bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract C56: High αV integrin level is associated with metastasis to the brain in athymic rat hematogenous models
Authors: Wu Jeffrey , Pagel Michael , Muldoon Leslie L. , Neuwelt Edward A.
Published: August 2014

Brain metastases from systemic cancers occur in approximately 20% of lung cancer patients including both small cell lung carcinoma (SCLC) and non–small-cell lung cancer (NSCLC), and in 5-8% of breast cancer and melanoma. Brain metastases tend to present as multiple brain lesions with high morbidity and poor prognosis. We investigated the expression of αV integrin cell adhesion protein in multiple human cancer cell lines (3 SCLC, 4 NSCLC, 2 melanoma, 1 breast cancer), and its correlation with migration in vitro and brain metastatic ability in vivo using a hematogenous metastasis technique in which cancer cells are infused into the internal carotid artery in athymic nude rats. Expression of αV integrin protein was highly variable among human cancer cell lines. Four cell lines tested had high αV integrin expression and formed multiple brain metastases. H460 NSCLC cells formed 13 ± 6 metastases per brain (mean volume 3 ± 9 mm3) with median survival 34 days (n = 4). A549 lung adenocarcinoma cells formed 9 ± 5 metastases per brain (8 ± 21 mm3 volume, 62 day survival, n = 6). MDA-MB231BR-HER2 breast cancer and A2058 melanoma cells also formed multiple brain metastases. Low αV expressing lung cancer cells, including H146 and DMS79 SCLC, and H520 squamous NSCLC cells failed to form brain metastases by 12-16 weeks after intracarotid cell infusion. In an in vitro gap closure migration assay, A549 cells (high αV integrin) showed a higher rate of migration than H520 cells (low αV integrin). Regardless of αV integrin level, incubation with intetumumab anti-αV integrin monoclonal antibody significantly inhibited cell migration of both A549 and H520 cells. We conclude that cancer cellular αV integrin level is positively correlated to the development of brain metastases and may be a potential biomarker of patients likely to develop brain metastases. We suggest that blocking αV integrin with intetumumab has prophylactic potential to prevent distant metastasis and improve survival in cancer patients.

Citation Format: Jeffrey Wu, Michael Pagel, Leslie L. Muldoon, Edward A. Neuwelt. High αV integrin level is associated with metastasis to the brain in athymic rat hematogenous models. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr C56.
Abstract C61: GRP78 as a regulator of liver steatosis and cancer progression mediated by loss of the tumor suppressor PTEN
Authors: Chen Wan-Ting , Zhu Genyuan , Pfaffenbach Kyle , Kanel Gary , Stiles Bangyan , Lee Amy S.
Published: August 2014

Glucose-regulated protein 78 (GRP78), a molecular chaperone widely elevated in human cancers, is critical for endoplasmic reticulum (ER) protein folding, stress signaling, and PI3K/AKT activation. Genetic knockout models of GRP78 revealed that GRP78 maintains homeostasis of metabolic organs, including liver, pancreas, and adipose tissues. Hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC) are the most common liver cancers. There is a lack of effective therapeutics for HCC and CC, highlighting the need to further understand liver tumorigenic mechanisms. Phosphatase and tensin homolog (PTEN), a tumor suppressor that antagonizes the PI3K/AKT pathway, is inactivated in a wide range of tumors, including 40-50% of human liver cancers. To elucidate the role of GRP78 in liver cancer, we created a mouse model with biallelic liver-specific deletion of Pten and Grp78 mediated by Albumin-Cre-recombinase (cPf/f78f/f). Interestingly, in contrast to PTEN, deletion of GRP78 was progressive but incomplete. At 3 months cPf/f78f/f livers showed hepatomegaly, activation of lipogenic genes, exacerbated steatosis, and liver injury, implying that GRP78 protects the liver against PTEN-null mediated pathogenesis. Furthermore, in response to liver injury, we observed increased proliferation and expansion of bile duct and liver progenitor cells in cPf/f78f/f livers. Strikingly, bile duct cells in cPf/f78f/f livers maintained wild-type GRP78 level while adjacent areas showed GRP78 reduction. Analysis of signaling pathways revealed selective c-Jun N-terminal protein kinase activation, β-catenin downregulation, along with platelet-derived growth factor receptor α upregulation, which was unique to cPf/f78f/f livers at 6 months. Development of both HCC and CC were accelerated and evident in cPf/f78f/f livers at 8-9 months, coinciding with intense GRP78 expression in the cancer lesions, and GRP78 expression in adjacent normal areas reverted back to the WT level. In contrast, livers of Albumin-Cre; Grp78f/f (c78f/f) mice showed no malignancy even at 14 months. These studies reveal GRP78 is a novel regulator for PTEN-loss mediated liver injury and cancer progression.

