bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1880: Polymeric implants enhance bioavailability of curcumin by providing a continous (“24/7”) delivery system
Authors: Bansal Shyam S. , Kausar Hina , Aqil Farrukh , Vadhanam Manicka V. , Gupta Ramesh C.
Published: February 2011

Curcumin, a potent natural chemopreventive agent, has shown significant anti-oxidant, anti-inflammatory, anti-proliferative, anti-metastatic, anti-angiogenic properties and is found to be effective against many ailments of inflammatory origin. However, it has elicited only limited success in various pre-clinical and clinical studies presumably due to its poor oral bioavailability. To increase the bioavailability of curcumin, we have developed novel polymeric implants using polycaprolactone (Mol. Wt. 65,000) (P-65) to achieve continuous (“24/7”) controlled delivery of curcumin directly into the systemic circulation. In this study, we investigated rate of curcumin release from the polymeric implants and tissue levels of curcumin in vivo by subcutaneously grafting 2-cm implants containing 20% (w/w) curcumin in female rats. Animals were euthanized at different intervals, and blood, different tissues and implants were collected. Measurement of the residual curcumin in the implants showed a burst release of 4.66 (11.7%) and 7.34 mg (18.4%) after 7 and 14 days, respectively, followed by a more controlled release. After 70 weeks nearly 76% curcumin was released. In another study, implants containing 10% curcumin showed similar rate of release from the implants. The in vivo release patterns mimicked the rate of release observed in vitro under ‘sink’ conditions. A solvent extraction and HPLC methodology was developed to measure tissue levels of curcumin. After 4 days, curcumin was detected at a level of 60 ± 20 ng/g liver and declined to almost constant levels of 12-14 ng/g tissue after 22 days. Analysis of CYP1A1 and GSTM levels as the biomarkers of curcumin efficacy showed a 13- and 16-fold induction in CYP1A1 levels after 4 and 7 days, respectively. GSTM levels were also found to be increased by 3- and 4-fold after 4 and 7 days, respectively. These studies showed that polymeric implants can deliver controlled amounts of curcumin systemically over an extended period of time increasing its bioavailability and elicit modulation of phase I and phase II enzymes (Supported from the USPHS grant CA-118114 and Agnes Brown Duggan Endowment.).

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 1880.
Abstract 1889: Curcumin potentiates the cancer-preventive actions of selenium by inactivating selenoprotein thioredoxin reductase in prostate cancer cells
Authors: Gopalakrishna Rayudu , Schiffman Jason E. , Man Joshua , Lei Lok , Wu Adela , Gundimeda Usha
Published: February 2011

Our previous studies have shown that the cancer-preventive actions of selenium may be negated by an over-expression of selenoprotein thioredoxin reductase (TR). This reductase reverses selenocompound-induced sulfhydryl redox inactivation of protein kinase C (PKC), a step which is intimately involved in the cancer-preventive actions of selenium. Consistent with this observation, the TR-specific inhibitor auranofin increases the sensitivity of prostate cancer cells to methylseleninic acid (MSA). This is particularly significant for advanced prostate cancer cells which require higher concentrations of MSA than early stage cancer cells. Selenium prevents cancer in some cases but fails to do so in others. Selenium's failure in this respect may be due to the development of TR-mediated resistance to its chemopreventive actions. Therefore, there is a potential role for agents that inhibit TR and thereby sensitize cancer cells to selenium. Several studies have shown that cancer prevention by a single agent may be less effective than multiple agents administered simultaneously. Here we report that curcumin, another cancer -preventive agent, at 5 to 10 μM concentrations potentiates the action of MSA in inhibiting growth and inducing apoptosis of both androgen-dependent LNCaP cells and androgen-independent DU145 cells. Nearly 2- to 4-fold lower concentrations of MSA were needed for 50% inhibition of cell growth in the presence of curcumin than in its absence. Under these conditions, curcumin inactivated TR in these cell types. There was also a concomitant increase in MSA-induced sulfhydryl oxidative inactivation of PKC. Furthermore, we found a correlation between curcumin-induced inactivation of TR and curcumin-induced potentiation of MSA-associated growth inhibition and apoptosis. These results suggest TR may be a molecular target for cancer prevention. While auranofin, the prototypical TR inhibitor, is toxic, curcumin appears to be a safe alternative. Our results provide a mechanistic rationale to the combination of selenium and curcumin for cancer prevention. Just as resistance develops to cancer chemotherapy, cancer prevention may also be counteracted by resistance mechanisms. Similarly, a combination of agents may prevent the development of resistance in cancer prevention just as it does in chemotherapy. Instead of the random mixing of agents in an empiric manner, a mechanistically informed choice of agents as has been proposed here is a more rational approach to identify effective combination of agents for cancer prevention.

