bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 762: Prevention and reversion of multidrug resistance (MDR) in cancer by NSC23925
Authors: Yang Xiaoqian , Hornicek Francis , Duan Zhenfeng
Published: May 2015

The major limitation to the success of chemotherapy in cancer is the development of multidrug resistance (MDR). Preventing the emergence of MDR during chemotherapy treatment has been a high priority of clinical and investigational oncology, but remains an elusive goal. NSC23925 has recently been identified as a novel and potent MDR reversal agent. However, whether NSC23925 can prevent the development of MDR in cancer is unknown. Therefore, this study was to evaluate the effects of NSC23925 on prevention and reversion of MDR. Human osteosarcoma cell line U-2OS and Saos were exposed to increasing concentrations of taxol alone or in combination with NSC23925 for 6 months. The results showed that cells selected with increasing concentrations of taxol alone developed MDR with resistance to taxol and other Pgp substrates, while cells cultured with taxol-NSC23925 simultaneously prevented the development of MDR and cells remained sensitive to chemotherapeutic agents. Taxol selected resistant cells showed high expression and activity of drug transporter P-glycoprotein (Pgp), whereas taxol-NSC23925 combination treatment prevented the expression of Pgp. Our findings suggest that NSC23925 may prevent the development of MDR by specifically preventing the overexpression of Pgp. Moreover, NSC23925 can reverse MDR in drug resistant cells by modulating Pgp activity. Given the significant incidence of MDR in cancer and the lack of effective agents for prevention and revision of MDR, NSC23925 and derivatives hold the potential to improve the outcome of cancer patients with poor prognosis due to drug resistance.

Citation Format: Xiaoqian Yang, Francis Hornicek, Zhenfeng Duan. Prevention and reversion of multidrug resistance (MDR) in cancer by NSC23925. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 762. doi:10.1158/1538-7445.AM2014-762
Abstract 768: CB-PIC sensitizes chemoresistant cancer cells to drugs via suppression of MDR1 and its upstream signal molecules, AKT and p38-MAPK
Authors: Lee Duckgue , Yun Miyong
Published: May 2015

ATP-binding cassette (ABC) transporters including MDR1-/P-gp have been majorly targeted for drug resistant tumor therapies. In this study, we screened the compound that negatively affects MDR-1 expression in chomoresistant cancer cell lines. We found that CB-PIC, the novel synthetic compound, could decrease P-glycoprotein expression via disrupt AKT/p-38 signal pathway in paclitaxel resistant lung cancer cell, H460/PR and doxorubicin resistant breast cancer cell, MCF7/Adr with highly expressed P-glycoprotein. However, CB-PIC did not regulate enzyme activity of P-glycoprotein, while EGCG and tannic acid accumulated Rhodamine 123 intracellular region of cell via short time efflux assay. CB-PIC treatment resulted in disruption of proliferation of drug resistant cells. In addition, CB-PIC dramatically induced apoptosis in time and dose dependent manner in both paclitaxel and doxorubicin resistant cancer cells. Furthermore, CB-PIC sensitizes drug-resistant cancer cells to paclitaxel and doxorubicin by decrease of survival proteins, survivin, Mcl-1 and Bcl-2, as well as MDR1 suppression leading to accumulate the treated drug intracellular region. Taken together, CB-PIC suppresses the P-glycoprotein expression through inhibition of AKT/p-38 MAPK signaling resulting in the increase of drug sensitivity in chemoresistant cancer cells.

Citation Format: Duckgue Lee, Miyong Yun. CB-PIC sensitizes chemoresistant cancer cells to drugs via suppression of MDR1 and its upstream signal molecules, AKT and p38-MAPK. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 768. doi:10.1158/1538-7445.AM2014-768
Abstract 81: Deregulated expression of HDAC9 in B cells leads to the development of lymphoproliferative disease and lymphoma
Authors: Gil Veronica S.
Published: May 2015

Aberrant epigenetics leading to changes in chromatin structure and patterns of gene expression is an important factor in cancer pathogenesis. Histone Deacetylase 9 (HDAC9) is a class IIa chromatin-modifying enzyme that, within the haematopoietic system, is preferentially expressed in the B-cell lineage. Furthermore, HDAC9 expression is deregulated in B-lymphoid malignancies. To examine the role of HDAC9 in malignant B-cell transformation, we generated an in vivo mouse model that constitutively expresses human HDAC9 from early stages of B-cell development under the control of the immunoglobulin heavy chain (IgH) enhancer (Eµ). In this study, we demonstrate that from an average of six months of age, Eµ-HDAC9 transgenic mice displayed splenomegaly and immunophenotypic features that resemble lymphoproliferative disease (LPD) in spleen. Later on in life, up to twenty-three months of age, 24% (19 out of 78) of Eµ-HDAC9 mice developed mature B-cell neoplasms such as splenic marginal zone lymphoma (SMZL) and aggressive diffuse large B-cell lymphoma (DLBCL) that could represent a progression stage of the observed lymphoproliferative disease (LPD) and monoclonal B-cell lymphocytosis (MBL). In contrast, a low background incidence of these malignancies was observed in control mice, 2% (1 out of 46) (p = 0.0008). The molecular pathogenesis of the Eµ-HDAC9 B-cell tumors seems to involve activation-deactivation of the BCL6-p53 axis by HDAC9 influenced deacetylation thus affecting DNA repair and cell cyle progression in germinal centre (GC) during immune response. Consistently, ZFN-mediated HDAC9-knockout results in cell cycle changes leading to strong inhibition of cell growth in vitro. In summary, aberrant expression of HDAC9 leads to the development of lymphoproliferative disorders, including GC and post-GC lymphomas, demonstrating an oncogenic role for HDAC9 in B-cells and highlighting its importance as a therapeutic target.