Citation Format: Wan-Ting Chen, Genyuan Zhu, Kyle Pfaffenbach, Gary Kanel, Bangyan Stiles, Amy S. Lee. GRP78 as a regulator of liver steatosis and cancer progression mediated by loss of the tumor suppressor PTEN. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr C61.
Abstract IA12: The noncoding RNA revolution in cancer research
Authors: Pandolfi Pier Paolo
Published: August 2014

We will discuss exciting new data regarding the role of pseudogenes, lincRNAs and miRNAs in the pathogenesis of human cancer as also studied in vivo in the mouse. We will also focus on competing endogenous RNAs (ceRNAs), circular(circ)-ceRNAs and pseudo-ceRNAs and give important attention to how understanding the ceRNA language will facilitate efforts to deconvolute ceRNA networks and their role in tumorigenesis.

Citation Format: Pier Paolo Pandolfi. The noncoding RNA revolution in cancer research. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr IA12.
Abstract IA22: PI3K and cancer metabolism
Authors: Cantley Lewis C.
Published: August 2014

Phosphoinositide 3-Kinase (PI3K) is a central enzyme in a signaling pathway that mediates cellular responses to growth factors. The signaling pathway downstream of PI3K is highly conserved from worms and flies to humans and genetic analysis of the pathway has revealed a conserved role in regulating glucose metabolism and cell growth. Mutational events that lead to hyperactivation of the PI3K pathway result in hamartoma syndromes and cancers. Activating mutations in PIK3CA, encoding the p110alpha catalytic subunit of PI3K or inactivating mutations in PTEN, a phosphoinositide 3-phosphatases that reverses the effects of PI3K, are among the most common events in solid tumors. Drugs that target PI3K are in clinical trials for a variety of cancers. It is likely that PI3K pathway inhibitors will need to be combined with other drugs to be broadly effective. We have employed genetically engineered mouse models that develop cancers due to mutations in genes in the PI3K pathway and are using these models to explore the efficacy of PI3K pathway inhibitors as single agents or in combination with other drugs. The role of PI3K inhibitors for treating cancers in these mouse models and in human trials will be discussed.

Citation Format: Lewis C. Cantley. PI3K and cancer metabolism. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr IA22.
Abstract IA4: Deconstructing p53 pathways in vivo
Authors: Brady Colleen A. , Broz Daniela Kenzelmann , Jiang Dadi , Mello Stephano Spano , Bieging Kathryn , Johnson Thomas M. , Jarvis Leslie A. , Kozak Margaret M. , Basak Shashwati , Attardi Laura D.
Published: August 2014

The activation of the p53 protein by cellular stress signals is fundamental for tumor suppression but also promotes pathological states, such as provoking the side effects of genotoxic cancer therapies. To better understand the mechanisms of p53 action in different contexts, we have leveraged both mouse genetic and genomic approaches. First, we have used mouse genetics to define transcriptional programs involved in p53 function in different in vivo settings, specifically by generating a panel of p53 transcriptional activation domain mutant knock-in mouse strains. These include strains expressing p53 mutants in the first (p5325,26), second (p5353,54), or both transactivation domains (p5325,26,53,54). We have observed that p5325,26 is severely compromised for transactivation of most classical p53 target genes, but retains the ability to activate a subset of p53 targets, while p5325,26,53,54 lacks transactivation activity completely. Interestingly, although unable to induce apoptosis or cell cycle arrest in response to acute DNA damage signals, p5325,26 retains full activity in suppressing cancers of a wide range of types, indicating that robust transactivation of most canonical p53 targets is dispensable for tumor suppression. Importantly, as p5325,26 activates only a subset of p53-dependent genes, yet retains tumor suppressor activity, it has helped to define a small set of novel p53-inducible tumor suppression-associated genes, which we are currently analyzing in detail. Second, we have utilized genomic approaches to better understand p53 function. Using ChIP-sequencing and RNA-sequencing to analyze transcriptional programs in acute DNA damage-treated mouse embryo fibroblasts, our studies have revealed an extensive p53-regulated autophagy program that contributes to p53 responses. Together, these approaches will help better define the transcriptional networks important for p53 action in different settings.

