bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 4497: Biomarkers of HPV in head and neck squamous cell carcinoma
Authors: Liang Caihua , Marsit Carmen J. , McClean Michael D. , Nelson Heather H. , Christensen Brock C. , Haddad Robert I. , Clark John R. , Wein Richard O. , Grillone Gregory A. , Houseman E. Andres , Halec Gordana , Waterboer Tim , Pawlita Michael , Krane Jeffrey , Kelsey Karl T.
Published: August 2012

Human papillomavirus (HPV) is an accepted cause of head and neck squamous cell carcinoma (HNSCC) and patients with HPV-associated disease have a favorable prognosis. Currently there is no general guidance on the most appropriate biomarkers for clinical assessment of HPV. We measured DNA-based and serological markers in a single population-based study to assess their associations with disease outcome and to determine the most relevant biomarker for clinical use. HPV16 serology was determined using an immunoassay for the capsid protein L1, and multiplex serology based on the capsid protein L1 and the early oncoproteins E6/E7 for 488 patients. Immunohistochemical detection of p16 expression in tumors was performed in a subset of 233 cases, and we used two different methods to assess the presence of HPV16 DNA in a subset of 179 cases’ tumors. Seropositivity for the E6 and E7 proteins (individually) was significantly associated with enhanced survival (HRE6 or HRE7=0.3, 95%CI=0.2-0.5). Neither the presence of HPV16 DNA or p16 overexpression was associated with significant enhanced overall survival (HRDNA=0.8, 95%CI=0.5-1.3; HRp16=0.8, 95%CI=0.5-1.4). However the combination of HPV positive DNA and E6/E7 serology was associated with similarly enhanced overall survival (HRDNA+/E6+/E7+=0.1, 95%CI=0.01-0.6), while E6/E7 seronegative patients with evidence of HPV in tumor DNA did not show any evidence of favorable survival (HRDNA+/E6-/E7-=1.2, 95%CI = 0.7-2.2). Further, the combination of p16 staining and early gene seropositivity showed a favorable survival (HRp16+/E6+/E7+=0.3, 95%CI=0.1-0.9), while those who were p16 positive and early gene seronegative had significantly increased hazard of death (HRp16+/E6-/E7-=2.1, 95%CI=1.1-4.2). Determination of only HPV16 DNA status or p16 immunostaining is not an effective prognostic biomarker for HNSCC. However the combination of measurement of HPV16 DNA and p16 immunostaining with E6/E7 antibodies has high predictive clinical value, strongly suggesting that a combined biomarker approach is necessary for the assessment of the prognosis of HNSCC patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4497. doi:1538-7445.AM2012-4497
Abstract 4511: The effect of obesity-related adipokines at diagnosis on breast cancer survival
Authors: Lee Sang-Ah , Noh Dong-Young , Choi Ji-Yeob , Ahn Sei-Hyun , Kang Deahee
Published: August 2012

Background: Although a number of experimental studies have suggested the effect of obesity-related adipokines in breast cancer progression, limited numbers of epidemiological studies have examined the relationship between the adipokines and breast cancer survival. Patients and methods: Preoperative serum levels of obesity-related adipokines including leptin, adiponectine and hepatocyte growth factor (HGF) were measured in 370 breast cancer patients recruited between 2004 and 2007. We examined the association between those adipokines and disease-free survival (DFS) using Cox proportional hazard regression model. Results: There was no difference in age distribution, menopausal status, BMI, PR status, tumor size, and lymph node status between survivor and recurrence or death subjects. On the other hand, the TNM stage, ER status and histological grade were observed the association with breast cancer survival. Elevated adiponectin were associated with reduced DFS of breast cancer (P for trend = 0.047) with adjustment for age, ER/PR status, histological grade, nuclear grade and TNM stage among patient with less than 4 year survival time. There was no association of leptin and HGF with the breast cancer survival. Conclusions: Our study suggests that the elevated levels of adiponectine at diagnosis are associated with breast cancer survival, but the leptin and HGF level in blood not associated.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4511. doi:1538-7445.AM2012-4511
Abstract 4521: Differential whole genome methylation profile in a responder and a non-responder to chemotherapy in colorectal cancer patients
Authors: Ashktorab Hassan , Tbaishat Rana , Rahi Hamed , Lee Edward , Smoot Duane T. , Ajayi Olaide , Sun Xueguang , Jia Xi-Yu , Chung Hunter , Bacon Eliza , Leavitt Ron , Brim Hassan
Published: August 2012

