bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4319: Clinical significance and possible role of GPNMB in patients with breast cancer
Authors: Futamura Manabu , Kanematsu Masako , Yamada Atsuko , Morimitsu Kasumi , Morikawa Akemi , Mori Ryutaro , Yoshida Kazuhiro
Published: August 2015

Glycoprotein non-metastatic B (GPNMB) is a type I transmembrane protein, which is isolated from differential expression assay using metastatic melanoma cells. The physiological function is very little known but may be supposed to be associated with cell invasion and motility particularly in breast cancer cells. Here we investigated the role of GPNMB in breast cancer. First we checked expression of GPNMB by RT-PCR and western blot in several cancer cell lines including breast, gastric, and colon cancer followed by establishment of GPNMB measurement by ELISA because it's reported that GPNMB is shed at extracellular domain by sheddase such as ADAM10. GPMNB expressed in breast (5/6:83%), gastric (3/6:50%), colon (1/7:14.3%) cancer cell lines. Of breast cancer cell lines, GPNMB was highly expressed in SK-BR3 (HER2 positive), BT474 (HER2/ER positive), MDA-MB-157 (Triple negative) cells. Shed GPNMB in culture medium was measurable and correlated with expression of each cell line. Next we evaluated serum GPNMB in patients with breast (n = 164; primary 119, metastatic 43), gastric (n = 38), and colorectal (n = 50) cancer in our institute from 2011.9-2014.2.) This study was approved by the central ethics committee of Gifu University. Serum GPNMBs were 9.403, 5.751, 6.550 ng/ml, respectively. GPNMB for breast cancer patients was statistically higher than those by colorectal cancer patients (p = 0.018). Of breast cancer patients, GPNMB for HER2-type patients was higher than those for Luminal type and DCIS patients (p = 0.0386, p = 0.0195, respectively). Those for triple negative patients was also higher than those for DCIS patients (p = 0.0459). Interestingly, serum GPNMB was dramatically reduced in accordance with chemotherapy in some patients. Based on these clinical observations, we further investigated relationship between GPNMP and HER2 in vitro. Blockage of GPNMB induced not only HER2 but also EGFR expression. On the other hand, inhibition of HER2 by trastuzumab increased expression of GPNMB. Depletion of GPNMB increased sensitivity of trastuzumab, suggesting that GPNMB may play an important role in crosstalk of signal transduction for breast cancer. These notions may suggest a novel therapeutic strategy to overcome HER2 positive breast cancer.

Citation Format: Manabu Futamura, Masako Kanematsu, Atsuko Yamada, Kasumi Morimitsu, Akemi Morikawa, Ryutaro Mori, Kazuhiro Yoshida. Clinical significance and possible role of GPNMB in patients with breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4319. doi:10.1158/1538-7445.AM2015-4319
Abstract 432: Macrophage promotes EMT in gastric cancer through activating CXCR2 signaling
Authors: Zhang Chang-Hua , Xia Guangkai
Published: August 2015

Background and aims: Macrophage as a member of the tumor microenvironment and chronic inflammation was considered as an important pro-tumoral factor in cancer. CXCR2 was an inflammation receptor and expressed in gastric cancer. But its’ association with macrophage remains unknown. We aimed to investigate the role of macrophage in the development of gastric cancer from inflammation.

Methods: Human GC cell lines AGS, BGC-823 were studied. Western Blot was used to detect the protein expression in gastric cancer cells. Migration of gastric cancer cells were tested by Transwell.

Results: Macrophages and gastric cancer cell lines can secret CXCR2′ ligands when coculutred with each other. CXCR2 knock down in gastric cancer cells can decrease migration and inhibit EMT. Macrophage promotes EMT of gastric cancer though CXCR2 signaling. CXCR2's ligands CXCL1 and CXCL5 can activate stat3 and promote the migration of gastric cancer cells. CXCL1 and CXCL5 are responsible for the EMT of gastric cancer.

Conclusion: Macrophage can promote gastric cancer EMT through activating CXCR2 signaling. CXCR2 ligands CXCL1 and CXCL5 are the crucial factors for promoting EMT in gastric cancer.

Note: This abstract was not presented at the meeting.

Citation Format: Chang-Hua Zhang, Guangkai Xia. Macrophage promotes EMT in gastric cancer through activating CXCR2 signaling. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 432. doi:10.1158/1538-7445.AM2015-432
Abstract 4333: Association of TP53 mutation status with clinical outcomes in patients with EGFR mutated non-small cell lung cancer (NSCLC)
Authors: Ahmed Saman , Aggarwal Charu , Mick Rosemarie , Bauml Joshua , Cohen Roger , Kadauke Stephan , Sterlicchi Trisha , Evans Tracey L. , Kucharczuk John C. , Desphande Charu , Morrissette Jennifer J D , Daber Robert , Langer Corey J.
Published: August 2015

Introduction:

Molecular characterization of tumors is critical in the modern treatment of NSCLC. Epidermal Growth Factor Receptor (EGFR) and p53 are commonly mutated genes in NSCLC with independent prognostic implications, but the significance of the co-existence of these mutations inpatients (pts) with Stage IV NSCLC is not well established. Herein, we describe the relationship of EGFR and TP53 mutations (MT) in pts with Stage 4 or recurrent NSCLC and their correlation with pt demographics and therapeutic outcomes.

