bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 556: Extracellular matrix protein mixture isolated from cultures of human tumor and stromal cells supports growth of human tumor cells in vitro and in vivo
Authors: Mishra Pravin J. , Mishra Prasun J. , Gao Hui , Glod John , Banerjee Debabrata
Published: February 2011

Carcinoma Associated Fibroblasts (CAFs) along with other cells present in the tumor milieu play a major role in tumor progression. We have previously demonstrated differentiation of human bone marrow derived mesenchymal stem cells to CAF like phenotype upon exposure to tumor conditioned medium. The present study evaluates the ability of these stromal components to support tumor progression. An in vitro system was used to study impact of the components of the tumor milieu on growth of luciferase expressing MDA MB 231 cells, representing basal subtype of human breast cancer. CAFs along with phorbol ester induced HL60 cells representing a macrophage like phenotype supported growth of MDAMB231 cells in vitro as measured by a luciferase assay. When injected subcutaneously in vivo in nu/nu mice, tumor cells comingled with CAFs or with differentiated HL60 cells showed almost similar tumor growth rate, whereas CAFs and differentiated HL60s when mixed together showed more than 4 fold increased tumor growth rate compared to the tumor cells alone and 2 fold compared to tumor cells plus MatrigelTM.

Electrophoretic analysis of extracellular matrix protein mixture isolated from mixed cultures of CAFs plus MDAMB231 cells and differentiated HL60 cells (3 cells) using previously published methods showed presence of laminins and collagens, and demonstrated robust tumor cell growth compared to MatrigelTM. The ECM protein mixture derived from the three cell system offers a completely humanized substitute for MatrigelTM useful for study of tumor growth and response to therapy in vitro as well as in vivo.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 556.
Abstract 5563: The anti-cancer and anti-hyperlipidemia effects of triterpenoids from Gynostemma pentaphyllum in the Apcmin/+ mouse model
Authors: Tai William Chi-Shing , Wong Wing Yan , Wang Yu , Hsiao Wen-Luan Wendy
Published: February 2011

Apcmin/+ mice, a commonly used mouse model for colorectal cancer, manifest both high number of intestinal polyps and hyperlipidemia, which make the min/+ mouse an ideal animal model for drug intervention studies for both hyperlipidemia and colorectal cancer Triterpenoids, produced in many plants, are widely used in Asian medicine. Gynostemma pentaphyllum (Gp), also called jiaogulan, is a rich source of dammarane-type triterpenoids and is used as a traditional Chinese herbal medicine for the treatment of various diseases including cancer and hyperlipidemia. However, the proven efficacy and the underlying mechanism have not been investigated systematically. Using the Apcmin/+ mouse as the animal model, treatment with Gp total triterpenoids (GpMix) markedly reduce the numbers and sizes of polyps. In conjunction with 5-fluorouracil (5-FU), the size and numbers of polyps of the treated animals were further reduced. Besides the anti-cancer effect of GpMix, we have also shown that the GpMix can effectively suppress the plasma triglycerides and cholesterol levels in Apcmin/+ mice. To delineate the precise mechanisms of the anti-cancer and anti-hyperlipidemia effects of GpMix, we performed molecular and proteomic analysis of the plasma obtained from animals treated with and without GpMix.

Results showed that lipoprotein lipase, PPARα and adiponectin are upregulated and HMG-CoA reductase is down-regulated upon GpMix treatment. Plasma protein profiling revealed that several downregulated proteins are directly related to cancer, inflammatory, cell proliferation, including serum amyloid A1 and A2, serum amyloid P-component, major urinary proteins, glutathione peroxidase 3, complement C3 and serotransferrin. In the same treated animals, four lipogenesis proteins including apolipoprotein A-I, apolipoprotein A-II, apolipoprotein A-IV, and apolipoprotein C-III by which the activity of lipoprotein lipase is tightly regulated are downregulated. These protein profiling data, together with the adipocyte-specific signaling data shown above provide solid evidence of the anti-cancer and anti-hyperlipidemia effects of GpMix. [This study was financially supported by Research Grants Council of Hong Kong under HKBU2/07C and FRG/07-08/II-50 and FRG/08-09/II-61 Grants to WLWH.]

