bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2466: Elafin, a serine protease inhibitor, is deregulated during breast cancer progression
Authors: Caruso Joseph A. , Karakas Cansu , Hunt Kelly , Keyomarsi Khandan
Published: August 2012

Deregulated protease activity promotes diverse aspects of tumorigenesis, including tumor growth, invasion, inflammation, and angiogenesis. Elafin is an endogenous inhibitor of the serine protease, Neutrophil Elastase (NE). Imbalance between NE and its inhibitors is implicated in the pathogenesis of many chronic inflammatory diseases. We hypothesize that elafin is deregulated during breast cancer progression, resulting in increased NE activity, accelerated tumor progression, and overall poor patient outcome. To test this hypothesis, we examined elafin expression by immunohistochemistry (IHC) in normal breast tissue from reduction mammoplasty, pre-malignant Ductal Carcinoma In Situ (DCIS), and invasive breast carcinoma (n>1000, using Tissue Microarray [TMA]). We observed that elafin is intracellularly (predominate nuclear localization) expressed in normal breast tissue. In the majority (>80%) of DCIS cases, intracellular elafin is expressed in a manner consistent with normal breast tissue. However, in cases of invasive breast cancer, intracellular elafin is largely downregulated (>80% of cases) suggesting elafin loss may have a role in progression to invasive carcinoma. Interestingly, examination of elafin expression in invasive breast cancer also revealed a subset (10-20%) of tumors with robust cytoplasmic overexpression of elafin, correlating with poor outcome. Similar results were seen in TMAs of high-grade ovarian carcinoma (n>200). Examination of publically available microarray data (TCGA) demonstrates that elafin measured at the mRNA level is correlates with poor outcome in ovarian cancer, consistent with our IHC results, and good outcome in breast cancer, incompatible with our IHC results. To further investigate the expression of elafin in breast cancer we examined by IHC a patient cohort (>200 patients) for whom full sections (not TMA) were available. Using these patient samples we found a unique subset of breast cancer patients that demonstrate high-level secretion of elafin into the tumor stroma, significantly correlating with increased survival especially in ER negative patients. Collectively, these data suggest that the prognostic role of extracellular elafin is related to the well-characterized ability of elafin to modulate inflammation and immunity.

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 2466. doi:1538-7445.AM2012-2466
Abstract 2480: High-level activation of the Notch pathway in diffuse intrinsic pontine glioma
Authors: Hutt Marianne , Lim Kah Jing , Warren Katherine , Chang Howard T. , Eberhart Charles G. , Raabe Eric H.
Published: August 2012

Diffuse intrinsic pontine glioma (DIPG) is a universally fatal pediatric brain tumor. Due to its critical location within the brainstem, and its characteristic MRI signature, biopsy is rarely performed. Since many patients die at home in hospice care, autopsy specimens have traditionally been rare. Because of this difficulty in obtaining tissue, three institutions (Johns Hopkins, Children's National Medical Center, and the National Institutes of Health - Pediatric Oncology Branch) have formed the Mid-Atlantic DIPG Consortium (MADC), to collect and disseminate rapid-autopsy material from patients with DIPG. Using a fresh tissue specimen obtained from a rapid autopsy and disseminated through the MADC, we have established a human DIPG xenograft and neurosphere cell line, JHH-DIPG1. In vitro, these cells are GFAP positive and express human neural stem cell markers often observed in glioblastoma, including human specific SOX2 and NESTIN. They also show marked invasive properties in vitro as measured by transwell invasion assay. In vivo, JHH-DIPG1 forms tumor xenografts in approximately six months in immunocompromised mice. Tumor cells diffusely infiltrate distant brain structures, recapitulating the invasive phenotype of DIPG. Due to their high level expression of neural stem cell markers, and the well-known roles of the Notch pathway in stem cell biology and aggressive brain tumors, we examined the expression of members of the Notch pathway in DIPG. Notch downstream effectors, specifically Hes1, Hes5 and Hey1, are highly expressed in JHH-DIPG1, at levels equal to or exceeding those observed in neurosphere cell lines derived from adult supratentorial glioblastoma. The JHH-DIPG1 tumor cell line represents one of only a handful of pediatric DIPG cell lines extant, and the high level of Notch pathway activation in these cells suggests that blockade of the Notch pathway using gamma secretase inhibitors and shRNAs against the canonical Notch effector CBF may be efficacious in DIPG. Our laboratory is actively investigating the role of the Notch pathway in DIPG using this and other DIPG cell lines, with the goal of moving Notch inhibition into clinical trials for these currently highly lethal tumors.

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 2480. doi:1538-7445.AM2012-2480
Abstract 2481: Exploiting BRAF mutation for therapy of childhood low-grade astrocytoma
Authors: Kibler Aaron , Manap Sagymbek , Bid Hemant K. , Phelps Doris A. , Kurmasheva Raushan T. , Pelloski Christopher E. , Houghton Peter J.
Published: August 2012