Study Design:

We conducted a retrospective analysis of Stage 4 and recurrent EGFR MT (+) NSCLC pts with TP53 MT status determined by the Gene Sequence Analysis Panel obtained at the Abramson Cancer Center between 6/1/2012 and 2/1/2014. Overall survival (OS) and progression-free survival (PFS) were estimated for the TP53 MT (+) and wild type (WT) cohorts from the date of diagnosis.

Results:

Thirty-six EGFR (+) pts were identified, of whom 26 (72%) were TP53 MT (+). Pt characteristics were similar between cohorts, except for a higher rate of bone metastasis in pts with p53 WT (p = 0.07, Table 1). Median potential follow-up for OS was 15.1 mos. 60% of patients in TP53 WT group received TKI compared to 69% in TP53 MT (+). Of the 36 pts, 9 (25%). have died. A trend towards shorter median OS was seen in pts with p53 MT. (p = 0.08). PFS did not correlate with p53 MT status (p = 0.53).

Conclusion:

Coexistence of p53 MT and EGFR MT may be associated with shorter survival. Larger sample size and longer clinical follow up will be necessary to determine if these differences are statistically significant and whether TP53 MT status is an independent prognostic factor in EGFR MT NSCLC.

Citation Format: Saman Ahmed, Charu Aggarwal, Rosemarie Mick, Joshua Bauml, Roger Cohen, Stephan Kadauke, Trisha Sterlicchi, Tracey L. Evans, John C. Kucharczuk, Charu Desphande, Jennifer J D Morrissette, Robert Daber, Corey J. Langer. Association of TP53 mutation status with clinical outcomes in patients with EGFR mutated non-small cell lung cancer (NSCLC). [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4333. doi:10.1158/1538-7445.AM2015-4333
Abstract 4336: Direct genotyping of CYP2A6 whole gene deletion: A new biomarker for prognosis of lung adenocarcinoma in Japanese smoking male
Authors: Okano Yasuko , Ichikawa Yasushi , Miyagi Yohei , Chishima Takashi , Natsumeda Yutaka
Published: August 2015

Purpose: Cytochrome P450 (CYP2A6) is known as an enzyme responsible for the metabolism of activation of tobacco-specific nitrosamines to their ultimate carcinogenic forms and metabolism of nicotine. We here report that the genetic polymorphisms of human CYP2A6*4 is associated with primary lung cancer and their overall survival.

Experimental Design: We prepared genomic DNA samples from 387 Japanese patients with primary lung cancer and the genetic polymorphisms of human CYP2A6*4 (whole gene deletion) by the rapid genetic testing method we developed in this study. We analyzed the association between the CYP2A6*4 gene, NRF2 gene and overall survival.

Results: Patients harboring wild-type CYP2A6*1 and whole gene deletion CYP2A6*4/*4 numbered 380 (98.2%) and 7 (1.8%), respectively. Its frequency was four-fold higher in male patients than in female (P = 0.038).Multivariate logistic regression models revealed that the genetic polymorphisms of human CYP2A6*4 were significantly related to gender and smoking behavior. Interestingly, lung cancer patients carrying CYP2A6*4/*4 (whole gene deletion), the NRF2 SNP heterozygous (c.-617C/A) and wild-type homozygous (c.-617C/C) exhibited remarkable survival over 1,500 days after surgical operation.

Conclusion: CYP2A6*4/*4 whole gene is considered to be a prognostic biomarker for assessing both the gender-related risk of lung adenocarcinoma in the smoking sub-population and their overall survival.

Citation Format: Yasuko Okano, Yasushi Ichikawa, Yohei Miyagi, Takashi Chishima, Yutaka Natsumeda. Direct genotyping of CYP2A6 whole gene deletion: A new biomarker for prognosis of lung adenocarcinoma in Japanese smoking male. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4336. doi:10.1158/1538-7445.AM2015-4336
Abstract 4337: Comprehensive analysis of genetic variations of genes involved in DNA damage response pathways or cell cycle checkpoints and treatment outcome in non-small cell lung cancer patients treated with platinum doublets
Authors: Ryu Jeong Seon , Yi Bo-Rim , Lee Seul-Ki , Hong Soon-Sun
Published: August 2015

Genes involved in DNA damage response (DDR) pathways or cell cycles checkpoints (CCC) are thought to be closely related to effect of platinum doublets, backbone in treatment for lung cancer patients. We tried to evaluate validity of germ-line variations of all genes related to DDR or CCC as potential predictive biomarker of treatment outcome in non-small-cell lung cancer (NSCLC) patients treated with platinum doublets. A total of 967 NSCLC patients treated with platinum doublets as front line chemotherapy was enrolled and divided randomly into three sets: screening, training and validation. For screening set, whole exons, -1,000 bps upstreams, and splicing junctions in the 170 genes related to DDR or CCC were analyzed using next generation sequencing, then genetic variations were selected with several criterions such as minor allele frequency > 5%, call rate > 90%, p value for HWE > 0.001, and impact of genetic variation. Their effects on progression free survival or response to platinum doublets were evaluated in two phases: training and validation process. Clinical characteristics were not different between training (n = 372) and validation (n = 381) cohorts. A total of 4,697 genetic variations and 7 In/Del were identified in screening set, then 232 genetic variations and 7 In/Del from 118 genes were finally selected with predefined criterions. In training cohort, 29 genetic variations were found to be associated with progression free survival or response. However, the effects of these genetic variations were not failed to be validated. In subgroup analysis with non-adenocarcinoma patients, CLSPN (rs79083467), MSH3 (rs32952), and RAD51C (rs12946397) were shown to be associated with progression free survival of the patients (cox p values for CLSPN, MSH3, RAD51C: 0.028, 0.022, 0.035 in training cohort; 0.046, 0.026, 0.008, respectively in validation cohort). In this study, we have failed to show validity of the germ-line variations in predicting treatment outcome of NSCLC patients treated with platinum doublets. This result show germ-line variations related to pharmacodynamics have a limited value in predicting platinum effect.