Background: Colorectal adenoma (CRA) to cancer (CRC) progression involves in epigenetic changes including DNA methylation. Here we performed whole genome Bisulfite Next-Generation sequencing (WGBSNGS) on a normal, a tubular adenoma and a tumor patients’ tissue DNA to elucidate epigenetic drivers in normal to cancer progression. Patients and Methods: Genomic DNA was isolated from fresh frozen tissues from a patient with normal colon, from a tubular adenoma lesion (<0.5 cm) and from a carcinoma. A WGBSNGS at >20X depth was performed on these DNA samples., Alignment, mapping and CpG methylation analyses were done. Pyrosequencing and immunohistochemistry (IHC) was used to confirm selected CpG methylation. Results: We identified 94 unique CpG sites hypermethylated in 4 novel genes in adenoma compared to normal and 180 unique CpG sites in 7 genes in cancer. Ingenuity pathway analysis (IPA) showed that the methylated genes including a novel gene ATXN7L1 were involved in WNT pathway, ubiquotlation process and cross-talk with B-catenin, in the adenoma. Three of the hypermethylated genes in cancer were, Loc100506436: a novel gene with 17 unique CpG sites (EGFR pathway, cRNA CpG site hypermethylated is part of MEK1/2), GPNMB with 7 unique CpG sites (a G protein) and TNFAIP2 with 8 unique CpG sites (its role was noted in Head and neck cancer). IHC confirmed the lack of expression in cancer. Conclusion: This work provides insight into differential CpG island methylation profiles in CRA, CRC vs. normal colon tissue and provides a window into the more complex factors, including alteration of known and novel genes as a multiple CpG methylation drivers in colorectal carcinogenesis. Investigations into the possible roles of the novel gene targets in the context of early and prognostic methylation biomarker are underway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4521. doi:1538-7445.AM2012-4521
Abstract 4530: A novel blood-based method for evaluating KRAS in circulating microvesicles from colorectal cancer patients
Authors: Yeatts Kimberly , Millis Meredith , Smith Shannon , Duncan Janet , Moran Brittany , Ortiz Gerri , Little Tanya , Conrad Amber , Spetzler David B. , Pawlowksi Traci
Published: August 2012

Circulating microvesicles (cMV) are lipid-encapsulated bodies that are secreted from various tissues and can be detected in a number of body fluids, including plasma. Once collected from plasma, they can be exploited diagnostically for their protein and RNA signatures. Mutations in KRAS are diagnostically important for predicting response to chemotherapy and prognosis. A blood-based method of assessing KRAS mutation status would be helpful for patients with colorectal cancer (CRC). Traditional methods of KRAS detection examine the genomic DNA sequence. We developed a method to sequence exon 3 from KRAS using Pyrosequencing. The limit of detection for the assay was 0.78 ng/uL of mutant cDNA for both Pyrosequencing and Sanger sequencing. To further enhance mutation detection, we sorted cMV from CRC patients by first capturing cMV with a CRC associated membrane protein and then sorting for CD63 and CD24 positive events. A Taqman gene expression assay was not sensitive enough to detect KRAS in the sorted samples. However, Pyrosequencing was sufficient to identify mutant and wild-type sequences in patient plasma samples.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4530. doi:1538-7445.AM2012-4530
Abstract 4569: Amplicon-targeted ultra-deep sequencing for the detection of KRAS, BRAF, and PIK3CA actionable mutations
Authors: Donahue Amber C. , Sferruzza Anthony D.
Published: August 2012

Background: Next-generation deep sequencing techniques have the potential to revolutionize clinical testing, especially in the arena of mutation detection. Molecular diagnostic testing for KRAS, BRAF, and PIK3CA mutations has become an integral part of clinical practice for the determination of treatment eligibility for anti-EGFR therapy. Purpose: Here we investigate the feasibility of ultra-deep advanced sequencing with the 454 GS Junior instrument (454 Life Sciences/Roche, Branford CT) for the simultaneous detection of actionable mutations in KRAS, BRAF, and PIK3CA in formalin-fixed paraffin-embedded (FFPE) colorectal cancer tissue. Methods: Advanced sequencing with a 4-amplicon panel was performed on 17 de-identified residual FFPE colorectal cancer biopsy specimens originally submitted for mutation analysis of one or more genes by standard sequencing. The 4-amplicon panel was multiplexed for up to 4 samples through “barcoding” with molecular identifier (MID) sequences, for a total of 16 amplicons per run. Results: Sequence analysis with ultra-deep coverage of 3000-5000 reads detected KRAS mutations in 5 (29%) tumor specimens, BRAF mutations in 3 (18%), and PIK3CA mutations in 4 (24%). Ten cases had a single mutation and one had two: PIK3CA E545K and BRAF D594G. Coverage was equivalent for each of the four amplicons, and in both the forward and reverse directions, providing a high level of confidence for each mutation detected. Sequencing data was obtained for each sample tested. When sample quantity allowed, Sanger sequencing and/or castPCR (Life Technologies, Grand Island, NY) were performed as method comparisons, and results were concordant. Conclusions: The use of MIDs with ultra-deep advanced sequencing allowed the simultaneous detection of mutations in multiple amplicons from multiple patients in a single sequencing run. This proof-of-concept study confirmed that using advanced sequencing to simultaneously test for multiple biomarkers in one FFPE sample is feasible, and that amplicon-based advanced sequencing is a robust and high-throughput platform for the detection of clinically actionable mutations.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4569. doi:1538-7445.AM2012-4569
Abstract 4579: The novel MEK Inhibitor GSK1120212b inhibits HMCL growth in vitro and synergises with a range of conventional and novel anti-myeloma therapies
Authors: Khong Tiffany T. , Leow Christopher , Monaghan Katherine , Spencer Andrew
Published: August 2012

