bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4124: Pre-treatment complement system serum protein levels predict pathological response to neoadjuvant chemoradiotherapy for esophageal cancer
Authors: Maher Stephen G. , McDowell Dermot T. , Murphy Sinead A. , Collins Ben C. , Muldoon Cian , Gallagher William M. , Reynolds John V.
Published: August 2014

Introduction: Esophageal cancer has one of the worst prognosis of all cancers, and its incidence is increasing rapidly in the Western world. The standard of care for patients with esophageal cancer involves neoadjuvant chemoradiotherapy (neo-CRT) followed by surgery. Unfortunately, only ∼30% of esophageal cancer patients respond to neo-CRT. The aim of this study was to analyse the serum proteome early into neo-CRT to reveal biomarkers indicative of response to therapy.

Methods: 31 esophageal cancer patients received a standard neo-CRT regimen. Pathological response to therapy was assessed using the Mandard Tumour Regression Grade system. Serum was collected at pre-treatment, 24 h and 48 h timepoints into treatment. Albumin/IgG-depleted serum samples were analyzed using SELDI-TOF-MS. Protein/peptide expression difference mapping was performed and the data applied to the EPO-KB database. Subsequently, high-scoring candidates were assayed by ELISA. A leave-one-out cross validation (LOOCV) predictive algorithm was employed to assess the ability of candidate biomarkers to correctly predict therapeutic outcome.

Results: 16 patients had a poor response to therapy, while 15 had a good response. On CM10 chips, 9 protein peaks were significantly different between the two treatment response groups, some at baseline, others at 24 h and 48 h into neo-CRT. SELDI spectral peaks m/z_8617.685 and m/z_8928.422 were identified as the complement proteins C4a and C3a, respectively, via the EPO-KB. ELISA revealed significantly higher pre-treatment levels of C4a and C3a in serum from poor versus good responders. Subdivision of the response groups by TRG indicated an inverse correlation between levels of C4a and C3a and pathological response to neo-CRT. In a pre-treatment setting, LOOCV analysis revealed that these two serum proteins could predict patient response to neo-CRT with a sensitivity and specificity of 78.6% and 83.3%, respectively.

Conclusion: In a relatively non-invasive approach, this study has identified that pre-treatment levels of serum C4a and C3a are potential pre-treatment biomarkers predicting response to neo-CRT for esophageal cancer. Additional serum proteins are being validated to further improve the sensitivity and specificity of predicting esophageal patient response to treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4124. doi:10.1158/1538-7445.AM2011-4124
Abstract 4126: Optimization of clinical sample collection for detection of c-Met phosphorylation
Authors: Nakagawa Takayuki , Kumar Namita , Kuznetsov Galina
Published: August 2014

The receptor tyrosine kinase c-Met is being investigated as a target for cancer chemotherapy. c-Met and its ligand HGF (hepatocyte growth factor) play important roles in cell growth, survival and migration. Dysregulation of the HGF-c-Met pathway (overexpression, constitutive activation, gene amplification, ligand-dependent activation, mutation or epigenetic mechanisms) can lead to oncogenic changes, and has been described in multiple types of cancer including lung, gastric, and colorectal cancer. For cancer therapies targeting the HGF-c-Met signaling pathway, elevated levels of c-Met expression and increased phosphorylation of c-Met have been explored as both the predictive and PD biomarkers. However, detection of c-Met phosphorylation by IHC in human clinical cancer samples has proven to be extremely challenging.

We have conducted a study aimed at finding a way to detect phosphorylation of c-Met in clinical samples. In the first attempt, we have acquired a set of 30 gastric cancer samples which were fixed in formalin in the presence of phosphatase inhibitors (PhosphoGuardTM fixation kit). While this approach has helped us to improve detection of several other phospho-proteins in tissues, it did not help with the detection of phospho-c-Met. We then hypothesized that this is a very labile phosphorylation which is rapidly lost during the cold ischemia time occurring between the surgical excision of the tissue and its placement in formalin for fixation (which could last 30-60 minutes or longer in the operating room). Studies with xenograft tissues showed that a 15 minute delay in fixation resulted in approximately 50% decrease in c-Met phosphorylation, while a 30 minute delay resulted in almost complete disappearance of phospho-c-Met. We therefore acquired a set of colorectal cancer samples which were placed into formalin within 5 minutes of surgical excision. Using these rapidly fixed samples, we were able to detect strong phosphorylation of c-Met in a small number of samples. The staining pattern for phospho-c-Met correlated with that of phospho-ERK (a downstream signaling molecule in the c-Met pathway) suggesting that the relatively low occurrence of c-Met phosphorylation may reflect the status of activation of this pathway in vivo at a given time point. This work suggests that rapid fixation of human tissues facilitates detection of phospho-c-met and its use as a PD and predictive biomarker in clinical trials.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4126. doi:10.1158/1538-7445.AM2011-4126
Abstract 4127: Levels of circulating CD45dimCD34+VEGFR2+ progenitor cells correlate with outcome in metastatic renal cell carcinoma patients treated with targeted therapies
Authors: Farace Françoise , Cropet Claire , Blanc Ellen , Pérol David , Négrier Sylvie , Escudier Bernard
Published: August 2014

Background: A biomarker allowing to predict the efficacy of targeted therapies would be of great clinical value in patients with metastatic renal cell carcinoma (mRCC). We tested the hypothesis that circulating endothelial cell (CEC), bone marrow-derived CD45dimCD34+VEGFR2+ progenitor cell levels are associated with clinical outcome in mRCC patients undergoing targeted treatments.