Citation Format: Colleen A. Brady, Daniela Kenzelmann Broz, Dadi Jiang, Stephano Spano Mello, Kathryn Bieging, Thomas M. Johnson, Leslie A. Jarvis, Margaret M. Kozak, Shashwati Basak, Laura D. Attardi. Deconstructing p53 pathways in vivo. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr IA4.
Abstract PR08: Autophagic degradation of Δ133p53 during replicative cellular senescence: An isoform-specific protein degradation mechanism for p53
Authors: Horikawa Izumi , Fujita Kaori , Jenkins Lisa Miller , Hiyoshi Yukiharu , Mondal Abdul M. , Vojtesek Borivoj , Lane David P. , Appella Ettore , Harris Curtis C.
Published: August 2014

Δ133p53, a p53 isoform that dominant-negatively inhibits full-length p53, is downregulated at replicative senescence in a manner independent of mRNA regulation and proteasome-mediated degradation. We here demonstrate that, in contrast to proteasomal degradation of full-length p53, Δ133p53 is degraded by autophagy. Pharmacological inhibition of autophagy restores the reduced expression of Δ133p53 in replicatively senescent fibroblasts. The siRNA-mediated knockdown of pro-autophagic proteins (ATG5, ATG7 and Beclin-1) also results in restored expression of Δ133p53. The degradation of Δ133p53 via selective autophagy is indicated by its ubiquitination at the C-terminal lysine residues, its colocalization in the cytoplasm with a ubiquitin-binding protein p62/SQSTM1 and an autophagosome component LC3-II upon autophagy induction, and its stabilization by siRNA knockdown of p62/SQSTM1. A chaperone-associated E3 ubiquitin ligase STUB1, rather than MDM2, physically interacts with Δ133p53 and, consistent with the downregulation of endogenous STUB1 at replicative senescence, siRNA knockdown of STUB1 induces autophagic degradation of Δ133p53 and thereby senescence. Our data reveal that Δ133p53 degradation is controlled specifically by STUB1, rather than overall activity of autophagy. This study has identified for the first time a p53 isoform-specific protein turnover mechanism that orchestrates p53-mediated senescence and supports the hypothesis that Δ133p53 functions at the crossroads of senescence and autophagy.

This abstract is also presented as poster B19.

Citation Format: Izumi Horikawa, Kaori Fujita, Lisa Miller Jenkins, Yukiharu Hiyoshi, Abdul M. Mondal, Borivoj Vojtesek, David P. Lane, Ettore Appella, Curtis C. Harris. Autophagic degradation of Δ133p53 during replicative cellular senescence: An isoform-specific protein degradation mechanism for p53. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr PR08.
Abstract PR12: Bringing the outside in: Macropinocytosis and cancer therapeutics
Authors: Commisso Cosimo , Chenna Venugopal , Grabocka Elda , Ramirez Craig , Anirban Maitra , Bar-Sagi Dafna
Published: August 2014

Oncogenic Ras mutations are prevalent in a variety of tumor types, including adenocarcinomas of the pancreas, colon, and lung. One of the most overt phenotypes associated with the expression of oncogenic Ras mutants is the stimulation of macropinocytosis, an endocytic process that involves extensive membrane remodeling and the internalization of extracellular fluid via large membrane-bound vesicles called macropinosomes. The functional consequences of this stimulation in oncogenic Ras-expressing cancer cells were unknown prior to our recent work where we linked macropinocytic uptake to nutrient delivery and amino acid supply in tumor cells. We found that Ras-transformed cells utilize macropinocytosis to internalize extracellular albumin, which is then lysosomally degraded releasing the constituent amino acids intracellularly. These protein-derived amino acids have the capacity to enter central carbon metabolism and fuel tumor cell proliferation even in a nutrient-depleted environment. Of particular relevance is the finding that the pharmacological inhibition of macropinocytosis compromises the growth of Ras-driven pancreatic xenograft tumors. This discovery raises the question of whether the inhibition of macropinocytosis can be utilized as a therapeutic intervention in a subset of cancers. Moreover, because macropinocytosis is an established mechanism of drug delivery for nanoparticles, our work highlights the possibility of exploiting this feature of Ras-induced tumors for the effective delivery of nanoscale therapeutics. Our recent studies focused on identifying novel modulators of macropinocytosis and examining macropinocytosis as a delivery mechanism for nab-paclitaxel, an albumin-based nanoparticle therapy, will be discussed.

