsAb of irrelevant target antigen specificity (PD-L1xMock). Importantly, treatment with PD-L1xEGFR selectively enhanced activation status, INF-γ production, and activity of anti-CD3xanti-EpCAM-redirected T cells in presence of EGFR+/EpCAM+, but not EGFR-/EpCAM+ carcinoma cells. Similarly, capacity of PD-L1xEGFR to promote proliferation and IFN-γ production by CMVpp65-specific CD8+ effector T cells was enhanced in the presence of EGFR+/CMVpp65+ cancer cells. In contrast, the clinically-used PD-L1-blocking antibody MEDI4736 (durvalumab) promoted T cell activation indiscriminate of EGFR expression on cancer cells. Additionally, in tumor-bearing mice 111In-PD-L1xEGFR showed a significantly higher tumor uptake compared to 111In-PD-L1xMock. In conclusion, PD-L1xEGFR blocks the PD-1/PD-L1 immune checkpoint in an EGFR-directed manner, which promotes a more selective reactivation of anticancer T cells. This novel targeted approach may be useful to enhance efficacy and safety of PD-1/PD-L1 checkpoint blockade in EGFR-overexpressing malignancies.

Citation Format: Iris Koopmans, Douwe F. Samplonius, Robert J. van Ginkel, Peter J. Wierstra, Sandra Heskamp, Edwin Bremer, Wijnand Helfrich. A novel bispecific antibody for EGFR-directed blockade of the PD-1/PD-L1 immune checkpoint [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 4553.
Abstract 4600: Biomarker analysis in circulating cell-free DNA in patients treated with sorafenib for advanced hepatocellular carcinoma
Authors: Park Sook Ryun , Oh Chung Ryul , Kong Sun-Young , Kim Min Kyeong , Yoon Kyong-Ah , Cho Eun-Hae , Lee Junnam , Kang Jihoon , Ryoo Baek-Yeol
Published: August 2018

Purpose: We evaluated the potential role of Vascular Endothelial Growth Factor-A (VEGFA) amplification and genome-wide copy number variations (CNVs) using circulating cell-free DNA (cfDNA) as predictors of treatment outcome in hepatocellular carcinoma (HCC) patients treated with first-line sorafenib. Methods: Among 184 patients from a prospective biomarker cohort, who had started sorafenib between April 2015 and May 2016, 151 eligible patients were included in the analysis. Plasma cfDNA was extracted from peripheral blood in patients before starting sorafenib or healthy donors. Plasma VEGFA-to-EIF2C1 ratios (the VEGFA ratios) were determined using droplet digital polymerase chain reaction. We applied low depth whole genome sequencing in cfDNA to find CNVs and developed I-score to express genomic instability, which was defined as the sum of absolute Z-scores of sequenced reads on each chromosome. Results: The cfDNA concentrations were significantly higher in HCC patients than in healthy controls (0.71 vs. 0.34 ng/μL; p<0.0001). Patients who had progressive disease with sorafenib as best tumor response had significantly higher cfDNA levels (0.82 vs. 0.63 ng/μL; p=0.006) and I-scores (3,405 vs.1,024; p=0.0017) than those who achieved disease control. The disease control rate was 52.0% for the cfDNA-high group (above the median) vs. 75.0% for the cfDNA-low group (p=0.003), and 49.3% for the I-score-high group (above the median) vs. 77.6% for the I-score-low group (p=0.0003). The cfDNA-high group also had significantly worse TTP (median, 2.2 vs. 4.1 months; HR=1.71; p=0.002) and OS (median, 4.1 vs. 14.8 months; HR=3.50; p<0.0001) than the cfDNA-low group. Similarly, the I-score-high group had poorer TTP (median, 2.2 vs. 4.1 months; HR=2.09; p<0.0001) and OS (median, 4.6 vs. 14.8 months; HR=3.35; p<0.0001). In the multivariate analyses, cfDNA remained an independent prognostic factor for OS (p<0.0001), and I-score for both TTP (p=0.011) and OS (p=0.010). Although the VEGFA ratio was significantly higher in HCC patients than in healthy controls (2.50 vs. 2.17; p<0.0001), it was not significantly associated with treatment outcomes. Conclusions: Pretreatment cfDNA concentration and genome-wide CNVs in cfDNA are potential biomarkers predicting treatment outcomes in advanced HCC patients receiving first-line sorafenib.