Activating mutations of BRAF (predominantly V600E) are common in low-grade astrocytomas (∼23%), and xanthoastrocytomas (60%), with or without loss of the CDKN2A locus. Surgically inoperable astrocytoma has a poor prognosis with greater than half of these patients dying from disease. We have developed two models of low-grade astrocytoma (BT-35, BT-40) by direct heterografting patient tumor into immune-deficient mice. Both tumor lines have multiple copies of BRAF, but only BT-40 has an activating mutation (V600E), and is exquisitely sensitive to the MEK inhibitor AZD6244, whereas BT-35 is intrinsically resistant (Kolb et al. Ped Blood Cancer, 2010). Biochemical characterization showed that MEK1/2 were equally inhibited by AZD6244 treatment in both BT-35 and BT-40 xenografts, as demonstrated by the level and duration of inhibition of phospho-ERK1/2. However, inhibition of MEK1/2 had no effect on TORC1 signaling in BT-35 (insensitive) tumors whereas in BT-40 xenografts TORC1 signaling was completely inhibited by AZD6244. The pathway through which mutant BRAF controls TORC1 signaling is currently under evaluation. However, control of TORC1 by mutant BRAF potentially exposes these cells to therapeutic exploitation. We have shown previously that the TOR kinase inhibitor AZD8055 potently suppresses FANCD2, a protein involved in interstrand cross-link (ICL) repair of DNA damage. Similar to the effect of AZD8055, within 4 days treatment of mice with AZD6244, FANCD2 levels are dramatically reduced and were no longer detected in BT-40 xenografts. Preliminary results show that inhibition of TOR signaling by AZD8055 is synergistic with daily radiation in rhabdomyosarcoma xenografts. We hypothesize that because MEK inhibition leads to suppression of TORC1 signaling and FANCD2 in BT-40 astrocytoma xenografts, that combination of AZD6244 with ionizing radiation or drugs that induce ICL will be selectively synergistic in the context of BRAF mutation. Experiments to test the hypothesis are ongoing. Supported by a Peletonia grant from the Ohio State University.

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 2481. doi:1538-7445.AM2012-2481
Abstract 2488: Reovirus (Reolysin) as a potential therapy for malignant peripheral nerve sheath tumors
Authors: Sampson Valerie , Kamara Davida , Calio Brian , Kolb Edward A.
Published: August 2012

Malignant peripheral nerve sheath tumors (MPNST) are a rare form of soft tissue sarcoma which may occur sporadically in the general population or in association with neurofibromatosis type 1 (NF1), and are highly aggressive and often fatal. NF1 patients are at a greater risk for the development of malignant peripheral nerve sheath tumors than the general population. Current therapy relies upon surgical resection with adjuvant chemotherapy and/or radiotherapy. Clinical trials using Reolysin (Respiratory Enteric Orphan Virus) which is developed from the naturally-occurring human reovirus have demonstrated the efficacy of this reovirus as an oncolytic virotherapy. Reolysin selectively replicates in cancer cells with activated Ras (Ras-GTP), resulting in virus-mediated apoptotic cell death. Hence Reolysin demonstrates promising anticancer activity and has minimal toxicity in patients. Since MPNST is associated with neurofibromin (NF1) deficiency and active Ras we tested these tumors as therapeutic targets for Reolysin treatment using two NF1 deficient human MPNST-derived cell lines (CRL-2884, ST8814) with activated Ras and one sporadic MPNST cell line (STS26T) which expresses NF1 but not active Ras. These were compared with normal human Schwann cells (NHSC) which do not show active Ras and mammary gland cells derived from an NF1 patient (CRL-7329). Reolysin inhibited the proliferation and viability of the three MPNST cell lines at a dose of 0.1 to 10 virus particles/cell, including sporadic MPNST cells which do not have active Ras. In contrast, the NHSC and CRL-7329 cells were more resistant to viral treatment, with apoptosis induced at higher viral concentrations (10,000 and 100,000 virus particles/cell respectively). Furthermore, we examined the effects of Reolysin on the Ras-signaling pathway in a bladder cancer cell line (HTB-4) expressing NF1 and Ras and found that virus induced cytotoxicity was similar to the control cell lines (100,000 virus particles/cell). This study shows that MPNST-derived cell lines including sporadic MPNST without active Ras were efficiently transduced, promoted virus replication and were killed by the oncolytic reovirus. In contrast to the highly sensitive MPNST cell lines, NHSC, CRL-7329 and HTB-4 cells were resistant at similar viral concentrations. Our data suggests the use of oncolytic Reolysin may represent a novel treatment approach for patients with MPNST and provides the basis for preclinical testing of the antitumor effects. Therapeutic efficacy of Reolysin in a mouse model for MPNST is underway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2488. doi:1538-7445.AM2012-2488
Abstract 2489: A phase II trial of poly-ICLC in the management of recurrent or progressive pediatric low grade gliomas
Authors: Hartman Lisa R. , Durden Donald L.
Published: August 2012