Citation Format: Veronica S. Gil. Deregulated expression of HDAC9 in B cells leads to the development of lymphoproliferative disease and lymphoma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 81. doi:10.1158/1538-7445.AM2014-81
Abstract 812: Assessing combinations of FDA approved chemotherapy in childhood sarcoma cell lines
Authors: Yu Diana , Cubitt Christopher , Sullivan Daniel , Reed Damon
Published: May 2015

Background: Sarcomas are a heterogeneous group of mesenchymal malignant tumors that make up approximately 15% of all childhood tumors and represent the fourth most common group of malignancies in children, remaining common in young adults with cancer as well. While over 35 agents have been FDA approved for adult carcinomas in the past 3 years, little progress has been made in incorporating new agents into the chemotherapy plans for pediatric sarcomas in the past few decades. A system to rapidly evaluate many agents and combinations of agents which can immediately be translated into clinical trials in a clinically relevant manner is needed.

Methods: Here, we seek to examine the therapeutic activity of single agent and combinations of FDA approved chemotherapy agents using high-throughput screening platforms, based on well-established childhood cancer cell line models for most prevalent forms of pediatric bone sarcomas (osteosarcoma and Ewing sarcoma). Taking into consideration past lessons learned from in vitro experiments, we designed stringent screening conditions that assess the candidate compounds at clinically-relevant concentrations and time of exposures that mimic the in vivo pharmacokinetics in an effort to maximize the translational potential of these results to the clinical setting. Fifty-four agents, mainly FDA approved but also including agents with promising mechanism of action or preclinical data were screened across 5 cell lines for each histology. Caspace glo and cell titer glo assays were used to determine apoptosis and cell viability at the maximal achievable serum concentration from phase I studies and at two lower concentrations. Promising agents were then studied in 2 drug combinations to evaluate for synergy with careful attention to metabolism pathways, existing clinical data for combinations, and non-overlapping toxicities. Combinations of FDA approved agents with promising therapeutic efficacy and predicated non-overlapping toxicities will be translated to the clinic using the established Sunshine Project network.

Results: Our screening results indicate that a number of FDA approved chemotherapy agents, evaluated at clinically relevant concentrations and exposure times, significantly reduced cell viability and induced up regulation of caspase3/7 activities across all cell lines. These findings suggest promising opportunities for developing combinational approach to treating childhood bone sarcomas and can be utilized to explore other pediatric tumors with existing cell lines.

Citation Format: Diana Yu, Christopher Cubitt, Daniel Sullivan, Damon Reed. Assessing combinations of FDA approved chemotherapy in childhood sarcoma cell lines. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 812. doi:10.1158/1538-7445.AM2014-812
Abstract 813: A combination of cisplatin, irinotecan, and paclitaxel (CIP) as frontline treatment of patients with metastatic esophageal cancer (mEC)
Authors: Giever Thomas A. , Ritch Paul S. , Thomas James P. , Wiebe Lauren A. , Haasler George B. , Gasparri Mario G. , Johnstone David , Johnstone Candice A. , Gore Elizabeth M. , George Ben
Published: May 2015

Introduction: Median survival associated with mEC is 9-12 months. While systemic therapy is the mainstay of treatment for mEC, there is no consensus regarding the best frontline chemotherapy combination. We evaluated the efficacy of CIP (Cisplatin 30 mg/m2, Irinotecan 50 mg/m2 and Paclitaxel 50 mg/m2 on days 1 and 8 of a 21 day cycle) in patients with mEC.

Methods: We performed a retrospective review to identify mEC patients at the Medical College of Wisconsin (MCW) who were treated with frontline CIP between January 1, 2005 and December 31, 2010. Relevant clinical information was collected through a review of the electronic medical record. This study was approved by the Institutional Review Board at MCW.

Results: We identified 13 patients who were treated with CIP as frontline therapy for mEC. The median age at diagnosis was 59 years (range 48-71 years), 12 (92%) patients were male, and 9 patients (69%) had adenocarcinoma with the remaining being squamous cell carcinoma. Eleven patients (85%) presented with metastatic disease, while 2 patients (15%) were diagnosed with metastatic disease after having undergone initial surgical resection with disease free intervals of 39 and 14 months respectively. The most common sites of metastases were lymph nodes (11 patients), liver (6 patients), lung (4 patients), bone (3 patients), and peritoneum (2 patients); 77% of patients had at least 2 sites of metastases. The median number of chemotherapy cycles administered was 7 (range 1-14). Four patients (31%) developed grade 3/4 toxicities including neutropenia in 3 patients, nausea in 1 patient, and anemia in 1 patient. One patient (8%) had a complete response (CR), 7 patients (54%) had a partial response (PR), 2 patients (15%) had stable disease (SD), and 3 patients (23%) developed progressive disease (PD). The median progression free survival (PFS) was 5 months (range 0-29 months) and the median overall survival (OS) was 10 months (range 3-36 months).