Citation Format: Sook Ryun Park, Chung Ryul Oh, Sun-Young Kong, Min Kyeong Kim, Kyong-Ah Yoon, Eun-Hae Cho, Junnam Lee, Jihoon Kang, Baek-Yeol Ryoo. Biomarker analysis in circulating cell-free DNA in patients treated with sorafenib for advanced hepatocellular 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 4600.
Abstract 4658: Novel paclitaxel medical nanoparticle formulation for breast cancer
Authors: Chowdhury Pallabita , Chauhan Sumeet S. , Nagesh Prashanth K. , Hatami Elham , Hafeez Bilal B. , Khan Sheema , Jaggi Meena , Yallapu Murali M.
Published: August 2018

Objectives: Paclitaxel (Ptxl) is currently the most common first-line therapeutic option for breast cancer (BrCa). However, adverse side effects and problems associated with chemo-resistance, limits its use in clinical settings. Nanoparticle mediated delivery can improve Ptxl delivery and its activity at the tumor site, and considered to be an attractive strategy for BrCa therapy. Therefore, we aim to generate a novel Ptx medical nanoparticle formulation (PMNPs) using a polyphenol as a carrier, and evaluate its efficacy against BrCa cells. Methods: Physico-chemical characterization of this PMNPs was performed using TEM, DLS, FT-IR, TGA, X-RD, and SAXS methods. Its internalization and cellular availability was examined using HPLC method. Cell proliferation and colony formation assays were utilized to evaluate therapeutic efficacy of this unique nanoformulation in clinically relevant cell line models (MCF7 and MDA-MB-231). Additionally, molecular effects of this formulation on apoptosis, anti-apoptosis, and drug resistant associated proteins were evaluated using immunoblotting assays. A tubulin stabilization study was performed using confocal immunofluorescence microscopy analysis. Results: The PMNPs formulation showed optimal particle size and zeta potential, which can be efficiently internalized in BrCa cells. PMNPs exhibited potent anti-cancer efficacy via induction of the expression of apoptosis associated proteins (Bax and Bad, cleaved PARP, and caspase 3) and downregulation of anti-apoptotic proteins (Bcl-2 and Bcl-xL) in BrCa ell lines. Further, the expression of chemoresistance-associated proteins were also decreased with the treatment of PMNPs.

Conclusion: PMNPs formulation can efficiently be internalized in BrCa cells and induce enhanced therapeutic potential of Ptxl. Development of a targeted drug delivery system would not only reduce the dose of paclitaxel required to kill cancer cells but also minimize long-term paclitaxel associated systemic toxicity and drug-resistance.

Citation Format: Pallabita Chowdhury, Sumeet S. Chauhan, Prashanth K. Nagesh, Elham Hatami, Bilal B. Hafeez, Sheema Khan, Meena Jaggi, Murali M. Yallapu. Novel paclitaxel medical nanoparticle formulation for breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4658.
Abstract 4660: Tumor multicomponent targeting nanoparticle library for personalized cancer therapy & imaging
Authors: Sau Samaresh , Alsaab Hashem , Tatiparti Katyayani , Bhise Ketki , Rishi Arun , Iyer Arun K.
Published: August 2018