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5563.
Abstract 5565: Quantitative proteomics identifies phosphorylation targets, modulators and pathway signatures of oncogenic RAS-mediated MAPK signaling in lung adenocarcinoma
Authors: Sudhir Putty-Reddy , Hsu Chia-Lang , Wang Yi-Ting , Sung Ting-Yi , Hsu Wen-Lian , Chen Yu-Ju , Yang Ueng-Cheng , Chen Jeou-Yuan
Published: February 2011

Functional phosphoproteomics is applied to gain the molecular insights into oncogenic RAS-mediated signaling pathway in lung adenocarcinoma. By means of immobilized metal affinity chromatography, mass spectrometry and a robust label-free quantitative system, we compared the phosphorylation profiles of five cell lines including immortalized normal human bronchial epithelial cells (HBECs) (wt KRAS), 3KTR (KRASG12V-transfected HBECs), A549 (KRASG12D), H322 (amplified KRAS), and H1299 (NRASQ61K). This resulted in the quantification of 1508 phosphorylation events and detection of regulated events across the cell lines. NetworKIN analysis of 2-fold regulated events (n=77) induced by oncogenic RAS in HBECs revealed 20 novel site-specific phosphorylation targets of MAP kinases. By browsing through STRING database, we identified 44 known substrates of MAP kinases in the interactome of phosphoproteins in this study. Further analysis of the site-specific upstream kinases of regulated phosphorylation events revealed the significant increase in basal kinases such as PAK, AKT/PKB and PKA in H1299 (large cell carcinoma) relative to A549 and H322 (adenocarcinoma). This increase in basal kinases may in-turn modulate the output of RAS-mediated MAPK signaling in large cell carcinoma cells (H1299). Supporting these results, majority of the regulated phosphorylation events (n=77) including MAPK, identified in HBECs expressing oncogenic RAS, were also differentially regulated in A549 and H322 but not in H1299. Further, inferring pathway signatures by integrating expression data of phosphorylation profiles to pathway interaction database, revealed the activation of MAPK signaling in 3KTR, A549 and H322 but not in H1299, whereas the activation of AKT and inactivation of mTOR signaling in H1299. Taken together, we demonstrate the phosphorylation targets, molecular regulation, and pathway activity signatures of onogenic RAS-mediated MAPK signaling in lung adenocarcinoma. Furthermore, the method introduced here to infer the pathway signatures is adaptable for future studies.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5565.
Abstract 5570: An affinity lectin chromatography approach to characterize N-linked glycoproteome in metastatic osteosarcoma
Authors: Flores Ricardo J. , Li Yiting , Lau Serrine S. , Leung Eastwood , Lau Ching C. , Man Tsz-Kwong
Published: February 2011

Osteosarcoma (OS) is the most common bone cancer in children and adolescents. Approximately 20% of OS patients have clinically detectable metastasis at the time of diagnosis, which is the most consistent indicator of poor outcome. Therefore, an easy-to-use and sensitive method is urgently needed to detect and monitor metastatic patients in order to offer an intensified or personalized therapy. By analyzing glycoproteins in OS cells, we may identify biomarkers that would help detect early metastasis and shed light on how this cancer develops. Thus, we used a tumor-targeted glycoproteomic approach in order to identify metastasis-associated glycoproteins in OS cells, and test if these can be used as blood biomarkers to indicate metastasis or spread of disease at diagnosis. In this pilot study, we have analyzed a previously established human OS metastatic cell line (LM7). We captured a specific subset of N-linked glycoproteins from LM7 cells by Wheat Germ Agglutinin (WGA) affinity chromatography. Subsequently, we identified the captured glycoproteins by a multidimensional protein identification technology (MudPIT). In addition, two samples with or without PNGase treatment were tested to estimate the impact of protein glycosylation on the MS/MS analysis. A total of 121 and 368 unique peptides were identified in the non-treated and PNGase treated samples, respectively. All of the proteins identified with two or more distinct peptides were predicted to have potential N-linked glycosylation sites, and 57% of these were found to have one or more typical Asn-X-Ser/Thr N-glycosylation sites. Many of the identified glycoproteins are known to be involved in tumorigenesis. To this end, we have optimized the affinity lectin chromatography and mass spectrometry to capture and identify N-linked glycoproteins in metastatic osteosarcoma cells. Differentially expressed glycoproteins in metastatic OS will be further identified by comparing LM7 with a non-metastatic control. The candidate biomarkers will be validated in the plasma and tumor samples collected from OS patients.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5570.
Abstract 5571: Identification of autoantibody biomarkers in metastatic osteosarcoma using a high-density protein microarray approach
Authors: Li Yiting , Yu Alexander , Okcu Fatih , Hicks John , Chintagumpala Murali , Lau Ching C. , Man Tsz-Kwong
Published: February 2011

Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. A leading cause of death in OS is the development of metastasis. Therefore, it is important to detect metastatic patients as early as possible, so these patients can be stratified and treated while the disease is still in its early stages and potentially curable. Autoantibodies represent a promising class of plasma proteins that can be used for the early detection of cancer. These immunoglobulins can recognize aberrant proteins in the tumor cell during the development of metastasis. We have used a new high-density protein microarray platform, ProtoArray Human Protein Array (HPA), to identify significant autoantibody biomarkers that are associated with metastatic OS at the time of diagnosis. ProtoArray HPA consists of over 8,000 purified human proteins from different functional classes. Plasma samples from metastatic and localized OS patients, and children with non-cancerous diseases were analyzed by the microarrays. The results showed that the amounts of 136 distinct autoantibodies were significantly different between the plasma samples of metastatic and localized OS cases (p < 0.01). Among these autoantibodies, 66 were higher while 70 were lower in metastatic patients. Two of these autoantibodies, which target CTTN and PFKFB1, were selected for further investigation. We found that the phosphorylated form (p85) of the CTTN protein in two metastatic OS cell lines was expressed higher relative to the non-metastatic parental cells. This result is consistent with the previous finding that increased phosphorylation of CTTN is important for the metastatic phenotype of cancer cells. The PFKFB1 autoantibody in the localized patients was higher than the metastatic patients; however, both groups of OS patients had a higher PFKFB1 level relative to the control group. When compared to normal osteoblasts, Western blotting showed that PFKFB1 protein was overexpressed in seven of the eight OS cell lines. PFKFB1 is involved in the Warburg effect, which is important to the energy metabolism of cancer cells. Our results suggest that the high-density protein microarray approach is useful for identifying metastasis-associated autoantibodies in cancer patients. Further validation and investigation of the candidate autoantibody biomarkers may lead to discovery of novel biomarkers and unravel the underlying mechanism in metastatic OS.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5571.
Abstract 5574: Altered glycerophospholipid metabolism in non-small cell lung carcinoma
Authors: Hoang Chuong D. , Wu Manhong , Xu Yue , Merrit Robert , Whyte Richard , Shrager Joseph , Peltz Gary
Published: February 2011

Objective: It is hypothesized that accumulation of specific metabolites precedes morphologic changes in human lung tumors during malignant transformation, and that this may be exploited to develop a lung cancer biomarker.

Methods: We performed metabolomic profiling on 24 fresh-frozen specimens from patients with biopsy proven lung adenocarcinoma. The following tissues from each of the patients were analyzed: tumor, adjacent-to-tumor tissues, and normal lung. The metabolites were extracted by a methanol chloroform technique and separated by high resolution liquid chromatography. They were analyzed using the 6520 hybrid quadrupole time-of-flight mass spectrometer (Agilent). Mass Profiler Professional software (Agilent) was used for statistical analysis and metabolite profiling. To identify a metabolite of interest, the retention time and elemental composition data were searched against databases: METLIN Personal Metabolite (Agilent). After multiple data filtering steps we derived a non-small cell lung carcinoma-specific metabolomic fingerprint. We performed metabolic network analysis to assess which gene(s) may be associated with the metabolomic biosignatures.

Results: We detected an average of 2232 unique metabolites in the tumor specimens, 1795 in the adjacent-to-tumor specimens, and 1613 in the normal lung. When tumor and normal lung samples were compared, 201 metabolites were significantly over-expressed and 105 were under-expressed in the tumor tissue. The metabolic biosignature for the tumor group was highly distinct from the other lung regions (several fold change). The metabolites within the tumor-specific signature included the following families: lactate, amino acids, phospholipids, eicosanoids, and glycine derivatives. In particular, multiple phosphatidylethanolamines showed dramatic differential expression in tumor tissues compared to normal lung. Metabolic network analysis suggests that the altered metabolite levels could affect critical cellular functions, including: cell signaling, growth and invasiveness.

Conclusion: Metabolomic analysis has identified metabolites that were uniquely found in human lung adenocarcinomas. The specific metabolic biosignature of lung adenocarcinoma could suggest novel biomarkers that could be used for diagnosis, as well as suggest altered molecular pathways contributing to the pathogenesis of lung cancer.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5574.
Abstract 5577: Global analysis of protein kinase activity during hematopoietic differentiation
Authors: Nomanbhoy Tyzoon K. , Ng Dana M. , Patricelli Matthew P.
Published: February 2011