Note: This abstract was not presented at the meeting.

Citation Format: Jeong Seon Ryu, Bo-Rim Yi, Seul-Ki Lee, Soon-Sun Hong. Comprehensive analysis of genetic variations of genes involved in DNA damage response pathways or cell cycle checkpoints and treatment outcome in non-small cell lung cancer patients treated with platinum doublets. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4337. doi:10.1158/1538-7445.AM2015-4337
Abstract 4351: Identification of molecular signature associated with the castration-resistant prostate cancer (CRPC) development by next-generation sequencing (NGS)-based transcriptomic profiling
Authors: Kim Seon-Kyu , Kim Jeong-Hwan , Jeong Pildu , Kim Wun-Jae , Kim Yong Sung , Yun Seok-Joong , Kim Seon-Young
Published: August 2015

Backgrounds:

Castration-resistant prostate cancer (CRPC) is an advanced form of prostate cancer produced by disease progression following surgical or pharmaceutical castration. The development process of CRPC becoming castrate resistant is unclear. The objective of this study is to identify a gene set and putative signaling pathway for mediating the development of CRPC.

Methods:

RNA-seq data was generated using 54 prostate samples [primary prostate cancer (PCa), CRPC, and benign prostatic hyperplasia (BPH)], among which 8 were paired samples of PCa and CRPC obtained from 4 patients. Diverse statistical methods, including a generalized linear model likelihood ratio test, were applied to identify the signature associated with CRPC development. The interconnectivity between genes in the signature was verified by Ingenuity Pathway Analysis TM. We also confirmed a relevance of the signature with experimental assays using prostate cancer cell lines.

Results:

Unsupervised hierarchical cluster analysis of RNA-seq data revealed a clear gene expression pattern of tumorigenesis in PCa with heterogeneous molecular characteristics of CRPC. Comparing paired cancer samples, 309 genes were differentially expressed between PCa and CRPC samples. We also tested unpaired sample groups (PCa vs. CRPC) and found that 182 genes had significantly differences of expression (a generalized linear model likelihood ratio test, P < 0.001, a 2-fold or greater relative difference). When comparing two gene sets, a total of 15 genes was common in both paired and unpaired CRPC groups. Among them, AR and SPINK1 were significantly increased in the CRPC than in the PCa groups. Gene network analyses and experimental assays of AR and SPINK1 revealed that CRPC development could be mediated by SPINK1-AR pathway.

Conclusion:

The molecular signature defined by activation of SPINK1-AR may be a genetic determinant of progression from PCa to CRPC.

Note: This abstract was not presented at the meeting.

Citation Format: Seon-Kyu Kim, Jeong-Hwan Kim, Pildu Jeong, Wun-Jae Kim, Yong Sung Kim, Seok-Joong Yun, Seon-Young Kim. Identification of molecular signature associated with the castration-resistant prostate cancer (CRPC) development by next-generation sequencing (NGS)-based transcriptomic profiling. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4351. doi:10.1158/1538-7445.AM2015-4351
Abstract 4358: Targeting connexin 43 with α-connexin carboxyl-terminal (ACT1) peptide in breast cancer
Authors: Grek Christina , Rhett Joshua Matthew , Abt Melissa , Bruce Jaclynn , Ghatnekar Gautam , Yeh Elizabeth S.
Published: August 2015

Expression and functional studies have identified connexins (Cxs), a family of gap junction proteins, as potential tumor suppressors. Studies suggest that connexin 43 (Cx43) has a role in breast cancer cell proliferation, differentiation, and migration. Although pan-gap junction drugs are available, the lack of specificity of these agents increases the opportunity for off target effects. Consequently, a therapeutic agent that specifically modulates Cx43 would be beneficial and has not been tested in breast cancer. In this study, we now test a Cx43 specific agent, called α-connexin carboxyl-terminal (ACT1), in breast cancer. Our studies demonstrate that modulation of Cx43 activity in breast cancer can be effectively achieved with the agent ACT1 to sustain Cx43-mediated gap junctional activity resulting in impaired malignant progression. Furthermore, using ER+ and HER2+ breast cancer cell models, we show that ACT1 treatment enhances the activity of lapatinib and tamoxifen.

Citation Format: Christina Grek, Joshua Matthew Rhett, Melissa Abt, Jaclynn Bruce, Gautam Ghatnekar, Elizabeth S. Yeh. Targeting connexin 43 with α-connexin carboxyl-terminal (ACT1) peptide in breast cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4358. doi:10.1158/1538-7445.AM2015-4358
Abstract 4362: Screening of investigational antimalarials for anticancer activity in high risk N-MYC amplified neuroblastoma (NB)
Authors: Coulter Don W. , Vennerstrom Jonathan , Sharp John G. , Dong Yuxiang , Wang Xiaofang , McIntyre Erin , McGuire Tim
Published: August 2015

Introduction: NB is a pediatric tumor of neural crest origin. Patients who have N-MYC amplified metastatic tumors have poor outcomes. The importance of autophagy in protecting NB cells from chemotherapy has been postulated. In the following study we screened bisquinoline (BQ) and ozonide (OZ) antimalarials (potential autophagy inhibitors) in a highly N-MYC amplified NB cell line (BE-2) and a moderately amplified N-MYC NB cell line (IMR-32). A combination of monolayer cell culture and neurospheres were used to screen activity.

