bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 5549: Recipient-donor contradictory genotype with impact on anticancer drug pharmacogenetics after liver transplant: A deadly gift
Authors: Serdjebi Cindy , Fanciullino Raphaelle , Fina Frederic , Verschuur Arnaud , Roquelaure Bertrand , Lacarelle Bruno , Ouafik L'Houcine , Ciccolini Joseph , André Nicolas
Published: May 2015

A 5-year old girl with liver transplant was scheduled for a therapy including high-dose cytarabine for her Burkitt lymphoma. Cytarabine pharmacogenetics focus on cytidine deaminase (CDA) , the polymorphic liver enzyme responsible for its detoxification. Preliminary double testing for her CDA status on DNA lymphocytes showed none of the germline polymorphisms usually associated with CDA deficiency (ie, 79A>C (K27Q), 208G>A), but surprisingly with a functional mild deficiency syndrom. Despite 30% reduction in cytarabine dosing, life-threatening toxicities showed quickly and treatment was discontinued. Genetic HRM-based investigations performed in the patient on liver biopsy failed in generating a clear genotype. Further HRM+sequencing retrospective investigations on liver biopsy collected from the donor showed that he was actually bearing the homozygous CDA*2 allelic variant (ie, 79AA), a genotype usually associated with severe CDA deficiency. Based upon the donor liver genotype, and not the recipient genotype, treatement was resumed with further dose reduction, close monitoring and better tolerance eventually. The patient is now in complete remission more than a year after completion of the treatment. This case report illustrates the limits and risks of searching germline polymorphisms in patients with liver transplant when the story plays in the liver.

Citation Format: Cindy Serdjebi, Raphaelle Fanciullino, Frederic Fina, Arnaud Verschuur, Bertrand Roquelaure, Bruno Lacarelle, L'Houcine Ouafik, Joseph Ciccolini, Nicolas André. Recipient-donor contradictory genotype with impact on anticancer drug pharmacogenetics after liver transplant: A deadly gift. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5549. doi:10.1158/1538-7445.AM2014-5549
Abstract 5553: Monotherapy efficacy of catalytic and allosteric inhibitors of AKT in breast cancer models with AKT1E17K mutations
Authors: Davies Barry R. , Guan Nin , Logie Armelle , Palmer Amy , Jacobs Vivien , James Neil , Jenkins Emma L.
Published: May 2015

AKT1E17K mutations occur at low frequency in a variety of solid tumors, including those of the breast and urinary bladder. Although this mutation has been shown to transform rodent cells in culture, it was found to be less oncogenic than PIK3CA mutations in breast epithelial cells. Moreover, the therapeutic potential of AKT inhibitors in human tumors with an endogenous AKT1E17K mutation is not known. Expression of exogenous copies of AKT1E17K in MCF10A breast epithelial cells increased phosphorylation of AKT and its substrates, and induced colony formation in soft agar. These effects were inhibited by the allosteric and catalytic AKT inhibitors MK-2206 and AZD5363, respectively. We identified two breast cancer explant models with AKT1E17K mutation (HBC-x2 and HBCx-31). Dosing of AZD5363 at 150 mg/kg twice daily or MK-2206 at 120 mg/kg 3 times a week significantly inhibited tumor growth in both models, but AZD5363 was significantly more active than MK-2206 at the dosing schedules employed, in both models. Pharmacodynamics were assessed in the HBC-x2 model at 2 h after the final dose. Both compounds significantly inhibited phosphorylation of PRAS40 and S6, caused relocalisation of FOXO3a to the nucleus, and reduced Ki67 staining. The data indicate that breast cancers with AKT1E17K mutations are rational targets for monotherapy treatment with AKT inhibitors.

Citation Format: Barry R. Davies, Nin Guan, Armelle Logie, Amy Palmer, Vivien Jacobs, Neil James, Emma L. Jenkins. Monotherapy efficacy of catalytic and allosteric inhibitors of AKT in breast cancer models with AKT1E17K mutations. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5553. doi:10.1158/1538-7445.AM2014-5553
Abstract 5563: Docetaxel-induced mucositis in breast cancer patients: Association with plasma alpha-1-acid glycoprotein level and SLCO1B3 genotype
Authors: Jabir Rafid S. , HO Gwo F. , Anuar Muhammad Azrif A. , Stanslas Johnson
Published: May 2015

Mucositis is one of the major non-hematological adverse effects (AEs) of docetaxel, besides fatigue, nausea, vomiting, diarrhea and rash which limit its use in cancer patients. Alpha-1-acid glycoprotein (AAG) is an acute phase reactant glycoprotein and is a primary carrier of docetaxel in the blood. Docetaxel demonstrates extensive binding to plasma proteins (>98%) including AAG, lipoproteins and albumin. However, there is a remarkable inter-individual variation of AAG level among cancer patients. Low plasma AAG levels will result in excessive free drug in the circulation, which in turn causes more AEs. Consequently, either the dose is reduced or treatment modality is diverted. In addition, single nucleotide polymorphisms (SNPs) of genes encoding for proteins involved in the transport of docetaxel (SLCO1B3) could lead to reduced clearance and accumulation in the circulation and subsequent AEs. Hence, we investigated 91 Malaysian breast cancer patients receiving docetaxel as a single agent chemotherapy at 90-100 mg/m2 and 75 mg/m2 in the adjuvant and metastatic settings respectively, to determine the association between the SNP (SLCO1B3 334TG), plasma AAG level and docetaxel AEs. The plasma AAG level was determined using Enzyme Linked Immunosorbent assay (ELISA) technique, while the SNPs were analyzed using PCR-RFLP technique followed by DNA sequencing of selected samples. The patients consisted of 49 Chinese (54%), 34 Malays (37%) and 8 Indians (9%). We found a significant difference in plasma AAG level between patients with mucositis and those without (178±74 mg/dl vs 225±97 mg/dl,P<0.05). Moreover, patients with mutant SLCO1B3 334GG who developed diarrhea also had mucositis (p<0.05). In conclusion, patients with lower plasma AAG level are more susceptible to develop mucositis. In addition, patients carrying mutant SLCO1B3 334GG who have diarrhea tend to develop mucositis concurrently. Plasma AAG level and SNPs could serve as potential predictive biomarkers of docetaxel-induced mucositis.We would like to thank Malaysian Ministry of Higher Education (MOHE) and University Malaya for funding this project through FRGS (04-04-10-848FR) and HIR (UM.C/625/1/HIR) grants.