Cell proliferation and differentiation are highly regulated processes, and the deregulation of these processes can lead to a number of diseases including cancer. Protein kinases are key regulators of cell proliferation and differentiation, and consequently, potential targets for a number of oncology indications. A detailed understanding of the changes in kinase activity/expression during cell proliferation and differentiation may be one of the keys to the identification of novel biomarkers and targets for the diagnosis and treatment of these diseases. The HL-60 cell line is promyeloblastic, and protocols for the differentiation of these cells into either macrophages by 12-O-tetradecanoylphorbol-13-acetate (PMA), or granulocytes by retinoic acid (RA) are well-established. Importantly, HL-60 cell differentiation is a model system for the treatment of myelocytic leukemia, and identifying the mechanisms that induce terminal differentiation of these cells may reveal effective targets for treatment of this disease. With this in mind, we evaluated the kinase activity and expression profiles of HL-60 cells throughout the differentiation process using desthiobiotin acyl phosphate ATP and ADP probes and liquid chromatography-mass spectrometry. Greater than 150 protein kinases in resting, RA, and PMA HL-60 cells were analyzed. Significant changes in activity/expression were seen for a number of protein kinases in both differentiation pathways. These changes in kinase activity were generally consistent with changes in protein expression levels, as determined by Western blot. In both RA and PMA-induced HL60 differentiation, the kinetics for the changes in activity of the kinases was determined, and the results suggest that there is a well-defined sequence during which these changes occur. In summary, using activity based protein profiling, we identified a subset of kinases whose activity changed significantly during RA and PMA-induced HL-60 differentiation. Importantly, this methodology is widely applicable to other cellular systems and disease models, and has applications in the identification of biomarkers and targets for the management of these diseases.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5577.
Abstract 5579: Novel proteins highly expressed in tumor identified by a high throughput immunoproteomic approach
Authors: Grifantini Renata Maria , Pileri Piero , Pierleoni Andrea , Campagnoli Susanna , Grandi Alberto , Nogarotto Renzo , Parri Matteo , Canidio Elena , Cattaneo Davide , Pagani Massimiliano , Camilli Elisa De , Abrignani Sergio , Grandi Guido , Viale Giuseppe , Sarmientos Paolo
Published: February 2011

Cancer biomarker discovery is an extremely active research track in both academia and pharma industries. Several high throughput technologies were applied through the years to identify proteins specifically related to cancerous phenotype. However, most putative biomarkers identified from conventional proteomic/transcriptomic strategies still need to be clinically validated. Here we present a cancer biomarker discovery approach based on the use of a large library of antibodies raised against recombinant human proteins to detect tumor-associated proteins by immune-histochemistry analysis of clinical tumor tissues. Starting from the whole human genome, genes encoding proteins predicted as membrane- or secreted were selected and high through-put cloned and expressed. Recombinant proteins were used to build a polyclonal antibody library (YOMICS®) currently comprising more than 1700 murine immune sera. The ability of sera to recognize specific targets predominantly present in tumors was assessed by Tissue MicroArray (TMA, a miniaturized immunohistochemistry analysis) of tumor tissue samples and healthy controls from pedigreed patients affected by the most common human tumor types, including colon, ovary, breast, lung, ovary and prostate cancers.

While the screening is still in progress, six antibodies were identified showing high reactivity on a high percentage (ranging from 40 to 95%) of tumor tissues on one or more cancer type, with negative staining on normal tissues. The corresponding protein targets, being novel tumor-associated proteins, were validated and characterized at cellular and molecular level by analyzing their expression, cellular localization and biological role in a panel of tumor cell lines. Interestingly, two of the six proteins are localized at the plasma membrane, while the other proteins are intracellular. Data so far available indicate that one of the six proteins confers an invasive phenotype to tumor cell lines. The newly identified proteins are promising candidates as new biomarkers and could be exploited to develop target-specific drug therapies.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5579.
Abstract 5584: DNA vaccine encoding prostatic acid phosphatase (PAP) elicits long-term T-cell responses in patients with recurrent prostate cancer
Authors: Becker Jordan T. , Johnson Laura E. , McNeel Douglas G.
Published: February 2011

Prostatic acid phosphatase (PAP) is a tumor antigen in prostate cancer, and several clinical trials have used this as a tissue-specific target for anti-tumor vaccines. Ultimately the goal of anti-tumor vaccines is to elicit a sustainable immune response able to eradicate a tumor or at least restrain its growth. We have investigated plasmid DNA vaccines encoding PAP and have previously reported the results of a phase I trial in which 22 patients with biochemically recurrent prostate cancer (clinical stage D0) were immunized six times at biweekly intervals using a DNA vaccine encoding PAP (pTVG-HP). In previous reports we identified that PAP-specific T-cells were elicited, including PAP-specific cytolytic T cells, two weeks following the six immunizations. In the current study we investigated the immunological efficacy of subsequent booster immunizations in two individuals, and conducted more detailed immunological analysis by IFNγ ELISPOT and T cell proliferation, with cells obtained prior to immunization, after every two immunizations, and quarterly for one year following immunization. These studies were conducted to determine when immune responses were elicited, and whether these responses were durable over time, to guide the schedule of vaccine administration to be used in future clinical trials with this particular DNA vaccine. We found that the majority of immune responses elicited with vaccination were only detectable after four to six immunizations. In addition, PAP-specific T-cell responses did persist in several individuals for many months, or were detectable only after several months. Finally, we identified that immune responses previously elicited could be further amplified by subsequent booster immunizations months later. These results suggest that future trials using this DNA vaccine, and potentially other anti-tumor DNA vaccines, should investigate ongoing, rather than limited, schedules of administration with periodic booster immunizations. Moreover, these results suggest that DNA vaccines targeting PAP could potentially be combined with other vaccine strategies targeting this particular antigen to augment PAP-specific T-cell immunity. A pilot trial to evaluate different schedules of DNA vaccine administration for the establishment of persistent PAP-specific memory T cells is currently underway.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5584.
Abstract 559: Human ovarian cancer mesenchymal stem cells demonstrate abnormal BMP signaling and promote tumorigenesis
Authors: Buckanovich Ronald J. , McLean Karen , Silva Ines , Gong Yusong , Pennington Kathryn
Published: February 2011