Methods: Using a 96 well MMT based assay and 16 hour treatment with compounds dissolved in DMSO, cytotoxicity was calculated as a percent decline in absorbance compared to DMSO controls. 32 BQ and 13 OZ antimalarials were tested. All compounds were tested at a concentration of 1 mcg/ml; an achievable concentration in patients. Selected compounds that were cytotoxic in monolayer culture will be tested in neurosphere culture and then in a mouse xenograft model alone and combined with chemotherapy.

Results: The bisquinolone antimalarials were consistently active in both BE-2 and IMR cell culture. Among the bisquinolones, Q1-9, Q2-5, Q2-61, and Q3-75 had the greatest cytotoxicity. There was no clear structure-activity-relationship for this small bisquinoline library. Among the ozonides, OZ465, OZ513, and OZ521 were the most active. From these data, we could infer that the presence of a weak base functional group was required, but was insufficient for cytotoxicity. Initial studies in BE-2 and IMR neurosphere culture using an LDH assay for cytotoxicity suggests similar activity. Monolayer results are shown in the Table below:

SymbolDrug ClassBE-2*IMR-32*Q1-9BQ38.535.8Q2-5BQ29.844.4Q2-61BQ40.731.4Q3-75BQ40.619.3OZ465OZ3820.6OZ513OZ73.627.7OZ521OZ4415.9

*Percent decline in absorbance compared to DMSO control

Conclusion: The above data indicates that two classes of antimalarials had moderate cytotoxicity in both 96-well monolayer assays as well as in a neurosphere assay (data not shown). The results suggest cytotoxic activity in high and moderately amplified N-MYC NB in both monolayers and neurospheres.

Note: This abstract was not presented at the meeting.

Citation Format: Don W. Coulter, Jonathan Vennerstrom, John G. Sharp, Yuxiang Dong, Xiaofang Wang, Erin McIntyre, Tim McGuire. Screening of investigational antimalarials for anticancer activity in high risk N-MYC amplified neuroblastoma (NB). [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4362. doi:10.1158/1538-7445.AM2015-4362
Abstract 4421: GPER-mediated activation of the cAMP response element-binding protein (CREB) and subsequent IGFBP-1 expression is altered during the development of tamoxifen resistance
Authors: Vaziri-Gohar Ali , Houston Kevin D.
Published: August 2015

Tamoxifen is a commonly prescribed anti-estrogen treatment for breast cancer patients; this selective estrogen receptor modulator functions as an ERα-antagonist in breast tissue and inhibits cell proliferation and survival of breast cancer cells. More recently, the active metabolite of tamoxifen, 4-OHT (Tam), has been shown to activate the G protein-coupled estrogen receptor (GPER). Previous studies from our laboratory identified a novel, GPER-dependent mechanism of Tam action in breast cancer cells that requires the extracellular accumulation of the insulin-like growth factor binding protein-1 (IGFBP-1). The molecular mechanisms of Tam-induced IGFBP-1 production mediated by GPER have not been determined. GPER activation has been shown to stimulate cAMP production and PKA activation in several cell types. We hypothesized the cAMP response element-binding protein (CREB) activation is required for Tam-induced IGFBP-1 production in breast caner cells. To determine if CREB activation may be involved in IGFBP-1 transcription, CREB phosphorylation (S133) status and nuclear accumulation were determined in MCF-7 cells after Tam treatment. The phosphorylation and nuclear accumulation of CREB were observed in Tam-treated MCF-7 cells suggesting that CREB activation may be required for Tam-induced IGFBP-1 transcription. Increased CREB phosphorylation and IGFBP-1 transcription were inhibited by pretreatment with the PKA inhibitor H-89 supporting a role for CREB in Tam-induced IGFBP-1 expression. H-89 pretreatment also significantly decreased the previously reported inhibition of IGF-1-stimulated signaling by Tam treatment in breast cancer cells. We have previously reported that Tam-induced IGFBP-1 expression is dysregulated in Tam-resistant MCF-7 cells (MCF-7TamR). To determine if aberrant CREB signaling results in the observed dysregulation, similar experiments were performed in MCF-7TamR cells. Our results show that CREB phosphorylation and nuclear accumulation are decreased in Tam-resistant cells compared to Tam-sensitive cells. Taken together, these results suggest that activation of GPER results in CREB-dependent IGFBP-1 production and subsequent inhibition of IGF-1-stimulated signaling in breast cancer cells treated with Tam. Furthermore, dysregulation of GPER-mediated CREB activation may be involved in the development of Tam resistance in breast cancer.