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 1889.
Abstract 1892: Curcumin suppresses prostate cancer cells through modulation of β-catenin and protein kinase D1
Authors: Sundram Vasudha , Chauhan Subhash C. , Jaggi Meena
Published: February 2011

OBJECTIVE: Prostate cancer is the most commonly diagnosed cancer in males in the US. One in every 6th male will be diagnosed with prostate cancer this year. Understanding the molecular basis of prostate cancer progression can serve as a tool for early stage diagnosis and development of novel treatment strategies for this disease. Inappropriate activation of β-catenin signaling is linked to a wide range of cancers, including prostate cancer, resulting in transcription of genes involved in cancer progression such as c-Myc, c-Jun and cyclin D1. However, strategies to regulate this specific signaling pathway are very limited or not well developed. Synthetic or natural products which regulate this signaling pathway can be developed as a novel therapeutic modality for prostate cancer treatment. Curcumin, the active ingredient of turmeric, has shown anti-cancer properties via modulation of a number of different molecular pathways. Herein, we have investigated a putative molecular mechanism involved in curcumin mediated suppression of prostate cancer cell growth and the effect of curcumin on prostate cancer cells growth and its effect on β-catenin transcription activity.

MATERIALS AND METHODS: Cellular characteristics of the C4-2 prostate cancer cells were measured using anchorage independent growth, anchorage dependent growth, motility, and proliferation assays. β-catenin/T cell factor (TCF) transcription activity was measured by luciferase reporter assays using reporter constructs Topflash or Fopflash and pRL-TK (Renilla luciferase). The Luciferase activities were assayed using the Dual Glo reagent (dual reporter assay, Promega, Madison, WI). Subcellular localization of Protein Kinase D1 (PKD1) and β-catenin was determined by confocal microscopy.

RESULTS: Our results demonstrate that curcumin effectively suppresses prostate cancer cell growth, proliferation and cellular motility. In the luciferase assays, curcumin treatment diminished β-catenin transcription activity in C4-2 prostate cancer cells. Moreover, curcumin treatment influences the translocation of β-catenin from the nucleus to the cell surface membrane. Interestingly, the suppression of β-catenin transcription activity was correlated with the modulation PKD1 expression/phosphorylation within 1-3 hrs post-curcumin treatment.

CONCLUSION: These results suggest that curcumin treatment attenuates

β-catenin transcription activity in prostate cancer cells via modulating the expression/activation of PKD1. These results suggest a novel molecular mechanism of curcumin related suppression of prostate 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 1892.
Abstract 190: p53 target gene expression, DNA hypomethylation, and clinical response in MDS patients undergoing decitabine therapy
Authors: Link Petra A. , Zhang Wa , Gold David , Wetzler Meir , Baer Maria R. , Karpf Adam R.
Published: February 2011

The myelodysplastic syndromes (MDS) are a heterogeneous group of blood disorders characterized by peripheral cytopenias, a cellular bone marrow with dysplasia and increased intramedullary apoptosis, and are associated with risk of progression to acute myelogenous leukemia (AML). MDS are often fatal due to complications from cytopenias and/or progression to AML. DNA methyltransferase inhibitors (DNMTi), including 5-azacytidine and 5-aza-2’-deoxycytidine (decitabine) have recently assumed a major role in the clinical management of MDS, and are two of only three FDA-approved treatments for these disorders. The mechanistic basis for the activity of DNMTi in MDS is unclear. We recently reported that decitabine treatment induces the expression of p53-inducible ribonucleotide reductase p53R2/RRM2B in tumor cell lines and MDS patient bone marrow (Link et al., Cancer Research 68; 9358-9366, 2008). Importantly, p53R2 induction was independent of DNA hypomethylation at its promoter region, and the level of p53R2 induction correlated with clinical response in a cohort of 13 patients. These data suggest that important pharmacodynamic responses to DNMTi may be associated with DNA damage and/or p53 activation. To better understand the overall nature of the molecular responses occurring in the bone marrow of MDS patients treated with decitabine, in the current study we examined a variety of pharmacodynamic endpoints including: p53 target gene expression, global DNA methylation, tumor antigen gene expression and methylation, and tumor suppressor gene expression. Specifically, we measured the expression of three classical p53 target genes (p21, GADD45-α, NOXA), LINE-1 repetitive element methylation, the expression and methylation of cancer-germline (CG) antigens including MAGE-A1, NY-ESO-1, XAGE-1, and CTCFL/BORIS, and the expression of the tumor suppressor genes (TSGs) p15INK4b and PDLIM4. Our data reveal that decitabine treatment causes robust induction of p53 target genes in MDS patient bone marrow, which coincides with LINE-1 hypomethylation and clinical responses. In contrast, while CG antigen genes showed some evidence for DNA hypomethylation post-therapy, this did not coincide with CG antigen gene induction. Surprisingly, TSGs were also not induced by decitabine therapy in MDS patient bone marrow. Ongoing studies seek to define p53 mutational status in MDS patients and to determine how this relates to p53 target gene induction and/or clinical response. In summary, our data reveal a connection between p53 target gene activation, global DNA hypomethylation, and clinical response in MDS patients undergoing decitabine 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 190.
Abstract 1900: Antioxidant studies on the Peruvian cherimoya fruit give an insight into the healthy lifestyle of the Incas
Authors: Elera Gaytri D. Gupta , Whitaker John , Chauca Jorge , Robison Richard A. , Neill Kim L. O’
Published: February 2011