Conclusion: CIP is an active and well tolerated regimen in patients with mEC that merits prospective evaluation.

Citation Format: Thomas A. Giever, Paul S. Ritch, James P. Thomas, Lauren A. Wiebe, George B. Haasler, Mario G. Gasparri, David Johnstone, Candice A. Johnstone, Elizabeth M. Gore, Ben George. A combination of cisplatin, irinotecan, and paclitaxel (CIP) as frontline treatment of patients with metastatic esophageal cancer (mEC). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 813. doi:10.1158/1538-7445.AM2014-813
Abstract 829: The novel camptothecin derivative and IAP inhibitor FL118 is an effective treatment for irinotecan-refractory colorectal cancer
Authors: Westover David , Ling Xiang , Liu Xiaojun , Lam Hong , Gongora Celine , Rio Maguy Del , Li Fengzhi
Published: May 2015

The current standard care for metastatic colorectal cancer is combination of leucovorin, 5-fluorouracil, and irinotecan. Irinotecan is a semisynthetic analogue of the alkaloid camptothecin and functions by inducing DNA double-strand breaks through inhibition of topoisomerase I (Top1) activity. The camptothecin-derived Top1 inhibitors irinotecan and topotecan are effective anti-cancer agents that are currently utilized as first-line therapies in several human cancers. Unfortunately, resistance to these agents usually develops during treatment, often through upregulation of the drug efflux pump ABCG2, an ATP-binding cassette transporter. By increasing the rate of drug export, ABCG2 decreases the amount of irinotecan or topotecan that accumulates intracellularly, thereby protecting cancer cells from the cytotoxic effects of these drugs. Therefore, a camptothecin derivative that is also a poor substrate for ABCG2 could potentially overcome ABCG2-induced irinotecan resistance and restore therapeutic efficacy. Our lab recently reported on a novel camptothecin derivative, FL118, which shows strong anti-cancer activity both in vitro and in vivo. Here we report the ability of FL118 to overcome irinotecan resistance. Several irinotecan-resistant colon cancer cell lines derived from HCT116 were treated with either FL118 or SN38, the active metabolite of irinotecan. We observed a decrease in the potency of SN38 in irinotecan-resistant cells that overexpressed ABCG2 compared to cells that did not overexpress ABCG2. In contrast, this loss of potency was not observed for FL118. To confirm that the decrease in potency was ABCG2-dependent, HCT116-A2, an ABCG2 overexpressing cell line, was treated with SN38 or FL118 in the presence or absence of Ko143, an ABCG2 inhibitor. We observed that Ko143 could restore potency to SN38 in HCT116-A2 cells, confirming that irinotecan resistance in HCT116-A2 cells is mediated by ABCG2. However, Ko143 could not modulate the potency of FL118, further suggesting that FL118 is not affected by ABCG2 activity. Similar results were observed in studies with anti-ABCG2 shRNA. Based on these observations, we conclude that FL118 is not a substrate for ABCG2, and that further testing is warranted to determine whether FL118 would be an effective anti-cancer agent for patients with irinotecan-resistant colorectal cancer overexpressing ABCG2.

Citation Format: David Westover, Xiang Ling, Xiaojun Liu, Hong Lam, Celine Gongora, Maguy Del Rio, Fengzhi Li. The novel camptothecin derivative and IAP inhibitor FL118 is an effective treatment for irinotecan-refractory colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 829. doi:10.1158/1538-7445.AM2014-829
Abstract 84: Functional evaluation of synchronous inactivation of PTEN and P53 in a murine model of prostate cancer
Authors: Uemura Hirotsugu , Kura Yurie , Ando Naomi , Fukushima Emiko , Hatanaka Yuji , Yamamoto Yutaka , Shimizu Nobutaka , Yoshimura Kazuhiro , Nozawa Masahiro , Yoshikawa Kazuhiro , Nishio Kazuto , Velasco Marco A. De
Published: May 2015

The prostate-specific PTEN conditional knockout (K/O) mouse model has been extremely useful as tool to investigate the biology of the tumorigenesis process, particularly during early tumor development. However, cancer progression is gradual with a low metastatic rate. This feature poses a challenge to this model when used for survival studies and end-stage biological characterization of prostate cancer. Inactivation of P53 is a characteristic that is frequent in advanced prostate cancer. To address this issue, we generated series of double K/O mice based on either P53 loss or gain of function in combination with PTEN inactivation. Conditional inactivation of both PTEN and P53 results in the development of aggressive tumors compared to PTEN K/O mice. Homozygous PTEN/P53-mutants mice grow prostate tumors at a faster rate and experience dramatically reduced survival. Homozygous double K/O mice also developed a higher frequency and number of metastases. Furthermore, haploinsufficient P53 gain of function mutations contributed to an invasive cancer phenotype compared to P53 loss of function haploinsufficient tumors. In conclusion, our analyses demonstrate that synchronous inactivation of PTEN and P53 promotes an aggressive prostate cancer and provides rationale that supports that use of such models is better suited for studies requiring long-term survival and end-stage cancer characterization.