Citation Format: Ali Vaziri-Gohar, Kevin D. Houston. GPER-mediated activation of the cAMP response element-binding protein (CREB) and subsequent IGFBP-1 expression is altered during the development of tamoxifen resistance. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4421. doi:10.1158/1538-7445.AM2015-4421
Abstract 4422: TTT-28, a newly synthesized thiazole-valine peptide, antagonizes multidrug resistance by inhibiting the efflux activity of the ABCB1 transporter
Authors: Wang Yi-Jun , Anreddy Nagaraju , Patel Bhargav A. , Chufan Eduardo E. , Singh Satyakam , Zhang Guan-Nan , Zhang Yun-Kai , Barbuti Anna Maria , Ambudkar Suresh V. , Talele Tanaji T. , Chen Zhe-Sheng
Published: August 2015

Cancer cells often exhibit either intrinsic or acquired resistance to chemotherapy through a phenomenon known as multidrug resistance (MDR). Different mechanisms contribute to the development of MDR, preeminent among them being the accelerated drug efflux mediated by overexpression of ATP-binding cassette (ABC) transporters. Currently, it has been found that some small molecule tyrosine kinase inhibitors (TKIs), such as motesanib, linsitinib, masitinib and nilotinib, were able to modulate the activity of ABC transporters. Thus, the aim of this study was to determine whether TTT-28, a newly synthesized thiazole-valine peptide, could reverse ABCB1-mediated MDR. The results showed that TTT-28 significantly sensitized both ABCB1-transfected and drug-selected cell lines overexpressing this transporter to its substrate anticancer drugs. Using calcein-AM efflux assay, we identified TTT-28 (IC50 = 1.0 μM) carrying 3,4,5-trimethoxybenzoyl and 2-aminobenzophenone groups, respectively, at the amino and carboxyl termini of the monothiazole zwitterion. TTT-28 significantly increased the accumulation of [3H]-paclitaxel in ABCB1 overexpressing cells by blocking the efflux function of ABCB1 transporter. Furthermore, TTT-28 inhibited the photolabeling of ABCB1 with [125I]-iodoarylazidoprazosin with IC50 = 0.75 μM and stimulated the basal ATP hydrolysis of ABCB1 in a concentration-dependent manner (EC50 ATPase = 0.027 μM). Consistent with these findings, biochemical and docking studies showed site-1 to be the preferable binding site for TTT-28 within the drug-binding pocket of human ABCB1. Therefore, we report that TTT-28 antagonizes MDR by inhibiting the efflux activity of the ABCB1 transporter. These findings reveal high clinical values for the co-administration of TTT-28 and ABCB1 substrate chemotherapeutic drugs in cancer patients that overexpress ABCB1 and stimulate further research on circumventing the ABCB1-mediated MDR in cancers.

Citation Format: Yi-Jun Wang, Nagaraju Anreddy, Bhargav A. Patel, Eduardo E. Chufan, Satyakam Singh, Guan-Nan Zhang, Yun-Kai Zhang, Anna Maria Barbuti, Suresh V. Ambudkar, Tanaji T. Talele, Zhe-Sheng Chen. TTT-28, a newly synthesized thiazole-valine peptide, antagonizes multidrug resistance by inhibiting the efflux activity of the ABCB1 transporter. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4422. doi:10.1158/1538-7445.AM2015-4422
Abstract 4424: Molecular diagnostics of drug resistant multiple myeloma cases using targeted next generation sequencing
Authors: Ikeda Hiroshi , Sasaki Yasushi , Igarashi Tetsuyuki , Aoki Yuka , Hayashi Toshiaki , Ishida Tadao , Tokino Takashi , Sinomura Yasuhisa
Published: August 2015

In multiple myeloma (MM), there are two distinct genetic subtypes based on copy number alterations and translocations. About half of all MM cases are a hyperdiploid type, which is characterized by multiple trisomies of chromosomes 3, 5, 7, 9 11, 15, 19 and 21 and a lower prevalence of primary translocations involving the immunoglobulin heavy chain (IgH) locus at 14q32. The remaining cases are the non-hyperdiploid type, and chromosomes 8, 13, 14, and 16 are frequently lost. Non-hyperdiploid myeloma is strongly association with translocations of IgH alleles with various partner chromosomes, such as t(11;14)(q13;q32), t(4;14)(p16.3; q32), t(14;16)(q32;q23), t(6;14)(p21;q32) or t(14;20) (q32;q11). The copy number alterations in other chromosome regions such as 1q, 6q, 8p, and 16q occur in both subtypes. Overall, non-hyperdiploid MM is associated with worse survival than hyperdiploid MM. In this study, we sequenced all exons of 409 cancer-related genes in matched tumor and normal DNA from 5 non-hyperdiploid MM patients using a next-generation semiconductor sequencing protocol. DNA was extracted from magnetic bead-enriched bone marrow CD138 positive tumor cells from the patients and CD138 negative cells were used as matched normal cells. We detected both point mutations and copy number variations (CNVs). One individual with refractory MM (IgG λ type, ISS stage II) displayed 8 non-synonymous somatic mutations in addition to numbers of CNVs including CCND1 and RB1. We observed 2 point mutations and 3 CNVs, affecting NF-kB pathway genes: IKBKB, CYLD, IKBKE, CD79B and SYK. Although the basis of NF-kB pathway activation is only partially understood, our findings greatly expand the mechanisms by which NF-kB may be activated in this patient. We also detected several gene alterations, which may be associated with poor prognosis and poor response to chemotherapy in patients with refractory MM. Although its value should be further confirmed in larger samples, the targeted next generation sequencing is a valuable tool for high-throughput genetic testing in clinical research.

