bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2514: CSPG4 as a target of antibody-based immunotherapy for malignant mesothelioma
Authors: Zeyana Rivera , Ferrone Soldano , Wang Xinhui , Sandro Jube , Yang Haining , Gaudino Giovanni , Carbone Michele
Published: August 2012

Malignant mesothelioma (MM) is an aggressive tumor, resistant to conventional therapies, with median survival of 1 year from diagnosis. The lack of effective therapies available prompted us to implement antibody-based immunotherapy against novel targets and to investigate possible synergies with chemotherapy on MM. CSPG4 is involved in the onset and progression of melanoma and other tumors, as well as in neo-angiogenesis. CSPG4-specific monoclonal antibodies (mAb) have shown anti-tumor effects in both preclinical and clinical trials of melanoma. We hypothesized that targeting CSPG4 with a CSPG4-specific mAb would have therapeutic efficacy against MM. We found that chondroitin sulphate proteoglycan 4 (CSPG4) was aberrantly expressed in 6 out of 8 MM cell lines and in 24 out of 40 MM biopsies, with minimal expression in surrounding non-malignant cells. The expression of CSPG4 in MM was higher upon engagement of extracellular matrix components (ECM) and was associated with increased MM cell viability and motility. Silencing of CSPG4 expression by siRNAs supported this functional significance of CSPG4 in MM. Consistently, the CSPG4-specific mAb TP41.2 inhibited MM cell attachment to the ECM, resulting in decreased phosphorylation of FAK and AKT, decreased expression of cyclin D1 and apoptosis. Moreover, TP41.2 significantly reduced MM cell motility, migration and invasiveness, and inhibited MM growth in soft agar. TP41.2 prevented the outgrowth of human MM tumors in SCID mice and inhibited the growth of established MM xenografts, resulting in significantly extended survival of tumor-bearing mice. These results represent a novel approach for CSPG4 mAb-based immunotherapy of MM. Combinatorial strategies with chemotherapeutic agents and with mAbs targeting c-Met related receptor are ongoing, aimed at translating CSPG4-specific mAb into clinical setting.

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 2514. doi:1538-7445.AM2012-2514
Abstract 2527: Antibody-dependent cellular cytotoxicity of cetuximab against colorectal cancer cells with wild-type and mutant KRAS
Authors: Nakadate Yusuke , Kitamura Yuka , Tamura Tomohide , Koizumi Fumiaki
Published: August 2012

Cetuximab is a chimeric IgG1 monoclonal antibody (mAb) that targets the extracellular domain of epidermal growth factor receptor (EGFR). In addition to direct EGFR inhibition, antibody-dependent cellular cytotoxicity (ADCC) is considered to be an important mechanism of action of cetuximab. Recently, KRAS mutations in tumors have been shown to be a negative predictive factor for response to cetuximab treatment for colorectal cancer (CRC). KRAS activating mutations constitutively transduce RAS/RAF/MEK/ERK pathway signaling, which theoretically cannot be suppressed by inhibition of the upstream receptor through the use of cetuximab. Thus, KRAS mutations potentially contribute to the resistance to the direct inhibitory effect of cetuximab. However, the influence of KRAS mutations on cetuximab induced ADCC is not fully understood. In this study, we investigated cetuximab-mediated ADCC against human CRC lines, HCT116 and DLD-1, which carry mutated KRAS, and their derivative cell lines, HKh2 and DKO-4 respectively, in which the mutated KRAS allele was deleted by targeted disruption (KRAS wild-type). Peripheral blood mononuclear cells (PBMCs) from healthy volunteers and NK92, a natural killer cell line, exogenously expressing Fcα3A (CD16a) were used as effector cells. We first determined the growth-inhibitory effect of cetuximab alone using a WST-8 assay. Growth inhibition was observed only in KRAS wild-type cell lines, but its effect was only slight. In a standard short-term (4hr) ADCC assay that mainly evaluated lytic activity via perforin/granzyme, ADCC activities were exhibited to a similar extent against CRC cell lines regardless of the KRAS mutation status. We also examined a long-term (24hr) ADCC assay under the perforin-inhibited condition to evaluate the death receptor ligand-induced apoptosis, which is another major mechanism of ADCC. In this assay, cetuximab-mediated ADCC was induced in wild-type but not mutant KRAS cell lines. Similar to a previous report, Hkh2 and DKO-4 displayed a higher sensitivity to recombinant human Fas ligand (FasL) and TRAIL than did HCT116 and DLD-1, respectively. Furthermore, blocking experiments with neutralizing antibodies revealed that induction of apoptosis via Fas-FasL interaction is involved in perforin-independent cetuximab-mediated ADCC against KRAS wild-type cells. In conclusion, these results suggest that mutated KRAS may contribute to the resistance to the antitumor effects of cetuximab not only through direct receptor inhibition but also through ADCC mediated by Fas-FasL interaction.