Low-grade astrocytomas and gliomas are the most common type of childhood brain tumor. Although children with low-grade gliomas often survive many years after conventional treatment with surgery and radiotherapy, there can be tremendous morbidity depending on the location of tumor and the effects of therapy. Newer forms of treatment with reduced side effects are much needed. We have designed a phase II trial to examine efficacy and safety of the biological response modifier poly-ICLC in children with low-grade gliomas, and to identify surrogate biomarkers for response to this agent. Poly-ICLC, a synthetic complex of polyinosinic and polycytidylic acid, stabilized with polylysine and carboxymethyl cellulose is a double stranded RNA (dsRNA) that is a ligand for the Toll-like Receptor 3 (TLR3), an important component of innate immunity. It has been shown to have direct antiviral and antineoplastic effects mediated by at least two interferon inducible nuclear enzyme systems of OAS and PKR. A previous Phase I/II trial of poly-ICLC conducted in pediatric patients with brain tumors (all histopathologic subtypes) suggested the low-grade glioma cohort had the best response to this therapy (50% PR rate). Additional clinical trials in adult patients have confirmed that the most responsive subgroup to poly-ICLC is low-grade glioma patients. We are currently conducting a multi-institutional phase II study of poly-ICLC administered at 20mcg/kg/dose intramuscularly twice weekly to children with recurrent or progressive low-grade gliomas. Simon's two-stage design will be used for statistical analysis. The primary goal of this study is determine the radiographic response rate. We will also characterize toxicity and determine if surrogate markers of tumor response and progression can be identified in patient serum, peripheral blood mononuclear cells (PBMC), and/or cerebrospinal fluid (CSF). To date, 4 pediatric subjects (ages 2-15) with recurrent low-grade gliomas have enrolled in the study. Poly-ICLC therapy has been well tolerated and none of the subjects have shown radiologic or clinical progressive disease. These results suggest that poly-ICLC may have a beneficial role in children with low-grade gliomas while minimizing treatment side effects. We have begun analyzing blood and CSF samples to attempt biomarker identification and correlation with subject responses, in hopes to yield a basis towards stratification and personalized therapy with recurrent disease.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2489. doi:1538-7445.AM2012-2489
Abstract 2490: High AXL promotes migration in non-MYCN amplified neuroblastoma cell lines
Authors: van Noesel Max M. , Duijkers Floor A.M. , Meijerink Jules P.P. , Pieters Rob
Published: August 2012

Background: Neuroblastomas (NBL) are tumors with a high rate of metastases (50% of patients). In general, in NBL MYCN amplification (NMA) is the most important prognostic factor. However NMA is infrequent and 70% of NBL are non-NMA tumors and not much is known about the driving oncogenic event. Recently, we identified among 17 NBL cell lines a group of non-NMA NBL cell lines with a distinct gene expression profile (HU133plus2.0 arrays, Affymetrix) and high expression of the AXL gene. AXL is a tyrosine kinase receptor, associated with the metastatic process of several cancers. We hypothesized that AXL contributes to metastsizing potential in non-NMA neuroblastomas and tested if AXL silencing diminishes malignant properties of high AXL expressing cell lines. Results: High expression of AXL was detected in G-IM-EN, SK-N-AS and two subclonal cell lines SH-EP-2 and SH-EP-21N. AXL was silenced (lentiviral AXL shRNA expression plasmids, pLKO.1_shAXL, Sigma-Aldrich) in GI-M-EN and SH-EP-2 with an infection efficiency of >90% and 80-85% AXL-mRNA knockdown. We examined the silencing effect on migration and invasion. A significant reduction of migration (74.7%) and invasion (99.8%) was achieved in GI-M-EN. In the intermediate AXL expressing expressing cell line SH-EP-2 migration reduction was less (41.9%). Furthermore, GI-M-EN and SH-EP-2 were compared for cytoskeletal, morphological changes after silencing. Cells became rounded with small lamellipodia and filopodia and F-actin fluorescent staining showed induction of F-actin positive stress-fibers (17.3%), both most prominent in high AXL expressing GI-M-EN. In SH-EP-2 the number of cells with stress fibers was increased (9.9%). Expression of, matrix metalloproteinase 9 frequent down-stream target of EMT-like processes, was not changed upon silencing of AXL. Also, we observed no effects on cell proliferation, apoptosis or downstream pathways (PI3K-AKT, MAPK-ERK pathway). Conclusion: The AXL gene is highly expressed in a subset of non-NMA NBL cell lines and seems involved in migratory and invasion properties. AXL is a possible mediator of NBL metastasis.

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 2490. doi:1538-7445.AM2012-2490
Abstract 25: CBL enhances breast tumor formation by inhibiting tumor suppressive activity of TGF-beta signaling
Authors: Kang Jin Muk , Park Sujin , Kim Staci Jakyong , Kim Seong-Jin
Published: August 2012

CBL protein family functions as multifunctional adaptor proteins and E3 ubiquitin ligases that are implicated as regulators of signaling in various cell types. Recent discovery revealed mutations of proto-oncogenic CBL in the linker region and RING finger domain in human acute myeloid neoplasm and these transforming mutations induced carcinogenesis. However, the adaptor function of CBL mediated signaling pathway during tumorigenesis has not been well characterized. Here, we show that CBL highly expresses in breast cancer cells and significantly inhibits TGF-β tumor suppressive activity. Knockdown of CBL expression resulted in the increased expression of TGF-β target genes, SERPINE1 and CDK inhibitors such as p15INK4b and p21Cip1. Furthermore, we demonstrate that CBL is frequently overexpressed in human breast cancer tissues and the loss of CBL decreases a tumorigenic activity of breast cancer cells in vivo. CBL directly binds to Smad3 through its proline-rich motif, thereby preventing Smad3 from interacting with Smad4 and blocking nuclear translocation of Smad3. CBL-b, one of CBL protein family, also interacted with Smad3 and knockdown of both CBL and CBL-b further enhanced TGF-β transcriptional activity. Our findings provide evidence for a previously undescribed mechanism by which oncogenic CBL can block TGF-β tumor suppressor activity.