Annona cherimola, known as “cherimoya” was grown as an essential crop in the Incan empire, and was believed to prevent disease. Indigenous to Peru and Ecuador, this “pearl of the Andes” is consumed mainly for its exceptional taste. American writer Mark Twain called the cherimoya “the most delicious fruit known to men.” Recent interest in this fruit stems from its high concentration of acetogenins, which are bioactive polyketides. Recent research has accredited beneficial effects to acetogenins, including the exhibition of cytotoxic and anti-tumorous activity, particularly in mammary, pancreatic and colon cancer cells (Cheng et. al 1998). This project explores the health benefits of cherimoya consumption by examining the antioxidant effect of its components, including: skin, flesh and juice. These components were analyzed for antioxidant concentrations using the Oxygen Radical Absorbance Capacity (ORAC) assay.

A cellular model was also established that evaluated the antioxidant capacity of cherimoya components in the presence of Raji cells, a Burkitt lymphoma cell line, and HT-29 cells, a colon cancer cell line. After incubation with the cherimoya components, cell lysate samples were assessed for antioxidant capacity using the ORAC assay. Results for Raji cell lysate exposed to media, juice, skin and flesh were as follows: 886 Trolox Equivalents per Liter (TE/L), 1238 TE/L, 1018 TE/L, and 1045 TE/L respectively. Results for HT-29 cell lysate exposed to media, juice, skin and flesh were: 294 TE/L, 924 TE/L, 505 TE/L and 627 TE/L respectively. These data shows that cells treated with cherimoya juice exhibited a higher antioxidant uptake.

An alternate cellular model was established in which Raji cells and HT-29 cells were pre-exposed to oxidant AAPH, 2,2’-azobis-(2-amidinopropane) dihydrochloride, and then treated with cherimoya juice, skin and flesh. Results for Raji cells exposed to AAPH and then treated with media, juice, skin and flesh were as follows: 1054 TE/L, 1363 TE/L, 1183 TE/L, and 1254 TE/L respectively. Results for HT-29 cells exposed to AAPH and then treated with media, juice, skin and flesh were: 1075 TE/L, 1528 TE/L, 1350 TE/L and 1374 TE/L respectively. These data indicated that when cells were pre-exposed to an oxidant, thus simulating cells under oxidative stress, antioxidant cell uptake increased for both cell lines. These results suggest that cells can sense oxidative stress in their environment and can increase their ability to uptake available antioxidants.

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 1900.
Abstract 1905: Inhibition of hypoxia-inducible factor 1 (HIF-1) in tumor can augment anti-tumor immune responses in tumor-bearing mouse through suppression of transforming growth factor-beta (TGF-β)
Authors: Teramoto Koji , Kawaguchi You , Hori Tetsuo , Kitamura Shoji , Hanaoka Jun , Tezuka Noriaki , Kontani Keiichi
Published: February 2011

Hypoxia is one of distinctive features of the tumor microenvironment. In the tumor tissues under hypoxia, a transcription factor, hypoxia-inducible factor 1 (HIF-1) plays a critical role in regulating expressions of various genes associated with tumor growth. In order to understand the immunobiology of hypoxic tumor microenvironment, it is of great interest to elucidate the anti-tumor immune responses regulated by HIF-1. From these points of view, we focused on a relationship between HIF-1 and an immunosuppressive cytokine, transforming growth factor-beta (TGF-β) in the tumor. TGF-β which is derived from either tumor cells or tumor stromal cells has been reported to suppress anti-tumor immune responses in tumor-bearing hosts. In this report, we hypothesized that inhibition of HIF-1 would eliminate expression of TGF-β in tumor tissue and decreased level of TGF-β would contribute to augmentation of anti-tumor immune responses. We examined the possibilities in tumor-bearing mice models. C57BL/6 mice were inoculated subqutaneously with syngeneic Lewis lung carcinoma cell LLC. Five days after tumor inoculation, mice were administrated HIF-1 siRNA into the established tumor twice at 2-day interval. Three to 7 days after the last administration of HIF-1 siRNA, anti-tumor immune responses elicited in the mice were examined. As results, HIF-1 siRNA into the established tumor contributed to low expression of TGF-β mRNA as well as low level of TGF-β protein. In the tumor-draining lymph nodes, tumor antigen-specific CD8+ cells producing IFN-γ were increased and CD4+CD25+Foxp3+ (Treg) cells were decreased by administration of HIF-1 siRNA. Enhancement anti-tumor immune responses were associated with antigen-specific cytotoxic activity in spleen. Subsequently, tumor growth was effectively suppressed in mice administrated HIF-1 siRNA. Inhibition of HIF-1 in the tumor can eliminate expression of TGF-β there, and anti-tumor immune responses are augmented through eliminated TGF-β-mediated immunosuppression. These results demonstrate that hypoxia in the tumor tissue is associated with TGF-β-mediated immunosuppression and HIF-1 inhibitors can improve anti-tumor immune responses in tumor-bearing hosts. Our mouse models provide a new rationale with the molecular-targeting therapy for HIF-1.