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 2527. doi:1538-7445.AM2012-2527
Abstract 2529: A monoclonal anti-Globo H antibody, VK9 can mediate CDC/ADCC and inhibit adhesion of Globo H+ cancer cells to extracellular matrix
Authors: Hung Jung-Tung , Cheng Jing-Yan , Wu Chia-Cheng , Yu Alice L.
Published: August 2012

Globo H is a hexasaccharides expressed on many carcinomas including breast, pancreas, prostate, and stomach, with limited expression on normal tissue. Monoclonal antibodies against glycan, such as CH14.18 GD2-specific antibody, have been proven to be a promising strategy for cancer therapy. Here we evaluated the biological activity and potential application of VK9, a murine IgG3 anti-Globo H antibody. FACS analysis showed that VK9 bound to MCF7 cells in a dose dependent manner at 0.01 - 30 μg/mL. It does not bind to GH (-) cell lines, including AU565 and MDA-MB-231. The specificity of VK9 as determined by glycan array of 87 printed carbohydrates revealed binding of VK9 to three glycans (Globo H, BB4, and BB3) only, without reactivity to Gb5 or sialyl-Gb5. Investigation of the biological activity of VK9 showed no obvious effects on the proliferation of MCF7 cells. However, VK9 inhibited adhesion of MCF7 cell on fibronectin (83.5 ± 1.3%, p = 0.001) or laminin (70.5 ± 2.8%, p = 0.001), but not BSA (85.6 ± 0.2%, p = 0.08) or collagen IV (105.1 ± 2%, p = 0.37), when compared to IgG3 isotype control. Moreover, VK9 was able to mediate CDC against MCF7 cells. It could also induce ADCC of human IL-2 activated peripheral blood mononuclear cells at an E/T of 6.25-100. Furthermore, we cloned the Fab region of VK9 to generate a mouse-human chimeric anti-Globo H antibody, cVK9. The binding affinity of cVK9 was similar to VK9 at concentration range of 6.25 - 50 ug/mL by ELISA analysis. In conclusion, VK9 could not only mediate ADCC and CDC but also inhibit adhesion of MCF7 cell on fibronectin and laminin. Thus, VK9 have the potential as therapeutic antibody for Globo H expressing carcinoma.

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 2529. doi:1538-7445.AM2012-2529
Abstract 2530: Long-term tumor regression induced by a novel antibody drug conjugate that targets 5T4, an oncofetal antigen expressed on tumor-initiating cells
Authors: Sapra Puja , DiJoseph John , Damelin Marc , Dougher Maureen , Narayanan Bitha , Khandke Kiran , Lucas Judy , Golas Jon , Tchistiakova Lioudmila , Leal Mauricio , Hu George , Maderna Andreas , Marquette Kimberly , Loganzo Frank , Dushin Russell , O'Donnell Christopher J. , Abraham Robert , Gerber Hans-Peter
Published: August 2012

Antibody-drug conjugates (ADCs) represent a promising therapeutic modality for the clinical management of cancer. We have developed a novel ADC that targets 5T4 (also known as TPBG), a tumor-associated antigen that rapidly internalizes. In preclinical models of NSCLC, 5T4 was identified as a marker of undifferentiated tumorigenic cells that express properties of tumor-initiating cells (TICs) and was associated with a highly proliferating (Ki67 positive) cell phenotype. Based on the biological properties of 5T4, we designed the ADC by conjugating an internalizing humanized anti-5T4 antibody (Ab), A1, to the tubulin inhibitor monomethylauristatin F via a maleimidocaproyl (mc) linker (A1mcMMAF) with a drug/antibody ratio of ∼ 4. The A1 Ab has a Kd of 0.9 nM against the 5T4 antigen, binds to a broad range of 5T4 expressing cell lines, and rapidly internalizes (66% internalized within 4h). As an ADC, A1mcMMAF retains similar properties to the unconjugated Ab. In vitro, A1mcMMAF exhibited potent and target-specific cytotoxic activity against a panel of 5T4+ cell lines with an IC50 of 5 ng Ab/ml against high 5T4 expressing cells. In vivo, A1mcMMAF exhibited potent anti-tumor activity in all five tumor models tested (with a broad range of 5T4 expression) and caused long-term regressions monitored up to 100 days after the last dose. Strikingly, animals were cured of disease in every model with doses as low as 3 mg Ab/kg Q4d x 4. In a lung cancer patient-derived xenograft in which 5T4 is not uniformly expressed, but instead is preferentially expressed on the less differentiated sub-population of tumor cells, A1mcMMAF treatment resulted in sustained tumor regression. This result highlights the clinical potential of ADCs that target aggressive cell subpopulations such as TICs. In exploratory safety studies, A1mcMMAF exhibited no overt toxicities when administered at 10 mg Ab/kg/cycle x 2 to cynomolgus monkeys and had a half-life of ∼5 days. The encouraging efficacy and safety data of the conjugate resulting in high therapeutic index in preclinical models warrants further clinical testing of this ADC.

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 2530. doi:1538-7445.AM2012-2530
Abstract 2532: Integrated therapeutic antibody development at the National Research Council of Canada
Authors: Jaramillo Maria Luz , O'Connor-McCourt Maureen
Published: August 2012

