bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 5337: Laminin alpha 2 enables glioblastoma stem cell growth
Authors: LI Meizhang 
Published: August 2012

Abstract
Glioblastomas (GBMs) are lethal cancers that display a cellular hierarchy parallel to the normal brain and are resistant to conventional therapies. It has been suggested that interaction between GSCs and their adjacent perivascular niche plays the critical roles to maintain their malignancy and self-renewal. Extracellular matrix (ECM) cues instruct stem cell behavior in many niches and the objective of our study was to elucidate its composition and contribution to GSC maintenance in the perivascular niche. We utilized human tumor tissue for immunofluorescence analysis and derived GSC for subsequent functional studies from tumor tissues. We find that laminin family of ECM proteins are located in the perivascular GBM niche and are negative prognostic indicators of patient survival. To identify the source of laminins, we characterized cellular elements within the niche and found that key laminins were expressed by non-stem tumor cells and tumor-derived endothelial cells (ECs). Targeting the laminin alpha 2 chain by RNA interference inhibited GSC growth and self-renewal. In co-culture studies of GSCs and non-stem tumor cells, laminin alpha 2 knockdown in non-stem tumor cells resulted in decreased GSC growth. These findings indicate that growth/proliferation of GSCs depends on the direct connection between extrinsic laminin alpha 2 and environmental matrix. Our studies highlight the contribution of non-stem tumor cell-derived laminin juxtracrine signaling. As laminin alpha 2 has recently been identified as a molecular marker of aggressive ependymoma, we propose that the brain vascular ECM promotes tumor malignancy through maintenance of the GSC compartment providing not only a molecular fingerprint but also a possible therapeutic target.
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 5337. doi:1538-7445.AM2012-5337
Abstract 5349: Cancer stem cells isolation and characterization from breast tumor of a germline carrier of TP53 p.R337H (Brazilian founder) mutation
Authors: Mambelli Lisley I. , Kerkis Irina , Hainaut Pierre , Achatz Maria Isabel W. 
Published: August 2012

Abstract
Li-Fraumeni syndrome (LFS), an inherited cancer predisposition syndrome, is associated with germ line mutations in TP53. It is characterized by high risk of multiple, early cancers. In Brazil, a variant form of LFS is exceedingly frequent due to a widespread founder TP53 mutation, p.R337H, detected in about 0.3% of the general population in Southern Brazil. This mutation occurs in p53 oligomerization domain and its effect on p53 oligomerization is supposed to be dependent upon pH conditions. We have isolated and characterized Cancer Stem Cells from tumors of p.R337H mutation carriers. After informed consent and surgical resection fragments were dissociated and brought in culture. Adherent cells and spheroids were derived from different tumor types. Spheroids derived from a breast cancer (BC) were further analyzed to demonstrate positive immunolabeling for CD44, CD24, Oct4 and Sox2 antibodies. Time-lapse videomicroscopy showed rapid growth, frequent asymmetric division and absence of senescent phenotypes for least 17 passages. At confluence, some cells spontaneously differentiated into adipogenic and muscle-like cells. Floating and adherent spheroids contained self-renewing cells capable of long-term proliferation without losing their primary characteristics. These properties are similar to those of stem cells with inactivated p53 protein. However, treatment of adherent cells with the DNA damaging agent doxorubicin elicited p53 accumulation, p21 induction, cell cycle arrest at low doses of Dox and cell death/apoptosis at higher doses. These results are consistent with the notion that p.R337H may retain a quasi-normal p53 function in response to stress while expressing properties similar to mutant p53 in long-term CSC culture. These p.R337H CSC may provide a very useful model for studying the defects underlying inherited cancer in p.R337H carriers.
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 5349. doi:1538-7445.AM2012-5349
Abstract 5361: Enriching and characterizing cancer stem cell grown on collagen type IV-coated plate in primary pediatric brain tumors
Authors: Kim Dong Ok , Park Jun Bum , Ra Young Shin 
Published: August 2012