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 25. doi:1538-7445.AM2012-25
Abstract 2510: Increased Fc-FcR interaction of human phosphatidylserine targeting antibody enhances pro-inflammatory and ADCC anti-tumor mechanisms
Authors: Doerr Jeanette R. , Larson Gary , Archer Rich , Gong Jian , Nguyen Van , Iyengar Aditi , Johnson Amy E. , Wakabayashi Mary , Peacock Matthew , Hughes Christopher , Chang Connie , Freimark Bruce
Published: August 2012

Anionic phospholipids, principally phosphatidylserine (PS), become exposed on the external surface of vascular endothelial cells in tumors, providing an excellent marker for tumor vascular targeting. Binding of antibodies targeting PS on the tumor endothelial cells and tumors recruits immune cells and engages the immune system to destroy tumor vasculature. The antibodies also enhance anti-tumor immunity by blocking the immunosuppressive action of PS. A chimeric anti-PS antibody, bavituximab, is being used in combination with chemotherapy to treat patients with solid tumors in Phase II trials. We have engineered mutations in the Fc region of fully human PS-targeting antibody PGN635 that enhance the interaction with FcRgammaIIIa and have characterized their ability to exert anti-tumor effects through various mechanisms, including ADCC and cytokine induction. Mutations that enhance Fc-FcR interaction of PS targeting antibodies enhance production of pro-inflammatory chemokines such as MCP-1 (CCL2), MIP-1alpha (CCL3) and MIP-1beta (CCL4). In addition, the same Fc mutations enhance ADCC of PS expressing tumor targets. Our data indicate that improvement of multiple immune effector functions via enhanced interaction of Fc-FcR interaction of PS-targeting antibodies may improve the treatment of cancer.

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 2510. doi:1538-7445.AM2012-2510
Abstract 2515: Anti- GP88 (progranulin) antibody to GP88 (progranulin) inhibits breast tumor growth and restores sensitivity to tamoxifen
Authors: Serrero Ginette , Dong Jianping , Hayashi Jun
Published: August 2012

GP88 (Progranulin, PCDGF, acrogranin) is an 88 kDa glycoprotein that is the largest member of the granulin/epithelin family of growth modulators characterized by 7.5 repeats of a highly conserved 12-cysteine granulin/epithelin motif. Our laboratory demonstrated the role of GP88 as an autocrine growth and survival factor in breast cancer: 1) GP88 expression increases with tumorigenesis; 2) inhibition of GP88 expression by antisense transfection in MDA-MB-468 cells inhibited tumor formation in nude mice by 90%; 3) in ER+ breast cancer cells, GP88 stimulates proliferation and its overexpression confers estrogen independence; 4) GP88 is a strong survival factor that confers resistance to anti-estrogen and aromatase inhibitors in ER+ breast cancer; 5) GP88 is expressed in 80% invasive ductal carcinoma and 60% of ductal carcinoma whereas it is negative in benign lesions and normal mammary epithelial cells; 6) pathological studies with 530 cases of ER+ invasive ductal carcinoma showed that GP88 tumor expression is a prognostic indicator of recurrence in early stage breast cancer patients; 7) GP88 is secreted and can be detected in the serum of breast cancer patients at an increased level when compared to healthy subjects. Based on these results GP88 represents an ideal therapeutic and diagnostic target in breast cancer. We developed a neutralizing anti-GP88 antibody AG1. AG1 inhibits GP88 biological effect in a dose-dependent fashion. In the present study, we investigated the effect of AG1 on the tumor development of tamoxifen sensitive (MCF-7) and resistant (TamR) estrogen receptor positive breast cancer cells lines injected in nude mice. We show that AG1 alone (5mg/kg) inhibited tumor growth of MCF-7 cells injected into nude mice in a similar efficacy as tamoxifen. Interestingly, AG1 in combination with tamoxifen inhibited by more than 50% tumor formation of TamR cells, whereas tamoxifen only had no effect. These data suggest that inhibiting GP88 provide a novel and alternate pathway for restoration of tamoxifen sensitivity in ER+ breast cancer. This works is supported by grant 2R44CA124179 from 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 2515. doi:1538-7445.AM2012-2515
Abstract 2520: CHC promotes tumor growth and angiogenesis through regulation of HIF-1α and VEGF signaling
Authors: Tung Kuo-Hua , Wu Han-Chung
Published: August 2012

Pancreatic adenocarcinoma is an aggressive disease with a high mortality rate. Currently, treatment options are limited. In an effort to improve the efficacy of treatments for pancreatic adenocarcinoma, we have newly generated a monoclonal antibody (mAb), Pa65-2, which specifically binds to various cancer cells and tumor blood vessels but not to normal cells. According to immunoaffinity chromatography, LC-MS/MS analysis, co-immunoprecipitation and RNA interference studies, the target protein of Pa65-2 is human clathrin heavy chain (CHC). Knocking down CHC with shRNA reduced tumor cell growth, colony formation and invasion in vitro, and inhibited tumor angiogenesis and growth. One of the key functions of CHC was to bind with the transcription factor subunit HIF-1α, increasing the stability of this protein and facilitating its nuclear translocation, thereby regulating the expression of VEGF. In animal studies using NOD/SCID mice bearing pancreatic cancer xenografts, Pa65-2 and CHC shRNA could each inhibit tumor growth and angiogenesis by regulating HIF-1α and VEGF. Furthermore, the expression of CHC, HIF-1α and VEGF was significantly correlated with tumor stage, suggesting that CHC might be a useful prognostic indicator for cancer. Considered together, our results indicate that CHC interacts with and stabilizes HIF-1α, and plays a role in HIF-1α nuclear localization and HRE promoter binding, leading to an increased production of VEGF. These results suggest that the CHC mAb, Pa65-2, can potentially be used to inhibit tumor angiogenesis and growth.