Citation Format: Hirotsugu Uemura, Yurie Kura, Naomi Ando, Emiko Fukushima, Yuji Hatanaka, Yutaka Yamamoto, Nobutaka Shimizu, Kazuhiro Yoshimura, Masahiro Nozawa, Kazuhiro Yoshikawa, Kazuto Nishio, Marco A. De Velasco. Functional evaluation of synchronous inactivation of PTEN and P53 in a murine model of prostate cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 84. doi:10.1158/1538-7445.AM2014-84
Abstract 861: Survivin-mediated adaptive response: a risk factor for IGRT
Authors: Grdina David J. , Murley Jeffrey S. , Miller Richard C. , Woloschak Gayle E. , Li Jian Jian , Weichselbaum Ralph R.
Published: May 2015

Exposure of cells to very low doses of ionizing radiation can induce an enhanced resistance or adaptive response to a subsequent larger radiation dose as demonstrated by an increase in cell survival. Expression of a radio-adaptive response has been attributed to pro-survival signaling processes induced by very low radiation doses in the range of 5 to 100 mGy. The radiation-induced adaptive response is gaining considerable attention due in part to the expanding use of imaging technologies such as computerized axial tomography and portal imaging to monitor tumor response and positioning during multi-dose standard radiation therapy protocols. The focus of this study was to investigate whether this process could occur when the very small imaging radiation dose was administered prior to or following the administration of the first of two typically used radiation therapy level doses. SA-NH mouse sarcoma cells and SA-NH cells stably transfected with mutated IκBα (SA-NH+mIκBα1) to inhibit the activation of NFκB were grown to confluence in vitro or as tumors in the right hind leg of C3H mice. Percentage of cell survival, apoptosis frequencies and survivin protein levels were determined as a function of radiation conditions and transfection of cells with survivin siRNA. A significant radio-adaptive response was observed in cells exposed to 5 mGy - 100 mGy 15 min to 6 h prior to or after irradiation with the first of two 2 Gy therapeutic radiation doses and was associated with a concomitant reduction in apoptosis frequencies and a rapid elevation in survivin protein levels. The very low dose radiation-induced elevation in survivin levels, the reduction in apoptosis frequencies and enhanced cell survival were not observed in SA-NH+mIκBα1 cells or SA-NH cells transfected with survivin siRNA. We have identified a very low radiation dose-induced survivin-mediated adaptive response that could compromise therapeutic outcomes associated with the increased usage of imaging procedures in radiation therapy. This work was supported by NIH/NCI grant R01 CA132998 and DOE Low Dose Program/Project Grant DE-SC0001271 (D.J.G.)

Citation Format: David J. Grdina, Jeffrey S. Murley, Richard C. Miller, Gayle E. Woloschak, Jian Jian Li, Ralph R. Weichselbaum. Survivin-mediated adaptive response: a risk factor for IGRT. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 861. doi:10.1158/1538-7445.AM2014-861
Abstract 864: Increased rates of radiation pneumonitis in patients receiving stereotactic ablative radiotherapy for central versus peripheral lung tumors
Authors: Chaudhuri Aadel A. , Tang Chad , Trakul Nicholas , Wynne Jacob , Loo Billy , Diehn Maximilian
Published: May 2015

Purpose: Treatment of central lung tumors with stereotactic ablative radiotherapy (SABR) has been associated with higher rates of toxicities than patients with peripheral tumors. Here we report our institution's experience in treating central lung tumor patients with SABR, compared to patients treated for peripheral lung tumors.

Methods: We retrospectively reviewed outcomes in 82 patients with a total of 96 primary or metastatic lung tumors treated by SABR. The cohort included 47 central and 49 peripheral tumors and all were treated with 50 Gy in 4-5 fractions. Outcomes and radiation-associated toxicities were compared between the two groups, not including patients in whom central and peripheral tumors were concurrently treated (8 such patients). Dosimetric analysis was performed on patients with central tumors to determine max point dose (MPD) and volumetric max dose (Vmax) to central structures.

Results: Median follow-up time was 15 months (range 1 to 59 months). Median overall survival was 34 and 40 months (p=0.75) for patients with central and peripheral tumors, respectively. We observed no hemoptysis and no grade 5 toxicities. Rates of symptomatic toxicity were higher in patients with central tumors (grade 2+, 38% vs. 11%, p=0.0056) but were equivalent for severe toxicity (grade 3+, 5.4% vs. 5.4%, p=0.93). The vast majority of toxicities were due to radiation pneumonitis, incidence of which was significantly greater in patients with central tumors (grade 2+, 30% vs. 2.7%, p=0.0021; grade 3+, 5.4% vs. 0%, p=0.15). Dosimetric analysis of 36 central tumor treatment plans revealed that 23 (64%) exceeded at least one of the dose constraints used in RTOG 0813, most commonly for central airway. Rates of grade 2+ pneumonitis were not significantly different between patients exceeding RTOG 0813 central airway dosing constraints vs. those meeting them (24% vs. 20%, p=0.69), but trended towards significance for grade 3+ radiation pneumonitis, with patients exceeding RTOG 0813 central airway dosing constraints experiencing higher rates (0% vs. 13%, p=0.095).