Citation Format: Rafid S. Jabir, Gwo F. HO, Muhammad Azrif A. Anuar, Johnson Stanslas. Docetaxel-induced mucositis in breast cancer patients: Association with plasma alpha-1-acid glycoprotein level and SLCO1B3 genotype. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5563. doi:10.1158/1538-7445.AM2014-5563
Abstract 5567: Impact of progesterone receptor (PR) isotype-specific immunohistochemistry (IHC) on the diagnosis of triple negative breast cancer (BC)
Authors: Gilles Erard , Bosq Jacques , Caplier Laura , Bexon Alice , Zukiwski Alexander , Bonneterre Jacques
Published: May 2015

Background: Given the poor therapeutic outcomes for triple negative BC, diagnostic accuracy is vital. In routine IHC, PR is measured using bispecific antibodies (Ab) that recognize epitopes common to PRA and PRB. PR A and B expression is imbalanced in BC. Relative expression of the PR isotypes appears to be prognostic in tumors evaluated with a bispecific Ab (Hopp 2004). Tumors can express PRA or PRB on different cells in the same tumor (Mote 2008), which differ from those expressing ERα (Zukiwski 2013). A true triple negative phenotype might escape detection with the use of one single Ab to detect both PR A and B epitopes, depending on its sensitivity/specificity. This study evaluated the use of two isotype-specific PR Abs to fully characterize PR status.

Methods: 312 archived BC specimens with clinical data came from Oscar Lambret Cancer Center, Lille. HER2 status was previously determined. IHC was performed using anti-ERα, anti-PRA and PRB Abs. ERα and PR tumor positivity was defined as ≥ 1% stained tumor cells.

Results: 276 cases had complete data. Table 1 shows ERα, PRA and PRB status. 38/276 cases (14%) were ERα and HER2 negative: 22 were PR A or B negative, 9 were PR A and B positive, 2 were positive only for PRA and 5 only for PRB. Discussion: Bispecific PR Abs rarely report PRA and PRB similarly (Mote 2001). PR Phosphorylation and activation may vary across tumors. Potentially only one PR isotype may drive BC tumor growth. Triple-negative BC status determination could be PR Ab-dependent. These cases are currently being tested with a standard bispecific PR Ab to understand which would be considered triple-negative with routine IHC.

Conclusion: Further evaluation of IHC with specific PRA and PRB Abs is warranted to determine if certain patients, currently classified as having triple-negative BC, could respond to antiprogestins, which have shown activity in BC.

Table 1% of276 casesPR (PRA or PRB)PRAPRBERαNegativePositiveTotalNegativePositiveTotalNegativePositiveTotalNegative148221742115722Positive77178146579106868Total217910031691002575100

Citation Format: Erard Gilles, Jacques Bosq, Laura Caplier, Alice Bexon, Alexander Zukiwski, Jacques Bonneterre. Impact of progesterone receptor (PR) isotype-specific immunohistochemistry (IHC) on the diagnosis of triple negative breast cancer (BC). [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5567. doi:10.1158/1538-7445.AM2014-5567
Abstract 5568: CYP17A1 and abiraterone: Implications for breast cancer endocrine therapy
Authors: Capper Cameron P. , Johnson Michael D. , Larios José M. , Rae James M.
Published: May 2015

The cytochrome P450 17A1 (CYP17A1) inhibitor abiraterone, which blocks synthesis of all steroid hormones including androgens and estrogens, is an effective treatment for castrate resistant prostate cancer. Its utility for treating hormone receptor-positive breast cancer has yet to be established. In prostate cancer, the anti-proliferative effects of abiraterone are mediated by inhibiting CYP17A1 activity resulting in a significant reduction in circulating concentrations of the sex steroids dehydroepiandrosterone (DHEA), androstenedione, testosterone, and dihydrotestosterone (DHT). Recently, it has been proposed that the reduction in aromatizable androgens by abiraterone may make this drug useful for the management of hormone-dependent breast cancer. However, data from our lab suggests that abiraterone has estrogenic properties and can induce proliferation in breast cancer cells expressing the estrogen receptor (ER). Crystal violet assays were used to assess abiraterone-induced proliferation in the ER-positive, estrogen-dependent breast cancer cell lines MCF-7 and T47D. We show that abiraterone induced a dose dependent increase in cell proliferation with an IC50 of 3.7 μM and maximal stimulation (200%) observed at 8μM, compared to vehicle treated cells. Abiraterone also induced the expression of the ER response gene, GREB1. Abiraterone-induced proliferation and gene expression was blocked by increasing doses of the selective estrogen receptor down-regulator (SERD) fulvestrant. Abiraterone induced the dose-dependent expression of luciferase in MCF-7 cells transfected with an estrogen responsive luciferase reporter construct. In conclusion, our data suggest that abiraterone can induce ER-positive breast cancer cell proliferation in vitro by acting as a weak ER agonist. Further studies are underway to determine whether estrogenic effects of abiraterone are also observed using in vivo models of ER response. If abiraterone exhibits estrogenic effects in vivo then these data would suggest that abiraterone should be combined with an ER antagonist if used for the clinical management of women with ER+ tumors.