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 1905.
Abstract 1910: Infiltration of ovarian carcinoma by altered plasmacytoid dendritic cells could contribute to local immune tolerance
Authors: Labidi-Galy Sana Intidhar , Ray-Coquard Isabelle , Sisirak Vanja , Treilleux Isabelle , Meeus Pierre , Mithieux François , Cassignol Alexandre , Bouchard François , Gabelle Philippe , Guastalla Jean-Paul , Ménetrier-Caux Christine , Caux Christophe , Bendriss-Vermare Nathalie
Published: February 2011

Ovarian carcinomas are usually considered as immunogenic tumors, as they are capable of inducing tumor-associated antigen immunity. However, immune system fails to eradicate established tumors partly due to alteration of immunosurveillance and induction of an active process of « tolerization » by the tumor microenvironment. We prospectively collected blood, tumors, and ascites samples from 38 patients with ovarian carcinomas (localized and advanced stages) and 35 healthy donors that gave written informed consent. Lin−CD4+CD11c−CD123+BDCA2+ pDC are the most abundant DC subset within ovarian carcinomas and neoplasic ascitis while they are profoundly depleted in peripheral blood of ovarian carcinoma patients compared to healthy donors. The depletion of pDC in patient's blood is partially restored when they are in complete remission (after chemotherapy and surgery). These observations suggest preferential pDC recruitment into tumor tissues that may involve CCR7 and CD62L which are downregulated in Tumor-associated (TA)-pDC compared to ascitis or blood pDC. Furthermore, TApDC express a partially mature phenotype compared to patient's blood and ascitis pDC that may reflect an in situ activation. Importantly, ex vivo TApDC produce lower amounts of IFN-α, TNF-α, IL-6 and RANTES in response to TLR ligands than ascitis or healthy donor pDC, suggesting a specific inhibition by ovarian tumor environment. In vitro, tumor or ascitis-derived surnageants (SNTUM) induce inhibition of the production of IFN-α and TNF-α by healthy pDC in response to TLR activation, suggesting the implication of soluble factors in this functional inhibition. In addition, TApDC but not ascitis pDC or healthy donors pDC direct allogeneic naive CD4+ T lymphocytes to produce high amounts of IL-10. Thus, TApDC may contribute to the tumor environmental immunosuppressive network. Elucidation of the mechanisms of dysfunctions of TApDC could have clinically relevant implications for future therapeutic strategies.

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 1910.
Abstract 1916: Robust pro-inflammatory cytokine secretion by tumor-infiltrating t cells despite altered Stat1/Stat3 signaling responses
Authors: O'Donnell Erika A. , Nolan Garry P. , Engleman Edgar G. , Mende Ines
Published: February 2011

The mechanisms whereby tumors reap the pro-growth benefits of an inflammatory context while still evading the immune response are not fully understood. Using phospho-specific flow cytometry, we demonstrate differential biasing of STAT phosphorylation responses within the tumor microenvironment, with B16 and CT26 cancer cells responding preferentially to pro-inflammatory IL-6 and IFN-γ and tumor-infiltrating T cells responding preferentially to anti-inflammatory IL-10. Investigation of cytokine responsiveness biasing within tumor-infiltrating T cells revealed that the intratumoral T cell population is shaped by non antigen-specific accumulation of CD62L− T cells, which lack responsiveness to pro-inflammatory cytokines despite their robust secretion of these factors. The unchecked production of pro-inflammatory factors by antigen-experienced T cells within the tumor suggests a novel role for these cells in supporting tumor growth by contributing to the inflammatory milieu within the tumor microenvironment.

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 1916.
Abstract 1917: Immunological Tumor Maps: a Landscape of Infiltrating Immune Cells in Colorectal Cancer Based on Complete Tissue Section Analyses
Authors: Halama Niels , Zoernig Inka , Foged Niels T. , Schirmacher Peter , Jaeger Dirk , Grabe Niels
Published: February 2011

