bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract B17: Heat shock-induced danger signals in tumor cells are crucial for a rapid differentiation of monocyte to therapeutic dendritic cells
Authors: Aguilera Raquel , Saffie Carlos , González Fermin , Tittarelli Andres , Reyes Diego , Ferrada Carlos , López Mercedes N. , Salazar-Onfray Flavio
Published: January 2011

Dendritic cells (DCs) are potent antigen-presenting cells (APCs) that initiate a cell-mediated antitumor immune response. Recently, we showed that monocyte-derived DCs loaded with a heat-shocked melanoma cell lysate (TRIMEL®) induce melanoma-specific immune responses, thus prolonging patient survival. Here, we describe the rapid effect of TRIMEL on human monocytes differentiation to tumor-antigen presenting cells (TAPCells®). TAPCells showed a mature DC-like phenotype and effectively activated melanoma associated antigen (MAA)-specific CD4+ and CD8+ T lymphocytes. Moreover, heat shock (HS) pre-conditioning of TRIMEL composing cells increased calreticulin (CRT) cell surface translocation and released high-mobility group box 1 protein (HMGB1), which was closely associated to the induction of APC maturation and efficient MAAs cross-presentation. Finally, 26 out of 43 patients vaccinated with TAPCells showed a TRIMEL-specific delayed type hypersensitivity reaction (DTH) associated to anti-melanoma T cell recognition, reduced rates of progression and prolonged patient survival. Our results suggest that heat-shocked tumor cell lysates are optimal source of MAAs, inducing activated APCs with improved MAAs cross-presentation capacity and clinically effective immunogenicity.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B17
Abstract B18: Induction of Th1/Th17 and inhibition of Th3 cellular profile by DC-based immunotherapy is associated with immunological and clinical anti-tumor responses in melanoma patients
Authors: Durán Claudia , Segal Gabriela , Salazar Lorena , Pereda Cristián , Fabián Tempio , Salazar-Onfray Flavio , López Mercedes N.
Published: January 2011

The ability of DCs to induce a primary immune response makes them ideal candidates for cancer vaccines against tumors. Recently, in a serie of phase I/II clinical studies, we showed the effectiveness of autologous DCs loaded with a conditioned allogeneic melanoma cell lysate named TRIMEL. Our results demonstrated a correlation between positive tumor-specific delayed type hypersensitivity (DTH) reaction induced by DC-vaccination, and improved long-term patient survival in late-stage melanoma patients. In fact, 60% of patients showed a positive DTH reaction showing a three-fold prolonged survival compared to non-responder patients (33 and 11 months respectively). Herein, we investigated whether cellular and cytokine profiles in patients PBL before and after vaccination reflects differential responses associated to the clinical outcome. Analysis of T cell populations in blood showed that melanoma patients (n=28) have an increased proportion of Th3 (CD4+TGF-β+) regulatory T lymphocytes (2.64 % ± 0.16) compared to normal donors (n=14) (1.72 % ± 0.22). However, we didn't observed differences in TGF-β concentrations at serum level. Moreover, we analyzed vaccine effect on cytokine and cellular responses in PBL. Interestling, although there was not differences in Th3 population proportions between responder (n=14) and non-responder patients (n=14) before vaccination, after treatment responder patients showed a three-fold reduction of CD4+TGf-β+ T cells compared to non-responders (1.65 % ± 0.19 and 5.54 % ± 0.28 respectively). Furthermore, our results show that DC-vaccination resulted in a three-fold augment of IFN-γ releasing by Th1 population and a two-fold increase of IL-17 producing Th17 lymphocyte population in responder patients, but not in non-responders. A direct correlation between IFN-γ and IL-17 production in responder patients blood was observed (p < 0.0001) suggesting that those cytokines may favour an anti melanoma response. Taken together our results indicates that despite undetectable differences in the cellular and cytokine profiles in melanoma patients before DC-vaccination, immunization resulted in two different pattern of response associated to the immunological and clinical outcome. Our study may contribute to a better understanding of clinical immunological responses and to an earlier prognosis of patient outcome.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B18
Abstract B2: Patterns of serum biomarkers for lung cancer are different for men and women
Authors: Streeper Robert , Izbicka Elzbieta , Campos David , Diaz Armando , Baek Sung , Mussman Rhys
Published: January 2011

Introduction: Lung cancer has one of the highest mortality rates among all human cancers. Higher mortality has been noted in the past in men with the history of smoking but no significant differences in mortality exist in never smoking men and women. Some gender differences to date have been linked to genes located on sex chromosomes.

Methods: We quantified levels of 57 cytokines, chemokines and growth factors using multiplexed immunoassays to determine patterns of serum biomarkers in men and women with non-small cell lung cancer (NSCLC; 250 men, 116 women) and compared these with serum marker patterns in subjects with asthma (AST; 68 men, 132 women) and normal controls (NOR; 250 men, 116 women). Data were reduced using inter-pathology comparisons with statistical significance determined using the Kruskall-Wallis U test with p < 0.05 considered significant. Scalar sums of inter-pathology percentage differences (NSCLC vs. NOR, AST vs. NOR and NSCLC vs. AST) in marker expression were used for hierarchically ranking the markers. We selected markers based on at least one statistically significant difference was present for a given marker; those showing marginally significant differences were discarded.