Methods: The TORAVA trial was a randomized phase II study aimed to determine the efficacy and safety of temsirolimus and bevacizumab combination (arm A) in comparison to 2 control arms, sunitinib (arm B) or bevacizumab and α-interferon (arm C). Samples were collected at baseline and day 14 for each treatment arm. CEC levels were analysed in 117 patients (pts) using the CellSearch™ (Veridex, Warren, NJ) platform. Bone marrow-derived CD45dimCD34+VEGFR2+ progenitor cell levels were measured in 10ml blood samples of 42 pts by four-color flow cytometry after progenitor cell enrichment. Relation between CEC and CD45dimCD34+VEGFR2+ progenitor cell levels and clinico-biological characteristics, response to treatment (determined by RECIST criteria after 2 cycles) and progression free survival (PFS) was examined.

Results: CEC levels were significantly higher in patients without nephrectomy (p<0.001). A trend between the association of high CEC levels and a PS of 2 was observed (p=0.08). CEC and CD45dimCD34+VEGFR2+ progenitor cell levels at both baseline and day 14 were not correlated to response to treatment. No correlation was found between baseline or day 14 CEC values and PFS. However, pre-treatment CD45dimCD34+VEGFR2+ progenitor cell levels were significantly correlated to PFS (p=0.003). Patients with elevated pre-treatment levels of CD45dimCD34+VEGFR2+ progenitor cell had an increased risk of tumor progression.

Conclusion: Our results indicate that pre-treatment CD45dimCD34+VEGFR2+ progenitor cell levels could accurately predict PFS in MRCC patients receiving targeted therapies. The association between CEC and CD45dimCD34+VEGFR2+ progenitor cell levels and overall survival will be presented. These data suggested that measurement of CD45dimCD34+VEGFR2+ progenitor cell levels before targeted therapy in mRCC patients will allow to identify at an early stage patients who will benefit most from treatment.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4127. doi:10.1158/1538-7445.AM2011-4127
Abstract 4128: Serum insulin-like growth factor (IGF)-1 levels predict treatment efficacy of anti-angiogenic therapy for patients with advanced hepatocellular carcinoma (HCC)
Authors: Shao Yu-Yun , Hsu Chih-Hung , Huang Chien-Chung , Cheng Ann-Lii
Published: August 2014

Background Patients with liver cirrhosis or HCC were found to have decreased serum IGF-1 levels. Serum IGF-1 levels have been shown to predict the prognosis of patients with various types of cancer. Whether the IGF-1 levels can predict the treatment efficacy of anti-angiogenic therapy for advanced HCC remained unknown.

Methods Advanced HCC patients, enrolled in a phase II study (Hsu et al: J Hepatol 2010) and its extended cohort, were treated with sorafenib plus metronomic oral tegafur/uracil until disease progression. Serum samples were collected before treatment and 4 weeks after the start of treatment. The levels of IGF-1 and IGF-2 were determined by ELISA. The baseline levels and post-treatment changes of these factors were analyzed for their associations with treatment outcomes.

Results Among 68 pts treated with the protocol treatment, 64 were enrolled to this study. Among them, 57 (89%) were male, 49 (72%) were chronically infected with hepatitis B virus, and 40 (63%) had extrahepatic metastasis. All patients had liver reserve graded as Child-Pugh class A. There were 4 partial responder (PR) and 33 patients with stable disease (SD) according to RECIST 1.0. The disease control rate (DCR, defined as PR+SD) was 58%. The median PFS and OS were 3.7 and 7.4 months, respectively. Both IGF-1 and IGF-2 levels significantly decreased after treatment (p < 0.001 and p < 0.001, respectively), but the post-treatment changes were not associated with treatment response or disease control. Patients with higher (≥ median) baseline IGF-1 levels, compared to those with lower levels, had significantly better disease control rates (DCR, 72% vs. 44%, p = 0.023) and longer PFS (median, 5.6 vs. 2.0 months, p = 0.034). OS was also longer in patients with higher baseline IGF-1 levels, but with borderline significance only (median, 8.1 vs. 5.0 months, p = 0.052). In multivariate analysis including all clinicopathologic variables with potential predictive values, high baseline IGF levels remained as an independent predictor for longer PFS (p = 0.008). Baseline IGF-2 levels were not found to be associated with treatment outcomes.

Conclusions High baseline IGF-1 levels can predict better treatment efficacy of anti-angiogenic therapy for patients with advanced HCC. (This study was supported by the grant of NSC 98-3112-B-002-038.)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4128. doi:10.1158/1538-7445.AM2011-4128
Abstract 4131: Genetic variant associations with treatment outcome and safety in metastatic colorectal cancer patients treated with sorafenib and bevacizumab (BEV) as salvage therapy
Authors: Lafky Jacqueline M. , Bot Brian M. , Morlan Bruce W. , Grothey Axel
Published: August 2014

Purpose: Polymorphisms in genes involved in the angiogenesis pathway have been investigated as predictive markers for efficacy and tolerability to anti-angiogenesis treatment, such as BEV. This study examines single nucleotide polymorphisms (SNPs) in VEGF, VEGFR2, and HIF-1α for their potential association with outcome and safety in metastatic colorectal cancer (mCRC) patients treated with the combined vertical blockade of VEGF signaling (i.e., sorafenib and BEV) as salvage therapy.