Citation Format: Cameron P. Capper, Michael D. Johnson, José M. Larios, James M. Rae. CYP17A1 and abiraterone: Implications for breast cancer endocrine therapy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5568. doi:10.1158/1538-7445.AM2014-5568
Abstract 5569: Ki67 is an independent prognostic marker in breast cancer even after accounting for molecular subtype
Authors: Mahmoud Abeer M. , Macias Virgilia , Al-alem Umaima , Kresovich Jacob K. , Khramtsova Galina , Kajdacsy-Balla Andre , Wiley Elizabeth L. , Rauscher Garth H.
Published: May 2015

Ki67 labeling index (LI) has been demonstrated to be a prognostic marker in invasive breast cancer; a high Ki67 LI is associated with poorer survival among breast cancer patients. The goal of these analyses was to examine the extent to which Ki67 was associated with specific breast cancer subtypes, and whether it could serve as an additional prognostic marker above and beyond breast cancer subtype, in an ethnically diverse sample of 287 patients (86 Non-Hispanic White, 84 Hispanic and 116 African American).

IHC analysis was performed on tissue microarrays constructed from invasive breast cancer samples obtained from the Breast Cancer Care in Chicago study (patients diagnosed between 2005-2008). Tissue samples were tested for the expression of estrogen receptor (ER), progesterone receptor (PR), HER2, EGFR and CK5/6. From these results, breast cancers were classified as Luminal A (ER+/PR+/HER2-), Luminal B (ER+/PR+/HER2+), HER2 enriched (ER-/PR-/HER2+), and triple negative (TN) (ER-/PR-/HER2-). TN tumors were subclassified into basal like (BL) if they expressed either EGFR or CK5/6 or otherwise classified as unspecified (US) if negative for both EGFR and CK5/6. Ki67 LI was classified as low (<14%) or high (≥14%). We used multivariable logistic regression to examine associations between Ki67, breast cancer subtype, and histological grade, controlling for race/ethnicity and age at diagnosis in all models.

In adjusted models, the proportion of tumors with a high Ki67 LI was much lower for luminal A (28%) than for luminal B, HER2, TN-US, and TN-BL subtypes (91,66, 77 and 89%, respectively, p<0.0001). The proportion of tumors that were high histologic grade was also much lower for luminal A (29%) than for luminal B, HER2, TN-US, and TN-BL subtypes (85, 86, 94 and 68%, respectively, p<0.0001). Before accounting for subtype, 68% of high Ki67 tumors were high grade, compared with only 26% of low Ki67 tumors (p<0.0001); after controlling for breast cancer subtype these percentages were 58% and 37%, respectively, p=0.001).

In this multi-ethnic sample of breast cancer patients, we found that Ki67 was a significant independent predictor of more aggressive, higher grade disease, even after accounting for molecular subtype. Ki67 might serve as a useful additional marker for the classification of breast cancer into molecular subtypes.

Citation Format: Abeer M. Mahmoud, Virgilia Macias, Umaima Al-alem, Jacob K. Kresovich, Galina Khramtsova, Andre Kajdacsy-Balla, Elizabeth L. Wiley, Garth H. Rauscher. Ki67 is an independent prognostic marker in breast cancer even after accounting for molecular subtype. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5569. doi:10.1158/1538-7445.AM2014-5569
Abstract 5571: Genome-wide crosstalk of estrogen and progesterone signaling in breast cancer
Authors: Singhal Hari , Greene Geoffrey
Published: May 2015

Estrogen receptor alpha (ER) and progesterone receptor (PR) are established biomarkers in breast cancer and receptor-targeted therapies are routinely used in breast cancer treatment. Estrogen receptor exists as two subtypes, ERα and ERβ, and PR has two isoforms PRA and PRB. Furthermore, millions of women worldwide use estrogen and progesterone formulations as oral contraceptives and hormone replacement therapies. Although critical crosstalk occurs between estrogen and progesterone signaling, the global nature and mechanistic details of this interaction are not known. Our central hypothesis is that there is genome-wide dominance of progesterone over estrogen signaling in breast cancer. This hypothesis is based on our novel findings that gene expression and proliferation of T47D and ZR75 breast cancer cells upon co-treatment with estrogen and progesterone is similar to progesterone-only treatment and is significantly different from estrogen-only treatment. Furthermore, four derivative T47D sublines that lack either of the PR isoforms, both the isoforms of PR or ER along with PR are being used to study ERα-, PRA- and PRB-specific regulatory networks. ER- and PR-regulated gene networks are defined by correlating ER and PR genome binding data obtained from ChIP-seq with gene expression data of T47D cells acquired using RNA-seq. For instance, genes that are known to be important in breast carcinogenesis, such as GATA3, GREB1, PDZK1, FOXA1 are found to be co-regulated by ER and PR. Subsequently the intersection of identified ER- and PR-regulated gene networks will be studied and their clinical value will be assessed. Transcription cofactors FOXA1 and NF1C are critical for transactivation by ER and PR. Knockdown of these cofactors in T47D cells alters the ER/PR crosstalk in these cells. To validate the clinical value of genes co-regulated by ER and PR in breast cancer, meta-analysis for their enrichment in established breast cancer signatures will be done. Cluster analysis of the expression of these genes in patient tumor samples that differ in clinical outcomes will also be performed. Finally, a Kaplan-Meier survival analysis will be done to study a correlation between expression of these genes and patient survival. These innovative studies will determine the global influence of progesterone over estrogen signaling in breast cancer. The resulting map of ER- and PR-regulated gene networks will lead to the optimization of breast cancer treatments, hormonal contraceptives and hormone replacement therapies.