The fundamental limitations of conventional cancer treatment have promoted the development and application of nanoengineering platforms for achieving effective patient outcomes. Over a time, considerable clinical success has been accomplished in nanotechnology field but there are many challenges limiting the progress of nanomedicine from bench to the beside. The highly heterogeneous tumor environment, an inadequate understanding of interactions between tumor biomarkers and nanomedicine, and reproducibility in synthesis required for clinical translation are some of the major challenges. Thus, there is an urgent need to redirect our focus of nano-drug carrier that can target multiple components of tumor including, tumor epithelial cells, tumor angiogenic vessels, tumor hypoxia and stromal cells. To achieve this, we developed a library of various tumor biomarker targeting ligands, and drug and imaging agents encapsulated in nanoparticles (NPs). Finally, the library of targeting ligands was conjugated with NPs using reagent free ‘click' chemistry. This strategy will serve as an “off-the-shelf ready to configure drug formulation" that can be engineered based on a mix-and-match strategy for individual patient needs. It is envisaged that the patient can walk in the clinic and be given the targeted drug that is personalized to their need. For the proof-of-concept, we evaluated tumor selectivity of multiple cancer biomarker targeting oligo-micelles (OMs), iron oxide and lipid NPs in various patient derived tumor xenograft (PDx) models, such as non-small cell lung and triple negative breast cancer. We observed that the dual tumor hypoxia and cancer stem cell (CSC) targeting rod shaped OM has 8-fold higher tumor core penetration compared to non-targeted OM in these PDx tumors, as obtained from NIR-fluorescence imaging. This OM has significantly higher tumor vs. liver ratio and they follow tumor specific receptor competition. The tumor growth inhibition of drug loaded OM is superior compared to free drug. This approach demonstrated significant improvement of tumor stroma penetration with low healthy organ uptake of NPs portending more promising potentials for selective cancer treatment.

Citation Format: Samaresh Sau, Hashem Alsaab, Katyayani Tatiparti, Ketki Bhise, Arun Rishi, Arun K. Iyer. Tumor multicomponent targeting nanoparticle library for personalized cancer therapy & imaging [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 4660.
Abstract 4721: Combining STING agonists with an anti-PD-1 antagonist results in marked antitumor activity in immune-excluded tumors
Authors: Perera Samanthi A. , Kopinja Johnny E. , Ma Yanhong , Laskey Jason , Chakravarthy Kalyan , Chen Yiping , Cui Long , Presland Jeremy , Zhao Shuxia , Minnihan Ellen , Ferguson Heidi , Piesvaux Jennifer , Pan Bo-Sheng , Woo Hyun Chong , Knemeyer Ian , Cemerski Saso , Cumming Jared , Trotter Wesley , Tse Archie , Addona George H. , Long Brian J.
Published: August 2018

The innate immune agonist STING (STimulator of INterferon Genes) binds its natural ligand 2'3'-cGAMP (cyclic guanosine-adenosine monophosphate) and initiates type I interferon production. This promotes systemic antigen-specific CD8+ T-cell priming that eventually provides potent anti-tumor activity. To exploit this mechanism we synthesized a novel STING agonist, MSA-1, that activates both mouse and human STING with higher in vitro potency than cGAMP. MSA-1 was administered to immune-competent mice bearing MC38 syngeneic tumors to monitor pharmacodynamics, pharmacokinetics and in vivo efficacy. Intratumoral (IT) dosed MSA-1 demonstrated robust tumor and plasma cytokine upregulation and effective anti-tumor activity. The highest tolerated doses provided complete responses (CRs) in 100% of MC38 tumors. Tumor models such as CT26 and B16-F10 that are intrinsically resistant to single-agent therapy with a fully murinized mouse anti-PD-1 antibody (mDX400) also demonstrated long-term tumor regressions or CRs. Mechanistic studies in immune-deficient mice suggested that anti-tumor activity of IT dosed STING agonists are in part due to cytotoxicity and/or innate immune responses rather than development of robust adaptive anti-tumor immunity. To enhance the adaptive immune response, we combined MSA-1 with mDX400 in mouse syngeneic tumor models previously characterized to be unresponsive to anti-PD-1 blockade. This combination restored T-cell responses in both blood and tumors of the treated mice and provided long-lived immunologic memory in a majority of the animals. Taken together, these data strongly support the development of STING agonists in combination with Keytruda for patients with tumors that are partially responsive or non-responsive to single agent anti-PD-1 therapy.