Experimental procedures: Genomic DNA was extracted from peripheral blood for 78/79 evaluable patients in the North Central Cancer Treatment Group (NCCTG) Phase II study, N054C (Grothey, et al., ASCO 2010). TaqMan or direct sequencing was used for genotyping VEGF (rs25648, rs3025039, rs2010963, rs1005230, rs833061, rs699947, rs1570360), VEGFR2 (rs2305948, rs2219471, rs2071559, rs1870377), and HIF-1α (rs11549465, rs11549467) SNPs. Chi-square or Fisher's exact test was used to determine genotype associations with the study's primary endpoint, progression-free rate at 3 months of follow-up, as well as genotype associations with the most common grade 3+ adverse events at least possibly related (AE rels) to treatment (i.e., fatigue, hypertension, skin reaction).

Results summary: The presence of a mutant T allele in the 5’ UTR of VEGF (rs25648; C534T) was associated with fewer successes in progression-free rate at 3 month follow-up (Fisher's exact test, p=0.0187). In addition, the presence of a C allele in the promoter region of VEGFR2 (rs2071559, T-604C) was associated with fewer grade 3+ hypertension AE rels (Fisher's exact test, p=0.0101). However, after correcting for multiple comparisons using the method of Benjamini & Hochberg none of the p-values were significant at a 0.05 level. At the same time, no differences were observed between the genotype subgroups and either progression-free rate at 3 months or the most common grade 3+ AE rels for the remaining SNPs that were analyzed, before or after correction.

Conclusions: This study provides preliminary evidence that mCRC patients with the VEGF C534T (rs25648) variant may not benefit from combined treatment of sorafenib and BEV, whereas patients with the VEGFR2 T-604C (rs2071559) polymorphism may experience fewer grade 3+ AE rels. Further prospective validation of these SNPs and their association with this treatment combination in mCRC is warranted.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4131. doi:10.1158/1538-7445.AM2011-4131
Abstract 4138: Five prognostic subgroups differ in expression of Akt pathway genes: Biomarkers for therapy selection
Authors: Joy Anna M. , Smirnov Ivan , Ramesh Archana , Kim Seungchan , Bachoo Robert , Sarkaria Jann , Ram Prahlad , Shapiro William , Mills Gordon , Feuerstein Burt
Published: August 2014

Biomarkers that select patients for therapeutics would benefit clinical trial design and patient care. The Akt pathway is a therapeutic target in Glioblastoma Multiforme (GBM) and an important determinant of patient outcome. However, it is not known whether activity of this pathway varies among GBM tumors. To investigate we used Akt pathway genes from published GBM expression datasets. We detected 5 patterns of Akt pathway gene expression and these Akt subgroups correlated with prognosis. We analyzed networks within subgroups using both gene set enrichment analysis and a novel method that scores relevance based on both expression and local network connectivity (Komurov et al., PLoS Comput Biol. 2010 6:8, 1-10). The results suggest therapeutic targets within the individual subgroups. Furthermore, preliminary analysis indicates that human GBM xenograft models and primary human GBM manifest similar Akt subgroups. We are investigating rodent xenograft models of Akt subgroups to evaluate subgroup-specific drug sensitivity, and will present a method and examples of analysis integrating gene expression data from human GBM xenografts and primary human GBM. We hypothesize that Akt subgroups will help personalize treatment for GBM therapeutics. Supported by Barrow Neurological Foundation, Halle Family Foundation, and NIH 1 K01 NS064952-01A1 (AMJ).

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4138. doi:10.1158/1538-7445.AM2011-4138
Abstract 4143: Photodynamic detection using 5-aminolevulinic acid (ALA) of prostate cancer cells in the urine obtained following prostate massage
Authors: Anai Satoshi , Nakai Yasushi , Miyake Makito , Kuwada Masaomi , Chihara Yoshitomo , Tanaka Nobumichi , Fujimoto Kiyohide , Hirao Yoshihiko
Published: August 2014

Objectives: We evaluated the feasibility of photodynamic diagnosis (PDD) using 5-aminolevulinic acid (ALA) for detecting of prostate cancer (PCa) cells in the urine obtained following prostate massage.

Material and methods: Urine samples were collected from 97 men who had prostate massage (attentive digital rectal examination) before prostate needle biopsy. Urine sediments were exposed to 5-ALA (1mM) for 2 hours and cytospun onto glass slides. Protoporphyrin IX (PPIX), which is the metabolite of 5-ALA in the heme biosynthetic pathway, was observed using the fluorescent microscope (excitation filter 380-420 nm, emission filter 590 nm) as a positive signal of cancer cell. Any fluorescent signal in the samples was evaluated as positive. After histological diagnosis, the result of 5-ALA detection was compared with the histological diagnosis.