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 2520. doi:1538-7445.AM2012-2520
Abstract 2523: The in vitro effects of immobilized epratuzumab, a non-blocking anti-CD22 humanized monoclonal antibody, on malignant B cells
Authors: Gupta Pankaj , Michel Rosana B. , Goldenberg David M. , Chang Chien-Hsing
Published: August 2012

Epratuzumab (hLL2), a humanized anti-CD22 monoclonal antibody, is currently in advanced trials in non-Hodgkin lymphoma (NHL) and systemic lupus erythematosus (SLE) patients. As a single agent, hLL2 is well tolerated and depletes 30 to 50% of circulating B cells in patients with NHL and SLE. The mechanisms by which hLL2 depletes B cells in vivo is still largely unknown and continues to be a subject of current research interest. In vitro, hLL2 shows moderate antibody-dependent cell-mediated cytotoxicity, but no detectable complement-dependent toxicity, and displays cytotoxicity to CD22-expressing cells, for example, Ramos and Daudi human Burkitt lymphomas, only when immobilized onto plastic plates or in combination with a suboptimal amount (1 μg/mL) of anti-IgM plus a crosslinking secondary antibody. We have previously reported that hLL2, despite a lack of direct cytotoxicity, is effective in inducing the phosphorylation of CD22, translocation of CD22 to lipid rafts, and internalization of CD22. In this study, we evaluated the various effects of immobilized hLL2 on D(1-1), a subclone of Daudi selected for a higher expression of membrane IgM (mIgM). Our findings demonstrate for the first time that many of the intracellular events induced by binding CD22 to immobilized hLL2 were similar to those induced by binding mIgM with a sufficient high concentration (10 μg/mL) of anti-IgM; for example, both were found to induce phosphorylation of CD22, CD79a and CD79b, and the translocation of all three into the lipid rafts, which is critical for the observed cytotoxicity. Immunofluorescence analysis revealed CD22 colocalized with BCR and formed caps before being internalized. Like anti-IgM, immobilized hLL2 induced substantial apoptosis (25 to 40%) in D(1-1), which was caspase-dependent, accompanied by a pronounced increase in both pERKs and pJNKs with a concurrent decrease in the constitutively-activated p38, and preventable by inhibitors to JNKs or caspases. The involvement of mitochondria in the apoptotic pathway was evidenced by changes in mitochondrial membrane potential and generation of reactive oxygen species, as well as upregulation of Bax (pro-apoptotic) and downregulation of Bcl-xl, Mcl-1 and Bcl-2 (anti-apoptotic). Further studies are underway to elucidate how hLL2 interferes with B-cell survival in vivo.

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 2523. doi:1538-7445.AM2012-2523
Abstract 2534: Anti-LOXL2 conjugated gold nanoparticles: therapeutic probes for pancreatic cancer
Authors: Guo Yang , Zhang Zhuoli , Li Weiguo , Nicolai Jodi , Omary Reed , Larson Andrew
Published: August 2012

PURPOSE: Multi-functional gold nanoparticles may play an important role in the treatment of pancreatic cancer. The purpose of this study was to demonstrate the feasibility of using these anti-LOXL2 conjugated gold nanoparticles (anti-LOXL2@GoldMag) as therapeutic probes for the treatment of human pancreatic cancer cells. MATERIALS & METHODS: Human pancreatic cancer cell lines (PANC-1, BxPC-3 and Capan-1 cells) was used in this study. Western blot of LOXL2 expression in three cell lines were analyzed and compared to EGFR proteins. GoldMag, the hybrid gold shell and iron oxide core nanoparticles were purchases and their imaging characteristics (R2 and R2* relativity) was measured. Anti-LOXL2@GoldMag probes were constructed according to the instruction and the coupling efficacy of anti-LOXL2 antibodies with GoldMag were tested using a mass spectrophotometer. The specific targeting efficacy of anti-LOXL2@GoldMag was tested using immunofluorescence staining and electronic microscope. Pancreatic cancer cells were treated with anti-LOXL2@GoldMag at the concentrations of 0, 35, 70, 140 and 280 μg/ml and compared to anti-LOXL2 treatment alone at the same dosing concentration. After allowing for 48 hours incubation, the remaining number of viable cells was counted using trypan blue staining; cell proliferation percentage was calculated based on the total number of viable cells after treatment compared to the controls. RESULTS: The therapeutic efficacy of anti-LOXL2@GoldMag demonstrated similar therapeutic efficacy at both lower dose (18-51% reduced total viable cells) and enhanced therapeutic efficacy at the higher dose (86-99% reduced total viable cells) when compared to LOXL2 antibody alone. This suggests that binding of the LOXL2 antibodies to the GoldMag nanoparticles doesn't inhibit immunotherapeutic efficacy. MRI studies demonstrated that R2 and R2* measurements were well correlated to GoldMag probe concentrations. CONCLUSION: Our findings demonstrated significant anti-neoplastic efficacy in three different human pancreatic cancer cell lines. MRI could serve as a non-invasive imaging method to monitor the biodistribution of gold nanoparticles. Further pre-clinical investigations are now warranted in animal models of pancreatic cancer.