Conclusions: Central lung tumor patients treated with SABR appear to have greater risk for symptomatic radiation pneumonitis than patients with peripheral tumors. Exceeding RTOG 0813 dosimetric constraints for central airway may predispose to higher risk for severe radiation pneumonitis.

Citation Format: Aadel A. Chaudhuri, Chad Tang, Nicholas Trakul, Jacob Wynne, Billy Loo, Maximilian Diehn. Increased rates of radiation pneumonitis in patients receiving stereotactic ablative radiotherapy for central versus peripheral lung tumors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 864. doi:10.1158/1538-7445.AM2014-864
Abstract 869: Evaluate breast cancer diagnostic antibody specificity by using high density protein microarray technology
Authors: Ma Donghui , Chen Jian , Fu Wei , McDowell Julie , Yuan Kehu
Published: May 2015

Detection sensitivity and target specificity are the two key features for a good diagnostic monoclonal antibody. There are a number of well-established technology platforms to quantitatively measure antibody affinity. However, there is no complete technology solution to evaluate antibody target specificity. Antibody cross-reactivity could lead to false diagnostic reports. Currently, high density protein microarray technology is the best technology platform available for antibody specificity evaluation. Here we will showcase how we utilized protein microarray technology to evaluate target specificity for a couple of commonly used breast cancer IHC diagnostic antibodies. Our data suggested that cross-reactivity is one of the reasons to create off-target staining for breast cancer diagnostics. We will also show how we can utilize OriGene protein microarray chip technology to develop UltraMABs, the Ultra-specific monoclonal antibodies for anatomic pathology application.

Citation Format: Donghui Ma, Jian Chen, Wei Fu, Julie McDowell, Kehu Yuan. Evaluate breast cancer diagnostic antibody specificity by using high density protein microarray technology. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 869. doi:10.1158/1538-7445.AM2014-869
Abstract 891: Urinary ADAM12 levels detect the presence of pancreatic cancer
Authors: Roy Roopali , Zurakowski David , Kulke Matthew , Moses Marsha A.
Published: May 2015

Pancreatic malignancies are the fourth leading cause of all cancer-related deaths of both men and women in the United States. Each year, approximately 32,000 new patients are diagnosed with this disease and nearly the same number die from it. A majority of patients with pancreatic malignancies, including both pancreatic ductal adenocarcinoma (PDAC) and pancreatic neuroendocrine tumors (pNET) present with advanced disease due to a lack of specific symptoms and current diagnostic limitations, making this disease extremely difficult to detect. The combination of poor prognosis and late presentation of pancreatic cancer patients highlights the urgent need for the development of effective, early detection strategies for this disease. Our laboratory has established a comprehensive biomarker discovery initiative whose objective is to identify proteins present in urine of cancer patients, to determine whether their presence might be relevant to disease status and stage and to validate their diagnostic and prognostic efficacy in large scale clinical trials. In the current study, our goal was to determine whether ADAM12 (a disintegrin and metalloprotease 12) could be detected in the urine of patients with pancreatic malignancies and whether ADAM12 levels might serve as an independent predictor of disease status. Retrospective analyses of urine samples (n=130) from PDAC and pNET patients as well as age- and sex-matched controls were conducted. Urinary ADAM12 levels were determined using a monospecific ELISA system. In addition, ADAM12 protein expression in tumor and normal pancreatic tissues was analyzed via immunohistochemistry (IHC). Multivariable logistic regression analyses indicated that, when controlling for age and sex, urinary ADAM12 could serve as significant independent predictor for distinguishing PDAC (P<0.001) and pNET (P<0.008) patients from healthy controls. Kaplan-Meier analysis of estimated patient survival stratified by urinary ADAM12 levels indicated a significantly shorter patient survival time for PDAC patients with high ADAM12 levels (P=0.015) compared to patients with lower urinary ADAM12. In addition, IHC analysis indicated that ADAM12 protein expression was upregulated ∼4-fold in Grade I-III PDAC compared to normal pancreatic tissue. Taken together, our results suggest that the measurement of ADAM12 levels may have diagnostic value in detection and/or clinical monitoring of disease status in patients with pancreatic malignancies. [Supported by: The Advanced Medical Foundation]

Citation Format: Roopali Roy, David Zurakowski, Matthew Kulke, Marsha A. Moses. Urinary ADAM12 levels detect the presence of pancreatic cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 891. doi:10.1158/1538-7445.AM2014-891
Abstract 893: Mining the circulating immune cell transcriptome for ovarian cancer-specific biomarkers: A proof of concept study
Authors: Uppal Shitanshu , Kapur Arvinder , Felder Mildred , Medlin Erin , Shojaei Hadi , Gonzalez-Bosquet Jesus , Patankar Manish S.
Published: May 2015