Numerous recent reports indicate a critical role for MSC in solid tumor growth. Unfortunately, very little is known about human solid tumor associated MSC. We analyzed over 15 samples of primary ovarian tumors for the presence MSC. MSC were isolated from over 90% of these samples. These cells had the morphologic appearance of CFU-F (colony forming unit-fibroblastic), a normal karyotype, and were non-tumorigenic. Phenotypic characterization of these cells indicates they express the surface markers normally associated with MSC; they express traditional MSC markers including SH2, SH3, and CD90, and do not express hematopoietic markers such CD34, CD14, or CD4. Functional characterization demonstrates that these cells are multipotent, capable of differentiating into adipose, cartilage, and bone. However, ovarian cancer-derived MSC appear to be somewhat less efficient at differentiation than the healthy donor control MSC. When combined with tumor cells in immune deficient mice, the promotion of tumor growth by ovarian cancer associated MSC was significantly greater than that seen with healthy donor control MSC. Gene expression analysis demonstrated that cancer associated MSC have significantly altered BMP family member expression. These studies suggest that cancer associated MSC are unique from their healthy tissue associated counterparts and play a critical role in cancer cell growth.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 559.
Abstract 5591: Detection of autoantibody responses to prostate cancer-associated antigens following treatment of patients with androgen deprivation therapy
Authors: Maricque Brett B. , McNeel Douglas G.
Published: February 2011

Over the last decade, immunotherapies have been developed for the treatment of prostate cancer with the goal of generating anti-tumor immune responses. Vaccine-based treatments target the immune system to a specific antigen, or set of antigens, expressed by the tumor. Other therapeutic modalities, including androgen deprivation therapy, can result in tissue damage and have been demonstrated to elicit prostate antigen-specific immune responses. The evaluation of immune responses to individual tumor antigens in the context of these non-antigen-specific immunomodulatory treatments has been notoriously difficult and time-consuming. We have previously reported a panel of 125 commonly recognized prostate cancer-associated antigens identified from multiple IgG screening approaches. This panel included multiple cancer-testis antigens and antigenic proteins recognized by IgG in patients with chronic prostatitis or prostate cancer. In the current study we questioned whether this panel could be used to identify antigen-specific immune responses elicited in patients with prostate cancer following androgen deprivation. 125 lambda phage encoding these unique antigens were spotted in triplicate using a high-throughput, high-density SEREX methodology. Sera were obtained from 33 patients with prostate cancer collected pretreatment and three months post-androgen deprivation therapy. These sera were used to probe this panel and IgG antibody responses were detected and objectively quantified by densitometry. 31 sera sets met pre-specified criteria as adequate for analysis. Results demonstrated that autoantibodies were induced to 62 of the 125 antigens. IgG responses were elicited to 21 of the antigens in multiple patients, including MAD-Pro-34, MAD-CT-1, and LAGE-1, each of which have previously been demonstrated to be the targets of autoantibody responses in prostate cancer patients. IgG responses were detected to at least one protein in 18 of 31 patients, and to multiple antigens in 11 of 31 patients. These data demonstrate that androgen deprivation elicits IgG responses to prostate-associated proteins, and that responses can be detected using a panel of commonly recognized prostate antigens. Further studies will aim to characterize this panel and autoantibody profiles in patients treated with other non-antigen-specific immunotherapies or anti-prostate tumor vaccines.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5591.
Abstract 5593: A combination of HDAC inhibitor Entinostat (MS-275), All Trans Retinoic Acid (ATRA) and low dose Doxorubicin causes regression of established xenografts of triple negative breast cancer
Authors: Nguyen Nguyen K. , Korangath Preethi , Sabnis Gauri , Brodie Angela , Ordentlich Peter , Stearns Vered , Sukumar Saraswati
Published: February 2011