Improved therapeutic interventions over the past decade has led to prolonged survival rates of patients with multiple myeloma (MM) but unfortunately, the subsequent development of progressive disease is inevitable. As such, the identification of novel compounds and strategies that improve therapeutic response is essential. GSK1120212b, a novel orally bio-available MEK inhibitor, has been shown to inhibit tumour growth and induce cell cycle arrest in some cancer cells. Here we report the efficacy of GSK1120212b in human myeloma cell lines (HMCL) and primary myeloma cells when used as a single agent and in combination with conventional and novel anti-myeloma therapies. HMCL response to variable doses of GSK1120212b (10nM-5µM) was determined by MTS assay. FACS and cell cycle analysis were used to evaluate the profiles of the cells post-MEK inhibition. Mechanistic studies using Western analysis with antibodies against p-MEK, MEK, p-ERK and ERK were performed. Subsequently, the anti-MM activity of GSK1120212b was evaluated in HMCL co-cultured with the human stromal cell line HS5. Primary samples from relapsed/refractory MM patients were subjected to GSK1120212b treatment and the level of apoptosis in CD45negCD38pos myeloma cells was assessed by Apo2.7-PE. Finally, combination studies with conventional and novel anti-MM therapies were also performed. GSK1120212b demonstrated an IC50 of 10-5000nM against 3 HMCL at 72h hours. Induction of both apoptosis and accumulation of cells in the G0/G1 phase occurred in a dose dependent manner. Western Blot analysis revealed increased p-MEK activity while total MEK levels decreased. Moreover, a decrease in p-ERK was observed with no change in total ERK levels. GSK1120212b demonstrated variable ability to induce HMCL apoptosis in stromal cell (HS5) co-culture, with or without transwells. Finally, GSK1120212b exhibited synergism with a variety of conventional and novel anti-MM therapies but was most pronounced when used in combination with GSK2110183b (Akt inhibitor) or LBH589 (HDAC inhibitor). Considering the potent anti-MM effects of GSK1120212b and its ability to synergise with other anti-MM agents, MEK inhibition demonstrates a promising future avenue for the management of MM and warrants further investigation.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4579. doi:1538-7445.AM2012-4579
Abstract 458: Implication of epithelial to mesenchymal transition (EMT) in pancreatic cancer prognosis
Authors: Yamada Suguru , Fujii Tsutomu , Fuchs Bryan C. , Sugimoto Hiroyuki , Nomoto Shuji , Takeda Shin , Kodera Yasuhiro , Nakao Akimasa
Published: August 2012

[Background] In recent years, epithelial-to-mesenchymal transition (EMT) has been shown to play an important role during cancer invasion and metastasis, and is now being actively studied in various cancers. Moreover, EMT is thought to be involved in resistance to chemotherapy and this elucidation could contribute to improved therapeutic effect. A thorough examination of EMT in pancreatic cancer, especially using resected surgical specimens, has not been conducted to date. [Methods] Between April 2004 and March 2010, 174 patients who underwent the resection of their pancreatic cancers were used for this study. Immunohistochemistry of E-cadherin and Vimentin were performed on their resected specimens, and the results of the staining were graded as follows; grade 1: negative, grade 2: positive, grade 3: strong positive. V/E ratio (Vimentin grade/E-cadherin grade) was determined and the patients were categorized into three group as follows: Epithelial(E)1. The correlation between EMT status and clinicopathological factors were analyzed. [Results] 1. Of the 174 cases, EMT status was determined as 37 Epithelial, 48 Borderline and 89 Mesenchymal. We found a significant correlation between EMT status and washing cytology (P = 0.025), tumor size (P = 0.014), portal vein invasion (P = 0.038) and lymphatic invasion (P = 0.047). 2. The median survival was 40.2 months in E, 63.1 months in B, and 13.7 months in M, respectively. There was a significant difference between M and E, and M and B (P < .0001). 3. According to univariate analysis, tumor location (head), CA19-9 level, tumor size (≥ 2cm), anterior tumor invasion, bile duct invasion, duodenal invasion, portal vein invasion, lymphatic or vascular invasion, positive perineural invasion, lymph node metastasis and EMT status (B · M / E) were found to be significant prognostic factors. Multivariate analysis showed perineural invasion (P = 0.024), lymph node metastasis (P = 0.033) and EMT status (P <0.0001) were significant prognostic factors. 4. Adjuvant chemotherapy (gemcitabine or S-1) improved the median survival from 19.9 to 63.1 months in group B, from 10.8 to 16.1 months in group M (P = 0.034, P = 0.002, respectively), however, no significant difference was seen in group E. [Discussion] EMT status was found to be one of the most important prognostic factors in pancreatic cancer and suspected to be associated with cancer invasion and metastasis. Further studies are needed to validate these results, but chemotherapy strategies targeted to epithelial and mesenchymal tumors might improve the outcomes of this disease by increasing the effect of chemotherapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 458. doi:1538-7445.AM2012-458
Abstract 4586: Use of triple combination array analysis to identify STEAP family member 4 (STEAP4), a novel tumor related gene in hepatocellular carcinoma
Authors: Nomoto Shuji , Hishida MItsuhiro , Hayashi Masamichi , Inokawa Kuniyoshi , Kodera Yasuhiro
Published: August 2012