Abstract
There is accumulating evidence that cancer stem cells (CSC) in adherent culture maintain and enrich their characteristics by extracellular matrix (ECM). This study focused on differential expression of not only CSC markers and differentiation markers, but also embryonic signaling pathways when cultured on various ECM-coated plates. Four cells from pediatric patients with brain tumor were grown in DMEM/F12 medium supplemented with 10ng/ml FGF, bFGF and 0.5x B-27. After forming of neurosphere, the cells were cultivated on each three kinds of ECM-coated plate including collagen type IV, fibronectin and poly-D-lysine. And the expression level of CD133, CD15, Nestin, Oct3/4, SOX2 as CSC markers, GFAP, β-tubuliln III for differentiation markers, Wnt, Notch, Shh, their transcription factors like β-catenin, Hes1, Gli1 respectively for embryonic signaling pathways was assessed by reverse transcriptase (RT)-PCR. As a result, two among four brain tumor cells grown on collagen type IV-coated plate expressed higher expression level of CD133, CD15, Nestin, SOX2, Oct3/4, Wnt5a and Notch than in the cells cultured on fibronectin or poly-D-lysine-coated plate. And residual two cells cultivated on collagen type IV and fibronectin, not on poly-D-lysine were shown similar expression level of almost all genes. There is no difference of Shh, Gli1, GFAP and β-tubuliln III expressions in all cells grown on each of three ECM-coated plates. In conclusion, collagen type IV is required for enriching and maintaining of CSC characteristics in primary pediatric brain 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 5361. doi:1538-7445.AM2012-5361
Abstract 5367: Long-term survival in a phase II atudy of belagenpumatucel-L (TGF-β antisense modified tumor cell vaccine) in non-small cell lung cancer (NSCLC).
Authors: Bazhenova Lyudmila , Carrier Ewa , Shawler Daniel , Fakhrai Habib 
Published: August 2012

Abstract
Background: Cancer therapeutics vaccines continue to be investigated in a variety of solid malignancies. Belagenpumatucel-L (Lucanix®), a therapeutic vaccine comprised of 4 TGF-β2 antisense gene-modified allogeneic NSCLC cell lines. Phase II trial finished its enrollment in March of 2004 and was published in 2006. We now report an updated survival analysis with over 5 years of follow up. Methods: Three arms, open label clinical trial enrolled seventy-five subjects. 2 stage II, 12 stage IIIA, 15 stage IIIB, and 46 stage IV patients were randomized into three dose cohorts of 1.25, 2.5, or 5 107 cells/injection. Several cellular (ELISPOT and cytoplasmic cytokine expression) and humoral (antibody ELISA) immunity assays were also performed and correlated with survival. Results: Median follow up for all patients was 14.5 months and for patients with stable disease 44 months. Median survival for all subjects was 14.5 months and one-year, two-years and five-year survival were respectively 55%, 35% and 20%. Stages IIIB/IV subjects enrolled into cohorts 2 and 3 (N=40) had a median survival of 15.9 months and one-year, two-year and five-year survivals were respectively 61%, 41% and 18%. For stage IIIB/IV patients with non progressive disease following frontline chemotherapy, median survival was 44.4 months and five-year survival was 50%. For subjects who progressed following frontline chemotherapy, median survival was 14.1 months and five-year survival was 9.1%. Subjects who demonstrated an increase in both cellular and humoral immune reactivity following treatment had a significant survival advantage over subjects who showed an increase in only one measure of immunity with a median survival of 32.5 months vs. 11.6 months (p = 0.015). Conclusion: Long term follow up of a phase II clinical trial confirmed encouraging survival for patients with stage IIIB/IV disease. Based on these data, we have instituted an international, randomized, pivotal Phase III trial to evaluate the efficacy of belagenpumatucel-L in a maintenance setting in stage III/IV NSCLC patients who have stable disease or better following frontline chemotherapy. The trial is designed to enroll 506 patients and is powered to measure a 3.5 month survival difference. There are two planned interim analyses. To date, 285 patients have been enrolled in 8 countries. Confirmation of the phase II data in a randomized, phase III setting would provide an important improvement for the treatment of non-small cell lung 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 5367. doi:1538-7445.AM2012-5367
Abstract 5406: Glioblastoma derived exosomes induce apoptosis in T cells
Authors: Sabin Keith Z. , Winn Robert J. 
Published: August 2012