Citation Format: Hari Singhal, Geoffrey Greene. Genome-wide crosstalk of estrogen and progesterone signaling in breast cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5571. doi:10.1158/1538-7445.AM2014-5571
Abstract 5580: TGF-β1 enhanced cell migration involving pluripotent transcription factor OCT4 in endometriosis
Authors: Chang Te-Sheng , Au Heng-Kien , Ling Thai-Yen , Chi Ching-Chi , Tzeng Chii-Ruey , Chang Jui-Hung , Huang Yen-Hua
Published: May 2015

Endometriosis is the eutopic and/or ectopic growth of endometrial tissues. High levels of TGF-β in peritoneal fluid and increased expression of OCT4 have been well documented in endometriosis. However, the molecular mechanism between TGF-β and OCT4 expression in endometriosis still remains largely unknown. In this study, we examined the roles of OCT4 in TGF-β-induced endometrial cell migration and the progression of human ectopic endometriosis. We analyzed the plasma levels of TGF-β (n = 80) and gene expression levels of TGF-βR1 in patient tissues (n = 60) by an ELISA assay and quantitative real-time RT-PCR. The serum TGF-β and tissue TGF-βR1 expressions were significantly increased in the ectopic endometriotic tissues (n = 54, 29 for adenomyosis tissues and 25 for chocolate cysts) comparing with the normal endometrium (n = 5). The TGFβR1 transcriptional level in endometriotic tissues was positively correlated with expressions of OCT4 and genes associated with cell migration, such as VIMENTIN, TWIST, SNAIL and SLUG. TGF-β dose-dependently increased the OCT4 expression both in gene and protein levels in human endometriotic stromal cells and, at a less intensity, in HEC1A endometrial carcinoma cell line. Additionally, TGF-βincreased the expression of migration-associated genes and proteins (SNAIL and N-CAD) dose-dependently both in gene and protein levels in human endometriotic stromal cells. Knockdown of OCT4 significantly suppressed the TGF-β-induced the expressions of migration-associated genes and proteins (N-CAD and SNAIL), and promoted the migration of endometrial cells as evidenced by wound-closure and transwell assays. Conclusions: TGF-β,TGF-βRI, and OCT4 are significantly upregulated in human ectopic endometriotic tissues. The significant OCT4 expression may be involved in the pathology of ectopic endometrial growth through TGF-β-induced migration of endometrial cells.

Citation Format: Te-Sheng Chang, Heng-Kien Au, Thai-Yen Ling, Ching-Chi Chi, Chii-Ruey Tzeng, Jui-Hung Chang, Yen-Hua Huang. TGF-β1 enhanced cell migration involving pluripotent transcription factor OCT4 in endometriosis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5580. doi:10.1158/1538-7445.AM2014-5580
Abstract 5590: Molecular characterization of pancreatic tumors arising in the background of germline BRCA mutations
Authors: Hsiao Susan , Gonzalez Anne Koehne de , Mansukhani Mahesh M. , Fine Robert L. , Remotti Helen E.
Published: May 2015

Background: BRCA1 and BRCA2 are tumor suppressor genes which play a role in the repair of DNA double stranded breaks and the maintenance of genomic stability. Inherited BRCA mutations increase the risk of development of pancreatic cancer. In sporadic tumors BRCA mutations occur late in pancreatic tumorigenesis. Few studies to date have examined molecular heterogeneity within pancreatic tumors with respect to founder and progressor mutations. This is the first study to examine heterogeneity of mutations in pancreatic carcinomas associated with germline BRCA mutations.

Design: Cases of pancreatic tumors arising in four patients with previously confirmed germline BRCA1 or BRCA2 mutations were identified from our case files. Representative sections of formalin-fixed, paraffin embedded tissue from surgical resection specimens were reviewed and 3 sections of tumor from different geographic regions and 1 section of non-tumor tissue were selected for molecular analysis for each case. Samples were manually macrodissected and genomic DNA was isolated. Genomic DNA was used for target capture and amplification of 212 amplicons targeting mutational hotspots in 48 genes using the TruSeq Amplicon Cancer Panel. Next generation sequencing was performed on the MiSeq Benchtop Sequencer and analyzed with NextGENe software.

Results: In all cases concordant results were observed in all three tumor samples, with no evidence of intratumoral heterogeneity for the genes tested. In one patient with a BRCA1 germline mutation, TP53 and SMAD4 mutations were identified. In two patients with germline BRCA2 mutations, two additional driver mutations were identified involving KRAS, TP53, or SMAD4. In one case with BRCA2 germline mutation, no additional mutations were identified.