The continuous interplay between cancer cells and immune system allows an opportunity to identify cancer-specific biomarkers in immune cells. Here, we compare the transcriptomes of peripheral blood mononuclear cells (PBMCs) obtained from advanced stage ovarian cancer patients and healthy donors in a proof-of-concept study to identify ovarian cancer-specific biomarkers. PBMCs were collected from patients undergoing surgery with the gynecologic oncology service. RNA was extracted from the PBMCs of five patients (training set) with confirmed diagnosis of epithelial ovarian cancer and 4 health donor blood samples. Microarray analysis using Illumina platform was conducted and analyzed using the R statistical package. After data normalization, differentially expressed genes in the two cohorts were identified controlling for false discovery rate at 5 and 10%. The top ten up- and downregulated genes were selected. Gene ontology and pathway analysis led to identification of a Candidate Biomarker Panel (CBP) comprising of five of the ten genes that were most likely to serve as biomarkers for ovarian cancer. Validation of the CBP was achieved using PBMCs isolated from a separate group of six patients and healthy donors (test set) was performed using qPCR and flow cytometry and in vitro culture experiments. Bioinformatics analysis identified 391 genes that were up-regulated and 599 that were down-regulated. A set of five genes with high biomarker potential and were included in the CBP. All of the CBP genes were differentially expressed by 3-5 fold (p<0.03) in the cancer patient PBMCs as compared to healthy donors. Validation results for the CBP were similar between the training set and the test set. Co-culture experiments indicated differential expression of the CBP genes in healthy donor-derived PBMCs co-cultured with cancer cells. Continuous cross-talk between immune cells and abnormal tissues leads to changes in the immune cell transcriptome and proteome in response to a pathologic state. We demonstrate that the transcriptome and proteome of circulating immune cells is a repository for ovarian cancer-specific biomarkers. Our on-going studies are focused on validating the CBP in a larger cohort of samples obtained from healthy donors, ovarian cancer patients and subjects treated for benign gynecologic anomalies.

Citation Format: Shitanshu Uppal, Arvinder Kapur, Mildred Felder, Erin Medlin, Hadi Shojaei, Jesus Gonzalez-Bosquet, Manish S. Patankar. Mining the circulating immune cell transcriptome for ovarian cancer-specific biomarkers: A proof of concept study. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 893. doi:10.1158/1538-7445.AM2014-893
Abstract 904: EGFR expression predicts recurrence in patients undergoing adjuvant chemotherapy for advanced bladder cancer
Authors: Mansour Ahmed , Abdulreheem Mona , Elsherbeeny Mamdouh , Sultan Mohammed , Shokeir Ahmed , Mosbah Ahmed , Abol-Enein Hassan , Park Taeeun , Kim Hyung , Kim Jayoung
Published: May 2015

Epidermal growth factor receptor (EGFR) overexpression has been associated with bladder cancer progression and poor clinical outcomes. In vivo studies have linked EGFR subcellular trafficking and chemo-resistance to cisplatin-based protocols in other tumor types. This has not been studied in the clinical adjuvant setting in bladder cancer. In this study, we aimed to investigate the prognostic significance of EGFR expression in patients receiving adjuvant chemotherapy following radical cystectomy for advanced bladder cancer. Mansoura Urology and Nephrology Center database was reviewed. Patients treated with radical cystectomy and thereafter completed adjuvant chemotherapy for adverse pathological features or node positive diseases were identified. Patients who underwent palliative cystectomy, those with histological diagnosis other than pure transitional cell carcinoma and patients who received adjuvant radiotherapy were excluded. Immunohistochemical analysis for EGFR expression was performed on archived bladder specimens. The relationship of EGFR expression and clinical outcomes was assessed. In vitro studies were performed to determine whether EGFR expression associates with resistance to chemotherapeutic reagents. The study included 58 patients. Mean age was 57 years. All patients had pathologic stage T2 or greater. Majority of patients had node positive disease (n=53, 91%). Mean follow up was 26.61 months. EGFR was overexpressed in 25 cystectomy specimens (43%). Kaplan Meyer analysis revealed that EGFR over-expression significantly correlated with disease recurrence (p=0.021). Cox proportional hazard modeling identified EGFR overexpression as an independent predictor for disease recurrence (p=0.04). In vitro functional analysis demonstrated that EGFR expression level altered response to cisplatin-induced apoptosis, and that inhibition of EGFR enhanced sensitization of bladder cancer cells to cisplatin-induced apoptosis. In summary, our findings suggest that EGFR overexpression is associated with disease recurrence following adjuvant chemotherapy for advanced bladder cancer. EGFR status may help predict prognosis and select patients for adjuvant chemotherapy.

Citation Format: Ahmed Mansour, Mona Abdulreheem, Mamdouh Elsherbeeny, Mohammed Sultan, Ahmed Shokeir, Ahmed Mosbah, Hassan Abol-Enein, Taeeun Park, Hyung Kim, Jayoung Kim. EGFR expression predicts recurrence in patients undergoing adjuvant chemotherapy for advanced bladder cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 904. doi:10.1158/1538-7445.AM2014-904
Abstract 916: A multidimensional analysis of predictive biomarkers in colorectal cancer
Authors: Mariani Marisa , He Shiquan , McHugh Mark , Andreoli Mirko , Pandya Deep , Sieber Steven , Wu Zheyang , Fiedler Paul , Shahabi Shohreh , Ferlini Cristiano
Published: May 2015

Background: CRC cancer is one of the deadliest diseases in Western countries. In order to develop predictive biomarkers for CRC (colorectal cancer) aggressiveness, we analyzed retrospectively 267 CRC patients via a novel, multidimensional biomarker platform.