Results: Prostate cancer cells were successfully visualized by the PPIX. Of the 65 cases with biopsy-proven prostate cancer, 45 were evaluated as positive by 5-ALA exposure (sensitivity: 75 %). Although 32 cases were diagnosed as no cancer with biopsy, 10 5-ALA positive cases were detected (specificity: 68 %). The positive predictive value of 5-ALA cancer detection was 83 %.

Conclusions: We have successfully demonstrated that PPIX in urine sediments treated with 5-ALA could be detected in the PCa patients. Furthermore, the false positive patients could be diagnosed as PCa in the future re-biopsy. So this cancer detecting system appears to have the useful clinical application.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4143. doi:10.1158/1538-7445.AM2011-4143
Abstract 4145: “Real-time” monitoring of oncolytic viral delivery to solid tumors using contrast-enhanced computed tomography
Authors: Penheiter Alan R. , Dingli David , Russell Stephen J. , Bender Claire E. , Carlson Stephanie K.
Published: August 2014

RATIONALE: The inability to deliver viruses homogeneously throughout solid tumors is a universal obstacle encountered in oncolytic virotherapy. The initial intratumoral viral dispersion volume is a critical determinant of virotherapy efficacy, and depends not only on the number of infected cells, but also on the spatial distribution of the infected cells. In this study, we determined the feasibility and usefulness of monitoring the initial intratumoral delivery of oncolytic viruses to solid tumors using contrast-enhanced computed tomography (CT).

EXPERIMENTAL PROCEDURES: Human BxPC-3 pancreatic tumor xenografts were established in nude mice. An engineered oncolytic measles virus expressing the sodium iodide symporter reporter (MV-NIS) was mixed with a 1:10 dilution of Omnipaque-300 contrast material and injected directly into mouse tumors. Mice were imaged with micro-CT immediately pre and post-injection of the virus + Omnipaque solution and on day 3 post-injection with 99mTcO4 micro-SPECT/CT. Tumors were harvested immediately after micro-SPECT/CT imaging for autoradiography and immunistochemistry analysis of radioactivity uptake and viral infection. Linear regression and Spearman correlation analyses were performed to determine the relationship between intratumoral contrast enhancement and subsequent intratumoral viral infection.

RESULTS: This dilution of Omnipaque was found to have no effect on viral infectivity or cell viability in vitro, yet was more than adequate for CT imaging of the virus + Omnipaque injectate distribution following intratumoral injection. Ex vivo autoradiography and immunohistochemistry confirmed a strong correlation between intratumoral CT contrast enhancement, SPECT activity, viral infection, and sodium iodide symporter activity.

CONCLUSIONS: This injection method is translatable to the clinic and should allow optimization of intratumoral injection in humans.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4145. doi:10.1158/1538-7445.AM2011-4145
Abstract 4146: Optical imaging of bladder cancer with cancer-specific molecular contrast agents
Authors: Pan Ying , Liu Jen-Jane , Mach Kathleen E. , Volkmer Jens-Peter , Weissman Irving L. , Liao Joseph C.
Published: August 2014

Introduction and Objectives: Current diagnosis of bladder cancer is by white light cystoscopy, which has suboptimal specificity for differentiating non-papillary cancer from inflammation. Probe-based confocal laser endomicroscopy (pCLE) provides dynamic in vivo imaging of the endoluminal tracts with micron-scale resolution. Real-time analysis of confocal images remains challenging with non-specific contrast agents such as intravesical fluorescein. Tumor imaging specificity may be enhanced by coupling pCLE with fluorescently labeled tumor-specific antibodies or peptides. We report our preliminarily efforts of ex vivo bladder tumor imaging by pCLE using a mouse antibody against a known human tumor-specific surface biomarker and a dodecapeptide that binds human epidermal growth factor receptor (EGFR) as contrast. Studies have shown that both the antibody and peptide bind surface markers abundant in cancer but not present on the superficial layer of normal urothelium.

METHODS: The antibody or peptide was instilled intravesically into a fresh cystectomy specimen (n=4 for antibody; n=1 for peptide). Excess antibody or peptide was drained, and the bladder was opened for pCLE. Imaging was performed of normal-appearing, tumor, and suspicious areas, followed by excision of imaged tissues for histopathology with H&E. Tumor-specific staining of the antibody was further confirmed with anti-mouse IgG.

RESULTS: All bladders had high-grade urothelial carcinoma on pathology. The tumor-specific antibody consistently showed greater fluorescent staining on pCLE in areas of tumor, compared to normal urothelium, in all 4 bladders. A suspicious area was not stained by the antibody and was confirmed to be inflammation by pathology. Immunostaining showed tumor-specific antibody on the most superficial layer of tumor but not on normal urothelium. Similarly, preferential staining of tumor over normal urothelium was also observed when using the EGFR peptide as a contrast agent.

CONCLUSIONS: Both the known tumor-specific antibody and EGFR-specific peptide show specificity for bladder cancer by pCLE ex vivo. Our study shows the feasibility of optical imaging of bladder cancer with pCLE and tumor-specific contrast agents.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4146. doi:10.1158/1538-7445.AM2011-4146
Abstract 4152: The detection of circulating tumor cells is a negative prognostic factor in pancreatic cancer
Authors: Srovnal Josef , Benedikova Andrea , Klos Dusan , Lovecek Martin , Havlik Roman , Radova Lenka , Cwiertka Karel , Hajduch Marian
Published: August 2014

Purpose: We tested the hypothesis that the presence of CTC is a negative prognostic factor in patients with pancreatic cancer. Pancreatic cancer is one of the most aggressive malignancies with the poor prognosis. Assessment of the circulating tumor cells (CTC) in patients with this highly malignant disease could prevent burdensome surgery in patients with systemic dissemination.