Citation Format: Hiroshi Ikeda, Yasushi Sasaki, Tetsuyuki Igarashi, Yuka Aoki, Toshiaki Hayashi, Tadao Ishida, Takashi Tokino, Yasuhisa Sinomura. Molecular diagnostics of drug resistant multiple myeloma cases using targeted next generation sequencing. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4424. doi:10.1158/1538-7445.AM2015-4424
Abstract 4449: A novel Porcupine inhibitor is effective in the treatment of cancers with RNF43 mutations
Authors: Ke Zhiyuan , Madan Babita , Lim Shermaine Q.y. , Wang Sifang , Alam Jenefer , Ho Soo Yei , Jeyaraj Duraiswamy Athisayamani , Ghosh Kakaly , Chew Yun Shan , Ding Li Jun , Monoharan Vithya , Pendharkar Vishal , Ong Esther , Hill Jeffrey , Sangthongpitag Kanda , Keller Thomas , Lee May Ann , Virshup David M.
Published: August 2015

Various mutations in the Wnt pathway contribute to aberrant activation of Wnt signaling, which is implicated in multiple cancers. Besides the mutation of Adenomatous polyposis coli (APC) or beta-catenin which causes the continuous activation of Wnt signaling by stabilizing beta-catenin, there are mutations of other genes such as RNF43 that regulate Wnt signaling at the level of the ligand or cell surface receptor. Inhibition of the secretion of all human Wnts by blocking their palmitoleoylation by the O-acyltransferase, Porcupine (PORCN), could be an alternative therapeutic approach. Here we have developed a compound with a novel pharmacophore that can inhibit PORCN activity and hence Wnt signaling in nanomolar concentration. In vivo efficacy study demonstrated that the compound is highly efficacious in preventing tumor growth in genetic modification model MMTV-WNT1 mouse. We have identified a subset of pancreatic cancer, cholangiocarcinoma and mucinous ovarian cancer cell lines that are sensitive to the compound. Most of these cell lines harbor mutations in RNF43. Xenograft tumor models derived from the cancer cell lines were found to be sensitive to the compound. Our results demonstrate that inhibiting the Wnt/beta-catenin pathway by targeting PORCN with small-molecule inhibitors in a subset of cancers with mutation in RNF43 is a feasible and nontoxic strategy to overcome the problem of redundancy of Wnts, thereby, providing new option for therapy in diseases with up regulated Wnt expression.

Citation Format: Zhiyuan Ke, Babita Madan, Shermaine Q.y. Lim, Sifang Wang, Jenefer Alam, Soo Yei Ho, Duraiswamy Athisayamani Jeyaraj, Kakaly Ghosh, Yun Shan Chew, Li Jun Ding, Vithya Monoharan, Vishal Pendharkar, Esther Ong, Jeffrey Hill, Kanda Sangthongpitag, Thomas Keller, May Ann Lee, David M. Virshup. A novel Porcupine inhibitor is effective in the treatment of cancers with RNF43 mutations. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4449. doi:10.1158/1538-7445.AM2015-4449
Abstract 4487: Design, synthesis, and biological activity of N-phenyl ureidobenzenesulfonates (PUB-SOs) as new and innovative small-molecule drugs inhibiting proteins involved in DNA repair/replication mechanisms
Authors: Fortin Sébastien , Moussa Hanane , Gagné-Boulet Mathieu , Lacroix Jacques , Côté Marie-France , Velic Denis , Pauty Joris , Masson Jean-Yves
Published: August 2015

Our research group has developed a new family of promising anticancer agents designated as N-phenyl ureidobenzenesulfonates (PUB-SOs). PUB-SOs are based on a genuinely new molecular scaffold and pharmacophore constituted by two key aromatic moieties linked by a sulfonate bridge. PUB-SOs block the cell cycle progression in the S-phase and cause DNA double-strand breaks as confirmed by the phosphorylation of H2AX. In addition, PUB-SOs exemplified by prototypical PUB-SO referred to as SFOM-0046 activate selectively ATR-Chk1 pathway in all cell lines studied while it does not activate the ATM-Chk2 pathway. Using immunofluorescence, cell cycle analysis and cell survival assays, we showed that the combination of the Chk1 inhibitor 7-hydroxystaurosporine (UCN-01) and SFOM-0046 provide a proof-of concept that the cytotoxicity of PUB-SOs can be synergized by relevant anticancer drugs. Accordingly, we hypothesized that PUB-SOs inhibit proteins or enzymes involved in critical DNA repair/replication mechanisms.

In such a context, we designed a new structure-activity relationship study to assess the effects of the nature and position of different substituents on the aromatic ring B. More than 50 new PUB-SO derivatives were prepared so far and they exhibit antiproliferative activity on the low micromolar range on HT-1080 fibrosarcoma, HT-29 colon carcinoma, M21 skin melanoma and MCF7 breast carcinoma. They block also the cell cycle progression in the S-phase. These results show that substituents on ring B can be modulated to optimize both the antiproliferative and as well as biopharmaceutical properties.

In conclusion, PUB-SOs are easily synthesized, purified and pharmacomodulated, and they are therefore promising new and innovative small-molecule drugs inhibiting proteins involved in DNA repair/replication mechanisms through a genuinely innovative molecular scaffold and pharmacophore.