Background: Hepatocellular carcinoma (HCC) is one of the most common cancers world-wide, but the molecular mechanisms underlying hepatocarcinogenesis are not clear. Array technology has made it possible to identify novel genes with alterations in liver cancer. In this study, we performed triple combination array analysis, expression profiling, karyotyping analysis using SNP array, and array-based DNA methylation profiling in the same HCC sample and attempted to find a novel tumor related gene as a prognostic marker. Materials & Methods: A 68-year-old woman with chronic hepatitis C was diagnosed with HCC. The excised tumor and corresponding normal tissues were used for extraction of DNA and RNA. SNP array experiments were performed using the DNA. mRNA expression profiles were determined using the total RNA by Affimetrix HGU133A and B Gene-Chips. Bisulphite-converted genomic DNA was analyzed using Illumina's Infinium Human Methylation27 Beadchip Kit. We consequently analyzed the identified gene using quantitative RT-PCR, methylation and un-methylation-specific PCR in nine HCC cell lines and in 48 primary HCC tissues and corresponding non-cancerous tissues collected at Nagoya University Hospital, and then compared these results with clinico-pathological data. Results: STEAP4 gene, is located on 7q21.12, and expression was decreased at a Log2 ratio of –4.2 to –2.0 in cancerous tissue. There were two copies of 7q21.12, indicating no loss of heterozygosity (LOH) in the SNP array. The methylation value (0-1.0) in the tumor sample was 0.870 compared to 0.160 in non-cancerous tissue, indicating high methylation in tumor tissue in the Methylation array. Re-activation of STEAP4 expression was seen after 5-aza-dC treatment in HCC cell lines. Moreover, we found that 32 of 48 (67%) HCC samples showed promoter hypermethylation. In 32 methylated cases, the expression values of STEAP4 gene in tumor tissues by real-time RT-PCR were significantly decreased compared to corresponding normal samples (p<0.0001). In addition, methylated cases were significantly correlated with worse recurrence free survival (p<0.0001) and overall survival (p=0.0093). Conclusion: The Promoter hypermethylation of STEAP4 gene was a marker of worse prognosis in HCC. The triple combination array was a useful technique to identify the novel tumor related gene in HCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4586. doi:1538-7445.AM2012-4586
Abstract 4589: Clinical significance of the EML4-ALK fusion gene and association with EGFR and KRAS gene mutation in 208 Chinese patients with non-small cell lung cancer
Authors: Chen Jun , Liu Hongyu , Yang tong , Wei Sen , Li Ying , Wang Yuli , Li Yongwen
Published: August 2012

Objective: EML4-ALK fusion gene is a potentially relevant oncogenic event in lung cancer, which represents a new subgroup of non-small cell lung cancer (NSCLC) patients who respond positively to ALK inhibitors. The characteristics of EML4-ALK fusion gene in Chinese Patients with NSCLC are poorly understood. In this study, we aimed to analyze the prevalence of EML4-ALK fusion gene and their correlation to epidermal growth factor receptor (EGFR) status, KRAS mutation, and clinico-pathological data in Chinese patients with NSCLC. Methods: EML4-ALK fusion gene was detected by nested RT-PCR and confirmed by sequencing. EGFR and KRAS gene status were evaluated by RT-PCR and sequencing. Survival analysis was performed. A meta-analysis of the EML4-ALK fusion gene in lung cancer patients was also presented. Results: Two hundred eight NSCLC cases were evaluated. There were 51 cases of mutations in EGFR at exons 18-21 (24.5%, 51/208). Among exons 18-21, EGFR mutations occur predominantly (92%) in exons 19 and 21. In concordance with previous reports, these mutations are identified at high frequencies in females (47.5%, 29/61 vs 15.0%, 22/147 in males; P = 0.000); never-smokers (42.3%, 33/78 vs 13.9%, 18/130 in smokers; P = 0.000), and adenocarcinoma patients (44.2%, 42/95 vs 8.0%, 9/113 in non-adenocarcinoma patients; P = 0.000). There were only 6 cases (6/208, 2.88%) of KRAS mutations in our study group. We identified 7 patients who harbored the EML4-ALK fusion gene (3.37%, 7/208) which all confirmed by DNA sequencing. Of these 7 patients, 2 cases displayed the EML4-ALK variant 1 (28.6%), 1 case exhibited variant 2 (14.3%) and 4 cases carried variant 3 (57.1%). All of the positive cases corresponded to female patients (11.5%, 7/61). Six of the positive cases were non-smokers (7.69%, 6/78). The incidence of EML4-ALK translocation in female non-smoking adenocarcinoma patients is as high as 15.2% (5/33). No EGFR/KRAS mutations were detected among EML4-ALK positive patients. The meta-analysis demonstrated that EML4-ALK translocation was 4.71% (125/2652) in unselected patients with NSCLC, and was also predominant in female patients with adenocarcinoma. Conclusion: EML4-ALK translocations are infrequent in the entire NSCLC patient population, but are frequent in the NSCLC patient subgroup of female, nonsmokers, and adenocarcinoma patients. Our results indicate that the detection of the EML4-ALK translocation in subgroups of patients with NSCLC is crucial for applying targeted therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4589. doi:1538-7445.AM2012-4589
Abstract 4591: Biotinylated derivatives of selective high affinity ligand (SHAL) therapeutics as companion diagnostics for prescreening non-Hodgkin's lymphoma patients
Authors: Balhorn Rod , Rebhun Robert B. , Hok Saphon , Baum Richard , Sanger Jorg , Balhorn Monique Cosman
Published: August 2012