Conclusion: Pancreatic tumors arising in the background of germline BRCA mutations had between 1-3 driver mutations per tumor, which is consistent with the reported numbers of driver mutations in sporadically arising pancreatic tumors. No mutual exclusivity among the mutated genes was observed. Mutations observed include KRAS codon 12 mutation, TP53 mutation (within the DNA binding domain), and SMAD4 mutation. Taken together, these results suggest that pancreatic tumors occurring in BRCA patients show similar mutational profiles of key oncogenes and tumor suppressor genes as seen in sporadic tumors with little heterogeneity with respect to these driver mutations.

Citation Format: Susan Hsiao, Anne Koehne de Gonzalez, Mahesh M. Mansukhani, Robert L. Fine, Helen E. Remotti. Molecular characterization of pancreatic tumors arising in the background of germline BRCA mutations. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5590. doi:10.1158/1538-7445.AM2014-5590
Abstract 5599: Serum biomarkers reflecting specific tumor tissue remodeling is a valuable diagnostic tool for lung cancer
Authors: Willumsen Nicholas , Bager Cecilie L. , Leeming Diana J. , Smith Victoria , Christiansen Claus , Karsdal Morten A. , Dornan David , Bay-Jensen Anne-Christine
Published: May 2015

Background: The extracellular matrix (ECM) that integrates and surrounds malignant tumors consists of several proteins, amongst other type I collagen and elastin. These structural proteins, as well as intermediate filament (IMF) proteins like vimentin, are remodeled and dysregulated as part of malignant changes. We investigated whether serum biomarkers reflecting ECM and IMF remodeling/dysregulation could be used to differentiate lung cancer patients from healthy controls, other lung pathologies, and other cancer types.

Methods: Levels of matrix metalloproteinase (MMP) generated fragments of type I collagen (C1M), of elastin (ELM) and of citrullinated vimentin (VICM) were measured in serum from patients with lung cancer (n=40), gastrointestinal cancer (n=25), prostate cancer (n=14), malignant melanoma (n=7), chronic obstructive pulmonary disease (COPD) (n=13), and idiopathic pulmonary fibrosis (IPF) (n=10), as well as in age-matched controls (n=33). The area under the receiver operating characteristics (AUROC) was calculated for the biomarkers and a diagnostic decision tree was generated from specific cut-off values.

Results: Circulating levels of C1M and VICM were significantly elevated in lung cancer patients as compared to healthy controls (AUROC=0.98, p<0.0001) and other cancers (AUROC=0.83 p<0.0001). A trend was detected when comparing lung cancer with COPD+IPF. No difference could be seen for ELM. Interestingly, C1M and VICM were able to identify patients with lung cancer with a positive predictive value of 0.9 and an odds ratio of 40 (95% CI=8.7-186, p<0.0001).

Conclusion: This study indentifies biomarkers specifically reflecting ECM and IMF remodeling as applicable for lung cancer patients. We speculate that these markers may aid in diagnosing and characterizing patients with lung cancer.

Citation Format: Nicholas Willumsen, Cecilie L. Bager, Diana J. Leeming, Victoria Smith, Claus Christiansen, Morten A. Karsdal, David Dornan, Anne-Christine Bay-Jensen. Serum biomarkers reflecting specific tumor tissue remodeling is a valuable diagnostic tool for lung cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5599. doi:10.1158/1538-7445.AM2014-5599
Abstract 5600: Wntless (GPR177) regulates WNT/NF-кB signaling in leukemia cells, and its expression correlates with poor prognosis in B-cell precursor acute lymphoblastic leukemia
Authors: Hsu Shih-Hsien , Wang Li-Ting , Huang Shih-Bo , Chiou Shyh-Shin
Published: May 2015

B-cell precursor acute lymphoblastic leukemia (BCP ALL) is the most common childhood leukemia, with a cure rate of 80%. Nevertheless, disease relapse is the most important prognostic factor for the disease outcome. We aimed to elucidate the role of Wnt secretion-regulating protein, Wntless (Wls)/GPR177, on disease outcome in pediatric patients with BCP ALL, and assess its pathogenetic role in the regulation of the disease. Wls expression was characterized and correlated with Wnt and NF-κB pathway signaling in the bone marrow leukemia cells isolated from 44 pediatric patients with BCP ALL. The overexpression of Wls was detected in leukemia cells and was significantly correlated with the disease relapse and poor survival in the patients. The high expression of Wls also correlated with the Wnt expression, particularly with the NF-κB signaling activation. The Wnt-activated NF-κB signaling induced by Wls was shown to provide essential proliferation, transformation, and anti-apoptotic activity during leukemogenesis. These results indicated that Wls played an essential role in disease relapse and poor survival in patients with BCP ALL. Therefore, Wls may provide a potential future therapeutic target, particularly for patients who do not respond to existing therapies and suffer relapse.

Citation Format: Shih-Hsien Hsu, Li-Ting Wang, Shih-Bo Huang, Shyh-Shin Chiou. Wntless (GPR177) regulates WNT/NF-кB signaling in leukemia cells, and its expression correlates with poor prognosis in B-cell precursor acute lymphoblastic leukemia. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 5600. doi:10.1158/1538-7445.AM2014-5600
Abstract 566: The clinicopathological features and prognostic impact of ALK-positivity in resected gastric or gastro-esophageal cancer patients
Authors: Jeon Hongjae , Kim HyeRyun , Noh Sung Hoon , Chung Hyun Cheol , Rha Sun Young
Published: May 2015

Background: To determine the frequency and prognostic impact of ALK-rearrangements in gastric cancer patients who underwent curative surgical resection with extensive (D2) lymph node dissection.