Results: The markers showed unique gender- and disease specific patterns. For unisex analysis of NSCLC, we identified 36 markers with the 25% change cutoff threshold and 32 markers with the 50% cutoff. For women, we found 32 markers with the 25% cutoff and 30 with the 50% cutoff. For men, 39 markers were found at the 25% cutoff and 37 at the 50% cutoff. Expression of four markers was unique for women with NSCLC compared to NOR: IL-8 and serum amyloid P (downregulated), serum amyloid A and C-reactive protein (all upregulated). Five markers were unique for men with NSCLC compared to NOR: Insulin (downregulated), matrix metalloproteinases-7 and -8, resistin and hepatocyte growth factor (all upregulated). Three markers showed opposite patterns of expression; VEGF was downregulated in women and upregulated in men with NSCLC compared to NOR. Leptin was upregulated in women and downregulated in men and MIP-1α were upregulated in men and downregulated in women with NSCLC compared to NOR.

Conclusion: Our study has revealed surprisingly large qualitative and quantitative differences in biomarker expression patterns and levels in serum specimens from men and women suffering from NSCLC. The markers appear to be products of genes residing on several chromosomes and are not limited to sex chromosomes. Our findings may have implications for the diagnosis of non-small cell lung cancer, early detection of the disease through simple blood tests, characterization of disease progression and differentiation among the pathologies. It may find application in discovery and development of novel therapeutic strategies for human diseases.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B2
Abstract B25: Autocrine action of VEGF/VEGFR pathway in human Glioblastomas in addition to the paracrine angiogenic role
Authors: Martinez Juan-Carlos , Campo Leticia , Val Toledo Maria , Sacristan Silvia , Gatter Kevin C.
Published: January 2011

Angiogenesis is an important determinant for glioma growth and there is a growing interest in anti-angiogenic therapies as a way of blocking glioma development.

To get insights into brain tumor angiogenesis, we have addressed the expression of a panel of angiogenesis regulatory proteins in 34 human Glioblasotmas (GBs) specimens.

Methods: Immunohistochemistry (IHC) detection of Vascular endothelial growth factor (VEGF), Phosphorylated VEGF Receptor (PKDR), Hypoxia Inducible Factor (HIF-1) and the hypoxia marker Carbonic Anhydrase IX (CAIX).

Results & Discussion: VEGF & PKDR cytoplasmic staining of variable intensity in tumoral cells and vascular endothelium was observed in all cases. The highest levels were seen around necrotic foci and in areas of endothelial proliferation. However, VEGF & PKDR expression by tumoral cells was shown also in non-hypoxic areas far away from necrotic foci.

Hypoxia induces FIH expression, which in turn activates VEGF expression by tumoral cells. This is followed by diffusion, and interaction through a paracrine process with endothelial VEGF Receptors (VEGFR).

Interaction between VEGF and VEGFR, induces activation and phosphorylation of VEGFR, triggering angiogenesis by the emission of vascular sprouts.

Our finding of high levels of PKDR by neoplastic cells indicates that interaction and activation of the VEGF/VEGFR pathway can be as well an autocrine process. As VEGF is a powerful growth factor, this support that in addition to angiogenic induction, VEGF may play an active role in neoplastic cell growth. The strong expression of VEGF & PKDR in non-hypoxic areas suggests that VEGF may be induced by other stimulus than hypoxia.

This possibility my have a therapeutic influence in that treatment with VEGF blockers and or inhibitors, aimed at an antiangiogenic effect, could additionally act against neoplastic proliferation on neoplastic cells themselves.

Supported by Grant to JCM, Fundación de Investigación Mutua Madrileña.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B25
Abstract B26: The role of G alpha12 and thromboxane receptor beta interaction in bladder cancer progression
Authors: Woolworth Julie Ann , Desouki Mohamed , Halushka Perry , Watson Dennis , Smith Michael T. , Moussa Omar
Published: January 2011

In the United States, it is predicted that there will be over 70,000 new cases of bladder cancer this year. Bladder cancer is the fourth most common cancer in men and eighth in women. Our lab has previously identified Thromboxane receptor-β (TP-β) as potentially playing a role in bladder cancer progression. TP-β is a G-protein coupled receptor that is over-expressed in bladder cancer tumors and high levels of TP-β in bladder cancer patients correlate with decreased survival. G-proteins are involved in second messenger cascades that influence a variety of cellular functions, including transcription and motility. In multiple cellular systems TP-β has been found to interact with multiple G proteins such as Gαi, Gαs, Gα12, and Gα13. We found high levels of mRNA and protein expression of the G-protein, Gα12, in bladder cancer cell lines. We also observe high levels of Gα12 expression in human bladder cancer tissues. Studies using a constitutively active Gα12 mutant, suggest that signaling through Gα12 can contribute to oncogenic transformation. In this study we aim to elucidate the TP-β associated Gα12 dependent pathways that contribute to bladder cancer progression. We found that TP agonist U46619 increases cell migration and that treatment with TP antagonist blocks this increase in cell migration induced by U46619. TP agonist stimulation also increases the phosphorylation of extracellular regulated kinase (ERK) and U46619-induced phosphorylation of ERK is blocked with TP agonist treatment. Using a shRNA to knockdown Gα12 we observe a reduction in U46619-induced cell migration and ERK phosphorylation. Together our results suggest that TP-β mediated cell migration and the MAPK signaling pathway is in part regulated through Gα12. This work could have clinical implications by targeting the TP-β and Gα12 interaction as a potential therapeutic approach for bladder cancer treatment.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B26
Abstract B41: Antitumoral and angiostatic activities in skin secretion of frog Phyllomedusa bicolor
Authors: van Zoggel Hanneke , Hamma-Kourbali Yamina , Galanth Cécile , Ladram Ali , Nicolas Pierre , Courty José , Amiche Mohamed , Delbé Jean
Published: January 2011