Methods: Using nanofluidic technology for qPCR analysis and quantitative fluorescent immunohistochemistry for protein analysis, we assessed 33 microRNAs, 124 mRNAs and 9 protein antigens. Analysis was conducted in each single dimension (microRNA, gene or protein) using both the multivariate Cox model and Kaplan-Meier method. Thereafter, we simplified the censored survival data into binary response data (aggressive vs. non aggressive cancer). Subsequently, we combined the data into multidimensional scores using inverse sliced regression for sufficient dimension reduction. Accuracy was assessed using the Area under the Curve (AUC) and the receiver operating characteristic (ROC) method.

Results: Single dimension analysis led to the discovery of individual factors that were significant predictors of outcome. These included seven specific microRNAs, four genes, and one protein. When these factors were quantified individually as predictors of aggressive disease, the highest demonstrable area under the curve (AUC) was 0.68. By contrast, when all results from single dimensions were combined into multidimensional biomarkers, AUCs were dramatically increased with values approaching and even exceeding 0.9. Conclusions: Single dimension analysis generates statistically significant predictors, but their predictive strengths are suboptimal for clinical utility. A novel, multidimensional approach overcomes these deficiencies. Newly derived multidimensional biomarkers have the potential to meaningfully guide the selection of therapeutic agents for individual patients while elucidating molecular mechanisms driving disease progression.

Citation Format: Marisa Mariani, Shiquan He, Mark McHugh, Mirko Andreoli, Deep Pandya, Steven Sieber, Zheyang Wu, Paul Fiedler, Shohreh Shahabi, Cristiano Ferlini. A multidimensional analysis of predictive biomarkers in colorectal cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 916. doi:10.1158/1538-7445.AM2014-916
Abstract 92: Generating new syngeneic models for oral squamous cell carcinoma metastasis study
Authors: Shi Zonggao , Johnson Jeff , Liu Yueying , Stack Sharon
Published: May 2015

Cancer metastasis is a complex problem that presents clinical challenges and is the cause of most cancer-related deaths. Mechanistic studies on metastasis hold promise for finding better approaches to control cancer mortality. Oral cavity cancer brings over 200,000 deaths annually and is one of the top eight most frequently diagnosed cancers worldwide. However there is a lack of appropriate experimental models to address oral cancer metastasis. An ideal experimental system should recapitulate the complete process of cancer metastasis in vivo while maintaining molecular-level manipulability. Our current efforts are directed at development of a series of murine oral squamous cell carcinoma (OSCC) cell lines that will be amenable for syngeneic transplantation studies in immuno-competent mice. The cell lines were created from C57BL/6J mice, a strain frequently used in targeted gene manipulation. This strategy ensures that the cells also have utility in orthotopic xenograft cancer models using currently available knockout mice made with a syngeneic background. As abundant knockout mice are available on the C57BL/6J background, our new syngeneic cancer cell lines will greatly facilitate studies on the interrelationship between host and tumor during the process of metastasis. Cell lines were generated from primary murine tongue epithelial cells by hTERT induced immortalization and oncogene induced transformation. They were screened for tumorigenicity and metastasis in both immunocompetent and immunocompromised mice. Properties of in vitro and vivo growth, morphology and cytogenetics were characterized. As proof of concept, one type of gene knockout mouse was used to test transplantation of such new cellular models in vivo to observe the impact of gene loss in the host on the growth and metastasis of tumors. The completion of this project will establish solid and currently unavailable tools for the oral cancer metastasis research community.

Citation Format: Zonggao Shi, Jeff Johnson, Yueying Liu, Sharon Stack. Generating new syngeneic models for oral squamous cell carcinoma metastasis study. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 92. doi:10.1158/1538-7445.AM2014-92
Abstract 93: Antagonizing pathways leading to differential dynamics in colonic carcinogenesis in azoxymethane (AOM)-treated Sgo1 (Shugoshin1)-haploinsufficient chromosome instability (CIN) model mice
Authors: Rao Chinthalapally V. , Sanghera Saira , Zhang Yuting , Biddick Laura , Lightfoot Stan , Dai Wei , Yamada Hiroshi Y.
Published: May 2015

(a) Colon cancer is the second most lethal cancer and is predicted to claim 50,830 lives in 2013. Chromosome Instability (CIN) is observed in 80-90% of colon cancers and is thought to contribute to colon cancer progression and recurrence. For drug development, animal models that reflect cancer-specific etiology is necessary. However, animal models focusing on CIN have not been validated for colon cancer development and for drug testing. In this study, we set out to validate a mitotic error-induced CIN model mice, Shugoshin1 (Sgo1) haploinsufficient mice, as a colon cancer study model.

(b) We treated control and Sgo1-/+ mice with colonic carcinogen Azoxymethane (AOM), and tracked colon cancer development over 12, 24 and 36 weeks after AOM treatments. The three endpoints provided information regarding the time course for colon cancer development.

(c) Although we predicted an increase in colon tumors in Sgo1-/+ mice at a later (24 or 36 weeks) endpoint, precancerous lesions the Aberrant Crypt Foci (ACF) did not develop to larger tumors straightforwardly. Unexpectedly, the numbers of gross colon tumors showed no difference (P>0.05). Immunohistological analyses indicated IL6, Bcl2 and p16INK4A were differentially expressed in Sgo1-/+ mice.