Citation Format: Samanthi A. Perera, Johnny E. Kopinja, Yanhong Ma, Jason Laskey, Kalyan Chakravarthy, Yiping Chen, Long Cui, Jeremy Presland, Shuxia Zhao, Ellen Minnihan, Heidi Ferguson, Jennifer Piesvaux, Bo-Sheng Pan, Hyun Chong Woo, Ian Knemeyer, Saso Cemerski, Jared Cumming, Wesley Trotter, Archie Tse, George H. Addona, Brian J. Long. Combining STING agonists with an anti-PD-1 antagonist results in marked antitumor activity in immune-excluded tumors [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 4721.
Abstract 4761: Joint model of longitudinal tumor size and overall survival (OS) in non-small cell lung cancer (NSCLC): A Bayesian approach
Authors: Al-Huniti Nidal , Xu Hongmei , Zhou Diansong , Edlund Helena , Aksenov Sergey , Helmlinger Gabriel , Dunyak James
Published: August 2018

Objective: The methodology of joint modeling longitudinal tumor size and OS data is very promising and provides the opportunity of survival prediction in individual patients. However, current joint modeling software are restricted in terms of distributions/parameterizations available and non-linear mixed effects models are not supported. The objective of this work was to develop a joint model in Bayesian framework to relax these restrictions and enable investigation of additional (continuous and/or discrete) biomarkers. Methods: Longitudinal tumor size and OS data from gefitinib Phase 3 study in NSCLC (‘IPASS', NCT00322452) was used in the modeling analysis. Joint model was developed in JM package in R and the Bayesian formulation of the joint modeling was implemented in STAN. Different parametrizations were investigated for both sub-models and the impact of lag-times was explored. The parameter estimates and model fit were compared between models by JM package and STAN. The final Bayesian joint model was validated on an independent dataset from a Phase 4 study (‘IFUM', NCT01203917, EGFR positive patients only) by simulating individual survival based on their tumor size data and compared with observed survival. Results: The estimated parameter values were similar to those from the fit from JM package. The different parametrizations provided similar fits for the tumor size model, while the survival model was improved by using a b-spline model. The association parameters indicate a strong association between tumor size and survival and introduction of lag-times did not impact the association parameters to a great extent. Posterior predictive simulations of OS in phase 4 study were consistent with observed data. Conclusions: Bayesian formulation further improved the joint model of tumor size and OS which would enable investigation of additional biomarkers (e.g. new lesions).

Citation Format: Nidal Al-Huniti, Hongmei Xu, Diansong Zhou, Helena Edlund, Sergey Aksenov, Gabriel Helmlinger, James Dunyak. Joint model of longitudinal tumor size and overall survival (OS) in non-small cell lung cancer (NSCLC): A Bayesian approach [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 4761.
Abstract 4798: Targeting hTERT for treatment of glioblastoma (GBM)
Authors: Bollam Saumya Reddy , Kang Hyun-Jin , Peng Sen , Gokhale Vijay , Hurley Laurence , Berens Michael , Dhruv Harshil
Published: August 2018

Approximately 86% of GBM tumors exhibit a mutation at -124 or -146 bp upstream from the ATG start site in the transcription activating promoter region of Human telomerase reverse transcriptase (hTERT). Mutations in the hTERT promoter are known to impair repression, leading to overexpression of hTERT and tumor maintenance. Yet, complete understanding of mechanistic implications is still necessary. While overexpressed hTERT is associated with oncogenesis and resistance to apoptosis, we also observe phenotypes unrelated to the reverse transcription function. We characterized long-term glioma cell lines and glioma PDX models by hTERT promoter mutation status, hTERT mRNA expression, and hTERT protein expression in subcellular fractions. The -124 and -146 mutations are located in the major 5-12 G-quadruplex and result in misfolding of the silencer element, thus causing over-expression of hTERT. Using a diverse small molecule library, we identified a small drug-like pharmacological chaperone (pharmacoperone) molecule, TG-4260, which binds to the 26 mer base-pair heteroduplex loop, which is the nucleation site for cooperative folding of the major 5-12 G-quadruplex. The chaperone effect of TG-4260 corrects DNA hTERT G-quadruplex misfolding resulting from the promoter mutations and restores the silencer function of the G-quadruplex. TG-4260 directly decreases the transcription activity of the −124, −124/125, −138/139, and −146 mutants to a similar extent and suppresses the downstream gene BCL2, which activates caspase-3 and produces cell-cycle arrest, leading to cell death. This is the first example of the use of a pharmacoperone molecule to correct the misfolding of a DNA G-quadruplex element resulting from mutations in an early folding intermediate. We screened GBM cell models against this novel small molecule inhibitor that interferes with mutated hTERT promoter and demonstrated that TG-4260 selectively suppresses glioma cell viability without affecting non-transformed normal human astrocytes. Finally, telomere phenotypes from treated cells indicate non-canonical functions of telomerase may also contribute to glioma pathogenesis.