The skin secretions of neotropical and South American frogs contains large amounts of a wide range of biological active peptides. Commonly studied are small peptides with antimicrobial activities as the Dermaseptin family. These peptides constitute a primitive immune defence mechanism and are able to disrupted the membranes of microbes and, subsequently, to inhibit their growth leading to cell death. Some of these antimicrobial peptides also present an antitumoral activity whereby they use a similar disrupting mechanism on the plasma membrane of the tumoral cells [1]. Therefore, intense research is now performed on the secretions from amphibian skin to discover new pharmacological agents, especially useful for drug-based cancer treatments.

In this study we have postulated that the skin secretions from the South American frog Phyllomedusa bicolor contain such molecules and we reported antitumoral and angiostatic activities from a crude extract of these secretions [2]. After a two steps chromatography procedure, we have identified one of the bioactive compounds in this crude extract as Dermaseptin B2 (Drs B2) [3]. Drs B2 is a well known antimicrobial peptide which is 33-amino acid residues long. This natural α-helical peptide consist of a cationic NH2-terminal part (residues 1 to 23) predicted to form an amphipathic α-helix that is followed by a hydrophobic tail (residues 24 to 33).

A synthetic Drs B2 peptide was prepared and tested for its antitumoral and angiostatic activities. Results obtained with this synthetic molecule were equal to those obtained by testing the crude extract in vitro. Thus, Drs B2 inhibited the growth of different human tumoral cell lines like the prostatic adenocarcinoma cell line PC3 and the mammary carcinoma cell line MDA-MB231 when cultured on plastic or in soft agar with an EC50 of 1–2 μM. Drs B2 was also able to inhibit non adherent human tumoral cells like the Raji and LB-EBV lymphoma cell lines with the same efficiency. It is noteworthy that Drs B2 showed an inhibitory effect on non tumoral cells like NIH-3T3 and primary mouse embryonic cells but only when concentrations higher than 10 μM where used. Furthermore, Drs B2 was evaluated for its effect on endothelial cells growth and was able to inhibit to growth of the adult bovine aortic endothelial cells (ABAE) in vitro. Moreover, Drs B2 could also block the capillary network formation induced by FGF-2 of these cells when cultured on collagen in a Montesano model.

In conclusion, Drs B2 could represent a new interesting pharmacological leader molecule against the proliferation of human tumoral cells that now need to be evaluated on different cancer cell types in vitro but also in vivo in animal models. Besides that, also the mechanism of action of Drs B2 on tumoral cells has to be clear before further pharmacological development.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B41
Abstract B43: Gene expression in breast tissue associated with mammographic density in healthy women
Authors: Haakensen Vilde D. , Biong Margarethe , Lingjærde Ole Christian , Holmen Marit Muri , Frantzen Jan Ole , Ying Chen , Navjord Dina , Romundstad Linda , Solvang Hiroko K. , Vessela Kristensen N. , Ursin Giske , Børresen-Dale Anne-Lise , Helland Åslaug
Published: January 2011

Introduction: Mammographic density (MD), as assessed from film screen mammograms, is determined by the relative content of adipose, connective and epithelial tissue in the female breast. In epidemiological studies, high MD confers a four to six fold risk elevation of developing breast cancer, even after adjustment for other known breast cancer risk factors. However, the biologic correlates of MD are virtually unknown.

Methods: Gene expression analysis using whole genome arrays was performed on breast biopsies from 143 women; 79 women with no malignancy (healthy women) recruited through mammographic centres and 64 newly diagnosed breast cancer patients. MD percentage was determined using a previously validated, computerized method on scanned mammograms. Unsupervised hierarchical clustering was used to investigate the presence of subgroups of samples within the normal biopsies. Significance analysis of microarrays (SAM) was performed to identify genes associated with MD.

Results: Hierarchical clustering of normal biopsies on the basis of the gene expression data split the individuals into two main groups. These were not significantly associated with MD or BMI. SAM identified 24 genes differentially expressed between high and low MD in the healthy women. Several single nucleotide polymorphisms (SNPs) associated with the expression of these 24 genes were identified.

Conclusions: Breast biopsies from healthy women show substantial variation in their gene-expression profiles. Twenty-four genes associated with MD and SNPs associated with their expression were identified. Several of these genes have previously been found to be linked to breast biology and oestrogen metabolism. Validation and further analysis of these genes is ongoing.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B43
Abstract B44: Value of 1H-magnetic resonance spectroscopy chemical shift imaging (CSI) for detection of anaplastic foci in diffusely infiltrating gliomas with non-significant contrast-enhancement
Authors: Widhalm Georg , Krssak Martin , Minchev Georgi , Woehrer Adelheid , Traub-Weidinger Tatjana , Czech Thomas , Asenbaum Susanne , Marosi Christine , Knosp Engelbert , Hainfellner Johannes A. , Prayer Daniela , Wolfsberger Stefan
Published: January 2011

Objective: In diffusely infiltrating gliomas (DIG), positron emission tomography (PET) imaging is a clinically powerful method for detection of anaplastic foci. Recently, 1H-magnetic resonance spectroscopy chemical shift imaging (CSI) using choline/creatine (Cho/Cr) or choline/N-acetylaspartate (Cho/NAA) ratios has emerged as new non-invasive and widely available alternative. We therefore correlated CSI with 11C-methionine (MET)-PET data in a series of DIG with non-significant contrast-enhancement (CE).