(d) We propose that formation of ACF in Sgo1-/+ mice are facilitated by IL6-STAT3-SOCS3 oncogenic pathway and Bcl2-anti-apoptotic pathway, yet further development of the ACF to tumors are inhibited by p16INK4A tumor suppressor pathway. Manipulating these pathways would be beneficial for inhibiting development of colon cancer with CIN. The CIN mice may be useful for investigating carcinogenesis in earlier stage and tumor regression in later stage.

Citation Format: Chinthalapally V. Rao, Saira Sanghera, Yuting Zhang, Laura Biddick, Stan Lightfoot, Wei Dai, Hiroshi Y. Yamada. Antagonizing pathways leading to differential dynamics in colonic carcinogenesis in azoxymethane (AOM)-treated Sgo1 (Shugoshin1)-haploinsufficient chromosome instability (CIN) model mice. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 93. doi:10.1158/1538-7445.AM2014-93
Abstract 931: Role of IKKα in K-ras-driven lung carcinogenesis
Authors: Song Na-Young , Willette-Brown Jami , Dalta Mahesh , Hu Yinling
Published: May 2015

IKKα, one of subunits of the IKK complex, is required for the epidermal homeostasis. We previously demonstrated that keratin 5-specific deletion of IKKα induces spontaneous skin squamous cell carcinomas (SCCs) in mice. Interestingly, IKKα kinase dead knockin (IKKαK44A/K44A, KA) mice spontaneously develop lung SCCs which is associated with increased inflammation and IKKα reduction. Thus, IKKα downregulation is crucial for the SCC formation in skin and lung. Here, we attempted to verify whether IKKα inactivation plays a role in the development of lung adenocarcinomas (ADCs) as well. We have crossed between KA mice and K-rasG12D oncogenic mutant (Kras) mice. Then, these IKKαK44A/K44A;K-rasG12D (KA;Kras) mice were intratracheally treated with adenovirus carrying the cre recombinase gene to activate K-ras oncogene. At 4.5 months after intratracheal injection, the lung weight was significantly increased in KA;Kras compared to Kras mice. Moreover, the lungs of KA;Kras mice showed increased bronchial epithelial cells and Ki67-positive cells in the bronchial regions. At 7 months after treatment, lung tumors in KA;Kras mice were larger compared to tumors in Kras mice. Finally, KA;Kras mice had shorter life span due to breathing problems. KA;Kras mice started to die at around 20 weeks after injection, while KR mice started to die from around 30 weeks. To confirm whether IKKα reduction promotes K-ras-driven lung carcinogenesis, we generated IKKαf/f (FF) mice crossed with Kras (FF;Kras) mice. Similar to KA;Kras, FF;Kras mice showed significant increase of lung weight, hyperproliferative bronchial abnormalities, Ki-67-positive cells in the bronchial regions and the number of large tumors at 4 months following treatment compared with Kras only group. Taken together, these data suggest that IKKα downregulation promotes K-ras-driven lung tumorigenesis and plays an important role in lung ADCs as well as in SCCs. The underlying molecular mechanism needs to be further investigated.

Citation Format: Na-Young Song, Jami Willette-Brown, Mahesh Dalta, Yinling Hu. Role of IKKα in K-ras-driven lung carcinogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 931. doi:10.1158/1538-7445.AM2014-931
Abstract 96: A novel in vivo preclinical model for the study of human tumor angiogenesis with human vessels
Authors: Holden Kirsty A. , Batey Michael A. , Bottomley Martyn , Booth Catherine
Published: May 2015

The preclinical assessment of anti-angiogenic drug efficacy is hampered by the lack of availability of suitable model systems to study anti-human therapies. Good in vitro assays exist for testing anti-human-specific therapies but these are let down by the lack of robust translatable in vivo models. Currently available preclinical models such as the matrigel plug assay and the window chamber assay focus on the angiogenesis of mouse vessels which may not be truly reflective of the influence of test items on human vasculature, and in vivo models to study the angiogenesis of human tumors by human vessels are unfortunately lacking. We have developed a sophisticated model to address this using adult human skin engrafted onto immunocompromised CB17-SCID mice. Angiogenesis of the skin follows, with human and host vessels anastomosing, resulting in a viable skin graft containing functioning human CD31-labelled vessels. This skin graft can be intradermally injected with a cell line of interest, such as the human melanoma cell line A375, and the ensuing tumors contain human vessels as evidenced by immunohistochemistry with a human-specific anti-CD31 antibody. Using the Aperio® Scanscope and FDA-approved microvessel analysis algorithm, 5-10 human vessels per mm2 of human tumor are observed. Furthermore, the model has the potential to be used in combination with non-invasive in vivo imaging techniques to assess tumor growth inhibition in response to therapies in a longitudinal fashion. This novel technique could potentially enable the rapid preclinical testing of anti-angiogenic anti-human therapies to help provide a go/no-go decision prior to costly clinical trials.

Citation Format: Kirsty A. Holden, Michael A. Batey, Martyn Bottomley, Catherine Booth. A novel in vivo preclinical model for the study of human tumor angiogenesis with human vessels. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 96. doi:10.1158/1538-7445.AM2014-96