Patients and methods: This was a prospective study to test for the presence of CTC in systemic blood, portal blood, bone marrow and peritoneal lavage in 90 pancreatic cancer patients at the time of surgery using real-time RT-PCR for carcinoembryonic antigen (CEA), epidermal growth factor receptor 1 (EGFR1) and human telomerase (hTERT). Absolute gene expressions of tested markers were correlated with clinical/pathological characteristics and survival parameters.

Results: Overall, 51 of 90 (56.6%) pancreatic cancer patients died, the overall survival median was 8.8 months. We found a statistically significant association between EGFR and hTERT expression levels in the portal blood and clinical stage. We also showed high expression of EGFR and CEA in the peritoneal lavage of patients with metastatic disease, in contrast to patients without the presence of the metastases.

CTC positive patients measured as hTERT in tumor draining blood and CEA in bone marrow showed significantly poorer overall survival (OS) (p=0.005/ p=0.001). We also found statistically significant shorter OS in patients with EGFR positive peritoneal lavage (p=0.01).

Conclusion: The results of this study demonstrate a high sensitivity and specificity of the real-time RT-PCR method for CTC detection in pancreatic cancer. Detection of CTC seems to be negative prognostic factor for overall survival in pancreatic cancer and can influence the radical surgery decision.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4152. doi:10.1158/1538-7445.AM2011-4152
Abstract 4160: Glycoprotein biomarkers for bladder cancer detection identified using LC-MS/MS and label free quantification
Authors: Goodison Steven , Yang Na , Xie Xiaolei , Liu Yashu , Urquidi Virginia , Rosser Charles J. , Lubman David
Published: August 2014

PURPOSE: Cancers of the urinary bladder are the fifth most commonly diagnosed malignancy in the US. Early clinical diagnosis of bladder cancer remains a major challenge and the development of noninvasive methods for detection and surveillance is desirable for both patients and health care providers.

METHODS: In order to identify urinary proteins with potential clinical utility we enriched and profiled the glycoprotein component of urine samples using a dual-lectin affinity chromatography and LC-MS/MS platform.

RESULTS: From a primary sample set obtained from 56 cancer patients and 47 healthy individuals, a total of 265 distinct glycoproteins were identified with high confidence, and changes in glycoprotein abundance between groups were quantified by a label-free spectral counting method. Validation of candidate biomarker disease association was performed on second set of 70 independent samples using an antibody-lectin sandwich microarray and ELISA. Increased levels of urinary alpha-1-antitrypsin (A1AT) glycoprotein were identified and confirmed as being indicative of the presence of bladder cancer (p value < 0.0001).

CONCLUSION: The described strategy can be used to discover biomarkers for disease status in other body fluids. Application of the strategy in this study has provided novel candidate biomarkers for the non-invasive molecular detection of bladder cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4160. doi:10.1158/1538-7445.AM2011-4160
Abstract 4168: PI3K/AKT pathway alterations in paired primary and recurrent endometrial carcinoma
Authors: Wethington Stephanie L. , Garg Karuna , Olvera Narciso , Dao Fanny , Bogomolniy Faina , Soslow Robert A. , Levine Douglas A.
Published: August 2014

Objectives: The PI3K/AKT pathway is frequently deregulated in endometrial cancer due to somatic mutations and other molecular mechanisms. New drugs directed toward these pathways are currently in clinical trials. Many patients have available primary tumor specimens for correlative studies or enrollment eligibility. It is unknown if the same genomic alterations found in primary disease are also present in recurrent specimens. We hypothesize the genomic landscapes of primary and recurrent endometrial cancers are similar.

Methods: We identified 18 patients with available paired primary and recurrent endometrial cancer specimens. Histologic diagnoses were confirmed and categorized based on current FIGO criteria. DNA was extracted from microdissected FFPE tissue sections. Mass spectrometry mutation detection was used to assess 40 hot spot mutations in AKT1, KRAS, NRAS, and PIK3CA. Sanger sequencing was used to identify mutations in PTEN. PTEN IHC was performed using validated antibodies.

Results: Specimens included 9 cases with endometrioid histology, 8 with serous histology and 1 with mixed serous-endometrioid histology. Five patients had specimens from 2 separate recurrences. Of the 18 primary tumors, 4 (22%) had PIK3CA mutations, 5 (28%) had PTEN mutations, and 1 (5.5%) had a KRAS mutation. One PIK3CA and two PTEN mutations were each present in a primary tumor but not identified in the paired recurrent specimen. One NRAS mutation was identified in a secondary recurrent specimen but not found in either the paired primary or initial recurrent specimen. One AKT1 mutation was identified in both an initial and secondary recurrent specimen but not found in the corresponding primary specimen. PTEN immunostaining was absent in 9 (50%) primary specimens and in one of these cases PTEN staining found to be present in the recurrence.