Citation Format: Saumya Reddy Bollam, Hyun-Jin Kang, Sen Peng, Vijay Gokhale, Laurence Hurley, Michael Berens, Harshil Dhruv. Targeting hTERT for treatment of glioblastoma (GBM) [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 4798.
Abstract 4865: BI 907828: A highly potent MDM2 inhibitor with low human dose estimation, designed for high-dose intermittent schedules in the clinic
Authors: Rinnenthal Joerg , Rudolph Dorothea , Blake Sophia , Gollner Andreas , Wernitznig Andreas , Weyer-Czernilofsky Ulrike , Haslinger Christian , Garin-Chesa Pilar , Moll Jürgen , Kraut Norbert , McConnell Darryl , Quant Jens
Published: August 2018

MDM2 inhibitors block the interaction between the Tumor Protein p53 (TP53) and MDM2, its key negative regulator, and represent a new therapeutic concept for cancer therapy. MDM2 inhibitors are designed to restore p53 activity in TP53 wild-type tumors. Several MDM2 inhibitors are currently being evaluated in early clinical development with mainly daily dosing regimens. However, recent clinical data suggest myelosuppression as an on-target, dose-limiting toxicity for this class of inhibitors. Particularly thrombocytopenia could limit the clinical utility of MDM2 inhibitors. Hence there is a need to mitigate these side effects and to improve the therapeutic window. One approach is less frequent dosing to allow bone marrow recovery while still maintaining efficacious exposure levels and clinical activity. Here we present pharmacokinetic data on BI 907828, a novel and potent MDM2 inhibitor with optimized drug-like properties. Due to its high permeability, good physiological solubility and low systemic clearance, the compound shows a high bioavailability in mice, rats, dogs and minipigs. BI 907828 is not absorption-limited, even at multiples of the efficacious dose, and shows a reliable, dose-linear PK with low variability in AUC and Cmax across species. Based on a good PK-PD correlation with efficacy in preclinical models and a robust human PK prediction based on in vivo PK data across species, a low human efficacious dose is predicted. The PK properties of BI 907828 enable a variety of different clinical dosing schedules with the potential to improve the therapeutic index with respect to on-target safety. In summary, BI 907828 is a novel, potent, orally available MDM2 inhibitor with excellent PK properties and low estimated human dose suitable for high-dose intermittent dose schedules in the clinic.

Citation Format: Joerg Rinnenthal, Dorothea Rudolph, Sophia Blake, Andreas Gollner, Andreas Wernitznig, Ulrike Weyer-Czernilofsky, Christian Haslinger, Pilar Garin-Chesa, Jürgen Moll, Norbert Kraut, Darryl McConnell, Jens Quant. BI 907828: A highly potent MDM2 inhibitor with low human dose estimation, designed for high-dose intermittent schedules in the clinic [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 4865.
Abstract 5052: ID3 mediated vascular reprogramming of PCB exposed endothelial cells and its potential contribution to lung tumorigenicity
Authors: Doke Mayur , Das Jayanta , Felty Quentin
Published: August 2018