Methods: Thirty-two patients with DIG were examined with CSI on 3T MRI scanner and MET-PET. Maximum pathologic intratumoral ratios of CSI (=CSImax) and maximum tumor-to-normal-brain PET ratios (=PETmax; T/N ratio) were determined. Co-registration of MRI with CSI and PET was performed and the topographic overlap of CSImax and PETmax was analyzed. Cell proliferation rate in tumor samples inside and outside of CSImax was assessed by MIB-1 labeling index (LI).

Results: CSI showed a pathologic ratio in all patients, whereas PET demonstrated a pathologic T/N ratio in 21/32 patients. Topographical correlation of CSImax and PETmax revealed a ≥50% overlap in 18/21 and <50% overlap in 3/21 patients, respectively. Cho/Crmax and Cho/NAAmax showed a ≥50% overlap in 24/32, a <50% overlap in 8/32 patients. MIB-1 LI was significantly higher inside than outside the CSImax (13.6% versus 6.9%, p<0.001).

Conclusion: Our results indicate that CSI is a promising method for detection of anaplastic foci within DIG with non-significant CE. Intraoperative use of CSI by multimodal neuronavigation may increase the reliability of detection of malignant areas in glioma surgery and therefore optimize allocation of patients to adjuvant treatments.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B44
Abstract B47: Biomarker discovery and evaluation of response to anticancer therapeutics in breast cancer using a novel nanofluidic immunoassay
Authors: Iacovides Demetris C. , Johnson Aimee B. , Gray Joe
Published: January 2011

Cancer subtype specific molecular variations dramatically affect patient responses to already existing treatments. For example, the phosphorylation status of many proteins that are involved in signal transduction pathways perturbed in cancer cells is extremely important in determining whether these cells are susceptible to killing by available cancer therapeutics. Therefore, differentially phosphorylated protein isoforms can be a particularly useful prognostic biomarker of drug response in the clinic. However, accurate detection and quantitative analysis of cancer-related phosphoproteins in tumors is limited by current technologies.

Using a novel, fully automated nanocapillary electrophoresis technology (CB1000TM) designed to separate protein molecules based on their isoelectric point, we are currently developing highly sensitive assays for reliable assessment of the phosphorylation status of cancer-related phosphoproteins in tumors, before and during drug treatment. We have already developed and optimized assays measuring all AKT, ERK and MEK isoforms and their respective phosphoisoforms using a single detection reagent method. Using these assays, we were able to measure levels of activated MEK1/2, ERK1/2 and AKT1/2/3 in cancer cell lines, mouse xenografts and patient FNAs using protein extracted from as few as 125 cells. We have already measured and characterized changes in the phosphorylation states of these targets before, during and after treatment with inhibitors of the PI3K pathway in breast cancer, and based on this data we are currently developing molecular profiles that predict response to available therapeutics.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B47
Abstract B50: Design, synthesis, and functional characterization of new cycle-peptides inhibitors for C-X-C chemokine receptor-4
Authors: Portella L. , Napolitano M. , Consales C. , Polimeno M. N. , D'Alterio C. , Arra C. , Vitale R. , Amodeo P. , De Luca S. , Monfregola L. , Castello G. , Scala S.
Published: January 2011

Inhibition of the axis CXCR4/CXCL12 can affect tumor growth and metastases. Moreover, CXCR4 inhibition increases hematopoietic stem cell mobility in patients undergoing autologous transplantation for myeloma and non-Hodgkin lymphoma. Although several inhibitors were described neither one reached satisfactory efficacy, bioavailability and acceptable toxicity.

To develop new CXCR4 inhibitors through a rationale design approach, comparative studies were undertaken evaluating the N-terminal structure of the ligand CXCL12 and v-MIP II, an inhibitory chemokine like produced by the herpes virus 8-Kaposi associated. A common amino acidic motif (RFF) was identified in both structures although in inverted orientation. Having as a core this small domain, a new library (20 units) of cycle-peptide molecules was generated. 20 units of cycle-peptide molecules were synthesized that consists of 5 and 7 amino-acid residues cycled by a S-S bridge designed to interact with the receptor CXCR4. The peptides were characterized for functionally CXCR4 interaction through: 1. inhibition of CXCR4 binding; 2. inhibition of migration CXCL12-induced; 3. inhibition of calcium CXCL12-induced; 4. Inhibition of P-Erk CXCL12-induced. Four peptides were identified as possible CXCR4 inhibitors. These four CXCR4 inhibitory peptides were further evaluated for the in vivo efficacy through inhibition of metastasis formation. B16 mouse melanoma cells transfected with CXCR4 were injected in the vein tail in C57 Bl mice and peptide treatment followed for 10 days. Dramatic reduction in number and size of lung metastases were registered in mice peptides-treated. Moreover, xenograft of human renal cancer cells SN12C-EGFP were subcutaneously injected and systemic peptides treatment followed for 10 days. Statistically significant decrease in tumor growth was showed in peptides treated mice.