Advances in genomics and antibody engineering have enabled the development of an innovative class of targeted therapies to provide new treatment options for diseases with significant unmet medical needs such as cancer. Therapeutic antibodies represent one of the fastest growing classes of medications with sales anticipated to exceed $30 billion globally by 2012. The NRC has built a chain of cutting edge technology platforms needed to discover, engineer and produce therapeutic monoclonal antibodies with the goal of partnering with industrial and academic centers to advance research and development of this important class of therapeutics. Target Identification- To establish and validate the technology platform, tumor targets for candidate therapeutic antibody production were identified using a combination of proteomics, transcriptomics and bioinformatic approaches. Out of these lists, approximately 40 tumor targets were selected (known therapeutic antibody targets were excluded), and over 2,000 antibodies of mouse and camelid origin were then generated against these targets. Antibody generation- Once identified, the recombinant target protein of interest can be produced using the NRC's high efficiency cell expression platforms in CHO or HEK293 cells and purified protein used for immunization or panning. For targets which are difficult to express or purify, the capabilities for direct immunization with plasmid DNA constructs provides another option. Clone selection is carried out by ELISA and typically 50 antibodies/target are identified for further characterization. Antibody characterization and validation- The affinities of the antibodies are determined by SPR biosensor analysis. Reverse phase protein arrays and Western blot analysis on protein mixes and cell lines allow the characterization of the specificity of the antibodies. Epitope mapping can be carried out so that representative antibodies from each epitope bin can be selected for further validation in appropriate cell-based assays (many of which are established at NRC) and animal models. Therapeutic antibody Optimization, Bioprocessing and Biomanufacturing- Therapeutic antibodies selected for development can be further optimized using antibody engineering technologies to humanize them and/or modify their glycosylation patterns to improve their effector function, pharmacokinetics, solubility and stability as well as reduce their immunogenicity. The NRC platform for large scale protein production has the capacity to manufacture up to 13.5 g of commercial grade antibody using serum free, low endotoxin media in a cGMP certified CHO cell line which is ready for transfer to CMOs or other industrial partners.

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 2532. doi:1538-7445.AM2012-2532
Abstract 2536: VEGF as a potential survival factor for keratinocytes following UV exposure in vivo
Authors: Nye Kelly E. , Lachey Monica K. , Parent Allison E. , Wilgus Traci A.
Published: August 2012

Non-melanoma skin cancer (NMSC) is the most prevalent type of cancer. These cancers, including basal cell carcinoma (BCC) and squamous cell carcinoma (SCC), are primarily caused by chronic exposure to ultraviolet light (UV) from the sun. Vascular endothelial growth factor (VEGF) is produced by the skin in response to UV and is known to promote NMSC indirectly through the induction of angiogenesis. Evidence is emerging that VEGF can also promote skin carcinogenesis by directly affecting epidermal keratinocytes and tumor cells. In addition, epidermal keratinocytes express VEGFR-1 and proliferate and migrate directly in response to VEGF. In endothelial cells, VEGF functions as a potent survival factor by inhibiting apoptosis; however, it is not known if VEGF promotes survival in keratinocytes. Keratinocytes undergo apoptosis in response to UV exposure, a process that can be disrupted during carcinogenesis. Here, the role of VEGF and VEGFR-1 in keratinocyte and SCC-13 tumor cell survival after UV exposure was explored. Neutralizing VEGF or VEGFR-1 activity or adding VEGF did not affect UV-induced apoptosis, suggesting that VEGF-VEGFR-1 signaling is not important for keratinocyte or SCC-13 survival in vitro. The role of VEGFR-1 in keratinocyte survival was also examined in vivo. Mice with VEGFR-1-deficient keratinocytes (K14-Cre/VEGFR-1fl/fl) were generated and exposed to a single acute dose of UV light. Keratinocytes in the epidermis of K14-Cre/VEGFR-1fl/fl mice showed a significant increase in apoptosis 24 hours following UV exposure compared to K14-Cre/VEGFR-1+/+ controls. Future studies will investigate skin carcinogenesis in K14-Cre/VEGFR-1fl/fl mice chronically exposed to UV light. Overall, these results suggest that VEGFR-1 mediates the survival of keratinocytes exposed to acute UV in vivo and points to a mechanism by which VEGF could directly promote skin carcinogenesis.

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 2536. doi:1538-7445.AM2012-2536
Abstract 2537: UVB irradiation induces nuclear translocation of TC-PTP in keratinocytes
Authors: Lee Young S. , Smith Liza Morales , Slaga Thomas J. , DiGiovanni John , Kim Dae J.
Published: August 2012

Ultraviolet B radiation (UVB) can cause DNA damage, induce gene mutations and modulate intracellular signal transduction contributing to the development of skin cancer. Signal transducer and activator of transcription 3 (Stat3) modulates various physiological functions, including apoptosis, cell cycle regulation, and tumor angiogenesis through regulation of gene expression, and its constitutive activation is associated with human epithelial cancers. Recent studies demonstrated that T-cell protein tyrosine phosphatase (TC-PTP) dephosphorylates activated Stat3 in keratinocytes after UVB irradiation. In the current work, we report that TC-PTP translocates from the cytosol to the nucleus and dephosphorylates Stat3 following UVB irradiation in keratinocytes. Western blot analysis showed that TC-PTP mainly localized in the cytoplasm of keratinocytes and translocated from cytoplasm to nucleus upon UVB irradiation. Immunofluorescence analysis of the exogenous expression of TC-PTP confirmed its nuclear translocation, and showed that nuclear translocation of TC-PTP occurs independently of its phosphatase activity. Nuclear translocation of TC-PTP was increased with increased dose of UVB exposure. Dephosphorylation of Stat3 by TC-PTP was enhanced after UVB irradiation of cultured keratinocytes with nuclear accumulation of TC-PTP. Knockdown of TC-PTP with siRNA suppressed dephoshorylation of Stat3 and enhanced cell viability in response to UVB irradiation. UVB irradiation also induced expression of 14-3-3α, a protein that has been shown to be involved in the transport of proteins. Further studies revealed that UVB-induced nuclear translocation of TC-PTP was effectively inhibited by knockdown of 14-3-3α, suggesting 14-3-3α is involved in nuclear import of TC-PTP. These results indicate that TC-PTP plays an important role as a regulator of Stat3 signaling in keratinocytes upon UVB irradiation. Nuclear translocation of TC-PTP is regulated by a 14-3-3α-dependent signaling pathway, resulting in the rapid dephosphorylation of activated Stat3 in the nucleus.