Citation Format: Sébastien Fortin, Hanane Moussa, Mathieu Gagné-Boulet, Jacques Lacroix, Marie-France Côté, Denis Velic, Joris Pauty, Jean-Yves Masson. Design, synthesis, and biological activity of N-phenyl ureidobenzenesulfonates (PUB-SOs) as new and innovative small-molecule drugs inhibiting proteins involved in DNA repair/replication mechanisms. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4487. doi:10.1158/1538-7445.AM2015-4487
Abstract 4495: Small molecule approach for targeting tumor-associated macrophages via a functional FRβ
Authors: Wheeler Leroy W. , Lu Yingjuan , Cross Vicky , Lloyd Alex , Parker Nikki , Qi Longwu , Wang Kevin , Wang Ian , Hahn Spencer , Vaughn Jeremy , Vlahov Iontcho P. , Low Philip S. , Leamon Christopher P.
Published: August 2015

Tumor-associated macrophages (TAMs) are considered an attractive target for oncotherapy because of their important role in tumor microenvironment. While TAMs mostly arise from inflammatory blood monocytes, they become quickly programmed to the pro-tumor phenotype. In response to tumor-derived factors, these M2-biased TAMs are essentially tumor helper cells, as both cancer cells and TAMs promote each another through a paracrine signaling loop involving epidermal growth factor and colony stimulating factor-1. This important cellular interaction is critical at all stages of tumor progression including angiogenesis, invasion, growth and metastasis.

In recent years, TAMs (CD68+, CD163+) have been found to express a functional folate receptor-β (FRβ) that displays high-affinity ligand binding and subsequent ligand-initiated endocytosis. FRβ-expressing TAMs are present in various cancer subtypes, the perivascular region, and the tumor invasive front. The abundance of FRβ+ TAMs has been linked to poor patient survival in pancreatic ductal adenocarcinoma. In the present study, we first characterized the phenotype of FRβ+ TAMs ex-vivo from both syngeneic and xenographic tumor models (breast, lung, melanoma, and pancreatic cancers). Using thioglycollate-elicited macrophages as positive controls, we estimated FRβ levels on TAMs with monoclonal antibodies and functional folate ligands. Using a ligand conjugation approach, we focused on targeting FRβ+ TAMs in tumors arising from cancer cells that do not express functional FR themselves. Preliminary proof-of-concept was established in the highly aggressive FR-negative 4T1 mammary tumor with a folate-targeted DNA alkylating agent. In addition, immunohistochemical analysis showed high FRβ expression in human anaplastic thyroid cancer, a TAM-rich disease with very poor prognosis. Collectively, our data suggest that targeting a TAM-rich cancer via FRβ may be a viable therapeutic approach with potential applications in FR-negative tumors.

Citation Format: Leroy W. Wheeler, Yingjuan Lu, Vicky Cross, Alex Lloyd, Nikki Parker, Longwu Qi, Kevin Wang, Ian Wang, Spencer Hahn, Jeremy Vaughn, Iontcho P. Vlahov, Philip S. Low, Christopher P. Leamon. Small molecule approach for targeting tumor-associated macrophages via a functional FRβ. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4495. doi:10.1158/1538-7445.AM2015-4495
Abstract 450: Suppression of innate and adaptive immune responses in the human lung tumor microenvironment via microRNA-183
Authors: Tejera Melba Marie , Tejera Melba Marie , Donatelli Sarah S. , Zhou Jun-Min , Gilvary Danielle L. , Chen Xianghong , A Erika
Published: August 2015

Innate and adaptive immunity plays a critical role in defense against cancer but can be undermined by the tumor microenvironment. To understand the underlying mechanism, we set out to analyze the molecular and cellular basis for natural killer (NK) and CD8 T cell suppression by human lung tumor cells. We found that transforming growth factor-beta (TGFb), abundantly produced by tumor cells, effectively reduced tumoricidal function in activated NK cells and CD8 T cells by specifically targeting a key adaptor protein, DAP12, which is required for surface anchoring of multiple NK receptors, including NKp44. For proof of the TGFb-miR-183-DAP12 circuit, flow cytometric analysis demonstrated that TGFb reduced surface levels of NKp44 on activated primary human NK and CD8 T cells, but western blot analysis demonstrated their intact presence in the cytoplasm. This was due to loss of DAP12, which is critical for surface expression of NKp44 and for downstream signaling to mobilize lytic granules to kill tumor cells. Mechanistically, TGFb readily induced miR-183 expression in both primary human NK cells and T cells which could bind specifically to the 3′UTR of human DAP12 to inhibit transcription and translation. Only miR-183 but not miR-185 or control (CT) transfection repressed luciferase activity of DAP12 3′UTR luciferase reporter constructs transferred into HELA cells. Moreover, we overexpressed lentiviral miR-183 in either primary NK and T cells and found that such expression effectively reduced DAP12 levels in both cell types. Flow cytometric analysis of these same cells indicated that loss of DAP12 was accompanied by loss of NKp44 surface expression and by loss of cytotoxic function against tumor cells. As further proof, we showed that lentiviral anti-sense miR-183 transfection into NK cells and T cells prevented TGFb from suppressing DAP12 expression. Thus, we have uncovered a seminal TGFb-miR-183-DAP12 pathway utilized by tumor cells to silence infiltrating NK and T cells, constituting the first report of miR control of an adaptor protein to control both NK and T cell immune function.