Taken together the data define three new CXCR4 inhibitory peptides, designed through innovative strategy, effective in “in vivo” deserving further development.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):B50
Abstract CN1-2: Mutations and expression profile to guide treatment in sarcomas and GIST
Authors: Blay Jean-Yves
Published: January 2011

Recent progress in the understanding of the biology of soft tissue sarcomas and locally aggressive connective tissue tumors have enabled the identification of distinct molecular and pathological entities: six molecular subgroups of connective tissue tumors may be distinguished: 1) sarcoma with specific translocations generating fusion gene whose protein products modulate transcription or may act as growth factors (e.g. EWS/Fli1 in Ewing sarcomas, PDGF-col1a1 in DFSP); 2) sarcomas with mutated activated kinases (KIT in GIST); 3) sarcomas with deletion of tumor suppressor genes such as NF1 sarcomas, or rhabdoid tumors (INI1); 4) Sarcomas with simple genetic alterations (mdm2/cdk4 amplification in WD/DD liposarcomas; 5) Sarcomas with gross genetic alterations (e.g. leiomyosarcomas); 6) Tumors with alterations of the intercellular adhesion pathways (aggressive fibromatosis with APC deletion or β catenin mutations). This classification is rapidly evolving.

The identification of the “driver” mutation in some of these diseases paved the way for the selection of efficient targeted therapies. One of the most demonstrating examples is the identification of KIT and PDGFRA kinase mutations in GIST which led to the development of imatinib, sunitinib, and more recently hsp90 inhibitors. Imatinib is also active in DFSP, a PDGF driven tumor and in giant cell tumor of the soft part (pigmented villonodular synovitis), an M-CSF driven tumor. Within the 1st molecular subgroup of tumors, in Ewing sarcoma, whose fusion gene product regulates IGFBP3, anti-IGF1R Ab yielded responses in now several phase I and II trials. Within the 4th molecular subgroup of tumors, CDK4 inhibitors and MDM2 inhibitors are currently being investigated in the clinical setting in well differentiated liposarcomas. In addition to these examples, more empiric approaches are also developed, with anti-angiogenics (pazopanib, sorafenib) and mTOR inhibitors (deforolimus) showing efficacy in phase II trials in subtypes of sarcomas; mTOR inhibitors are also currently evaluated in phase II and III trials.

The understanding of the driver molecular alterations of these tumors now guides the development of targeted treatment inhibiting driver mutations. Translational research is critical for the development of new treatments, and will be essential to understand the biological mechanisms of response and resistance in these tumor models

Citation Information: Clin Cancer Res 2010;16(7 Suppl):CN1-2
Abstract CN4-1: Genome-wide discovery and clinical validation of pharmacogenomic markers for chemotherapy
Authors: Huang R. Stephanie
Published: January 2011

Pharmacogenomics is emerging as an important component in facilitating both new drug development and in improving the utility of existing chemotherapeutic agents. However, the process of identifying these genetic variants is still evolving and there are a number of different approaches that can be employed. Typically, identification of the genetic contribution to drug response comes from clinical observations; but these clinical studies are limited by our inability to control environmental factors in vivo, the difficulty of manipulating the in vivo system to evaluate biological changes and the lack of single drug regimens in large oncology trials. Therefore, we have developed a whole genome, cell-based model for identifying genetic variation(s) important in chemotherapeutic agent-induced cytotoxicity using lymphoblastoid cell lines (LCLs) from samples included in the International HapMap project as well as extended CEPH pedigrees that have been subject to extensive genotyping for linkage mapping studies. We have identified that SNPs associated with chemotherapeutic sensitivity are enriched in expression quantitative trait locus (eQTLs). The presentation will include an overview of current cell-based models used in pharmacogenomic discovery, replication and translation into the clinical arena. As technology advances, a more comprehensive evaluation of the human genome, including the examination of rare SNPs, copy number variations, tandem repeats, epigenetic effects, will further improve our understanding of the relationship between genetics and drug response in cancers.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):CN4-1
Abstract CN4-2: Genetic polymorphisms of ABC transporters in cancer chemotherapy: Development of SmartAmp2 method for clinical genetic testing
Authors: Ishikawa Toshihisa , Aw Wanping , Lezhava Alexander , Hayashizaki Yoshihide
Published: January 2011

Pharmacogenomics data can facilitate our understanding of the sources of variability in drug response, which can potentially lead to improved safety and efficacy of chemotherapy for individual patients. Genetic polymorphisms of human ABC transporter genes are reportedly related to the risk of diseases or patients' responses to medication. However, a key requirement for the development of individualized medicine or personalized therapy is the ability to rapidly and conveniently test patients for genetic polymorphisms and/or mutations. For rapid genetic diagnosis, we have recently developed a rapid and cost-effective method named “SmartAmp2” that enables us to clinically test genotypes. Since ABCB1, ABCG2, ABCC4, and ABCC11 genes are known to be critically involved in cancer chemotherapy, we have examined whether the SmartAmp2 method could detect SNPs in those ABC transporter genes. In our study using clinical samples, the SmartAmp2 method has been proven to accurately detect and discriminate all possible homozygotes and heterozygotes of the SNPs we tested. Furthermore, we developed SmartAmp2 primers to detect mutations of K-RAS and EGFR genes in colon and lung cancers. Since the SmartAmp2 method enables us to detect genetic polymorphisms or mutations within 30 to 45 min under isothermal conditions without DNA isolation and PCR amplification, this method would provide a practical tool of genetic testing in cancer chemotherapy.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):CN4-2
Abstract ED3-1: Genome-wide association studies in cancer: Insights in cancer etiology
Authors: Chanock Stephen J.
Published: January 2011