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 2537. doi:1538-7445.AM2012-2537
Abstract 2591: Caffeic acid phenethyl ester (CAPE) suppresses melanoma tumor growth by inhibiting AKT/XIAP pathway
Authors: Pramanik Kartick C. , Kudugunti Shashi , Moridani Majid , Srivastava Sanjay K.
Published: August 2012

The incidence of melanoma is constantly on the rise indicating that current available therapies are not very effective. Moreover, melanoma is highly metastatic and resistant to chemotherapeutic drugs. Our previous studies have demonstrated that caffeic acid phenethyl ester (CAPE) suppress the growth of melanoma cells and induce ROS generation. However, the exact mechanism of the growth suppressive effects of CAPE was not clear. Here we determined the potential mechanism of CAPE against melanoma in vitro and in vivo. In a concentration and time-dependent study, CAPE treatment suppressed the activating phosphorylation of PI3K at Tyr-458, PDK1 at Ser-241, mTOR at Ser-2448 and AKT at Ser-473 in B16F0 and SK-MEL-28 melanoma cells. Furthermore, the expression of XIAP, survivin and BCl2 were down-regulated by CAPE treatment in both the cell lines. Significant apoptosis was observed in both the cell lines by CAPE treatment as indicated by cleavage of caspase-3 and PARP. AKT kinase activity was inhibited by CAPE in a concentration-dependent manner. Since Akt regulates XIAP, their interaction was examined by immunoprecipitation studies. Our results show that CAPE treatment decreased the interaction of AKT with XIAP. To establish the involvement of Akt in the apoptosis-inducing effects of CAPE, cells were transfected with Akt. Our results reveal that AKT over-expression attenuated the decrease in XIAP and significantly blocked CAPE-mediated apoptosis. Similarly, over-expression of XIAP further decreased CAPE-induced apoptosis. To confirm the involvement of ROS in the inhibition of AKT/XIAP pathway, cells were treated with antioxidant NAC prior to CAPE treatment. Our results indicate that NAC blocked CAPE mediated AKT/XIAP inhibition and protected the cells from apoptosis. In addition, oral administration of 10mg/kg/day CAPE substantially suppressed the growth of B16F0 tumor xenografts in C57BL/6 mice. Tumors from CAPE treated mice showed reduced phosphorylation of PI3K, AKT, mTOR and protein level of XIAP and enhanced the cleavage of caspase-3 and PARP. Taken together, our results suggest that CAPE suppresses AKT/XIAP pathway leading to apoptosis in melanoma tumor cells in vitro and in vivo. [Supported in part by R01 grant CA129038 (to S.K.S) awarded by the National Cancer Institute].

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 2591. doi:1538-7445.AM2012-2591
Abstract 2592: Vitamin E forms and 13′-carboxychromanol, a long-chain metabolite of α-tocopherol, induce cell death by interrupting de novo sphingolipid synthesis in human cancer cells
Authors: Jang Yumi , Rao Xiayu , Jiang Qing
Published: August 2012

Studies have shown that vitamin E forms including gamma-tocopherol (≥T), delta-tocopherol (≥T) and especially gamma-tocotrienol (≥TE), a major form of vitamin E in palm oil, exhibited potent anticancer activities in various types of cancer cells. 13′-carboxychromanol (13′-COOH), a long-chain metabolite of αT, has recently been shown to have anti-inflammatory activities and appears to induce apoptosis in certain type of cancer cells. However, the molecular mechanisms of the anticancer effects are not understood. In the current study, we found that 13′-COOH inhibited the growth and induced death of human colon and pancreatic cancer cells in a time- and dose-dependent manner. In these activities, 13′-COOH was more potent than αTE, which is much stronger than αT and αT. Since we have previously shown that αTE and αT induce prostate cancer cell death by modulation of sphingolipid metabolism, we investigated whether αTE and 13′-COOH have effects on sphingolipids using liquid chromatography tandem mass spectrometry. Treatment of HCT116 cells with αTE or 13′-COOH significantly increased dihydroceramides and dihydrosphingosine, two sphingoid bases in de novo synthesis pathway of sphingolipids. In contrast, sphingomyelins and ceramides remained largely unchanged after treatment. This accumulation of intracellular dihydroceramides and no apparent change of sphingomyelins suggest potential inhibition of dihydroceramide desaturase (DEGS) enzyme by these compounds. Consistently, we found that αTE inhibited the DEGS activity in a time- and dose-dependent manner in an in situ assay, while it did not have any effect on DEGS expression. These results indicate that αTE and 13′-COOH interrupt de novo synthesis of sphingolipids, which play a role in regulating cell death.