Conclusions: As clinical trials increasingly incorporate molecularly targeted agents, mutational and other genomic profiling of tumors will be helpful to identify patients most likely to benefit from these drugs. These data suggest that most recurrent endometrial carcinomas harbor similar genomic alterations in the PI3K/AKT pathway as their primary counterparts. Further studies will be required to expand upon these preliminary findings and determine if repeat biopsy of recurrent disease in clinical trial participants is necessary.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 4168. doi:10.1158/1538-7445.AM2011-4168
Abstract LB-187: Identification of CIT-0665, a novel inhibitor of LSD1
Authors: Stephens Bret J. , Warner Steven L. , Vankayalapati Hari , Bearss David J. , Sharma Sunil
Published: August 2014

Lysine-specific demethylase 1 (LSD1/AOF2/KDM1A) is a flavin-dependent histone demethylase that catalyzes the posttranslational oxidative demethylation of mono- and dimethylated lysines on histones. Methylation of lysine residues on histones can signal transcriptional activation or repression depending on the specific residue involved. H3K4me2 is a transcription-activating chromatin mark at gene promoters, and demethylation of this mark by LSD1 is thought to prevent expression of tumor suppressor genes important in human cancer. In contrast, methylation of H3K9 is a repressive mark and LSD1 activity has been shown to upregulate tumor promoting pathways. Thus, LSD1 is emerging as an important target for the development of specific inhibitors as a new class of antitumor drugs. Currently, only a few existing compounds, such as monoamine oxidase inhibitors, have been shown to act as inhibitors of LSD1. More potent and specific inhibitors of LSD1 are needed to advance LSD1 biology and test for potential efficacy in tumor models. We utilized the 3-D crystal structure of LSD1 to computationally dock more than 10 million virtual compounds into the active site of the protein. From the docking experiments, we physically screened 52 compounds in an LSD1 biochemical assay (resorufin-based assay measuring the amount of hydrogen peroxide generated by LSD1). The compound CIT-0665 was identified as a potent inhibitor of LSD1 enzymatic activity, with an IC50 of 290nM. Additionally, cell lines treated with CIT-0665 show increased levels of H3K4 methylation. A large panel of cancer cell lines was also tested for sensitivity to CIT-0665, using a cell viability assay. We are currently engaged in determining possible mechanisms of sensitivity from the cell line panel and making analogs of CIT-0665 with improved drug-like qualities. In conclusion, CIT-0665 is a novel LSD-1 inhibitor with activity in biochemical and cell-based assays.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-187. doi:10.1158/1538-7445.AM2011-LB-187
Abstract LB-188: Chk1 suppresses bypass of mitosis and tetraploidization in p53-deficient cells
Authors: Wilsker Deborah F. , Chung Jon H. , Bunz Fred
Published: August 2014

Many checkpoint-deficient cancer cells are unable to maintain a numerically stable chromosome complement. The Chk1 kinase has a well-established role in the DNA damage checkpoint responses and is also essential for viability; however little is known about its role throughout the unperturbed cell cycle. Here, we rigorously examine the role of Chk1 in the maintenance of cell ploidy and mitotic progression. Using recombinant adeno-associated virus-based gene targeting techniques, we disrupted one allele of CHK1 in human cancer cell lines that are proficient or deficient for the tumor suppressor p53. We assessed ploidy by fluorescence in situ hybridization and metaphase chromosome analysis, and examined cell cycle progression by live cell time-lapse fluorescent microscopy, DNA labeling, flow cytometry and biochemical analysis. Loss of one CHK1 allele caused an elevated frequency of mitotic bypass in p53-deficient cells. CHK1-haploinsufficient, p53-deficient cells frequently underwent sequential rounds of DNA synthesis without an intervening mitosis, which caused whole genome endoreduplication and tetraploidization. CHK1 haploinsufficient cells exhibited reduced Cdk1 activity, which has been shown to lead to the bypass of mitosis. This mitotic bypass phenotype could be suppressed by targeted reversion of a p53 mutation or by exogenous Cdk1 expression. Interestingly, the rate of mitotic bypass was independent of upstream signaling by ATR to Chk1 and was not associated with DNA damage. These findings demonstrate a previously unrecognized role for Chk1 in promoting the initiation of mitosis during the cancer cell cycle and thereby maintaining ploidy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-188. doi:10.1158/1538-7445.AM2011-LB-188
Abstract LB-19: The Smac mimetic LBW242 induces apoptosis in melanoma in a TLR-3-dependent and TNFR-independent manner
Authors: McHenry Kevin T. , Jacobson Elena , Donovan Jerry , Levine Kymberly , Zhang Jingxin , Fisher David E. , Zawel Leigh , Porter Dale
Published: August 2014