Genome-wide association studies (GWAS) in cancer have already discovered over 100 novel regions of the genome associated with cancer risk. For nearly all regions conclusively identified by GWAS, the estimated per allele effect sizes are less than 1.3 and the susceptibility alleles are common, with minor allele frequencies greater than 5%. Each common allele appears to confer a small effect overall, underscoring the complex nature of common genetic variants that contribute to primary carcinogenesis. Interestingly, none of the variant markers associated with etiology have been strongly associated with outcomes, such as metastatic disease or survival, which suggests that common genetic variation may influence distinct processes important for carcinogenesis as opposed to those associated with disease progression. GWAS have also demonstrated that the spectrum of cancers could have distinct genomic architectures (e.g., differential contributions of alleles with a range of frequencies and effect sizes). GWAS have identified perhaps as many as 30 regions in the genome associated with prostate cancer, whereas for lung cancer, only three so far, and one of them on chromosome 15q24/25 is also strongly associated with smoking behavior phenotypes. This observation could be related to the strong environmental effect of smoking on lung cancer. It is also notable that GWAS have mapped multiple cancers to two distinct regions, 8q24 and 5p15.33. Consequently, follow-up studies are needed to map the most promising variants, preferably in different populations that can point investigators towards the optimal variants for studies designed to investigate plausibility for the association. Consequently, parallel laboratory investigation of each region can lead to new insights into novel pathways and mechanisms associated with carcinogenesis. Though there has been considerable discussion of the promise of common variants as markers for disease risk, it is too early to apply the common variants to clinical and public health venues. The success of GWAS has opened new horizons for exploration and highlighted the complex genomic architecture of disease susceptibility.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):ED3-1
Abstract ED4-3: PET tracer for therapy monitoring
Authors: Wester Hans-Jürgen , Buck Andreas
Published: January 2011

Molecular or metabolic imaging with PET plays a crucial role not only for the individual patient management but also for further validation of new therapies. The concept of using PET and [18F]fluorodeoxyglucose (FDG) for assessment of response is based on early changes in tumor glucose use which correlate to the effectiveness of a specific treatment. However, FDG-PET is a suitable surrogate marker for response prediction only in intensely FDG-avid tumors. For imaging in the context of planning of a chemotherapeutic intervention and for therapy monitoring, tracers with higher specificity are required. With the advent of molecular imaging, tumor tissue can be non-invasively characterized by imaging probes targeting biological processes such as metabolism, cell growth and migration as well as vascularization and oxygen tension. Based on such measurements signals can be defined which detect and quantitatively describe pharmacologic effects in tumor tissues within a few days after initiation of therapy. Initial results suggest that therapeutic strategies can be successfully adapted based on molecular imaging results, which are able to identify responders with improved clinical outcome. These specific and sensitive imaging signals are needed to match therapeutic targets with imaging agents addressing the same or corresponding biologic processes. Inhibiting neoangiogenesis, for example, represents an attractive therapeutic option, which benefits from imaging probes addressing molecular targets such as integrins and VEGF receptors as well as measuring perfusion and blood volume. Prior to treatment, these tracers enable to define the expression of a specific target. Alternative probes for molecular cancer imaging address metabolic key processes, such as cell proliferation ([18F]FLT), protein biosynthesis ([18F]FET, [11C]MET), neoangiogenesis ([18F]Galacto-RGD), hypoxia ([18F]FAZA), somatostatin receptor expression ([68Ga]DOTATOC), estrogen receptor expression ([18F]FES) or new concepts, such as cytokine / chemokine receptor expression ([68Ga]CPCR4). For response evalu ation, the tracer uptake has to correlate with the therapeutic effect at all imaging time points. Here, tracers with high sensitivity are required, allowing for the early assessment of therapeutic effects and response evaluation.

Citation Information: Clin Cancer Res 2010;16(7 Suppl):ED4-3
Cell-Penetrating TAT-FOXO3 Fusion Proteins Induce Apoptotic Cell Death in Leukemic Cells
Authors: Essafi Makram , Baudot Alice D. , Mouska Xavier , Cassuto Jill-Patrice , Ticchioni Michel , Deckert Marcel
Published: January 2011

FOXO proteins are Akt-regulated transcription factors involved in the control of cell cycle, DNA repair, stress defense, apoptosis, and tumor suppression. We reported that plasmid-based overexpression of constitutively active FOXO3 in cells from chronic lymphocytic leukemia (CLL) reduced their survival, suggesting that increasing FOXO3 activity in hematologic malignancies may represent a promising therapeutic strategy. The transactivating transcription factor (TAT) protein transduction domain (PTD) derived from the HIV TAT protein was shown to efficiently deliver macromolecular cargo in various cell types. In this study, wild-type FOXO3 and FOXO3 mutated on Akt sites [FOXO3 T32A/S253A/S315A or TM (triple mutant)] were fused to the TAT-PTD. Using biochemical techniques, flow cytometry, and microscopy analysis, we found a rapid and dose-dependent cell penetration into leukemic cells of unlabeled and fluorescein isothiocyanate-labeled TAT-FOXO3 fusion proteins followed by their accumulation within nuclear and cytoplasmic compartments. Treatment with TAT-FOXO3 TM—but not wild-type TAT-FOXO3—proteins induced Jurkat and K562 leukemic cell death and affected cell viability of other hematologic malignancies including primary cells from CLL. Cell transduction with TAT-FOXO3 TM induced apoptotic cell death as shown by morphologic changes, Annexin V/7-AAD (7-amino-actinomycin D) staining, activation of effector caspases, and PARP cleavage, caspase blockade through the use of the inhibitor Z-VAD, and expression of Bim and p27KIP1. By contrast, TAT-FOXO3 TM blocked cell proliferation of primary T cells, without affecting their viability. Together, our data show that cell penetrating TAT-FOXO3 TM fusion proteins constitute novel potential therapeutic agents in the treatment of lymphoproliferative disorders and hematologic malignancies. Mol Cancer Ther; 10(1); 37–46. ©2011 AACR.
Therapeutic Targeting the Loss of the Birt-Hogg-Dubé Suppressor Gene
Authors: Lu Xiaohong , Wei Wenbin , Fenton Janine , Nahorski Michael S. , Rabai Erzsebet , Reiman Anne , Seabra Laurence , Nagy Zsuzsanna , Latif Farida , Maher Eamonn R.
Published: January 2011