Patients and methods: We concurrently analyzed the ALK-rearrangements and HER2 overexpression in whole tumor sections of 455 curatively resected gastric cancers. We performed immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH), and analyzed the relationship between ALK positivity and clinical survival outcome.

Results: Of the 455 tumors that were screened, 38 (8.4%) were ALK+ by IHC, which consisted of 1+ (24, 7.5%), 2+ (3, 0.7%), and 3+ (1, 0.2%), but no cases were confirmed by FISH. ALK+ patients were significantly younger (57 vs. 61 years, P=0.02) and showed similar patterns of ALK+ lung adenocarcinoma. ALK+ patients were significantly more likely to have abundant signet ring cells (defined as ≥ 10% of tumor cells). Moreover, as the ALK intensity (measured by IHC) increased, so did the density of signet ring cell in tumors (Ptrend = 0.02). Also, the disease free survival (DFS) and overall survival (OS) of ALK+ patients in stages were shorter than ALK- patients (DFS: 21.2 vs.100.5 months; P = 0.007, OS: 34.3 months vs. median survival time unattainable; P=0.016)

Conclusions: ALK+ is an independent negative prognostic factor in surgically resected gastric cancers and is associated with signet ring cell histology. Furthermore ALK+ might be a relevant therapeutic target in gastric cancer patients.

Citation Format: Hongjae Jeon, HyeRyun Kim, Sung Hoon Noh, Hyun Cheol Chung, Sun Young Rha. The clinicopathological features and prognostic impact of ALK-positivity in resected gastric or gastro-esophageal cancer patients. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 566. doi:10.1158/1538-7445.AM2014-566
Abstract 57: Blood parameters in renal cell carcinoma patients can have prognostic impact and differ from those of healthy blood donors
Authors: Goebel Steffen , Bluemke-Anbau Karen , Altermann Wolfgang , Bilkenroth Udo , Meye Axel , Fuessel Susanne , Lautenschlaeger Christine , Melchior Andres , Heynemann Hans , Fornara Paolo , Taubert Helge
Published: May 2015

The aim of this study was to evaluate the clinical relevance of blood parameters as circulating tumor cells (CTCs) and HLA subtype in renal cell carcinoma (RCC) patients.

233 peripheral blood samples from 154 RCC patients were tested for the presence of disseminated tumor cells (DTCs) by using the autoMACS technique and immunocytochemical (ICC) staining of cytokeratin. The frequency of CTCs was analyzed statistically and correlated with relevant clinical data. In addition, HLA subtype was analysed for 91 out of the 154 RCC patients and for 203 healthy blood donors. A statistical comparison was performed.

After following the CD45 autoMACS depletion protocol, we identified CTCs in 96 out of 233 peripheral blood samples (41%), which originated from 81 out of 154 (53%) RCC patients. Besides a significant correlation between the detection of CTCs before and after primary tumor resection a significant association with prognosis was found (p < 0.002; X2-test). RCC patients with CK+ CTCs in their peripheral blood had a 5 times increased risk of tumor-related death compared to patients without these cells. In a Kaplan-Meier-Analysis these results were confirmed (p < 0.001). On one hand, RCC patients had less alleles HLA-Cw*07 (p < 0.05; X2-test) and HLA-DRB1*04 (p < 0.05; X2-test) compared to healthy blood donors. On the other hand, RCC patients showed an increased occurrence of the allele HLA-Cw*12 compared to healthy blood donors. However, this difference was not significant.

In summary, for RCC patients the presence of CTCs was associated with a poor overall survival and significantly correlated with an increased risk of tumor-related death. Detection of CK+ CTCs in peripheral blood is a significant and independent prognostic factor for renal cell carcinoma patients. Occurrence of the alleles HLA-Cw*07, and HLA-DRB1*04 was significantly reduced in RCC patients compared to healthy blood donors and may reduce the risk to suffer from a RCC.

Citation Format: Steffen Goebel, Karen Bluemke-Anbau, Wolfgang Altermann, Udo Bilkenroth, Axel Meye, Susanne Fuessel, Christine Lautenschlaeger, Andres Melchior, Hans Heynemann, Paolo Fornara, Helge Taubert. Blood parameters in renal cell carcinoma patients can have prognostic impact and differ from those of healthy blood donors. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 57. doi:10.1158/1538-7445.AM2014-57
Abstract 577: Genetic and epigenetic alterations of the WWOX gene in head and neck cancer
Authors: BUYRU NUR , Ekizoglu Seda , Koseoglu Hikmet , Karaman Emin
Published: May 2015

Head and neck squamous cell carcinoma (HNSCC) is a heterogenous disease and the 6th most frequent cancer in the world. Despite advances in medical treatment over the past two decades, the overall survival rate for patients with HNSCC remains around 50%. Well known environmental risk factors for developing HNSCC are cigarette smoking, especially in combination with alcohol consumption or tobacco chewing. In addition, many genetic and epigenetic changes are associated with sporadic HNSCC including alterations in the p53, VEGF and cyclin D1genes. Yet the molecular mechanisms of HNSCC carcinogenesis are still undergoing intensive investigation.

The WWOX gene is located on a chromosomal fragile site 16q23.2 which spans the common fragile site FRA16D. It has been shown that exposure to environmental carcinogens such as smoking and alcohol consumption increases the potential for chromosomal breakage at the FRA16D site in esophageal and non-small cell lung cancers. On the other hand, genomic abnormalities affecting chromosome 16q are frequently reported in cytogenetic studies of various epithelial tumors, including prostate, breast, ovarian, esophageal, lung, gastric, and hepatocellular carcinomas. However, there is no data in the literature investigating the WWOX gene in the HNSCC. Therefore, the aim of this study was to determine the role of WWOX as a tumor suppressor gene in patients with HNSCC.