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 2592. doi:1538-7445.AM2012-2592
Abstract 2603: Subtype-specific ovarian cancer risk associated with SNPs in IL1A
Authors: Charbonneau Bridget , White Kristin L. , Schildkraut Joellen M. , Palmieri Rachel T. , Iversen Ed , Berchuck Andrew , Vierkant Robert A. , Rider David N. , Cicek Mine S. , Sutphen Rebecca , Birrer Michael J. , Pharoah Paul P.D. , Song Honglin , Tyrer Jonathan , Gayther Simon A. , Ramus Susan J. , Wentzensen Nicolas , Yang Hannah P. , Garcia-Closas Montserrat , Phelan Catherine M. , Cunningham Julie M. , Fridley Brooke L. , Sellers Thomas A. , Goode Ellen L.
Published: August 2012

As pathways of the immune system play an important role in multiple malignancies, we assessed the association between single-nucleotide polymorphisms (SNPs) in inflammation-related genes and risk of ovarian cancer in a multi-site case-control study. Using a custom array, we genotyped 930 epithelial ovarian cancer cases and 1,037 controls for SNPs in ALOX12, ALOX15, ALOX5, CCL11, CCL2, CCL3, CCR3, CRP, CXCL16, IL10, IL15RA, IL18, IL1A, IL1B, IL1RN, IL4R, IL6, IL6R, IL7R, IL8RA, IL8RB, IL9, NOS3, PTGS1, PTGS2, TLR2, and TNF. After analysis by logistic regression, SNPs with p<0.10 were evaluated in an independent data set of 3,143 cases and 2,102 controls from the Follow-up of Ovarian Cancer Genetic Association and Interaction Studies (FOCI) collaboration. Combined analysis revealed association with interleukin 1, alpha (IL1A) SNPs (rs17561 and rs4848300, r2=0.990, p=0.005) which varied by histological subtype (heterogeneity p=0.03). For example, IL1A rs17561, which results in a missense change in amino acid at position 114 and correlates with multiple inflammatory phenotypes, was inversely associated with risk of clear cell, mucinous, and endometrioid subtypes, but not with the serous subtype (Table 1). Additional data on IL1A rs17561 in an even larger collection of ovarian cancer cases and controls are pending and will be analyzed soon. We also found that genotype at rs1864414 in ALOX5 (arachidonate 5-lipoxygenase) was associated with decreased risk combined across subtypes (OR 0.86, 95% CI 0.78-0.95; p=0.002). Thus, inherited variation in IL1A and ALOX5 appears to have a role in ovarian cancer risk, though it is limited to rarer subtypes for IL1A. These data add to existing literature on the importance of inflammation in tumorigenesis and growing evidence of subtype-specific traits in ovarian cancer and may provide important clues on prevention.

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 2603. doi:1538-7445.AM2012-2603
Abstract 2604: Transforming growth factor beta signaling pathway variants and breast cancer risk: results from the Shanghai Breast Cancer Genetics Study
Authors: Ma Xiangyu , Beeghly-Fadiel Alicia , Cai Qiuyin , Lu Wei , Gao Yu-Tang , Long Jirong , Zhang Ben , Shi Jiajun , Zheng Ying , Shu Xiao-Ou , Zheng Wei
Published: August 2012

The transforming growth factor beta (TGF-β) signaling pathway is critical for several important cellular functions, and plays dual roles in the carcinogenic process. We systematically evaluated associations of common variants in TGF-β signaling pathway genes with breast cancer risk using a multi-staged, population-based, case-control study among Chinese women. In Stage I, 341 single nucleotide polymorphisms (SNPs) with MAFs ≤ 0.05 across 11 genes were evaluated among 2,926 cases and 2,380 controls. In Stage II, 20 SNPs with promising associations were evaluated among an additional 1,890 cases and 2,000 controls. Six SNPs from the TGFB2, TGFBR1, TGFBR2 and TGFBR3 genes were found to have consistent and significant associations in combined analyses with breast cancer risk in combined analyses of Stage I and Stage II data. One variant, TGFBR2 rs1078985, had not only consistent, but also highly significant associations with breast cancer risk in both study stages, as well as promising results from in silico analysis, and so was further evaluated. Results from additional genotyping in Stage III (2,475 cases and 2,343 controls) were also consistent; compared with the major allele homozygotes (GG) of rs1078985, reduced breast cancer risks were associated with AA(OR=0.75, 95%CI=0.60-0.94) and AG (OR=0.89, 95%CI=0.82-0.96) genotypes (Ptrend =0.0003). These data support a role for common genetic variation in TGF-β signaling pathway genes, specifically for a variant in TGFBR2, in breast cancer susceptibility.

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 2604. doi:1538-7445.AM2012-2604
Abstract 2605: Apolipoprotein E polymorphisms, plasma cholesterol, and prostate cancer
Authors: Shui Irene M. , Platz Elizabeth A. , Stampfer Meir , Mucci Lorelei A. , Giovannucci Edward
Published: August 2012