Abstract
Intro: Glioblastoma multiforme (GBM) is the most common primary brain tumor in adults and is associated with an average lifespan of 15 months after diagnosis. One contributing factor to this poor prognosis is the immune protection afforded by the tumor microenvironment. One method of immune evasion poorly characterized is the effect of tumor released exosomes. Methods: FasL gene expression was determined using reverse transcription polymerase chain reaction (RT-PCR) in three GBM cells lines: LN-229, U-138 MG and T98G. Exosomes were isolated from serum free supernatants by differential ultracentrifugation. Exosomes were quantified using a protein concentration assay. A3 Jurkat T cells were treated with quantified exosomes, recombinant FasL or untreated. After 24 hours a TUNEL assay was performed to investigate the potential apoptotic effects of GBM-derived exosomes. Results: FasL is expressed in LN-229, U-138 MG and T98G cell lines as determined by RT-PCR. After treating A3 Jurkat T cells with exosomes it appeared apoptosis was induced as compared to an untreated control as determined by DNA fragmentation. Discussion: Studies using prostate and colon cancer cell lines showed that FasL was expressed in tumor derived exosomes. When T cells were co-cultured with exosomes apoptosis was induced in a FasL-dependant manner. Our lab previously demonstrated that GBM-derived exosomes are able to inhibit T cell proliferation. These current data suggests that GBM-derived exosomes induce apoptosis in T cells.
Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5406. doi:1538-7445.AM2012-5406
Abstract 5409: Cancer immunosuppression: The role of human leukocyte antigen G expression
Authors: Elliott Robert L. , Jiang Xian P. , Phillips Jeffery T. , Head Jonathan F. 
Published: August 2012

Abstract
Human leukocyte antigen G (HLA-G) is an immunotolerant nonclassical major histocompatibility complex class Ib molecule. During pregnancy it is expressed by trophoblastic placental cells to protect the fetus from maternal alloreactivity. HLA-G is involved in cancer immune evasion and is overexpressed by many tumors. Reverse transcription-polymerase chain reaction and immunohistochemistry were used to examine HLA-G expression in human normal breast and breast cancer cell lines, and human normal breast and breast cancer tissues. Reverse transcription-polymerase chain reaction confirmed that normal epithelial MCF-12A cells had no HLA-G mRNA, but cancer cell lines MCF-7, T47D, and MDA-MB-231 and NCI/Adr-Res had various levels of HLA-G mRNA expression. Thirty-eight breast cancers and 12 normal breast tissues were examined by immunohistochemistry. Fifty-eight percent (22/38) of cancers had medium to strong staining for HLA-G, whereas only 8% (1/12) of normal breast tissues had medium to strong staining, and the difference was statistically significant (p&lt;0.05). HLA-G staining was found in the cytoplasm and membranes of cancer cells. In conclusion breast cancer cells overexpress HLA-G mRNA and protein, which probably contributes to immune evasion and treatment resistance. We believe HLA-G should be monitored in breast and other epithelial cancers as it may be both a prognostic indicator and a therapeutic target.
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 5409. doi:1538-7445.AM2012-5409
Abstract 5410: Tim-3 expression and function in natural killer cells from metastatic melanoma patients
Authors: da Silva Ines P. , Jimenez-Baranda Sonia , Gallois Anne , Kuchroo Vijay K. , Osman Iman , Bhardwaj Nina 
Published: August 2012