ID3, a transcription regulator, has been shown to contribute to the aggressive spread of certain cancers by facilitating the generation of tumor blood vessels. Tumor vascular cells characterized as CD133+ endothelial stem/progenitor cells are associated with poor prognosis in lung cancer. Previously, we have shown that ID3 overexpression reprograms adult endothelial cells to acquire vascular stem markers including CD133. However, little is known regarding the molecular contribution of ID3 to tumor vascular reprogramming and how this may impact the aggressive growth of lung cancer. In the current project, we evaluated ID3 mediated vascular reprogramming upon polychlorinated biphenyl (PCB) exposure. This idea is consistent with the literature, which shows that PCBs accumulate in the human lung; PCBs produce pathological vascular remodeling; high levels of PCBs are found in human lung tissue; and epidemiological studies associating lung toxicity with PCBs. Recent studies identify PCB153 as one of the largest contributors for total PCB body burden in humans. Using human lung endothelial and smooth muscle cells, we exposed ID3 overexpressing and vector control cells to PCB153. We observed a significant increase in cell proliferation as determined by the BrdU incorporation assay and FACS analysis. Similarly, a 3D HuBiogel model, which mimics in vivo conditions, showed a significant increase in size and number of vascular spheres upon PCB153 treatment. Pluripotent vascular stem cells showed the loss of VE-cadherin and gain of MMP9, N-cadherin, and vimentin, which are markers of endothelial-mesenchymal transition. RNA-seq analysis showed that PCB153 exposed vascular stem cells had a significant increase in lncRNAs MALAT1 (Metastasis Associated Lung Adenocarcinoma Transcript 1) and HIF1A-AS2. A combination of ChIP-seq and transcriptome analysis identified significantly up-regulated genes (HES1, WEE1, E2F2) and down-regulated genes (SMURF2, CDKN2C, SH2B3). Our findings suggest that ID3 mediates vascular reprogramming through transcriptional regulation of pluripotency factors and epigenetic regulating lncRNAs. ID3 may also serve as a novel diagnostic/prognostic indicator for evaluating chemical-induced vascular reprogramming. Its potential application may be useful in identifying individuals who are susceptible to aggressive lung cancer from exposure to vascular toxicants.

Citation Format: Mayur Doke, Jayanta Das, Quentin Felty. ID3 mediated vascular reprogramming of PCB exposed endothelial cells and its potential contribution to lung tumorigenicity [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 5052.
Abstract 5117: A murine hematogenous bone metastatic cancer to model spontaneous metastasis of triple negative breast cancer
Authors: Lee Hye Jin
Published: August 2018

The development of bone metastasis in breast cancer and its pathologic progression are known to be clinically important problem. It is known that immunologic response in microenvironment is important as soil in the bone metastasis process and progression. Thus, immunologic modulation is expected to play a role in the prevention and treatment of the disease in bone metastasis. However, most studies to date; researches in bone metastasis were carried out using human breast cancer cell lines in xenograft model of immuno-deficient mouse or in vitro co-culture system. These approaches have limitations in regard to interaction among immune cells, cancer cells, and other stroma cells. To overcoming these limitation, we used the C3(1)Tag mouse model to establish a spontaneous bone metastasis model. First, a cell line (M6) obtained from a spontaneously occurring tumor in C3(1)Tag mice was infected with lentiviral-luciferase. M6 cells with lentiviral-luciferase were administered using intracardiac injection and C3(1) Tag mice were monitored until tumors are detected under bioluminescence imaging. In 3 weeks, metastatic bone lesions were apparent in 85.7% of mice. To understand biologic characteristics of in vivo selected bone metastatic subpopulation, metastatic cancer cells were harvested and were analyzed using immunohistochemical staing in comparison with parent cells. Tumor cells metastasized to bone showed differential expression in HIF1A, c-MET, and EMT markers. Unexpectedly, bone metastatic cancers showed better sensitivity to paclitaxel and cisplatin than parent cells in cell viability assay. In addition, immune microenvironment in bone metastatic foci was assessed using multiplex immunohistochemical staining and showed an immune suppressive MDSCs infiltration. Taken together, we established a murine hematogenous bone metastatic cancer and characterized biologic characteristics to model human triple negative breast cancer. The potential targets identified in this study might be a future immunologic target in patients with bone metastasis.

Citation Format: Hye Jin Lee. A murine hematogenous bone metastatic cancer to model spontaneous metastasis of triple negative breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5117.