In colorectal cancer (CRC) large scale tissue microarray (TMA) based quantitative immune cell counts using immune cell surface molecules (CD3, CD8, Granzyme B, and CD45RO) have identified the number of infiltrating immune cells to be potentially better predictors for patient survival than the classical TNM system. The spatial heterogeneity of immune cells may not be well reflected in the highly selected, and typically small (0,6-1 mm2) tissue cores of the TMA. This represents an obstacle in the individual prognosis prediction or classification of a single patient. To investigate this aspect, the localization and distribution of immune cell subpopulations based on the analysis of complete tissue sections by a dedicated novel staining and imaging system were performed. Using a specialized staining platform and whole slide imaging & analysis by virtual microscopy (VM), immunological “tumor maps” were generated. These tumor maps are based on cell densities in fields of 1mm2 size, visualizing intratumoral heterogeneity for the surface markers CD3, CD8, Granzyme B, and CD45RO. In total, an area of 867 mm2 was automatically evaluated with an average of 48 mm2 of evaluated tumor tissue per patient slide. Cell counts varied within a patient significantly, ranging from 0 to up to 2550 cells / mm2. Further analyses revealed, that sampling of single field counts within the tumor can only yield clear diagnostic decisions for a fraction of the analyzed patients, with ambiguous decisions for 11 out of 20 patients. Interestingly, the overall degree of heterogeneity also varied between patients, with lower heterogeneity found only in samples with lower cell counts. No samples with a homogeneous high cell density distribution were observed. The observed variability has implications for the individual prognosis prediction and represents the first spatial quantitative study of immune cells in a set of CRC primary tumors. The presented tumor maps therefore are a suitable tool to visualize heterogeneity. Furthermore, whole slide imaging & analysis by VM is essential in the identification of prognostic markers as well as in their subsequent application. In the future, spatial marker signatures could contribute to individual patient classification.

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 1917.
Abstract 192: Restoration of an epigenetically silenced tumor suppressor gene RASSF1A in human prostate cancer cells by a plant alkaloid, mahanine
Authors: Jagadeesh Shankar , Pal Bikas C. , Bhattacharya Samir , Banerjee Partha P.
Published: February 2011

The etiology of human prostatic carcinoma remains largely undefined. However, it is becoming clear that epigenetic inactivation of various tumor suppressor genes could play pivotal role in the development of various cancers including prostate cancer. One of such tumor suppressor is Ras-association domain family 1 (RASSF1) gene. Epigenetic inactivation of RASSF1A is probably the most frequently methylated gene described thus far in human cancer. Ectopic expression of RASSF1A in cancer cells that lack endogenous RASSF1A transcripts resulted in reduced growth of cancer cells in vitro and in nude mice supporting a role for RASSF1A as a tumor suppressor gene. Since, the restoration of RASSF1A expression in tumor cells impairs their tumorigenicity, factors that restore RASSF1A expression have immense importance in preventing tumor growth. In this investigation, we demonstrated that, mahanine, a plant derived carbazole alkaloid, restores RASSF1A expression in both androgen-responsive (LNCaP) and androgen-negative (PC3) prostate cancer cells by inhibiting DNA methyltransferase (DNMT) activity. Mahanine-induced expression of RASSF1A in turn significantly reduces cyclin D1 but not other cyclins. As a result of down-regulation of cyclin D1, cancer cells arrest at G0/G1 phase. To understand the inverse relationship between RASSF1A and cyclin D1, we observed that mahanine treatment down-regulates cyclin D1 and addition of RASSF1A siRNA prevents this inhibition. Inactivation of both Akt and CREB by mahanine and RASSF1A is associated with the down-regulation and translocation of cyclin D1. This study shows that mahanine can reverse an epigenetically silenced gene, RASSF1A in prostate cancer cells by inhibiting DNMT activity that in turn down-regulates a key cell cycle regulator, cyclin D1. Mahanine, therefore, promises an encouraging therapeutic choice for advanced prostatic cancer (Supported by NIH grant R01 CA131123).

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 192.
Abstract 1926: Adoptive γδ T cell transfer therapy for non-small cell lung cancer
Authors: Kakimi Kazuhiro , Murakawa Tomohiro , Fukami Takeshi , Goto Shigenori , Kaneko Toru , Yoshida Yukihiro , Takamoto Shin-ichi , Nakajima Jun
Published: February 2011

Human peripheral blood γδ T cells can recognize and kill tumor cells by their Vγ9Vδ2 T cell receptor and/or NKG2D. We have established large-scale ex vivo expansion of γδ T cells from peripheral blood mononuclear cells (PBMC) by zoledronate and IL-2. A clinical study of adoptive γδ T cell transfer therapy has been conducted for the treatment of non-small-cell lung cancer (NSCLC) patients to evaluate the safety profile and potential anti-tumor effects of γδ T cells. Prior to beginning the study, it was registered on a University hospital Medical Information Network Clinical Trials Registry (UMIN-CTR : http://www.umin.ac.jp/ctr/index.htm) on March 1, 2006 (UMIN ID: C00000036).