Abstract
BACKGROUND: Tim-3 (T-cell immunoglobulin domain and mucin domain 3) is a type-I glycoprotein receptor with pivotal roles in immune regulation and tolerance induction. Tim-3 has opposing roles in innate and adaptive immunity. It synergizes with TLR signaling in DCs inducing the production of inflammatory cytokines, while it acts as a negative regulator of Th1/Tc1 cell function by triggering T cell apoptosis upon interaction with its ligand, galectin-9. Tim-3 is also responsible for the functional exhaustion of T cells observed in some chronic infectious diseases and tumors, such as metastatic melanoma (MM). Natural Killer (NK) cells, innate immune cells which eliminate tumors and infected cells through cytotoxicity and IFNγ production, constitutively express Tim-3. Moreover, it has been described that some tumor cells (including melanoma cells) express and secrete galectin-9. However, little is known about the role of Tim-3 on NK cells, particularly in the presence of galectin-9, in healthy or melanoma patients. METHODS: Purified NK cells from PBMCs of 20 melanoma donors (MD) and 40 healthy donors (HD) were characterized according to the expression of Tim-3 by flow cytometry. Lamp-1 expression on NK cell surface was used to measure NK cell cytotoxicity against the following target cells: i) K562 cells which don't express Galectin-9 and ii) Gmel galectin-9+ and Gmel galectin-9- sorted melanoma cells. IFNγ production was assessed by intracellular staining after 4 hours culture in presence of IL-12. RESULTS: MD NK cells express higher levels of Tim-3 compared to HD NK cells (p&lt;0.05). Moreover, MD NK cells have a defect in cytotoxicity, proliferation and IFNγ production, but it still remains unclear if Tim-3 is involved in this exhausted phenotype. So far, we found that Tim-3 expression by itself (without engagement of specific ligands) does not negatively affect NK cell cytotoxicity. However, the expression of Galectin-9 by the target cells induces a decrease in NK cell cytotoxicity (Gmel Gal-9+ vs Gmel Gal-9-). CONCLUSION: These data suggest that when engaged with Galectin-9, Tim-3 expression down-modulates NK cell cytotoxicity. Therefore, galectin-9 expression may be a possible mechanism for tumors to evade immune surveillance. Given that an increasing body of data supports an important role for Tim-3/Galectin-9 interaction in immune regulation in infectious diseases and cancer, deciphering the function of Tim-3 in different cell types will be critical for potentially targeting it therapeutically.
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 5410. doi:1538-7445.AM2012-5410
Abstract 5415: Expression of immunosuppressive leukocyte inhibitory immunoglobulin like transcript 3(ILT3) receptors increases with the development and progression of ovarian tumors
Authors: Khan Mohammad F. , Yellapa Aparna , Bahr Janice , Bitterman Pincas , Abramowicz Jacques , Basu Sanjib , Edassery Seby , Barua Animesh 
Published: August 2012