Background: Men who use cholesterol-lowering statin drugs and those with lower cholesterol levels may have a decreased risk of aggressive prostate cancer (PCa), providing support for further investigation of cholesterol metabolism in the etiology of PCa. Apolipoprotein E (APOE) plays an important role in cholesterol metabolism and may also affect tumor growth and angiogenesis. Compared to the APOE wild-type isoform (e3e3), the e4 allele has been associated with elevated cholesterol and the e2 allele with lower cholesterol. Our study aimed to determine if APOE polymorphisms were associated with PCa risk, especially aggressive disease, and whether this association varied by plasma total cholesterol level. Methods: We included 1193 total PCa cases and 1244 controls nested in the prospective Health Professionals Follow-up Study (1993-2004). APOE status was characterized by genotyping two SNPs (rs429358 and rs7412). Total plasma cholesterol was measured enzymatically. We calculated odds ratios and 95% confidence intervals for PCa risk (overall, high grade, and advanced stage/lethal) using unconditional logistic regression comparing e2 and e4 carriers to men homozygous for e3. Results: APOE genotype prevalence in controls was: e3e3 (0.60), e2e3 (0.14), e2e2 (0.007), e3e4 (0.22), e4e4 (0.016); those with e2e4 (0.027) were excluded from the analysis. APOE genotype was significantly associated with total plasma cholesterol: e2 carriers had significantly lower mean and standard deviation (188.5+/−34.7 mg/dL) compared to the e3e3 men (200.4+/−35.4 mg/dL) or the e4 carriers (201.2+/−36.3 mg/dL). However, we saw no significant associations with PCa by APOE genotype for total, advanced stage/lethal, or high-grade PCa (Table). There was no significant effect modification by plasma total cholesterol. Conclusions: In this large prospective cohort study we did not observe an association between APOE genotype and PCa. Future analyses will assess whether associations are modified by statin use.

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 2605. doi:1538-7445.AM2012-2605
Abstract 2608: Interplay between BRCA1 and RHAMM regulates epithelial apicobasal polarization and may influence risk of breast cancer
Authors: Pujana Miguel Angel , Maxwell Christopher A. ,   Consortium of Investigators of Modifiers of BRCA1/2 (CIMBA).
Published: August 2012

Differentiated mammary epithelium shows apicobasal polarity, and loss of tissue organization is an early hallmark of breast carcinogenesis. In BRCA1 mutation carriers, accumulation of stem and progenitor cells in normal breast tissue and increased risk of developing tumors of basal-like type suggest that BRCA1 regulates stem/progenitor cell proliferation and differentiation. However, the function of BRCA1 in this process and its link to carcinogenesis remain unknown. Here we depict a molecular mechanism involving BRCA1 and RHAMM that regulates apicobasal polarity and, when perturbed, may increase risk of breast cancer. Starting from complementary genetic analyses across families and populations, we identified common genetic variation at the low-penetrance susceptibility HMMR locus (encoding for RHAMM) that modifies breast cancer risk among BRCA1, but probably not BRCA2, mutation carriers: n = 7,584, weighted hazard ratio (wHR) = 1.09 (95% CI 1.02-1.16), ptrend = 0.017; and n = 3,965, wHR = 1.04 (95% CI 0.94-1.16), ptrend = 0.43; respectively. Subsequently, studies of MCF10A apicobasal polarization revealed a central role for BRCA1 and RHAMM, together with AURKA and TPX2, in essential reorganization of microtubules. Mechanistically, reorganization is facilitated by BRCA1 and impaired by AURKA, which is regulated by negative feedback involving RHAMM and TPX2. Taken together, our data provide fundamental insight into apicobasal polarization through BRCA1 function, which may explain the expanded cell subsets and characteristic tumor type accompanying BRCA1 mutation, while also linking this process to sporadic breast cancer through perturbation of HMMR/RHAMM.

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 2608. doi:1538-7445.AM2012-2608
Abstract 2609: Addition of ER and PTEN, but not cytokeratins, aids identification for BRCA1 carriers in Malaysian breast cancer patients
Authors: Phuah Sze-Yee , Looi Lai Meng , Rhodes Anthony , Dean Sarah , Taib Nur Aishah Mohd , Yip Cheng Har , Teo Soo-Hwang
Published: August 2012

Background: Given that BRCA1 carriers are more likely to develop triple negative breast cancers (TNBC) and that current risk assessment models underestimate BRCA carriers in Asian populations, we have sought to determine the prevalence of BRCA mutations amongst TNBC patients and whether addition of pathological features improves the sensitivity of the Manchester scoring method of BRCA risk assessment. Methods: A total of 399 patients from the Malaysian Breast Cancer Genetic Study were fully analysed for BRCA1 and BRCA2 mutations, including 101 patients with TNBC. Pathological features, including estrogen receptor (ER) and PTEN status were assessed and included in a modified Manchester scoring method. Results: Our study in an Asian series of triple negative breast cancer patients has demonstrated that 22% [22/101] of patients have germline mutations in BRCA1 [18/101] and BRCA2 [4/101]. We find that amongst patients who were diagnosed with breast cancer aged 36-50 but with no family history of breast or ovarian cancer, the prevalence of BRCA mutations was similar amongst those who developed triple negative breast cancer (7%) and non-TNBC (7%). By contrast, in two other groups of patients (a) those who were diagnosed with breast cancer β35 years old with no family history of these cancers, and (b) those with family history of breast cancer, the prevalence of mutations was higher in those with TNBC compared to those with non-TNBC [29% and 11%; p=0.07; and 40% and 14%; p=0.001 respectively]. Furthermore, we find that incorporation of estrogen-receptor status improves the sensitivity of the Manchester Scoring method [38% to 62%] and in particular, incorporation of the status of PTEN, further improves sensitivity [43% to 100%]. Conclusion: We find that TNBC is an important criteria for highlighting patients who may benefit from genetic testing, but that this may be most useful for individuals with early onset breast cancer (≥35 years old) or with family history of cancers. Furthermore, addition of ER and PTEN status improves the sensitivity of the Manchester scoring method and may be particularly important in the Asian context when family history is often inaccurately reported.