PBMC were isolated from 67.5 ml of peripheral blood and stimulated with zoledronateand IL-2 for 14 days to obtain γδ T cells. The cultured cells on day 14 consisted mostly of γδ T cells. The predominant populations in fresh peripheral blood were cells with the CD45RA−CD27+ memory phenotype or a CD45RA+CD27− naïve phenotype. In contrast, most γδ T cells were shown to have the CD45RA−CD27− effector memory phenotype after 14 days culture. They expressed NKG2D and up-regulated the expression of activation marker CD69, and had cytotoxicity against allogeneic tumor cell lines. Upon stimulation, γδ T cells produced IL-2, TNF-α and IFN-γ. Though certain patients were refractory to zoledronate stimulation, γδ T cells from patients demonstrated comparable cytotoxic effector activity once they were successfully expanded.

Cultured cells were intravenously administered every 2 weeks for 6 injections. Fifteen patients who were resistant to current standard therapy were enrolled this study. The treatment was well tolerated and no severe adverse events related to γδ T cell therapy were observed. Two patients came off the study at the early stage of the treatment due to the aspiration pneumonia (grade 3) and radiation pneumonitis (grade3). The responses according to RECIST guideline were SD for 7 patients, PD for 6 patients and 2 inevaluable for response due to early drop off the study (The disease control rate was 46.7 %).

Transferred γδ T cells gradually accumulated in peripheral blood in patients who received more than 1×109 γδ T cells per injection and accounted for 10 % of PBMC even 2 weeks after the final injection. When PBMC were stimulated with PMA/ionomycin or zoledronate-pulsed Daudi cells, IFN-γ production and surface expression of CD107 were detected on γδ T cells, suggesting that transferred γδ T cells sustained their effector function in vivo.

These results indicated that γδ T cell therapy was well tolerated and feasible for the treatment of NSCLC.

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 1926.
Abstract 1930: Early post-operative adoptive multiple-cellular immunotherapy in gastric cancer: A 3-year follow-up report
Authors: Xu Yingxin , Qiao Zhi , Peng Zheng , Xia Shaoyou , Li Li , Wu Xin , Zhang Yanjun , Ning Ning , Li Jie , Li Rong
Published: February 2011

Objective Adoptive immunotherapy is an active approach, but efficacy is limited in the clinic for most solid tumor. We hypothesized that there are three factors might lead to improved clinical outcome in the patients with gastric cancer that include: 1) the combination of multiple immune cells, 2) more effector cells reaching at the site of the tumor by infusion in peritoneal cavity, 3) performing of immunotherapy as soon as possible after operation.

Patients & Methods 51 gastric cancer patients (stage III-IV 42/51) who underwent operation from September 2005 to March 2008 were retrospectively analyzed. Data were censored in 39 months (January 2009),Group A patients (n=22) received a course of immunotherapy which included autologous tumor lysate-pulsed dendritic cell (DC) via intradermic injection, cytokine induced killer cells (CIK) by iv, as well as DC-CIK by intraperitoneal injection in 11-14 days after operation, then a standard chemotherapy (18/22) and immunotherapy again. Group B patients (n=29) only received postoperative chemotherapy. Overall survival (OS) and disease-free survival (DFS) were calculated by the Kaplan-Meier method.

Results In total, 24 patients died, giving a median survival time of 25 months. The estimated 2-year OS and DFS of group A were 63.6% and 67.3%, while that of group B were 25.2% and 33.2%, respectively. Both of the estimated 3-year OS and DFS were significantly superior in group A (log-rank test, P<0.01).

Conclusions Early post-operative adoptive multiple-cellular immunotherapy with infusion in peritoneal cavity significantly improved overall survival and disease-free survival in gastric cancer patients.

Key words gastric cancer; early; postoperative; adoptive cellular immunotherapy

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 1930.
Abstract 1940: Successful eradication of established peritoneal ovarian tumors in SCID-beige mice following adoptive transfer of T cells genetically targeted to the MUC16 antigen
Authors: Chekmasova Alena A. , Thapi Dharmarao , Nikhamin Yan , Park Kay J. , Spriggs David R. , Brentjens Renier J.
Published: February 2011

Most patients diagnosed with ovarian cancer will ultimately die from their disease. For this reason, novel approaches to the treatment of this malignancy are needed. Adoptive transfer of a patients own T cells, genetically modified ex vivo through the introduction of a gene encoding an chimeric antigen receptor (CAR), an artificial T cell receptor, targeted to a tumor associated antigen, is a novel and promising approach to cancer therapy applicable to the treatment of ovarian cancer.

We have generated several CARs targeted to the retained extracellular domain of MUC16, termed MUC-CD, an antigen highly expressed on a majority of ovarian carcinomas. We investigate the in vitro biology of human T cells retrovirally transduced to express these CARs by co-culture assays on artificial antigen presenting cells (AAPCs) generated from NIH3T3 fibroblasts genetically modified to express the target MUC-CD antigen, as well as by cytotoxicity and proliferation assays utilizing the target human OV-CAR3(MUC-CD) ovarian tumor cell line. Finally, we assess the in vivo anti-tumor efficacy of human MUC-CD targeted T cells in a SCID-Beige orthotopic, xenogeneic OV-CAR3(MUC-CD) murine tumor model. CAR modified MUC-CD targeted T cells exhibited efficient in vitro cytolytic activity against cell lines which expressed the MUC-CD antigen and could be further expanded ex vivo through multiple cycles of co-culture on 3T3(MUC-CD/B7.1) AAPCs. Expanded MUC-CD targeted T cells infused into SCID-Beige mice bearing intraperitoneal human OV-CAR3(MUC-CD) tumors either delayed progression or fully eradicated tumor even in the setting of advanced disease.