Brit-Hogg-Dubé (BHD) syndrome, an autosomal dominant familial cancer, is associated with increased risk of kidney cancer. BHD syndrome is caused by loss-of-function mutations in the folliculin (FLCN) protein. To develop therapeutic approaches for renal cell carcinoma (RCC) in BHD syndrome, we adopted a strategy to identify tumor-selective growth inhibition in a RCC cell line with FLCN inactivation. The COMPARE algorithm was used to identify candidate anticancer drugs tested against the NCI-60 cell lines that showed preferential toxicity to low FLCN expressing cell lines. Fifteen compounds were selected and detailed growth inhibition (SRB) assays were done in paired BHD RCC cell lines (UOK257 derived from a patient with BHD). Selective sensitivity of FLCN-null over FLCN-wt UOK257 cells was observed in seven compounds. The most selective growth-inhibitory sensitivity was induced by mithramycin, which showed an approximately 10-fold difference in GI50 values between FLCN-null (64.2 ± 7.9 nmol/L, n = 3) and FLCN-wt UOK257 cells (634.3 ± 147.9 nmol/L, n = 4). Differential ability to induce caspase 3/7 activity by mithramycin was also detected in a dose-dependent manner. Clonogenic survival studies showed mithramycin to be approximately 10-fold more cytotoxic to FLCN-null than FLCN-wt UOK257 cells (200 nmol/L). Following mithramycin exposure, UOK257-FLCN-null cells were mainly arrested and blocked in S and G2-M phases of the cell cycle and low dose of rapamycin (1 nmol/L) potentiated mithramycin sensitivity (1.5-fold in G2-M population and 2-fold in G2-M period time, 2xGI50, 48 hours). These results provide a basis for further evaluation of mithramycin as a potential therapeutic drug for RCC associated with BHD. Mol Cancer Ther; 10(1); 80–9. ©2011 AACR.
A Combined Preclinical Therapy of Cannabinoids and Temozolomide against Glioma
Authors: Torres Sofía , Lorente Mar , Rodríguez-Fornés Fátima , Hernández-Tiedra Sonia , Salazar María , García-Taboada Elena , Barcia Juan , Guzmán Manuel , Velasco Guillermo
Published: January 2011

Glioblastoma multiforme (GBM) is highly resistant to current anticancer treatments, which makes it crucial to find new therapeutic strategies aimed at improving the poor prognosis of patients suffering from this disease. Δ9-Tetrahydrocannabinol (THC), the major active ingredient of marijuana, and other cannabinoid receptor agonists inhibit tumor growth in animal models of cancer, including glioma, an effect that relies, at least in part, on the stimulation of autophagy-mediated apoptosis in tumor cells. Here, we show that the combined administration of THC and temozolomide (TMZ; the benchmark agent for the management of GBM) exerts a strong antitumoral action in glioma xenografts, an effect that is also observed in tumors that are resistant to TMZ treatment. Combined administration of THC and TMZ enhanced autophagy, whereas pharmacologic or genetic inhibition of this process prevented TMZ + THC-induced cell death, supporting that activation of autophagy plays a crucial role on the mechanism of action of this drug combination. Administration of submaximal doses of THC and cannabidiol (CBD; another plant-derived cannabinoid that also induces glioma cell death through a mechanism of action different from that of THC) remarkably reduces the growth of glioma xenografts. Moreover, treatment with TMZ and submaximal doses of THC and CBD produced a strong antitumoral action in both TMZ-sensitive and TMZ-resistant tumors. Altogether, our findings support that the combined administration of TMZ and cannabinoids could be therapeutically exploited for the management of GBM. Mol Cancer Ther; 10(1); 90–103. ©2011 AACR.
Glutathione-Conjugate Transport by RLIP76 Is Required for Clathrin-Dependent Endocytosis and Chemical Carcinogenesis
Authors: Singhal Sharad S. , Wickramarachchi Dilki , Yadav Sushma , Singhal Jyotsana , Leake Kathryn , Vatsyayan Rit , Chaudhary Pankaj , Lelsani Poorna , Suzuki Sumihiro , Yang Shaohua , Awasthi Yogesh C. , Awasthi Sanjay
Published: January 2011