Triple negative breast cancer (TNBC) is a subgroup of breast cancer that rapidly develops resistance to chemotherapy. Currently, TNBC patients are treated with anthracycline and taxane-based regimens. These treatments result in some improvements in the clinical outcomes; however, they are associated with poor prognosis. Also, endocrine therapies are ineffective since the tumors do not express the estrogen receptor. Recent studies have shown that histone deacetylase (HDAC) inhibitors could reverse the epigenetic profile of some genes, including ER, EGFR, and RARβ2. We have recently shown that treatments that combine the HDAC inhibitor, Entinostat (MS-275), with the aromatase inhibitor, Letrozole, result in regression of xenografts of MDA-MB-231 breast cancer cells. Since RARβ2 is re-expressed as well, we reasoned that combining epigenetic therapy using MS-275, with differentiation therapy using a RARβ2 agonist (ATRA) will provide an effective combination of drugs against TNBC. We also examined possible potentiation of anticancer activity of these agents by low nontoxic doses of chemotherapy (doxorubicin). In vitro, human TNBC cell lines, MDA-MB-231, HCC1954, HCC 1143, and BT20 responded significantly better to the triple combination compared to the drugs used singly or in combination with MS-275. Upon treatment, re-expression of the silenced RARβ2 and downstream effectors was observed in the cell lines in vitro and in tumor xenografts of MDA-MB-231 cells. Subcutaneous tumor growth in immunodeficient mice was strongly inhibited (10/14 significant regression) using a combination of MS-275, ATRA and low dose Doxorubicin, compared to the drugs administered singly or in combination with MS-275. No significant weight loss was observed in any of the groups. These results suggest that combination of HDAC inhibitor and RARβ2 agonist with low dose chemotherapy has the potential to be an effective treatment against TNBC. This study provides encouraging new information for a nontoxic, and novel combination of drugs which could be effective against TNBC.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5593.
Abstract 5597: NK cells and CD8+ T cells in orthotopic pancreatic cancer model: Mediation of anti-cancer immune response by 5-fluorouracil and interferon-α
Authors: Khallouf Hadeel , Serba Susanne , Märten Angela , Jäger Dirk , Schmidt Jan
Published: February 2011

Pancreatic cancer has the poorest prognosis of all cancers urging the necessity of new therapeutic approaches. Recent clinical data show encouraging results for combining Interferon-α (IFN-α) immunotherapy with 5-Fluorouacil (5-FU) chemotherapy. However, the role of IFN-α in this immunochemotherapy scheme is still elusive. Here we investigate in vivo the relevance of different immune subpopulations in the anti-cancer immune response mediated by IFN-α in combination with 5-FU.

Luciferase-transfected Panc02 cells were implanted in the pancreas of C57BL/6 mice. Five days later, mice were treated with 5-FU alone, 5-FU + IFN-α or with a vehicle control. In parallel, CD4+ T cells, CD8+ T cells, and NK cells were depleted by neutralizing antibodies. Depletion of CD11c+ dendritic cells (DCs) was performed using transgenic CD11c. DTR mice. On day 21, mice were sacrificed and tumors were measured and frozen for further immunohistochemical analysis to evaluate the immune cells infiltration. NK cells were isolated and enriched from spleens of mice and their lytic activity was tested against Panc02 cells using a standard chromium release assay.

Our data show that treatment with IFN-α + 5-FU has significantly decreased tumor volume in comparison with control or 5-FU treatments. On the other hand, this decrease in tumor volume was remarkably abolished by depleting CD8+ T cells or NK cells. Furthermore, NK cells isolated from spleens of IFN-α + 5-FU treated mice showed enhanced cytotoxicity towards pancreatic cancer cells (Panc02). Meanwhile, studying the role of CD4+ T cells and DCs besides immunohistochemical analysis is still ongoing. To conclude, this study gives more insight about the mechanism of action of INF-α in combination with 5-FU and introduces CD8+ T cells and NK cells as main players in the observed anti-cancer immune response.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5597.
Abstract 5603: Impact of combination of PSK with TGF-beta inhibitor on anti-tumor immunity
Authors: Hiroaki Tanaka , Yamazoe Sadaaki , Iwauchi Takehiko , Muguruma Kazuya , Kubo Naoshi , Kato Yukihiro , Morimoto Junya , Kashiwagi Shinichiro , Fuyuhiro Yuhiko , Noda Eiji , Amano Ryosuke , Inoue Toru , Yamada Nobuya , Maeda Kiyoshi , Ohira Masaichi , Hirakawa Kosei
Published: February 2011

The transforming growth factor-β (TGF-β) has a role in tumor progression and metastasis. Secretion of TGF-β by tumor cells also suppresses an antitumor immune response in which dendritic cells (DCs) play an important role to activate cytotoxic T lymphocytes (CTLs). Here we report that small molecule TGF-β signaling inhibitor, SB-431542 (SB), induce DC maturation in vitro when combined with Protein-bound polysaccharide K (PSK). PSK has been used clinically as an anti-tumor agent in Japan. It has been reported that PSK have potential to induce anti-tumor immune response. In this study, we examined the impact of PSK and SB-431542 on maturation of DCs under immunological tolerant status. At first, we added SB and PSK to cultures of human DCs generated from peripheral monocytes in the presence of TGF-β and examined expression of CD83, production of Interleukin-12 (IL-12) and capacity of DCs to induce T cell proliferation. SB-431542 with PSK induced upregulation of CD83 of human DCs and improved their abilities to produce interleukin 12. They also augmented capacity of human DCs to activate naïve T cells in allogeneic mixed lymphocyte reaction. These results suggested that TGF-β receptor I kinase inhibitor such as SB-431542 should be able to induce antitumor immune response in immuno-tolerant patients associated with TGF-β activity.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5603.
Abstract 5604: Modulation of Stat3 in the tumor microenvironment enhances adoptive T cell therapy
Authors: Kujawski Maciej , Zhang Chunyan , Herrmann Andreas , Reckamp Karen L. , Scuto Anna , Jensen Michael , Figlin Robert , Yu Hua
Published: February 2011