Citation Format: Melba Marie Tejera, Melba Marie Tejera, Sarah S. Donatelli, Jun-Min Zhou, Danielle L. Gilvary, Xianghong Chen, Erika A.. Suppression of innate and adaptive immune responses in the human lung tumor microenvironment via microRNA-183. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 450. doi:10.1158/1538-7445.AM2015-450
Abstract 4502: Identification of an aspirin-derived small molecule as a potential therapeutic for colorectal cancer
Authors: Karelia Deepkamal , Pandey Manoj , Plano Daniel , Narayan Satya , Jani Jitesh , Sharma Jay , Amin Shantu , Sharma Arun K.
Published: August 2015

In the US, colorectal cancer (CRC) remains the second leading cause of cancer-related death. The 5-year survival rate for patients with CRC is 64%, although for patients who present with distant metastases at diagnosis the 5 year survival is only 12%. Currently used standard care for CRC can only marginally extend the survival of patients and is associated with numerous adverse events. Hence, there remains an urgent need for more effective and less toxic treatments, which can increase CRC patient survival with better quality of life. Non-steroidal anti-inflammatory drugs (NSAIDs) (e.g. Aspirin, Ibuprofen, Naproxen), which are widely used for their anti-inflammatory and analgesic effects, have also been shown to be effective in reducing colon cancer incidences and are proposed to be effective chemopreventive agents. However, these NSAIDs are not potent enough to be used as cancer therapeutics and also carry a liability of gastrointestinal bleeding mainly associated with COX inhibition. Through extensive structure activity relationship (SAR) studies based on newly synthesized NSAID-analogs, we have identified a novel aspirin analog, ASD-49, as the most effective small molecule that showed efficacy in different cancer cell lines, including CRC cells with an EC50 ranging between 1 to 2.5 μM at 48 h treatment in culture. This unique compound selectively killed cancer cells and is >50-times more potent than the current therapy for CRC and >100-times more potent than the parent compound, aspirin. Our preliminary results show that ASD-49 induces G1 and G2 cell cycle arrest in HT29 cells. In addition, ASD-49 induced apoptosis as indicated by PARP cleavage in HCT116 and RKO cells. Furthermore, we observed down-regulation of anti-apoptotic protein Bcl-2 in a dose-dependent manner in both HCT116 and RKO cell lines. Unlike aspirin, the novel analog did not alter COX-2 protein expression, suggesting that ASD-49 may not possess adverse effects related to COX-2 inhibition. In addition, >100-times lower dose requirement as compared to aspirin is expected to further minimize side effects associated with aspirin structure. Taken together, our results suggest that ASD-49 may be a promising agent for CRC treatment. Detailed results regarding the efficacy and mechanism of action of ASD-49 will be presented.

Citation Format: Deepkamal Karelia, Manoj Pandey, Daniel Plano, Satya Narayan, Jitesh Jani, Jay Sharma, Shantu Amin, Arun K. Sharma. Identification of an aspirin-derived small molecule as a potential therapeutic for colorectal cancer. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4502. doi:10.1158/1538-7445.AM2015-4502
Abstract 4507: Intra-tumoral pharmacokinetic profiling for doxorubicin within solid tumor micromilieu using multicellular layers culture in-vitro
Authors: Al-Abd Ahmed M. , Al-Abbasi Fahad A. , Khedr Alaa , Atteiah Salah G.
Published: August 2015

In contrast to plasma pharmacokinetics, intratumoral pharmacokinetics of DOX determines its spatial anti-tumoral activity. Three dimensional multicellular layers (MCL) model for solid tumors present optimum experimental platform for studying the intratumoral pharmacokinetics of DOX. This might imply new insights for understanding intratumoral pharmacokinetic parameters with realistic clinical implications. Herein, we are presenting simplified method for the spatial in-situ concentration assessment of DOX within the avascular simulating MCL solid tumor model of DLD-1 and HT-29 cell lines. DLD-1 and HT-29 formed viable well structured MCL model abundant in extracellular matrix component (fibronectin). DOX (100 μM) showed stronger anti-proliferative effect against MCL of DLD-1 compared to HT-29 MCL (38.8% and 27.9%, respectively). The differential potencies of DOX closely correlate to the intratumoral pharmacokinetics within MCL's of both cell lines. DOX penetrated faster and washed out slower through the MCL of DLD-1 compared to HT-29 MCL. Distribution of DOX within MCL of DLD-1 was more homogenous compared to HT-29 MCL. Tissue concentration of DOX within MCL of DLD-1 was significantly higher than HT-29 MCL's after 96 h exposure (0.7 and 0.4 μmole/gm tissue, respectively). Concentration of DOX within MCL of both cell lines exceeded the IC50 under monolayer conditions (2.3±0.6 μM and 0.6±0.1 μM, respectively). In addition, DOX was extensively metabolized to less active metabolites (doxorubicinol and doxorubicinone) during traversing the thickness of both MCL’s. In conclusion, Intratumoral pharmacokinetic barriers to DOX might be key determinant in drug resistance on the tissue level, despite cellular and molecular events.

Citation Format: Ahmed M. Al-Abd, Fahad A. Al-Abbasi, Alaa Khedr, Salah G. Atteiah. Intra-tumoral pharmacokinetic profiling for doxorubicin within solid tumor micromilieu using multicellular layers culture in-vitro. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4507. doi:10.1158/1538-7445.AM2015-4507