Targeted depletion of the RALBP1-encoded 76-kDa splice variant, RLIP76, causes marked and sustained regression of human xenografts of lung, colon, prostate, and kidney cancers without toxicity in nude mouse models. We proposed that the remarkable efficacy and broad spectrum of RLIP76-targeted therapy is because its glutathione-conjugate (GS-E) transport activity is required for clathrin-dependent endocytosis (CDE), which regulates all ligand-receptor signaling, and that RLIP76 is required not only for survival of cancer cells but also for their very existence. We studied RLIP76 mutant proteins and the functional consequences of their expression into RLIP76−/− MEFs, identified key residues for GS-E binding in RLIP76, established the requirement of RLIP76-mediated GS-E transport for CDE, and showed a direct correlation between GS-E transport activities with CDE. Depletion of RLIP76 nearly completely blocked signaling downstream of EGF in a CDE-dependent manner and Wnt5a signaling in a CDE-independent manner. The seminal prediction of this hypothesis—RLIP76−/− mice will be deficient in chemical neoplasia—was confirmed. Benzo[a]pyrene, dimethylbenzanthracene, and phorbol esters are ineffective in causing neoplasia in RLIP76−/−. PMA-induced skin carcinogenesis in RLIP76+/+ mouse was suppressed completely by depletion of either PKCα or RLIP76 by siRNA or antisense and could be restored by topical application of RLIP76 protein in RLIP76−/− mouse skin. Likewise, chemical pulmonary carcinogenesis was absent in female and nearly absent in male RLIP76−/− mice. In RLIP76−/− mice, p53, p38, and JNK activation did not occur in response to either carcinogen. Our findings show a fundamental role of RLIP76 in chemical carcinogenesis. Mol Cancer Ther; 10(1); 16–28. ©2011 AACR.
Bortezomib Sensitizes Malignant Human Glioma Cells to TRAIL, Mediated by Inhibition of the NF-κB Signaling Pathway
Authors: Jane Esther P. , Premkumar Daniel R. , Pollack Ian F.
Published: January 2011

Previous studies have shown that the tumor necrosis factor–related apoptosis-inducing ligand (TRAIL) has significant apoptosis-inducing activity in some glioma cell lines, although many lines are either moderately or completely resistant, which has limited the therapeutic applicability of this agent. Because our recent studies showed that inhibition of proteasomal function may be independently active as an apoptosis-inducing stimulus in these tumors, we investigated the sensitivity of a panel of glioma cell lines (U87, T98G, U373, A172, LN18, LN229, LNZ308, and LNZ428) to TRAIL alone and in combination with the proteasome inhibitor bortezomib. Analysis of these cell lines revealed marked differences in their sensitivity to these treatments, with two (LNZ308 and U373) of the eight cell lines revealing no significant induction of cell death in response to TRAIL alone. No correlation was found between sensitivity of cells to TRAIL and expression of TRAIL receptors DR4, DR5, and decoy receptor DcR1, caspase 8, apoptosis inhibitory proteins XIAP, survivin, Mcl-1, Bcl-2, Bcl-Xl, and cFLIP. However, TRAIL-resistant cell lines exhibited a high level of basal NF-κB activity. Bortezomib was capable of potentiating TRAIL-induced apoptosis in TRAIL-resistant cells in a caspase-dependent fashion. Bortezomib abolished p65/NF-κB DNA-binding activity, supporting the hypothesis that inhibition of the NF-κB pathway is critical for the enhancement of TRAIL sensitization in glioma cells. Moreover, knockdown of p65/NF-κB by shRNA also enhanced TRAIL-induced apoptosis, indicating that p65/NF-κB may be important in mediating TRAIL sensitivity and the effect of bortezomib in promoting TRAIL sensitization and apoptosis induction. Mol Cancer Ther; 10(1); 198–208. ©2011 AACR.
Pancratistatin Selectively Targets Cancer Cell Mitochondria and Reduces Growth of Human Colon Tumor Xenografts
Authors: Griffin Carly , Karnik Aditya , McNulty James , Pandey Siyaram
Published: January 2011

The naturally occurring Amaryllidaceae alkaloid pancratistatin exhibits potent apoptotic activity against a large panel of cancer cells lines and has an insignificant effect on noncancerous cell lines, although with an elusive cellular target. Many current chemotherapeutics induce apoptosis via genotoxic mechanisms and thus have low selectivity. The observed selectivity of pancratistatin for cancer cells promoted us to consider the hypothesis that this alkaloid targets cancer cell mitochondria rather than DNA or its replicative machinery. In this study, we report that pancratistatin decreased mitochondrial membrane potential and induced apoptotic nuclear morphology in p53-mutant (HT-29) and wild-type p53 (HCT116) colorectal carcinoma cell lines, but not in noncancerous colon fibroblast (CCD-18Co) cells. Interestingly, pancratistatin was found to be ineffective against mtDNA-depleted (ρ0) cancer cells. Moreover, pancratistatin induced cell death in a manner independent of Bax and caspase activation, and did not alter β-tubulin polymerization rate nor cause double-stranded DNA breaks. For the first time we report the efficacy of pancratistatin in vivo against human colorectal adenocarcinoma xenografts. Intratumor administration of pancratistatin (3 mg/kg) caused significant reduction in the growth of subcutaneous HT-29 tumors in Nu/Nu mice (n = 6), with no apparent toxicity to the liver or kidneys as indicated by histopathologic analysis and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling. Altogether, this work suggests that pancratistatin may be a novel mitochondria-targeting compound that selectively induces apoptosis in cancer cells and significantly reduces tumor growth. Mol Cancer Ther; 10(1); 57–68. ©2011 AACR.