Selective High Affinity Ligands (SHALs) are a new class of small molecule targeting agents that can be designed to bind selectively and with high affinity to unique sites located on almost any protein. A series of SHAL-based therapeutics have been developed to target the cell surface antigen HLA-DR10 that is over-expressed on many B-cell derived malignancies. SHALs are created by first identifying subsets of potential ligands, or “recognition elements,” that bind to two or three neighboring unique sites on the surface of the target protein. Bidentate or tridentate SHALs are produced by linking two or three of these recognition ligands together using carbodiimide chemistry and a miniPEG and lysine scaffold. SHAL synthesis is rapid, uses relatively inexpensive, commercially available components, and is very versatile. Multiple forms of the SHAL can be created by combining different recognition elements (ligands) or by attaching a variety of tags (biotin, chelating groups, dyes, inhibitors) to the end of the linker. Using this approach, SH7139 has been developed as a promising therapeutic for treating non-Hodgkin's lymphoma. Previous studies have shown SH7139 to be effective in treating human B-cell lymphoma in a mouse xenograft model at extremely low doses (3.3μg/kg); 70% of the tumors disappeared within 30 days after treatment and did not reoccur during the remainder of the life of the animal. Here we report that a biotinylated derivative of SH7139 can be used as companion diagnostic to prescreen needle aspirates or biopsies from B-cell lymphomas and determine whether or not the tumor is likely to respond to SH7139 therapy. The staining protocol involves detecting the bound biotinylated SHAL using commercially available alkaline phosphatase reagents, and it works equally well with both fixed and unfixed biopsy sections. Biotinylated SH7139 has been used to identify potentially responsive B-cell lymphomas in both canine and human patients. The development of SHAL-based diagnostics in parallel with new SHAL-based therapeutics will help advance individualized therapies and minimize the systemic trauma and valuable time lost by patients given treatments that prove to be ineffective. Our results also suggest this diagnostic may also prove useful in determining whether malignancies over-expressing HLA-DR10 have metastasized and individual lymphoma cells have migrated out of the tumor into surrounding normal tissue. This research was supported, in part, by NIH/NCI SBIR grant 1R43CA159843-01 to Rod Balhorn.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4591. doi:1538-7445.AM2012-4591
Abstract 4594: Dual PI3K/mTOR blockade by NVP-BEZ235 sensitizes head and neck squamous cell carcinoma to radiotherapy
Authors: Gao Ming , Wang Chun-Wei , Fang Chun Sheng , Lu Yen-Shen , Cheng Ann-Lii
Published: August 2012

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide accounting for over 500,000 new cases annually. Chemoradiotherapy is a mainstay of treatment. However, long-term survival rate has only improved marginally; and the severe toxicity often associated with chemoradiotherapy advocates a need for better radiosensitization strategies. Akt/mTOR pathway is frequently activated in HNSCC cells. This study sought to explore whether NVP-BEZ235, a novel imidazo [4,5-c] quinoline derivative that inhibits PI3K and mTOR kinase activity, has radiosensitization effect in the treatment of HNSCC. MTT assays revealed that NVP-BEZ235 dose-dependently inhibited the growth of two HNSCC cell lines, SAS and OC3 at low nanomolar concentrations; IC50 values were 27.87 nmol/L in SAS cells and 42.63 nmol/L in OC3 cells. By using clonogenic assay and median effect analysis, we found that dual PI3K/mTOR inhibition effectively sensitized SAS and OC3 cell lines to radiation with CI (combination index) value <1, indicating a synergistic effect. By using flow cytometry analysis, when SAS and OC3 cells were sequentially exposed to NVP-BEZ235 and radiation, an increase in the subG1 population was observed, indicating that NVP-BEZ235 sensitized HNSCC cells to radiation by augmenting apoptotic cell death. In SAS xenograft nude mice model, combination therapy with oral administration of 45mg/kg/day of NVP-BEZ235 and 3 Gy radiation for 5 days was well tolerated and significantly enhanced the antitumor effect of radiation (p=0.005). Our results indicated that dual PI3K/mTOR blockade effectively sensitizes radiotherapy for HNSCC. Clinical trials along this line are warranted. (The study was supported by the grant DOH100-TD-B-111-001)

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4594. doi:1538-7445.AM2012-4594
Abstract 4596: Missense mutations in the p53 DNA-binding motifs (DBMs) are associated with shorter time to progression (TTP) in erlotinib-treated, EGFR-mutated patients
Authors: Molina Miguel Angel , Bertran-Alamillo Jordi , Mayo Clara , Massuti Bartomeu , Camps Carlos , Carcereny Enric , Viteri Santiago , Benlloch Susana , Taron Miquel , Rosell Rafael
Published: August 2012