For this purpose, tumor and corresponding normal tissue samples obtained from 75 patients with HNSCC were analyzed for mutations in the coding exons and the flanking intronic sequences by direct sequencing of the WWOX gene. mRNA expression levels were also analyzed by quantitative-real time-PCR and correlated with the methylation level of the WWOX gene promoter. The methylation status was determined by methylation-specific PCR(MSP).

mRNA expression levels were significantly lower in tumor tissue than normal tissue (p≤0.001) and this downregulation was associated with hypermethylation of the promoter of the WWOX gene (p≤0.05). We identified 8 different alterations in the coding sequence of the WWOX gene with a frequency varying between 1.56-58.93%. Additionally, 17 different alterations in the noncoding regions were identified in our study group.

Our results indicate that the WWOX gene may play an important role in the development of HNSCC.

Citation Format: NUR BUYRU, Seda Ekizoglu, Hikmet Koseoglu, Emin Karaman. Genetic and epigenetic alterations of the WWOX gene in head and neck cancer. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 577. doi:10.1158/1538-7445.AM2014-577
Abstract 581: Identification of PCDHB3 as a potential tumor suppressor in colorectal cancer by exome sequencing
Authors: Ye Wen , Liu Ranyi , Wu Jiangxue , Li Yingchang , Huang Wenlin
Published: May 2015

Colorectal cancer (CRC) is one of the most common cancers worldwide and is believed to arise from combinations of genetic and epigenetic disorders. However, besides APC, KRAS, and p53, which are highly targeted by somatic mutations, other genes altered in mutations in CRC patients are under interpreted in their effects and relevance with CRC tumorigenesis.

Out of the genes in which we identified 453 nonsynonymous somatic mutations in this exome sequencing study, PCDHB3 was mutated in 30% exome-sequenced cases and was screened in another 104 CRC cases with a 7.02% mutation rate. Meanwhile, PCDHB3 is lowly expressed in 83.7% tumor of CRC patients and CRC cell lines compared to normal controls, which is detected to be caused by loss of heterozygosity and CpG island hypermethylation. In vitro and in vivo experiments revealed that over-expression of wildtype PCDHB3 in DLD1 cells inhibited tumor cell proliferation and migration (56.8% and 47.5% of control), whereas mutant PCDHB3 lacked of the functions. Furthermore, SW480-shRNA cells, which demonstrated a lower level of endogenous PCDHB3, exhibited a larger growth and migration advantage (2.28-fold and 2.26-fold of control).

In summary, our study identified PCDHB3 as a tumor suppressor gene in CRC. Loss of PCDHB3 suppressor function results from uncommon genetic mutations affecting the protein sequence as well as decreased expression of functional protein.

Citation Format: Wen Ye, Ranyi Liu, Jiangxue Wu, Yingchang Li, Wenlin Huang. Identification of PCDHB3 as a potential tumor suppressor in colorectal cancer by exome sequencing. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 581. doi:10.1158/1538-7445.AM2014-581
Abstract 586: Prognostic significance of Tip60 expression in human glioma
Authors: Bai Rongpang , Weng Yu , Zhao Zhongsheng , Bai Jin , Chen Guangdi
Published: May 2015

The tumor suppressor Tip60 plays a major role in transcription, DNA damage response, apoptosis, autophagy and cancer development, but its role in glioma is unknown. In this study, we investigated the role of Tip60 in glioma pathogenesis and assessed its prognostic value. Using tissue microarrays consisting of 437 cases of glioma, including 236 cases with low grade (grade I and II) and 201 cases with high grade (grade III and IV), we found that Tip60 expression was significantly reduced in high grade glioma compared to low grade glioma (P = 0.004) although no significant difference in Tip60 expression was found between grade I and II, or grade III and IV glioma (P > 0.05 for either). Kaplan-Meier survival curve analysis showed that reduced Tip60 expression was associated with a poorer five-year overall survival in glioma patients (P = 0.015). Our data indicate that Tip60 may serve as a potential biomarker for glioma patient outcome as well as a potential therapeutic target.

Citation Format: Rongpang Bai, Yu Weng, Zhongsheng Zhao, Jin Bai, Guangdi Chen. Prognostic significance of Tip60 expression in human glioma. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 586. doi:10.1158/1538-7445.AM2014-586
Abstract 588: Lrig1 suppresses breast cancer stem cell expansion and promotes mesenchymal to epithelial transition
Authors: Yokdang Nucharee , Sweeney Colleen
Published: May 2015