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 2534. doi:1538-7445.AM2012-2534
Abstract 2597: Identification of new genes for hereditary breast cancer
Authors: Kuligina Ekaterina Sh. , Sokolenko Anna P. , Suspitsin Evgeny N. , Iyevleva Aglaya G. , Chekmariova Elena V. , Abysheva Svetlana N. , Imyanitov Evgeny N.
Published: August 2012

Major hereditary breast cancer (BC) genes, BRCA1 and BRCA2, contribute to no more than 20-25% of familial BC clustering; therefore the majority of BC-predisposing mutations remain to be identified. We have selected for the study 95 BC cases, which showed accumulation of clinical signs of the genetic disease (e.g., occurrence of BC among multiple relatives and/or bilaterality and/or young age at onset), but could not be explained either by BRCA1/2 sequence alterations or by Russian founder mutations in CHEK2 (c.1100delC, c.444+1G>A, del5395) or NBN (657del5) genes. The entire coding regions of DNA repair genes were examined using high resolution melting (HRM) analysis and DNA sequencing. Three of 16 genes selected for the study (CHEK2, NBN, PALB2, BRIP1, BARD1, RAD51C, BLM, FANCG, CHEK1, PARP1, PARP2, ERCC1, XPE, BRD7, RNF8, RAD51A) contained potentially deleterious mutations. Two patients carried inactivating lesion in the PALB2 gene (p.R414X and p.Q921X); both these carriers had the bilateral form of BC disease. 1 woman harbored mutation in the FANCG gene (c.520_524delTCTAinsC), which resulted in the loss of serine at position 174 and lysine to glutamine substitution at position 175. The analysis of the BLM gene revealed two heterozygous carriers of the c.1642 C>T (Q548X) allele. This mutation was subjected to the extended study, which confirmed its recurrent character and strong association with BC risk: BLM c.1642 C>T (Q548X) heterozygotes were detected in 17/1,498 (1.1%) BC patients as compared to 2/1,093 (0.2%) healthy females (p = 0.004). As expected for BC-predisposing gene, the BLM 1670 C>T allele tended to be associated with first-degree family history of BC, early onset and bilateral appearance of the disease. This is the first demonstration of the role of the BLM heterozygosity in the determination of BC risk. The impact of constitutional BLM mutations in hereditary cancer predisposition deserves to be tested in other ethnic groups as well as in other tumor types.

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 2597. doi:1538-7445.AM2012-2597
Abstract 2602: CHEK2 contribution to hereditary breast cancer in non-BRCA famillies
Authors: Desrichard Alexis , Uhrhammer Nancy , Bidet Yannick , Bignon Yves-Jean
Published: August 2012

Mutations in the BRCA1 and BRCA2 genes are responsible for only a part of hereditary breast cancer (HBC). The origins of “non BRCA” HBC families may be attributed in part to mutations in genes giving moderate risk, such as CHEK2. We investigated the contribution of CHEK2mutations to non-BRCA HBC by direct sequencing of its entire coding sequence. Fifteen mutations were discovered among 507 non-BRCAHBC cases and four among 513 controls. The frequency of CHEK2variants was significantly higher among cases (p= 0.0076), and gave an OR for breast cancer of 4.72 for deleterious mutation carriers. We then used both in silico tools and in vitro kinase activity to evaluate recombinant mutant proteins. Tumor characteristics and tumor grade of paraffin-embedded tissue blocks from 8 CHEK2 mutated patients were evaluated by histology. To further characterize those tumors, breast cancer immunohistochemical markers such as hormone receptors, HER2 and P53 were assessed. Because the mechanisms of tumorigenesis in association with CHEK2 variants are still unclear, we performed genetic and epigenetic analysis of those tumors. Three relevant SNPs spanning the CHEK2 gene locus were used to determine loss of heterozygosity (LOH). Also, the proximal CpG islands of the CHEK2 gene were investigated for hypermethylation. Our results suggest a contribution of CHEK2 mutations to non-BRCA HBC, though the usefulness of moderate penetrance genes for genetic counseling remains controversial.

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 2602. doi:1538-7445.AM2012-2602
Abstract 2610: Association of polymorphisms and haplotypes in the insulin-like growth factor 1 receptor (IGF1R) gene with breast cancer in Korean women
Authors: Kim Mi Kyung , Kang Han-Sung , Ahn Sei Hyun , Hong Kyeong-Man , Mishra Siddhartha Kumar , Shin Kyung Hwan , Ro Jungsil , Lee Keun Seok , Kim Seok Won
Published: August 2012