Background: Advanced non-small-cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR) mutations has an average time to progression (TTP) of 14 months (mo) and overall survival of 27 mo when treated with erlotinib. However, EGFR mutations can only imperfectly predict outcome. In NSCLC cell lines, tyrosine kinase inhibitors (TKIs) induce p53 translocation from the cytoplasm to the nucleus and subsequent up-regulation of Fas and caspase activation leading to apoptosis. This mechanism was defective in p53-null cells. We tested whether mutations in the TP53 gene influence outcome to erlotinib in EGFR-mutated patients (p). Expression levels of the p53 repressor MDM2 were also examined. Methods: We assesed p53 status in pretreatment paraffin-embedded tumor samples from 93 erlotinib-treated, EGFR mutated advanced NSCLC p. Mutations in exons 5, 6, 7 and 8 were screened by High Resolution Melting analysis (HRM) followed by sequencing of the amplified products with non-wild type melt curves. All mutant samples were re-confirmed by standard PCR and sequencing. Expression levels of MDM2 mRNA were determined by quantitative RT-PCR. Results: Mutations in exons 5-8 of the TP53 gene were detected in 26 of 93 p (28%). We found an unusually high frequency of in-frame and frameshift deletions (23% of mutations), indicating that the spectrum of p53 mutations might be different in EGFR-mutated NSCLC. Mutations in the TP53 gene were not associated with sex, age, smoking status, histology, exon 19 vs. 21 mutation, or response, but, within the study population, were significantly less frequent in p with ECOG 2 or above. The mutations were also associated with the presence of the pretreatment T790M mutation. 14 p had mutations in one of the p53 DNA binding motifs (DBMs), and showed TTP to erlotinib of only 9 mo, compared to 19 months for wt p and 27 mo for those carrying a non-DBM mutation. Survival was 24 mo vs. 31 mo, and not-reached, respectively. Finally, MDM2 mRNA levels were significantly lower in tumors with p53 mutations, especially when these affected DBMs. In the case of wt p, high MDM2 expression correlated with a better TTP and survival. Conclusions: TP53 mutations co-exist with EGFR mutations in a significant number of p; and those in the DBMs are associated with poorer response to erlotinib. This finding paves the way for the possibility of combining erlotinib with a drug restoring p53 function in those p harboring DBM mutations in the TP53 gene.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4596. doi:1538-7445.AM2012-4596
Abstract 4604: Some of oral squamous cell carcinomas might originate from minor salivary glands, and these tumors shows aggressive biological behavior and poor prognosis
Authors: Kawamata Hitoshi , Izumi Sayaka , Doi Yutaka , Imai Yutaka
Published: August 2012

Oral cancer massively invades the adjacent tissues and frequently metastasizes to the regional lymph nodes. Most of the oral cancer is histopathologically squamous cell carcinoma, and approximately 10 % of the oral floor cancer is salivary gland cancer, such as mucoepidermoid carcinoma and adenoid cystic carcinoma. According to the clinical behavior and histopathological features of the oral squamous cell carcinoma (OSCC), we hypothesize that OSCC might originate from either oral squamous epithelium or minor salivary glands. In this study, we attempted to clarify the possibility, in which some of the OSCC originated from minor salivary glands by microarray and clustering analysis. Furthermore, by using the eleven molecular markers which were identified by the microarray and clustering analysis, we determined the origin of OSCC, and compared their biological aggressiveness in the patients. The mRNA expression profile of the samples obtained from 6 patients with OSCC was analyzed by the AB1700 Chemiluminescent Microarray Analyzer on the Human Genome Survey Array containing 29,098 genes. Clustering analysis was performed by GeneSping GX 7.3. Clustering analysis by the expression level of the 917 genes, which were differentially expressed at least 3-folds in normal oral squamous epithelium and normal salivary glands, revealed that 6 samples can be divided into 2 groups. The expression profile of one group (4 cases) was similar to that of normal oral squamous epithelium, and that of another group (2 cases) was similar to that of normal salivary glands. Furthermore, we identified 11 genes from the 917 genes which can easily determine the origin of the OSCC. Subsequently, we examined the expression levels of 9 marker genes in 73 OSCC (45 men, 28 women) by RT-PCR and determined the origin of the OSCC. Twelve cases from 73 OSCCs (16%) were considered to originate from minor salivary glands, and these tumors showed highly metastatic potentials and poor prognosis. In conclusion, OSCC originates from either oral squamous epithelium or minor salivary glands. OSCCs originating from minor salivary glands showed aggressive biological behavior. Determination of the origin of the OSCC is helpful to make a treatment plan for the patients with OSCC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4604. doi:1538-7445.AM2012-4604
Abstract 4609: Epigenetic regulation of BRM
Authors: Kahali Bhaskar , Gramling Sarah , Reisman David N.
Published: August 2012