Adoptive T cell transfer has shown promise in several malignancies. However, the need to expand antigen-specific T cells for prolonged periods ex vivo and the immunosuppressive effects of the tumor microenvironment severely limit its antitumor efficacy. To overcome these challenges, we tested the feasibility of stimulating and expanding transferred T cells in vivo by targeting Stat3, an oncogenic transcription factor critical for mediating tumor immunosuppression. In contrast to Stat3+/+ CD8+ T cells, transferred naïve CD8+ T cells with genetic Stat3 inhibition, effectively infiltrated tumors where they proliferated robustly, resulting in increased tumor antigen-specific CD8+ T cell activity and tumor growth inhibition. To facilitate the clinical translation of these findings, we further tested sunitinib, an FDA-approved tyrosine kinase inhibitor that has been shown to block Stat3 in several cell types. Sunitinib treatment combined with adoptive T cell transfer of Stat3−/− CD8+ T cells resulted in significant tumor growth inhibition in both immunosuppressed and immunocompetent hosts. Furthermore, sunitinib enhanced tumor T cell infiltration, dendritic cell activation and decreased tumor conversion of T regulatory cells. Targeting Stat3 in tumor cells and immune cells with sunitinib represents a novel approach to overcoming the obstacles that currently face adoptive T cell transfer therapy, which can be translated for immediate use in the clinic for treating cancer.

Supported by R01CA146092-01, K12 CA 01727, Kure It Kidney Cancer Research Fund and the Richard and Nancy Bloch Research Fund.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5604.
Abstract 5615: Effects of corticosteroids on tumor immunity induced by anti-CTLA-4 mAb therapy in a mouse model
Authors: Maeda Yuka , Kato Takuma , Wang Linan , Muraoka Daisuke , Sato Eiichi , Wolchok Jedd D. , Old Lloyd J. , Shiku Hiroshi , Nishikawa Hiroyoshi
Published: February 2011

Blockade of CTLA-4 activates/augments CD4+ and CD8+ tumor-specific T cell responses and results in tumor rejection. However, concomitant immune related adverse events (irAEs) in some anti-CTLA-4 mAb treated patients, such as skin rash and gastrointestinal inflammation, suggest that the breaking of tolerance to some normal self-antigens may accompany anti-tumor responses. As a consequence, patients are treated with immunosuppressive drugs, such as corticosteroids. Surprisingly, corticosteroids have not been reported to affect the anti-tumor responses induced by anti-CTLA-4 mAb; indeed, complete and enduring tumor regression has been seen in patients treated for irAEs with various immunosuppressive medications. In this study, we investigated the impact of corticosteroids on anti-tumor immune responses elicited by anti-CTLA-4 mAb in BALB/c mice bearing CMS5a-NY-ESO-1, a transplantable fibrosarcoma expressing NY-ESO-1 (an extensively studied human cancer/testis antigen). After CMS5a-NY-ESO-1 tumor inoculation, graded doses of methylprednisolone injection were initiated. Low doses of corticosteroids, similar to those used to treat patients with irAEs, did not suppress anti-tumor effects of anti-CTLA-4 mAb. However, high doses of corticosteroids, exceeding those used to treat patients with irAEs, impaired tumor growth inhibition by anti-CTLA-4 mAb. Furthermore, when high doses of corticosteroids were injected after the complete regression of tumors, approximately 30% of mice exhibited recurrence of CMS5a-NY-ESO-1 tumors at the initial tumor inoculation sites. This was associated with suppression of CD4+ and CD8+ T cell responses, indicating that anti-CTLA-4 mAb treatment induced an equilibrium-state of anti-tumor immunity, but not tumor eradication in these mice. Analyses of NY-ESO-1-specific CD4+ and CD8+ T cell responses in these mice showed that corticosteroids decreased the number of NY-ESO-1 specific T cells. We propose that dose and timing are important considerations for use of immunosuppression to treat irAEs of CTLA-4 blockade.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5615.
Abstract 5617: Enhancement of antitumor radiation efficacy and the abscopal effect by ECI301 mediated TLR4 dependent innate immunity in mice
Authors: Shiraishi Kenshiro , Yokochi Shoji , Ishiwata Yoshiro , Ishihara Mariko , Nakagawa Keiichi , Matsushima Kouji
Published: February 2011