These promising pre-clinical studies justify further investigation of MUC-CD targeted T cells as a potential therapeutic approach in the clinical setting treating patients with high risk MUC-16+ ovarian carcinomas.

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 1940.
Abstract 1944: In vitro analysis of suicide gene expression and function in human T lymphocytes transduced to express a tumor targeted chimeric antigen receptor
Authors: Park Jae , Yeh Raymond , Riviere Isabelle , Sadelain Michel , Brentjens Renier
Published: February 2011

Adoptive infusion of T cells genetically modified to express chimeric antigen receptors (CARs) targeted to tumor associated antigens (TAAs) is a promising novel approach to cancer therapy. However, in light of the fact that most TAAs targeted by such modified T cells are expressed, at least to some degree, by normal tissues as well as the fact that clinical experience with this technology is limited, safeguards are needed in the form of additional expression of transduced suicide genes by these T cells to allow for the efficient in vivo abrogation of infused T cells in the setting of unanticipated adverse events which may develop in the context of planned clinical trials.

Herein we report the in vitro function of 3 different suicide genes each initially inserted distal to a CAR gene and an IRES (internal ribosome entry site) element cloned into the SFG retroviral vector. Specifically, in this study we evaluated the herpes simplex virus thymidine kinase (HSV-TK SR39), E. coli derived nitroreductase (NTR), and inducible caspase-9 (iCasp-9). We initially found that all 3 suicide genes functioned sub-optimally in the second position of these IRES based bicistronic vectors. In order to enhance suicide gene expression we deleted the IRES elements replacing them with 2A linker peptides, which significantly improved gene expression and the cell growth inhibition upon the addition of respective substrate. In PG13 cells transduced with SFG 1928z. IRES. NTR we found 40% inhibition at 5mM of metronidazole (MTZ), but 90% in PG13 cells transduced with SFG 1928.2A.NTR. Similarly, PG13 (1928z.IRES.HSV-TK) fibroblasts exhibited 60% inhibition at 1µM of ganciclovir compared to 85% inhibition of PG13 (1928z.2A.HSV-TK) fibroblasts. Consistent with these findings, we found that human T cells transduced with either 1928z.2A.NTR or 1928z.2A.HSV-TK demonstrated 90% and 88% inhibition respectively at similar substrate concentrations.

In light of this encouraging in vitro data, we are currently in the process of assessing respective suicide gene function in vivo using a syngeneic, immune-competent mouse model recently developed in our laboratory. Ultimately we anticipate that these studies will allow us to rationally select the optimal suicide gene-prodrug combination to be incorporated into future clinical trials utilizing gene modified tumor targeted T cells.

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 1944.
Abstract 1946: Identification of miRNAs that contribute to melanoma brain metastasis
Authors: Gaziel Avital , Menendez Silvia , Segura Miguel F. , Zakrzewski Jan , Rose Amy , Kerbel Robert S. , Darvishian Farbod , Cohen Dalia , Osman Iman , Hernando Eva
Published: February 2011

Brain metastasis occurs in a large proportion of metastatic melanoma patients and is associated with a dismal prognosis. However, the molecular mechanisms that govern melanoma tropism to the brain remain poorly understood. MicroRNAs (miRNAs) play essential roles in many physiological and pathological processes, and have been recently shown to exert key roles during cancer metastasis. In this study we find that specific miRNAs may be important mediators of melanoma dissemination to the brain. First, we conducted a miRNA microarray analysis of metastatic melanoma tissues that revealed a subset of miRNAs differentially expressed in brain metastases (n=11) relative to other sites (n=48). A brain-specific signature comprised of seven miRNAs was further validated in an independent cohort of metastatic melanoma samples (n=36; 9 brain metastatic specimens). Then, we analyzed the trend of expression of those miRNAs during tumor progression by comparing their levels in primary tumors and their paired metastasis from patients with or without recurrence in the brain (n=18). Differential expression of some miRNAs was already evident at diagnosis in primary tumors that recurred in the brain, while for others it was acquired in the transition from primary to metastasis, suggesting that it may be a later event in tumor progression. Furthermore, in vitro modulation of specific signature miRNAs significantly altered the ability of melanoma cells to execute processes such as adhesion and transmigration through human brain endothelial cells and proliferation in human astrocytes conditioned media. Additionally, using an in vivo model of melanoma brain metastasis, we confirmed the capacity of specific miRNA alterations to promote melanoma cells’ competence to reach the brain. Finally, the analysis of potential downstream mediators of select miRNAs revealed the involvement of immuno-suppressive molecules, as well as inflammatory and chemotactic mediators in this process. Collectively, our results expand our understanding of the mechanisms that control melanoma brain metastasis, potentially revealing novel therapeutic avenues for patients for whom no viable approaches are currently available.