Abstract The insulin-like growth factor (IGF) signaling axis plays an important role in cancer biology. The insulin-like growth factor 1 receptor (IGF1R) is a tyrosine kinase growth factor receptor that plays important roles in numerous cellular functions and signal transduction pathways. IGF1R may also play a role in the initiation and progression of breast cancer. However, studies of common variation in the IGF1R gene, involved in steroid hormone and IGF-1 metabolism, have yet to provide convincing evidence that such variants predict breast cancer risk. We genotyped 384 tagging single nucleotide polymorphisms (SNPs) in 1026 cases of breast cancer and 392 age-matched control in a population of Korean women. Of the 384 SNPs, fifty one SNPs in the IGF1R gene were examined for association with breast cancer in the study. All the SNPs investigated were in Hardy-Weinberg equilibrium. These SNPs tested were significantly associated with breast cancer risk, after correction for multiple comparisons stratified by adjusting for age at diagnosis, BMI, age at menarche, and age at first parturition. Among 51 IGF1R SNPs, three intron located SNPs (rs8032477, rs11635251, and rs12916884) with homozygous genotype (variant genotype) had increased risk of breast cancer (OR(CI 95%): 0.66(0.44-0.99), 0.54(0.37-0.80), and 0.59(0.37-0.94)). Seven of the 51 IGF1R SNPs were in LD and in one haplotype block, and were likely to be associated with breast cancer risk. Overall, this comprehensive case-control study found statistically significant associations between breast cancer risk and polymorphisms in IGF1R gene. Taking the status of the IGF1R polymorphism into consideration, our results may help in improving the ability to develop effective treatment modalities for those conditions in which IGF1R is involved. To the best of our knowledge, this study is the first to demonstrate the association between IGF1R polymorphisms with the risk of breast cancer.

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 2610. doi:1538-7445.AM2012-2610
Abstract 2616: Association of UGT2B17 gene copy number variation with breast cancer risk: The Nashville Breast Health Study
Authors: Li Guoliang , Deming Sandra L. , Cai Qiuyin , Fair Alecia M. , Shrubsole Martha J. , Shu Xiao Ou , Kelley Mark , Zheng Wei
Published: August 2012

UDP-glucuronosyltransferase 2B17 (UGT2B17) catalyzes the transfer of glucuronic acid from uridine diphosphoglucuronic acid to a variety of substrates, including estrogen, a hormone that plays an important role in the etiology of breast cancer. Glucuronidated estrogens are more polar and water soluble; thus, they are more easily excreted in the bile and urine. The UGT2B17 gene displays variations in copy number (0 copies, 1 copy, and 2 copies), and the null genotype (0 copies) is associated with lower glucuronidating activity in human liver microsomes. In this study, we evaluated the association of UGT2B17 gene copy number variation with breast cancer risk. Included in the study were 1,536 breast cancer cases and 1,205 matched controls recruited into the Nashville Breast Health Study, a population-based, case-control study based in Nashville, Tennessee. Copy number of the UGT2B17 gene was evaluated by real-time PCR by using ABI's TaqMan Copy Number Assays reagents. UGT2B17 gene copy number variation was associated with breast cancer risk with odds ratios of 1.17 (95% CI: 1.00-1.38) and 1.33 (95% CI: 1.02-1.74), respectively, for 1 and 0 copies of the UGT2B17 gene, compared with 2 copies of the gene. The association varied little by menopausal status. The association was consistent in both whites and African Americans. Our study suggests that UGT2B17 gene copy number variation may play a role in the etiology of breast cancer.

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 2616. doi:1538-7445.AM2012-2616
Abstract 2620: Contribution of risk variant in miR-125b binding site of BMPR1B in women of African ancestry with breast cancer
Authors: Yoshimatsu Toshio F. , Zheng Yonglan , Zhang Jing , Huo Dezheng , Olopade Olufunmilayo I.
Published: August 2012

MicroRNAs play a fundamental role in gene regulation by selective binding to the 3′ UTR of specific mRNAs. Dysregulation of this post-transcriptional regulatory mechanism, in turn, has emerged to be an important process in cancer etiology. Recently a particular single nucleotide polymorphism (SNP) at mir-125b binding locus of BMPR1B, rs1434536, was reported as significant risk factor for breast cancer pathogenesis in Caucasians. The BMPR1B gene encodes a member of the bone morphogenetic protein (BMP) receptor family of transmembrane serine/threonine kinases. The C/T allelic variation within the target site disrupts the regulation of miR-125b, whereby increased BMPR1B expression ultimately elevates disease risk. To validate whether rs1434536 also contributes to breast cancer risk in women of African ancestry, we performed a case-control study of 805 breast cancer cases (422 African Americans (AA) and 383 Nigerians (NG)) and 786 controls (402 AA and 384 NG). The average age (±SD) was 46.9 ± 11.9 years for cases and 45.1 ± 12.4 years for controls. Genotyping was conducted using polymerace chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Test for Hardy-Weinberg equilibrium (HWE) suggested that there was no deviation from HWE (P > 0.05) in AA, or NG. Pearson's chi-squared tests showed that this SNP was significantly associated with breast cancer in AA (p=0.03) but not in NG (p=0.96). After adjusting for age in the logistic regression, we found the heterozygous genotype CT was associated with nearly 30% lower odds of breast cancer compared to CC genotype (OR=0.74, 95% CI 0.55-0.99; p=0.04) in AA. In the pooled analysis adjusting for both age and sites, the CT genotype was not statistically significant (OR=0.85, 95% CI 0.68-1.06; p=0.15). The common allele T (allele frequency = 0.60) was the risk allele in the previous study conducted in Caucasians, whereas the T allele was the minor allele in subjects of African descent (allele frequency = 0.129 in NG and = 0.232 in AA). In conclusion, our preliminary study found the T allele to have a trend for a protective effect in AA, which is disagreement with the observation made in the original study. Thus, this study indicates the need for replication studies in diverse populations to validate whether rs1434536 is a risk variant for specific subtypes of breast cancer.