BRM is part of the SWI/SNF complex and is key anticancer protein which is silenced in variety of tumor types. SWI/SNF is a prerequisite for the function of many different anticancer proteins such as Rb and p53 and it has essential roles in growth control, differentiation, development, cell adhesion, and DNA repair. Consequently, inactivation or impairment of this complex can stimulate tumor development and tumor progression through the abrogation of a number of anticancer mechanisms. Unlike many tumor suppressor proteins, BRM is not mutated, but rather epigenetically suppressed, which is validated by the fact that HDAC inhibitors were the first compounds found to restore BRM expression. Unfortunately, these compounds are too broadly active and in addition to restoring BRM expression, they inactivate it by acetylation. To try to better understand how BRM is epigenetically regulated, we knocked-out each of the known 11 HDACs, using shRNAs in BRM-deficient and BRM-positive cell lines to determine which ones regulate or inactivate BRM. We found that that shRNA knockdown of only HDAC3 or HDAC9 restores both BRM expression and function, while suppression of HDAC2 induces BRM acetylation. Similarly, we tested the impact of over-expression of 25 different Histone Acetyltransferases (HATs) on BRM expression and acetylation. We found ectopic expression of KAT6A, KAT6B, and KAT7 in BRM-deficient cell lines induces BRM whereas ectopic expression of KAT2B and KAT8 causes BRM acetylation in BRM-positive cell lines. We also uncovered that HDAC9 is greatly over-expressed only in BRM-deficient cell lines and its expression is driven by MEF2D and GATA3. Expression and knockdown experiments directed at MEF2D and GATA3 showed that these transcription factors regulate BRM and HDAC9. Similarly, we found that GATA3 expression is only expressed in BRM-deficient cell lines. This work outlines the complex array of proteins which are tied to the expression and regulation of BRM. It is important to note this epigenetic mechanism is not likely specific to BRM, but rather likely regulates a cadre of anticancer and cellular proteins as well. Hence, determining how BRM is aberrantly regulated is the first step in determining how to target BRM and this mechanism for therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4609. doi:1538-7445.AM2012-4609
Abstract 4614: Karyometry in atypical endometrial hyperplasia: A Gynecologic Oncology Group study
Authors: Alberts David S. , Garcia Franciso AR , Trimble Cornelia L. , Kauderer James , Curtin John , Lim Peter C. , Hess Lisa M. , Silverberg Steven , Zaino Richard J. , Yozwiak Michael , Bartels Hubert G. , Bartels Peter
Published: August 2012

Treatment for atypical endometrial hyperplasia (AEH) is based on pathologic diagnosis. About 40% of AEH is found to be carcinoma at surgery. The histopathologic classification of endometrial lesions is a difficult task requiring extensive experience. The Gynecologic Oncology Group (GOG) conducted a study (GOG 167A) that was designed to estimate the frequency of endometrial cancer in hysterectomy specimens in subjects who had a biopsy diagnosis of atypical endometrial hyperplasia. This study's objective is to derive an objective characterization of nuclei from cases diagnosed as AEH or superficially invasive endometrial cancer (SIEC). Cases from GOG study 167A were classified by a central pathology committee as AEH (n=39) or SIEC (n=39). High resolution digitized images of cell nuclei were recorded. Features of the nuclear chromatin pattern were computed. Classification rules were derived by discriminant analysis. Nuclei from cases of AEH and SIEC occupy the same range on a progression curve for endometrial lesions. Cases of AEH and SIEC both comprise nuclei of two phenotypes: hyperplastic characteristics and premalignant/neoplastic characteristics. The principal difference between AEH and SIEC is percentage of premalignant/neoplastic nuclei. When this percentage approaches 50-60% superficial invasion is likely. SIEC may develop already from lesions at the low end of the progression curve. AEH comprises cases which may constitute a low risk group involving <40% of AEH cases. These cases hold a percentage of <20% of nuclei of a preneoplastic phenotype. AEH cases from the central and high end of progression have >40 % of nuclei of the preneoplastic phenotype. Nuclei of the preneoplastic phenotype in AEH lesions are almost indistinguishable from nuclei in SIEC, where this percentage exceeds 60%. While karyometric assessment does not provide a distinctive diagnostic criterion for an individual patient, it does provide a measure of risk for the development of invasive disease. Studies to support the diagnostic assessment of endometrial lesions by computer aided morphometric analyses go back over 30 years. These efforts were aimed at two objectives, the diagnostic distinction between benign and cancer, and procedures that allow a stratification of patients into risk groups for the development of myometrial invasion. The percentage of nuclei of preneoplastic phenotype in AEH lesions might serve as criterion for assessment of risk for the development of invasive disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4614. doi:1538-7445.AM2012-4614
Abstract 4615: Key miRNAs and co-targets in patients of OPSC and OCC
Authors: Cui Shiying , Zhao Henan , Ding Yanfang , Zhao Jinyao , Bi Tie , Jiang Jiyong
Published: August 2012

Objectives: To identify the key microRNAs (miRNAs) and their top co-targets in patients of elderly ovarian papillary serous carcinoma (OPSC) patients with advanced stage and ovarian clear cell carcinoma (OCC), for the insight of possible regulation gene and pathway of epithelial ovarian cancer (EOC) prognosis. Methods: The study was opened to patients with primary, previously untreated EOC referred to member institutions. Pathologic diagnosis was confirmed by central review. High-throughput analysis of the miRNA profile in a panel of OCC and OPSC cells was assessed using a microarray platform. The miRNA-targets and possible pathway were predicted by TargetScans, MicroCosm Targets version 5, miRBase and Ingenuity. Finally, the downstream predicated co-target, activating transcription factor 3 (ATF3) was validated by quantitative Real-Time Polymerase Chain Reaction (qRT-PCR) and immunohistochemical (IHC) assay. Results: Of the 667 miRNAs analyzed from human, we identified 83 as enriched in the OPSC, and 15 in OCC (P<0.01 for all). Seventeen of 667 miRNAs were significantly expressed with at least 3 fold differences in the OPSC versus OCC. The expression of tree miRNAs (miR-30a*, miR-30e*, miR-505*) were most significantly lower in OCC comparing to elderly OPSC patients with advanced stage. ATF3, the top co-target gene of three key miRNAs mentioned above was highly expressed in OCC lesions, and a stronger distribution was shown in protein level of ATF3 by immunohistochemistry in OCC than OPSC lesions. Conclusions: We identified the key miRNAs that differentially expressed in patients of elderly OPSC with advanced stage and OCC tumors, predicted and validated the most popular co-target shared by the key miRNAs, indicating the possible regulation gene and pathway for EOC prognosis.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4615. doi:1538-7445.AM2012-4615
Abstract 4623: Human anti-nucleolin antibodies with broad spectrum anticancer activity
Authors: Fernandes Daniel J. , Otake Yoko , Capizzi Robert L.
Published: August 2012