Toll-like Receptor-3 (TLR-3), a pattern recognition receptor family member, has been shown to activate immune system pathways; however, it has also been shown to induce apoptosis in certain cancer cells. Combination of the synthetic TLR-3 ligand, poly I:C, with the Smac mimetic LBW242, an inhibitor of apoptosis protein (IAP) antagonist, has been shown to potently induce apoptosis in melanoma cells which are insensitive to either single agent. To better elucidate the mechanism responsible for this combination induced cell death, we performed a pooled shRNA screen. We identified receptor interacting protein kinase-1 (RIPK1) and as an important component of this process. Constitutive knockdown of RIPK1 prevents the LBW242 and poly I:C combination induced apoptosis. The IAP proteins cIAP1 and cIAP2 have been shown previously to regulate Caspase-8 activation downstream of Tumor Necrosis Factor Receptor-1 (TNFR1). Interestingly, we found that the combination induced apoptosis was independent of TNF signaling. Utilizing both gain of function and loss of function studies, we found that expression of TLR-3 is necessary for the combination induced apoptosis in melanoma cells. Taken together, our data indicate that the combination of the Smac mimetic LBW242 and TLR-3 agonist poly I:C induces a novel type of TLR-3 dependent apoptosis in melanoma cells that is independent of TNFR1.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-19. doi:10.1158/1538-7445.AM2011-LB-19
Abstract LB-199: SN38-dextran prodrug synthesis and cell delivery in a murine metastatic pancreatic cancer model
Authors: Shrestha Tej B. , Basel Matthew T. , Balivada Sivasai , Seo Gwi Moon , Pyle Marla , Bossmann Stefan H. , Troyer Deryl L.
Published: August 2014

Enzyme activated prodrugs have been investigated and sought after as highly-specific, low side effect treatments, especially for cancer therapy. We synthesized an SN38-dextran prodrug and utilized it in a cell delivery system that can carry both the prodrug and an activating enzyme to the cancer site. For delivery of our prodrug, we engineered Raw264.7 cells (mouse monocyte/macrophage like cells, Mo/Ma) to express intracellular rabbit carboxylesterase (InCE), which cleaves the prodrug to activate SN38. InCE expression was regulated by the TetOn® system, which silences the gene unless a tetracycline, such as doxycycline is present. To test the system, a murine pancreatic cancer model was generated by intraperitoneal (i.p.) injection of Pan02 cells. Engineered Mo/Ma cells were loaded with the SN38-dextran prodrug and were injected i.p. into the mice. Two days following injection, doxycycline was given i.p. to activate the InCE, which in turn activates the prodrug into active SN38. A survival study demonstrated that this system significantly increased survival in a murine pancreatic cancer model, with an average post-diagnosis life expectancy increase of 20%. Thus, for the first time, a prodrug/activating enzyme system self-contained within tumor-homing cells has been demonstrated that can significantly prolong the life of i.p. pancreatic tumor bearing mice.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-199. doi:10.1158/1538-7445.AM2011-LB-199
Abstract LB-202: Therapeutic efficacy of coencapsulated cisplatin and arsenic trioxide nanobins in murine models of breast cancer
Authors: Ahn Richard W. , Swindell Elden P. , Doan Ninh B. , Ugolkov Andrey , Malin Dmitry , Raja Meera R. , Mazar Andrew P. , Cryns Vincent L. , O'Halloran Thomas V.
Published: August 2014

We have developed a novel strategy to encapsulate a combination of cisplatin and arsenic trioxide as a precipitate inside 100 nm liposomal nanobins, NB(Pt, As). Cisplatin is a widely used chemotherapeutic but has significant dose-limiting toxicity. Arsenic trioxide (ATO) is a potent FDA-approved therapy for acute promyelocytic leukemia; however, ATO has not been effective in clinical trials of solid tumors. Encapsulation in nanoliposomes can improve the anti-cancer efficacy of chemotherapeutic agents by extending serum half-life, increasing tumor drug delivery and attenuating toxicity. We have tested the therapeutic efficacy of NB(Pt, As) in orthotopic xenograft models of triple negative breast cancer using MDA-MB-231 and MDA-MB-435/LvBr1 cancer cell lines. Treatment with either NB(Pt, As) or cisplatin for three weeks inhibited growth of MDA-MB-231 mammary tumors in vivo. However, the NB(Pt, As) did not cause weight loss in treated animals and did not result in abnormal serum chemistries.

NB(Pt, As) was also evaluated for its ability to inhibit lung metastases. Mice inoculated intraductally with highly metastatic MDA-MB-435/LvBr1 cancer cells develop mammary tumors that metastasize and form macroscopic lesions in the lung. Treatment with NB(Pt, As) was initiated when primary tumors reached∼250 mm3. Both NB(Pt, As) and cisplatin inhibited primary tumor growth in this model, whereas only NB(Pt, As) treatment significantly reduced lung metastatic burden.

Our results suggest that nanobin encapsulated cisplatin and ATO is a promising nanoscale therapeutic agent for metastatic breast cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-202. doi:10.1158/1538-7445.AM2011-LB-202
Abstract LB-206: A novel scaffold that targets the IKK complex
Authors: Bryant Vashti C. , Radhakrishnan Prakash , Hollingsworth Michael A. , Natarajan Amar
Published: August 2014