Leucine-rich repeats and immunoglobulin-like domains-1 (Lrig1) is known as a negative regulator of oncogenic receptor tyrosine kinases (ErbB, Met and Ret); and low Lrig1 expression correlates with poor patient prognosis in breast cancer. Lrig1 has been found to maintain stem cell quiescence and marks a subset of quiescent intestinal stem cells where its loss leads to heightened ErbB expression and duodenal adenomas. Lrig1 is defined as a tumor suppressor in healthy tissue such as intestine and epidermis where Lrig1 loss leads to stem cell expansion and epidermal hyperplasia. However, the role of Lrig1 in cancer stem cells (CSCs) and as a metastatic suppressor is unknown. Lrig1 expression is lowest in the highly invasive breast cancer subtype MDA-MB-231, and Lrig1 expression is highest in the weakly invasive luminal A breast cancer subtype. In patients, high Lrig1 expression exhibits longer relapse-metastatic free survival suggesting that Lrig1 expression protects against metastatic relapse. We hypothesized that Lrig1 suppresses breast CSC expansion and promotes mesenchymal to epithelial transition. Our results indicated that restoration of Lrig1 in highly aggressive cell lines promoted mesenchymal to epithelial transition (MET) and a loss of invasive morphology in 3D matrigel cell culture. These results demonstrated that Lrig1 suppressed cell proliferation in both 2D and 3D matrigel cultures and suppressed tumorsphere formation 10-fold. Importantly, we have shown that Lrig1 is down-regulated in the hTERT-immortalized mammary epithelial cell line-twist ER, (HMLE-twist ER) which undergoes EMT. When Lrig1 is knocked-down in HMLE-twist ER by shRNA, the results showed that loss of Lrig1 enhanced mesenchymal and stem cell expansion. These observations suggest that Lrig1 may play a role in EMT transition in metastasis, as well as providing a whole new area of therapeutic targets in breast cancer along with an improved basic understanding in cancer stem cell biology.

Note: This abstract was not presented at the meeting.

Citation Format: Nucharee Yokdang, Colleen Sweeney. Lrig1 suppresses breast cancer stem cell expansion and promotes mesenchymal to epithelial transition. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 588. doi:10.1158/1538-7445.AM2014-588
Abstract 592: HOXD10 suppresses mammary tumor growth and metastasis coincident with decreased CD11b+Gr1+ leukocyte infiltration and angiogenesis
Authors: Kachgal Suraj , Chen Amy , Sidani Mazen , Afghani Shahrzad , Boudreau Nancy J.
Published: May 2015

The tumor suppressor gene HOXD10 is frequently lost in breast cancer, and we have previously shown that restoring expression of this master transcriptional regulator can revert mammary tumor cells to a normal, polarized phenotype and inhibit their growth in nude mice (Carrio et al., Cancer Res., 2005). In addition, we showed that restoring HOXD10 also reduced VEGFA expression and decreased endothelial cell migration in vitro (Chen et al., Cancer Res., 2009). Considering the critical role of the immune system in mammary tumor progression and metastasis, we investigated whether Hoxd10 could also prevent breast tumor progression in an immunocompetent model. Metastatic 4T1 cell lines were transduced with Hoxd10 and were either orthotopically or intravenously injected into four to eight week old, female Balb/c mice. We observed a significant reduction in tumor burden at both primary at metastatic sites in tumors expressing Hoxd10. Immunohistochemical staining for CD31 of primary tumors expressing Hoxd10 revealed decreased angiogenesis. Flow cytometry and immunohistochemical staining also revealed a decrease in infiltrating Cd11b+Gr1+ leukocytes in Hoxd10-expressing tissues. Gene and protein analysis revealed that in addition to decreased VEGFA, HOXD10 also reduced expression of CCL2, a key chemokine linked to recruitment of Cd11B+Gr1+ leukocytes. Using an in vitro migration assay, we confirmed that the HOXD10-mediated reduction in CCL2 expression was directly linked to reduced recruitment of monocytes to tumor cells. Together, these data show that HOXD10 can stabilize the tumor microenvironment by coordinately reducing tumor cell growth, angiogenesis, leukocyte recruitment, and metastasis, and suggest a novel and comprehensive therapeutic approach to treat mammary tumors.

Citation Format: Suraj Kachgal, Amy Chen, Mazen Sidani, Shahrzad Afghani, Nancy J. Boudreau. HOXD10 suppresses mammary tumor growth and metastasis coincident with decreased CD11b+Gr1+ leukocyte infiltration and angiogenesis. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 592. doi:10.1158/1538-7445.AM2014-592
Abstract 593: Rb association with Bax is regulated by phosphorylation
Authors: Antonucci Lisa , Egger Jacklynn V. , Krucher Nancy A.
Published: May 2015

The Retinoblastoma protein (Rb) is important in the control of cell proliferation and apoptosis. Its activity is controlled by reversible phosphorylation on several serine and threonine residues that mediate binding to various cellular proteins. The role of Rb phosphorylation in the control of apoptosis is largely unknown, although several apoptotic stimuli result in dephosphorylation of Rb. Our previous work identified Rb (threonine-821) dephosphorylation as required for induction of apoptosis caused by UV light and inhibitions of cyclin dependent kinases. These studies led to the identification of proteins that bind to Rb under apoptotic conditions. One such Rb binding protein is the pro-apoptotic protein Bax. Bax, a member of the Bcl-2 family, undergoes a conformational change during apoptosis that is associated with the permeabilization of the outer mitochondrial membrane. In this study, we examined the role of Rb phosphorylation in the regulation of binding to Bax. In MCF7 cells, by activating the phosphatase responsible for dephosphorylating Rb, PP1, apoptotic death is triggered. We utilize siRNA to block the expression of PNUTS (Phosphatase Nuclear Targeting Subunit) which causes Rb dephosphorylation, and cell death. We have found that Bax binds to Rb under normal cell conditions, but when Rb becomes dephosphorylated on specific amino acid sites, Bax dissociates from Rb. Our cell fractionation studies have shown that phosphorylated Rb co-localizes with the mitochondrial fraction. Further examination of the specific sites of Rb phosphorylation required for binding to Bax is underway.

Citation Format: Lisa Antonucci, Jacklynn V. Egger, Nancy A. Krucher. Rb association with Bax is regulated by phosphorylation. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 593. doi:10.1158/1538-7445.AM2014-593