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 2620. doi:1538-7445.AM2012-2620
Abstract 2635: association between CASP7 and CASP14 genetic polymorphisms and the risk of childhood leukemia
Authors: Park Chulbum , Song Nan , Han Sohee , Choi Ji-Yeob , Park Sue K , Jeon Sujee , Kang Daehee , Ahn Hyo Seop , Shin Hee Young , Kang Hyoung Jin , Koo Hong Hoe , Seo Jong Jin , Choi Ji Eun
Published: August 2012

Current evidence suggests that apoptosis and the cell cycle system play an important role in cancer development. To identify susceptible genetic markers in these mechanisms, we did an association study in 63 patients and 148 controls. A total of 304 SNPs in 31 gene regions were selected. We evaluated an association at a gene region level by computing the minimum p-value (minP) and doing the false discovery rate (FDR) test. Both SNP and gene-based analyses presented associations with the risk of childhood leukemia for 5 genes: CASP7, CASP14, CASP8AP2, MYC, and RIPK1 (Ptrend < 0.05). There were statistically significant associations for CASP7 (rs12416109 and rs3814231, Ptrend = 0.002 and 0.009, respectively, minP = 0.013, FDR = 0.042) and CASP14 (rs8110862, Ptrend < 0.001, minP = 0.002, FDR = 0.027). This study suggests that genetic polymorphisms in apoptosis and cell cycle related genes might play a role in childhood leukemia development.

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 2635. doi:1538-7445.AM2012-2635
Abstract 2639: A community-based study of nucleotide excision repair polymorphisms in relation to risk of non-melanoma skin cancer
Authors: Wheless Lee E. , Kistner-Griffin Emily , Jorgensen Timothy J. , Ruczinski Ingo , Berthier-Schaad Yvette , Kessing Bailey , Hoffman-Bolton Judith , Woodward Lesley , Yao Yin , Strickland Paul T. , Kao W.H Linda , Alani Rhoda M. , Smith Michael W. , Alberg Anthony J.
Published: August 2012

Nucleotide excision repair (NER) is responsible for protecting DNA in skin cells against ultraviolet radiation-induced damage. Using a candidate pathway approach, a matched case-control study nested within a prospective, community-based cohort was carried out to test the hypothesis that single nucleotide polymorphisms (SNPs) in NER genes are associated with susceptibility to non-melanoma skin cancer (NMSC). Histologically-confirmed cases of NMSC (n=900) were matched to controls (n=900) on age, gender, and skin type. Associations were measured between NMSC and 221 SNPs in 26 NER genes. Using the additive model, two tightly linked functional SNPs in ERCC6 were significantly associated with increased risk of NMSC: rs2228527 (odds ratio (OR) 1.57, 95% confidence interval (CI) 1.20 - 2.05), and rs2228529 (OR 1.57, 95% CI 1.20 - 2.05). These associations were confined to basal cell carcinoma of the skin (BCC) (rs2228529, OR 1.78, 95% CI 1.30 - 2.44; rs2228527 OR 1.78, 95% CI 1.31 - 2.43). These novel, hypothesis-generating findings suggest functional variants in ERCC6 may be associated with an increased risk of NMSC that may be specific to BCC.

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 2639. doi:1538-7445.AM2012-2639
Abstract 2640: Risk of cytochrome P450 1B1 missense polymorphisms in renal cell carcinoma
Authors: Tanaka Yuichiro , Chang Inik , Majid Shahana , Saini Sharanjot , Zaman Mohd S. , Yamamura Soichiro , Chiyomaru Takeshi , Arora Sumit , Singh Kamaldeep , Shahryari Varahram , Dahiya Angela V. , Deng Guoren
Published: August 2012

In male kidney, 4-hydroxy-estradiol has been shown to play a role in the carcinogenesis process. This metabolite is formed from estradiol and due to the hydroxylation activity of the cytochrome P450 1B1 (CYP1B1) enzyme. Prior reports show polymorphisms of CYP1B1 to result in higher catalytic activity and thus, we hypothesize that single nucleotide polymorphisms of the CYP1B1 gene are involved in the malignant transformation of the renal cell. To test this hypothesis, the genetic distributions of four missense CYP1B1 polymorphisms were analyzed in 383 normal healthy subjects and 384 sporadic renal cell carcinoma (RCC) cases by sequence-specific PCR and sequencing. All subjects were men and from a Caucasian population. The polymorphic sites evaluated were at codons 48 (ArgαGly, rs10012), 119 (AlaαSer, rs1056827), 432 (LeuαVal, rs1056836), and 453 (AsnαSer, rs1800440). Results of these experiments demonstrate a trend for the codon 119 Ser/Ser genotype to be a risk for RCC (χ2, P=0.09). Odds ratio for the Ser/Ser genotype was calculated as 1.85 with 95% confidence interval of 1.07-3.20 as compared to the Ala/Ala genotype. The Leu allele on codon 432 was also found to approach significance as a risk for RCC as compared to Val (χ2, P=0.09). Codons 119 and 432 were observed to be linked (R2=0.25) and haplotype analyses show Ala and Val, representing non-risk alleles at these sites, respectively, to be significantly reduced in RCC cases (P=0.06). No association was found however, when analyzing polymorphic sites with clinical stage of RCC. These results demonstrate missense polymorphisms of CYP1B1 to be associated with renal carcinogenesis and are of importance in understanding their role in the pathogenesis of RCC.

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 2640. doi:1538-7445.AM2012-2640