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 2609. doi:1538-7445.AM2012-2609
Abstract 261: The novel Akt inhibitor API-1 induces c-FLIP degradation and synergizes with TRAIL to augment apoptosis independent of Akt inhibition
Authors: Li Bo , Rentsch Hui , Yue Ping , Chen Mingwei , Khuri Fadlo R. , Sun Shiyong
Published: August 2012

API-1 is a novel small molecule inhibitor of Akt, which acts by binding to Akt and preventing its membrane translocation, and has promising preclinical antitumor activity. In this study, we reveal a novel function of API-1 in regulation of c-FLIP levels and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis, independent of Akt inhibition. API-1 effectively induced apoptosis in tested cancer cell lines including activation of caspase-8 and caspase-9. It reduced the levels of c-FLIP without increasing the expression of DR4 or DR5. Accordingly, it synergized with TRAIL to induce apoptosis. Enforced expression of ectopic c-FLIP did not attenuate API-1-induced apoptosis, but inhibited its ability to enhance TRAIL-induced apoptosis. These data indicate that downregulation of c-FLIP mediates enhancement of TRAIL-induced apoptosis by API-1, but is not sufficient for API-1-induced apoptosis. API-1-induced reduction of c-FLIP could be blocked by the proteasome inhibitor MG132. Moreover, API-1 increased c-FLIP ubiquitination and decreased c-FLIP stability. These data together suggest that API-1 downregulates c-FLIP by facilitating its ubiquitination and proteasome-mediated degradation. Since other Akt inhibitors including API-2 and MK2206 had minimal effects on reducing c-FLIP and enhancement of TRAIL-induced apoptosis, it is likely that API-1 reduces c-FLIP and enhances TRAIL-induced apoptosis independent of its Akt-inhibitory activity. (This study was supported by the Georgia Cancer Coalition Distinguished Cancer Scholar award and National Cancer Institute NIH SPORE P50 grant CA128613. F.R.K. and S-Y.S. are Georgia Cancer Coalition Distinguished Cancer Scholars)

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 261. doi:1538-7445.AM2012-261
Abstract 2625: Is familial risk of cancer limited to relatively early onset cases
Authors: Kharazmi Elham , Fallah Mahdi , Janquest Kristina , Hemminki Kari
Published: August 2012

Purpose: It is widely believed that familial cancers are of early onset. We aimed to find out whether familial risk of cancer is limited to relatively early onset cases. Methods and material: Standardized incidence ratios (SIR) were calculated for familial risk in offspring by parental age at diagnosis in the nation-wide Swedish Family-Cancer Database including all Swedes born after 1931 with their biological parents, totaling >12.2 million individuals and including >1.1 million first primary cancer cases. Results: Median parental age at diagnosis in those whose offspring were affected with a concordant cancer was somewhat over 70 years. Some 56-86% of familial cancers in parents occurred at age over 69 years. Even when parents were diagnosed at advanced age (90+), the risk of concordant cancer in offspring was significantly increased for skin (SIR=2.1), colorectal (1.7), breast (1.6), and prostate cancer (1.5). In offspring aged 60 to 76 years whose parents were affected with a concordant cancer, almost none of the cases had a parent affected at <40 years. Conclusion: Despite the common believe that familial cancers are of early onset, the majority of familial cancers occur at advanced ages. Familial cancers may not be early onset in those whose probands were affected at advanced ages, which may suggest concordance of age of onset in familial cancers. We identified reliable familial risks for skin, colorectal, breast, and prostate cancer even when parents were diagnosed at age 90+. The present study provides novel information, important for clinical counseling, cancer genetics and screening guidance.

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 2625. doi:1538-7445.AM2012-2625
Abstract 2626: Apoptosis-related single nucleotide polymorphisms and the risk of non-small cell lung cancer in a case-control study of women
Authors: Pathak Anand , Wenzlaff Angela S. , Cote Michele L. , Keele Greg R. , Land Susan , Schwartz Ann G.
Published: August 2012