The IκB kinase (IKK) complex is a central component of the NF-κB pathway. The expression of a number of gene products are controlled and regulated by NF-κB. Dysregulation of the NF-κB pathway proteins has been implicated in a number of cancers including pancreatic cancer. A cell-based luciferase reporter assay was used to screen in house chemical libraries and identify NF-κB inhibitors. This led to the identification of quinoxaline urea (QU) analog as a validated hit. We found that QU inhibits the nuclear translocation of p65 in response to TNF-α stimulation. Our studies also indicate that QU inhibits a constitutively active form of the IKK complex. QU induced growth inhibition and apoptosis in a panel of pancreatic cancer cell lines with low-μM potency. NF-κB is an important player in the invasion and metastases of pancreatic cancer. The in vitro migration and matrigel invasion assay results with QU shows that it inhibits the migratory and invasive ability of pancreatic cancer cell lines. QU has comparable activities in all our assays to known NF-κB inhibitors, parthenolide and curcumin, that are in clinical trials for pancreatic cancer therapy. In summary, we present a novel scaffold that targets the constitutively active IKK complex with the potential to be developed as pancreatic cancer therapeutic.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-206. doi:10.1158/1538-7445.AM2011-LB-206
Abstract LB-283: JX-594, a targeted multi-mechanistic oncolytic poxvirus, selectively infects tumor vasculature and causes acute tumor vascular disruption and necrosis in advanced cancer patients
Authors: Breitbach Caroline , Arulanandam Rozanne , Patt Richard , De Silva Naomi , Parato Kelley , Daneshmand Manijeh , Hickman Theresa , Bell John C. , Kirn David H.
Published: August 2014

JX-594 is a first-in-class targeted oncolytic poxvirus designed to selectively replicate in and destroy cancer cells with cell cycle abnormalities and epidermal growth factor receptor (EGFR)/ras pathway activation. Direct oncolysis plus granulocyte macrophage -colony stimulating factor (GM-CSF) expression also stimulates anti-tumoral immunity. JX-594 infection was evaluated in vitro on human umbilical vein endothelial cells (HUVECs) and by immunohistochemical analysis in tumor biopsies from JX-594 treated (intravenous administration) patients with advanced, treatment-refractory solid tumors. JX-594 associated changes in tumor perfusion were also assessed in patients by dynamic contrast-enhanced magnetic resonance imaging (dce-MRI; at baseline and Day 5 after intratumoral JX-594 administration). In vitro susceptibility of HUVECs to JX-594 infection was shown to be dependent on vascular endothelial growth factor (VEGF) stimulation. Furthermore, JX-594 was capable of infecting tumor-associated endothelial cells after intravenous infusion in patients with advanced solid tumors. No clinical evidence of normal vasculature infection or toxicity was noted. Tumor perfusion was significantly decreased within 5 days post JX-594 treatment, including in hepatocellular carcinoma and colorectal cancer metastases. Perfusion was markedly reduced in both directly injected and distant non-injected tumors. Choi (necrotic) responses at later timepoints were demonstrated. In addition to targeting cancers by direct infection and lysis of tumor cells, JX-594 is capable of directly infecting VEGF-stimulated/tumor-associated endothelial cells. By targeting tumor-associated vasculature, JX-594 acutely disrupts the tumor's blood supply leading to tumor destruction. Targeted oncolytic poxviruses such as JX-594 represent a novel and highly selective class of vascular disrupting agents.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-283. doi:10.1158/1538-7445.AM2011-LB-283
Abstract LB-290: Glycogen synthase kinase-3β inhibits tumor growth and angiogenesis of gastric cancer
Authors: Ko Young San , Cho Sung Jin , Yoon Jiyeon , Chang Mee Soo , Lee Byung Lan
Published: August 2014

Objective: The mechanisms underlying angiogenesis in cancer are diverse. Although in vitro studies have shown that glycogen synthase kinase-3β (GSK-3β) inhibits the expressions of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in several cancer cells, the implication of GSK-3β in cancer angiogenesis in vivo has not been shown. We examined the effects of GSK-3β on angiogenesis and human gastric cancer growth.

Methods: Human gastric cancer cell lines SNU-638 and SNU-484 were stably transfected with either human wild-type (WT)-GSK-3β or kinase-dead (KD)-GSK-3β, and the effects of GSK-3β on tumor growth and angiogenesis were examined in in vivo experiments. Gastric cancer cells were injected into nude mice subcutaneously and the size of xenografted tumors was measured. The effects of GSK-3β inhibition on angiogenesis and HIF-1α activation were assessed by immunohistochemistry, Western blotting and semiquantitative reverse transcription-polymerase chain reaction. In addition, GSK-3β effects on the HIF-1α degradation and synthesis were examined.

Results: Xenografts in nude mice derived from stable gastric cell lines over-expressing KD-GSK-3β showed larger tumor volumes and higher microvessel density than those over-expressing WT-GSK-3β. Immunohistochemistry and Western blot analysis using xenografted tumor tissues showed that KD-GSK-3β over-expression increased immunoreactivity for Ki-67, HIF-1α, VEGF or CD31 compared with WT-GSK-3β over-expression. In addition, KD-GSK-3β over-expression enhanced HIF-1α expression under hypoxic conditions at the translational level, but not under normoxic conditions.

Conclusion: These results indicate that GSK-3β inhibits gastric tumor growth and angiogenesis by regulating the expression of HIF-1α and VEGF in gastric cancer cells under hypoxic conditions. Our results suggest the potential usefulness of GSK-3β/HIF-1α/VEGF pathway to find molecular targets in gastric cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr LB-290. doi:10.1158/1538-7445.AM2011-LB-290