Abstract
Background: Immunosuppressive tumor microenvironment has been suggested as one of the barriers to anti-tumor immune response in patient with ovarian cancer (OVCA). Leukocyte Inhibitory Receptor Immunoglobulin-like Transcript 3 (ILT3) has been proposed as a factor involved in the suppression of immune response in several malignancies. However, the expression and role of ILT3 in the development and progression of ovarian tumors has not yet been studied. Objective: The goal of this study was to examine the expression and association of ILT3 in ovarian tumors in laying hen, a preclinical model of OVCA in humans. Methods: A cohort of mature White Leghorn laying hens with normal or low egg laying rates were evaluated by transvaginal ultrasound scan (15 normal hens and 30 hens with OVCA).All hens were euthanized and serum and ovarian tissues from all hens were collected and processed for immunohistochemistry (IHC), Western blotting (WB) and RT-PCR. Presence of tumors and tumor staging were confirmed by gross morphology and histology. Expression of ILT3 was examined by IHC, WB and RT-PCR. Results: Immune cell like cells in the ovarian stroma as well as epithelium of the normal ovaries and ovarian tumors expressed ILT3. The intensity of ILT3 expression was significantly higher in hens with OVCA than normal hens. Moreover, the intensity of ILT3 expression varied with histological subtypes of ovarian tumors and was significantly higher in serous OVCA than endometrioid OVCA. A band of approximately 55kDa of ILT3 protein was detected in the homogenates of normal ovarian surface epithelium or tumor epithelium as well as whole normal or ovarian tumor extracts. Similar to IHC observations, protein expression was stronger in hens with OVCA than in normal hens. Conclusion: This is the first study to show that immunosuppressive ILT3 is expressed by ovarian tumor epithelium and the intensity of expression increases as the disease progresses. Thus the results are consistent with the hypothesis that ILT3 may be involved in the suppression of anti-tumor immunity in OVCA. The laying hens may be a useful model to understand the OVCA-associated expression of ILT3. This animal model also offers the opportunity to develop and test anti-ILT3 therapy to enhance anti-tumor immunity against OVCA in humans. Support: Department of Defense (OC#93303) and Elmer Sylvia and Sramek Foundation.
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 5415. doi:1538-7445.AM2012-5415
Abstract 5424: Docosahexaenoic acid (DHA) induces gene expression of RASSF1A through epigenetics mechanisms in breast cancer cells
Authors: de Cassia Borges de Castro Rita , Almeida Danielle Fontes de , Ravacci Graziela Rosa , Roela Rosimeire Aparecida , Mazzotti Tatiane Katsue Furuya , Brentani Maria Mitzi , Conti Aline De , Ong Thomas Prates , Moreno Fernando Salvador , Waitzberg Dan Linetzky 
Published: August 2012

Abstract
Background: Epigenetic changes, such as DNA methylation and post-translational histones modifications have an important role in mammary tumorigenesis. Changes in DNA methylation include global hypomethylation and hypermethylation of promoter regions of specific genes. Several genes are hypermethylated in breast cancer, especially the tumor suppressor gene RASSF1A, that have potential prognostic value. Epigenetic events are important as therapeutic targets due to its reversible feature. Dietary compounds can modulate the epigenetic processes in cancer. Experimentally, docosahexaenoic acid (DHA), a member of the omega-3 fatty acids, can reduce proliferation, induce apoptosis and decrease the invasive potential of breast tumor cells. However, the antitumor mechanisms of DHA and its ability to modulate epigenetic events are not yet fully elucidated. Objectives: To verify DHA action on RASSF1A expression and promoter region methylation in different human breast cancer cell lines. Material and Methods: Three different breast cell lines (MCF-7, MDA-MB-231, SKBR-3) were treated or not with 100 μM of DHA for 72 hours. Real time quantitative PCR were performed for gene expression quantification of RASSF1A. DNA methylation of the promoter region of RASSF1A was evaluated by methylation specific PCR. The western blot will be held for analyze of the histone 3 lysine 9 acetylated (H3K9ac) protein. Moreover, we will evaluate the phases of the cell cycle and apoptosis by flow cytometry. Results and Conclusion: Compared to control cells (not treated), DHA increased RASSF1A expression on MCF-7, but not in MDA-MB and in SKBR-3. These three transformed breast cells lines show methylation in specific region of RASSF1A promoter, indicating its role as a tumour supressing gene. DHA treatment increased RASSF1A promoter region hypomethylation in MCF-7, but not in MDA-MB and in SKBR-3. From this perspective, we can suggest that DHA could act by epigenetic mechanisms and reactivate RASSF1A, previously silenced. This action occured specifically in MCF-7, a less aggressive breast tumor cell line. It is expected that the present results promote better understanding of DHA role as a potential anti-oncogenic compound 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 5424. doi:1538-7445.AM2012-5424