Extensive preclinical studies have validated nucleolin as a new therapeutic target in oncology. Confocal microscopy, immunohistochemistry, and cell fractionation studies have shown that this protein is highly overexpressed in the plasma membrane and cytoplasm of a wide variety of human hematological and solid tumor cells, but is usually undetectable on the cell surface or in the cytoplasm of the corresponding normal cells. We have utilized our licensed platform technology to generate a first-in-class panel of eight fully human monoclonal IgG1 antibodies (HuMAbs). These antibodies bind specifically to nucleolin on the tumor cell surface, which in turn elicits potent cytotoxicity to a variety of human tumor cell lines. One such example is CP101, which killed MV4-11 (AML), MCF-7 (breast), DU145 (prostate), PANC-1 and MIA PaCa-2 (pancreas) tumor cell lines with IC50 values ranging from (0.5-2.0 µg/ml; 3-12 nM) following exposure to the HuMAb for 96 hrs. These in vitro assays were performed in the absence of human complement and immune effector cells required for complement-dependent cellular cytotoxicity (CDCC) and antibody-dependent cellular cytotoxicity (ADCC), respectively. These results are consistent with published observations that anti-nucleolin HuMAbs can exploit the known shuttling function of cell surface nucleolin to gain intracellular access and induce direct tumor cytotoxicity. In contrast to its cytocidal effects on tumor cells, CP101 had no effect on the viability of either MCF-10A normal human breast epithelial cells or normal human CD19+ B cells, neither of which expresses nucleolin on the cell surface. In summary, the results suggest that anti-nucleolin HuMAbs are unique in that they can exert broad spectrum antitumor activity independently of the immune mechanism of CDCC and ADCC, while having no detectable effects on the viability of the corresponding normal cells.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4623. doi:1538-7445.AM2012-4623
Abstract 4624: High affinity CD3 RECRUIT-TandAbs for T cell-mediated lysis of malignant CD19+ B cells
Authors: Reusch Uwe , Burkhardt Carmen , Knackmuss Stefan H. , Fucek Ivica , Eser Markus , McAleese Fionnuala , Gronewold Thomas , Zhukovsky Eugene , Little Melvyn
Published: August 2012

The TandAb technology enables the development of innovative antibody therapeutics for improved treatment of various diseases. This platform has been primarily applied to oncology and comprises of CD3 RECRUIT- and CD16A RECRUIT-TandAbs for activation of T and NK effector cells, respectively, and lysis of target cells expressing specific surface antigens. The CD3 RECRUIT-TandAb AFM11 is a human bispecific tetravalent antibody with two binding sites for the α-chain of CD3 and two binding sites for CD19. CD19 is expressed at early stages of B cell development and persists until the final differentiation into plasma cells. Thus, CD19 represents an attractive target for the treatment of various B cell malignancies including leukemias and lymphomas that lack CD20 expression or are refractory to anti-CD20 antibody therapies. In vitro cytotoxicity assays employing tumor cell lines demonstrate high potency of AFM11 in mediating target cell lysis: EC50 values are in the low to sub-picomolar range depending on CD19+ target cell lines and primary B-CLL tumor cells. Cytotoxic activity of tetravalent AFM11 is superior to those of alternative bivalent antibody formats. High affinity binding of AFM11 to CD3 and CD19 (low to sub-nanomolar range) contributes to efficacious T cell recruitment. This binding was determined by flow cytometry and by a label-free biosensor that measures real-time binding kinetics on live cells. Therapeutic recruitment of T cells has a potential for a devastating cytokine release if it results not in a site-specific but rather systemic activation. Thus, functional in vitro assays were used to demonstrate that high affinity and bivalent binding of AFM11 to CD3 is not sufficient for triggering activation signals in T cells. AFM11 activates T cells only in the presence of CD19+ target cells, and specifically mediates lysis of these cells without affecting antigen-negative bystander cells. In the absence of target cells and at high concentrations AFM11 induces down-modulation of the CD3/TCR complex from the surface of T cells. The latter may contribute additional safety to therapeutic potential of CD3 RECRUIT-TandAbs without affecting the efficacy, since AFM11-treated T cells can be re-engaged for target cell lysis. In vivo AFM11 demonstrated a robust dose-dependent inhibition of tumor growth in a study with Raji xenografts in NOD/scid mice and reconstituted human PBMC. Altogether, these data suggest that AFM11 is a highly efficacious novel drug candidate for the treatment of hematological malignancies and displays an advantageous safety profile.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4624. doi:1538-7445.AM2012-4624