Background: Often, lung cancer is diagnosed at later stages, resulting in a 15% 5-year survival rate. Improved survival is seen for those with early stage disease, thus it is crucial to identify genetic markers that can identify those at the highest risk for early disease. In this study, we focused on understanding the risk of developing non-small cell lung cancer (NSCLC) associated with germline single nucleotide polymorphisms (SNPs) in apoptosis related genes among women in the Detroit metropolitan area. Methods: 453 cases of NSCLC and 478 control subjects were examined in a population based case-control study. Since the allele frequency differed significantly in over 74% of the SNPs between whites and African Americans, analysis was stratified by race. Eight SNPs found not to be in HWE were excluded from the study, leaving 97 SNPs in 32 apoptosis related genes examined for association with risk of lung cancer. Odds ratios and 95% confidence intervals were estimated from unconditional logistic regression models using a dominant model, stratified by race, and adjusted for age, pack years smoked, ever/never smoking status, family history of lung cancer, history of COPD, BMI and education. Results: Among whites, a polymorphism in APAF-1 (rs1007573) was associated with a 2.86 fold increase in risk of lung cancer among smokers (95% CI: 1.58-5.16). SNPS in CD40 rs3765459 and rs1535045 were associated with a reduced risk of lung cancer among white smokers. After carrying out a backwards elimination procedure, only APAF-1 rs1007573 and CD40 rs1535045 remained as significant predictors of risk. Both of these SNPs were found to be noteworthy, as per the False Discovery Reporting Percentage (FDRP) method. In African Americans, 7 SNPs were found to be significantly associated with NSCLC. After the backwards elimination procedure, only SNPs in ATM rs1801516 (OR=24.15, 95% CI: 3.50-166.55), TNF rs1800629 (OR= 0.42, 95% CI: 0.18-0.99) and p63 rs6790167 (OR: 2.85, 95% CI:1.33-6.09) were found to be significant. Testing these SNPs using the FDRP method revealed that only p63 (rs6790167) was noteworthy. Indeed, this locus may represent an important target in NSCLC, as previous SNPs in p63 have been reported to increase the risk of lung cancer in larger GWAS studies. Conclusions: We report two significant SNPs associated with NSCLC risk in whites (APAF-1 rs1007573 and CD40 rs1535045) and one significant SNP in African Americans (p63 rs6790167) as potentially important contributors to NSCLC.

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 2626. doi:1538-7445.AM2012-2626
Abstract 263: Silence of SOX2 induces apoptosis through both mitochondria and death receptor signal pathway by activating the RAS/MAPK signals in human lung cancer cells
Authors: Chen Si , Li Xuefei , Xu Yingxi , Wang Lina , Chen Yanan , Lv Dan , Reisfeld Ralph A. , Li Na , Xiang Rong
Published: August 2012

Previous study showed that the stem cell gene-SOX2 plays an important role in the anti-apoptotic property of tumor cells, but the regulatory mechanism of SOX2 in the apoptosis signals is still elusive. In this study we used lentivirus system to deliver the shRNA to human lung cancer cell line- A549 and H460 and found that silencing of SOX2 gene can effectively induce apoptosis. Proteomic array assay and western blotting showed that down-regulation of SOX2 activates both the mitochondria and death receptor signal pathway, which were marked by the overexpression and phosphorylation of p53, overexpression of Bax and Bad, and activation of caspase 3 and 8 in A549 cells with SOX2 silencing. Real-time RT-PCR showed that down-regulation of SOX2 leads to the activation of Ras/MAPK signal pathway, which activated the transcription of two key inducers for apoptosis signals-TNF-α and p53 and also down-regulates the expression of survivin. In vitro and in vivo experiment further showed that this apoptotic effect of SOX2 silencing was compromised by overexpression of survivin. Immunohistochemistry study of human lung tissues showed that the protein expression level of SOX2 is closely correlated with that of survivin. In addition, SOX2 and survivin double positive patients showed worse prognosis, suggesting these two proteins can be used as dual-diagnosis factors to evaluate the clinical progression and prognosis of lung cancer patient. These findings revealed the important mechanism for SOX2 to regulate the apoptosis signals and demonstrated that survivin is one of vital downstream molecules that contribute to the anti-apoptosis property of SOX2.

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 263. doi:1538-7445.AM2012-263
Abstract 2631: Analysis of PI3K pathway genes in bladder cancer
Authors: Seven Didem , Tigli Hatice , Buyru Nur
Published: August 2012

Bladder cancer is the most common cancer of the urinary tract and is the ninth the most common cancer among men. Its incidence directly increases with age, it rarely occurs before the age of 50 and it is three to four times more common in men compared to women. Although there is now exclusive information on bladder cancer genetics, epigenetics and gene expression, several questions remain unanswered. Global profiling via expression and genomic microarray technologies may provide answers to some of the outstanding questions. Many studies have assessed the status of known oncogenes and tumor suppressor genes and have investigated common chromosomal alterations to identify novel genomic events in bladder cancer. The phosphatidylinositol 3-kinase (PI3K) signaling pathway plays critical role in cell growth, proliferation, and apoptosis. As a result of genomic alterations and loss of some tumor suppressor genes signaling through this pathway is commonly altered in many types of cancer including bladder. In the present study, our interest was focused on the expression analysis of some critical genes in this pathway. For this purpose, expression of seven genes (mTOR, KRAS, PIK3CA, TSC1, STK11, Rheb, PTEN and AKT1) acting in this pathway was analyzed in 35 matched bladder tumor and non-cancerous tissue samples by quantitative RT-PCR. Expression of all genes except PIK3CA and PTEN were similar for the normal bladder and tumor tissue. We observed downregulated expression of PTEN and PIK3CA genes in bladder tumors. The median PTEN mRNA levels in non-cancerous and tumor tissue samples were 2.11 and 0.35, respectively and the difference between the two groups was statistically significant (p=0.00). PIK3CA was also downregulated in the tumor tissues. Tumor tissue samples displayed 6.62 fold decrease in PIK3CA expression compared to normal tissue samples (p=0.002). On the other hand, slightly decreased Rheb mRNA expression was also observed in tumor tissue compared to the non-cancerous tissue. However, the difference expression levels of Rheb between two groups was not statistically significant. Our result indicate that, PI3K pathway has an important function in bladder cancer progression with the contribution of more than one gene in this pathway.

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 2631. doi:1538-7445.AM2012-2631