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 1946.
Abstract 1953: Functional identification of microRNA targets by integrated proteomics and microarray profiling: miR-193b in breast cancer
Authors: Leivonen Suvi-Katri , Rokka Anne , Östling Päivi , Kohonen Pekka , Corthals Garry , Perälä Merja , Kallioniemi Olli
Published: February 2011

MicroRNAs (miRNAs) regulate protein expression by targeting messenger RNAs (mRNAs) for cleavage or by inducing translational repression. Despite advances in bioinformatic predictions, it is a significant challenge to identify protein targets that the miRNAs regulate. Here, we applied a novel strategy combining functional data from quantitative proteomics using iTRAQ (isobaric tag for relative and absolute quantitation), gene expression microarray profiling and bioinformatic target predictions in order to characterize targets regulated by miRNAs. We focused on exploring the anti-cancer effects of miR-193b. According to our high-throughput functional studies (Leivonen et al., Oncogene, 2009), miR-193b induces cell cycle arrest and inhibition of estrogen-depedent growth when overexpressed in breast cancer cell lines.

Pre-miR-193b was over-expressed in MCF-7 breast cancer cells for 48 h, and the cell lysates were subjected to iTRAQ isobaric labelling followed by mass-spectrometric analyses. We identified over 600 proteins, of which 11 were repeatedly and significantly (p<0.05) down-regulated, and 6 up-regulated by miR-193b as compared to controls. Three of the down-regulated proteins were predicted to be targeted by miR-193b by the TargetScan algorithm. Of the predicted miR-193b targets, two were also among the most prominently down-regulated genes at the mRNA level according to microarray profiling of miR-193b-transfected MCF-7 cells. These data were validated by western blotting and quantitative RT-PCR. The results indicated that miR-193b targets included proteins which have previously been linked to breast cancer progression and poor prognosis, such as 14-3-3ζ (YWHAZ), as well as mini-chromosome maintenance protein-7 (MCM7), which is associated with DNA replication and cell cycle control.

In conclusion, we demonstrate here that the combination of proteomic expression profiling and gene expression microarray profiling is a powerful strategy to functionally identify protein targets and mechanisms of action for miRNAs that have anti-cancer effects when transfected into breast cancer cells.

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 1953.
Abstract 1955: Bone-marrow-derived cells incorporate into the prostate during regrowth
Authors: Placencio Veronica R. , Sherrill Taylor P. , Yu Xiuping , Bhowmick Neil A.
Published: February 2011

It is necessary to understand mechanisms of castrate resistant prostate cancer development and progression. As prostate cancer transitions from androgen dependent to castrate resistant, subsequent recurrence involves increased growth of the cancer epithelial cells. Understanding prostate regrowth in a normal prostate will allow us to better understand growth in castrate resistant prostate cancer. We hypothesized that altered chemokine signaling which accompanies the loss and subsequent return of androgen signaling results in the recruitment of immune cells to the prostate microenvironment from the bone marrow. A chimeric mouse model with GFP-labeled bone marrow was used to allow us to identify bone marrow cells recruited to the prostate. We studied how bone marrow derived cells (BMDCs) contributed to an androgen refractory response, specifically prostate regrowth. In a similar mouse model we used GFP-labeled mesenchymal stem cells (MSCs) to study this specific subset of BMDCs in response to prostate regrowth. Host mice were castrated or left intact as a control. Testosterone was given to the chimeric mice. The intact and castrated control mice had a low number of BMDCs recruited to the prostate. However, three and seven days following exogenous testosterone administration resulted in a dramatic increase in BMDC recruitment during prostate regrowth. GFP labeled MSC cells were also recruited to the prostate at three days following treatment with exogenous testosterone. Interestingly, even after four weeks the fully regrown prostates retained BMDCs that appeared to be incorporated in the epithelial compartment. Double immunofluorescence staining showed that a subset of BMDCs gained the expression of androgen receptor; p63, a basal epithelial marker; and cytokeratin 8, a luminal epithelial marker, in the prostate. Rosa26 expressing MSCs injected into GFP mice showed prostate co-localization of ß-galactosidase and GFP, suggesting that MSCs fused with resident prostate epithelial cells during regrowth. The recruitment and eventual incorporation of BMDCs in the prostate suggests a mechanism for exogenous cells in castration-resistant prostate cancer growth. This result has implications for castrate-resistant prostate cancer growth, as it too may be a result of MSC recruitment, incorporation, and fusion. NIH grants F31 GM079879 (VRP), CA108646 and CA126505 (NAB).

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 1955.