[Background] We have previously demonstrated that ECI301, a variant form of human MIP-1α, induced marked inhibition of tumor growth at the irradiated site, with complete tumor eradication and consistent induction of prevention of tumor growth at a non-irradiated site, designated as an abscopal effect (Clin Cancer Res. 2008;14(4):1159-66). This abscopal effect was tumor type independent. In current study, we further studied the action of ECI301 using C3H/HeN, C3H/HeJ and athymic mice implanted with tumor cells. Using RT-PCR analysis, we had found that high mobility group box 1 (HMGB1) RNA level, an important mediator of local and systemic inflammation was up-regulated by ECI301 in tumor tissue. So we also carried out anti-HMGB1 antibody treatment in tumor-bearing mice with ECI301 and radiation.

[Methods] FM3A cells were subcutaneously implanted in the both flanks of C3H/HeN and C3H/HeJ mice. When tumor size reached approximately 10 mm in diameter, local irradiation (6 Gy) was delivered from just above the right tumor and ECI301 was administrated as a single dose intravenously once a day for 5 consecutive days, beginning at day 1 after irradiation. Athymic nude mice bearing subcutaneous colon26 tumor cells were treated with local irradiation and ECI301 was administrated as the same manner.

[Results] ECI301 inhibited the tumor growth not only in the right flank but also in the left flank (non-irradiation site) in C3H/HeN mice implanted with FM3A cells. In C3H/HeJ mice, however, ECI301 did not enhance the effect of irradiation on the tumor in both flanks. In C3H/HeN study, HMGB1 RNA level was up-regulated in both sides of tumor. Moreover, anti-HMGB1 antibody treatment eliminated the anti-tumor effect of ECI301 in both sides. Furthermore, ECI301 did not inhibit the tumor growth in athymic mice implanted with colon26 cells.

These results suggest that the ECI301-induced abscopal effect may be exerted via TLR4 and depend on the activation of T cells, and indicates that the effect of ECI301 may be promoted via inflammation in irradiated tumor tissue with HMGB1-dependent mechanism. [Conclusion] Ionizing radiation induces inflammation and immune response in irradiated tumor tissue. This immunological mechanism is greatly enhanced by ECI301, resulting in the abscopal effect that is essentially intriguing for clinical setting. Our data will encourage future therapeutic application of ECI301 in the treatment of advanced or metastatic cancer with radiation therapy.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 5617.
Abstract 562: Stromal TLR4 and IL 6 expression as potential prognostic inflammation-associated markers for colo-rectal cancer progression
Authors: Cammarota Rosaria , Bertolini Valentina , Pennesi Giuseppina , Bucci Eraldo O. , Garlanda Cecilia , Barberis Massimo C. , Sessa Fausto , Laghi Luigi , Noonan Douglas M. , Albini Adriana
Published: February 2011

Epidemiological studies suggest that chronic inflammatory conditions constitute pre-neoplastic conditions.

To study how chronic inflammation could affect tumor growth and progression, we quantified the presence of inflammatory cells and expression of inflammatory cytokines by immunohistochemistry on archival tissue samples from 116 patients. We considered 13 cases of ulcerative colitis, 34 tubular or tubulo-villous adenomas with low or high grade dysplasia, and 53 infiltrating adenocarcinomas with T1, T2, T3, and T4 staging. Sixteen specimens of healthy mucosa resected along with the cancerous tissue were used as controls. We investigated expression of CD68 (macrophages), CD15 (neutrophils), CD31 (endothelial cells), Interleukin 6 (IL-6), Hepatocyte Growth Factor (HGF), and Toll-Like Receptor 4 (TLR-4). We considered 3 specific tissue compartments: epithelial, stromal and endothelial (vessel associated). Statistical differences between individual cell groups were determined using an unpaired two way t-test (Mann-Whitney).

Progression from healthy tissue to ulcerative colitis was characterized by a massive inflammatory-angiogenic reaction, as for a significant increase in expression of inflammation markers, paralleled by down-regulation of a potential suppressive molecule, HGF. Subsequently, a progressive increase in cells expressing CD68, CD15, TLR-4, and IL6 were found in the passage from adenoma to adenocarcinoma.

Adenocarcinoma patients that expressed high levels of TLR-4 in the stromal compartment had a significantly increased risk of relapse within 40 months. Moreover, in the subgroup of adenocarcinoma patients with a diagnosis of pT3 colon cancer (the most frequent pT), those with higher levels of TLR-4 in the tumor stroma showed increased risk for a relapse within 15 months.

These data suggest that inflammation correlates with progression in colorectal cancer, and that TLR-4 represents a potential prognostic marker of colo-rectal cancer progression.

Note: This abstract was not presented at the AACR 101st Annual Meeting 2010 because the presenter was unable to attend.

Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 562.