Citation Format: Cosimo Commisso, Venugopal Chenna, Elda Grabocka, Craig Ramirez, Maitra Anirban, Dafna Bar-Sagi. Bringing the outside in: Macropinocytosis and cancer therapeutics. [abstract]. In: Proceedings of the Third AACR International Conference on Frontiers in Basic Cancer Research; Sep 18-22, 2013; National Harbor, MD. Philadelphia (PA): AACR; Cancer Res 2013;73(19 Suppl):Abstract nr PR12.
Abstract B99: Antimetastatic and apoptosis induction potential of supercritical CO2 extract of Nigella sativa seeds
Authors: Baharetha Hussein M. , Nassar Zeyad D. , Aisha Abdalrahim F. , Kadir Abd , Ismail Zhari , Majid Abdul
Published: August 2014

Nigella sativa Linn. is a famous spice that has been used as a medicinal plant for centuries. Traditionally, it is known to have a magic healing power for different varieties of illness, range from intestinal disturbances to fever to cancer. In this study, essential oil was extract from seeds of Nigella sativa using supercritical CO2 extraction system at extraction pressure of 2500 psi and temperature of 60°C (these extraction conditions of pressure and temperature were reported from our preliminary screening anti-proliferative study against various human cancerous cell lines). The anti-metastatic and anti-invasive properties of the extract were studied using a panel of assays against human breast cancer cell line (MCF 7). Results revealed that the extract showed a potent anti-proliferation activity against MCF 7 cells with IC50 value of 53.34 ± 2.15 μg/ml once tested using MTT assay. The extract also halted MCF 7 cells migration and invasion, where 40 μg/ml of the extract A3 represented 49.80 ± 5.89% of migration inhibition and 79.29 ± 2.09% of invasion inhibition. In addition, the extract significantly induced apoptosis of the MCF 7 cells by activating caspases 3/7 and 8. The result of this study revealed the role of supercritical CO2 extracted oil of Nigella sativa in prevention of metastasis and invasion of breast cancer.

Note: This abstract was not presented at the conference.

Citation Format: Hussein M. Baharetha, Zeyad D. Nassar, Abdalrahim F. Aisha, Abd Kadir, Zhari Ismail, Abdul Majid. Antimetastatic and apoptosis induction potential of supercritical CO2 extract of Nigella sativa seeds. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr B99.
Abstract C12: Aberrant promoter methylation of HOPX-β, MUC1 genes and global methylation in esophageal squamous cell carcinoma
Authors: Kao Yu Chun , Lin Sheng Hsiung , Yu Chun Chieh , Li Ruei Nian , Wu Ming Tsang
Published: August 2014

Esophageal cancer ranks as the sixth most deadly cancer and esophageal squamous cell carcinoma (ESCC) is the sixth most common cause of cancer deaths in males in worldwide. Therefore, it is important to identify molecular markers for clinical diagnosis. Homeodomain-only protein X (HOPX)-β is a tumor suppressor gene and its promoter methylation is shown to be frequent in human cancer. Mucin 1 (MUC1) is a transmembrane mucin that is highly expressed in various cancers and correlates with malignant potential. In addition, the degree of whole genome methylation decreases as the lesion progressing from a benign proliferation of cells to an invasive cancer. The aim of this study was to investigate HOPX-β and MUC1 promoter methylation status and global methylation level in different stages of ESCC patients. Methylation-specific PCR (MSP) revealed that the methylation rate of HOPX-β was 26.5% (27/102) in tumor samples and only 9% (9/100) in paired non-tumor samples (p<0.05). In HOPX-β promoter hypermethylation samples, the frequency of lymph node invasion and tumor metastasis was increased about 3-fold and 2-fold, respectively. Furthermore, HOPX-β promoter methylation frequency was increased about 3-fold in advanced stage. On the other hand, the methylation frequency of MUC1 was 90.7% (97/107) in paired non-tumor samples while the methylation frequency was decreased to 53.3% (57/107) in tumor samples. 78.5% (84/107) of tumor samples showed significantly decline in the degree of methylation compared with paired non-tumor samples (p<0.0001). We found a decrease tendency in methylation frequency of MUC1 correlated with clinical stage (early/advanced), the frequency of lymph node invasion and metastasis. Importantly, the methylation rate of MUC1 was inversely related with the tumor size (p=0.0105). The level of global long interspersed nuclear element 1 (LINE-1) methylation in ESCC tissues (mean 65.8%) was significantly lower than paired non-tumor samples (mean 79.5%). However, there is no relation between the level of global LINE-1 methylation and tumor stage or survival rate. Our findings indicate that LINE-1, MUC1 methylation level and HOPX-β hypermethylation in ESCC are useful markers in clinical diagnosis.

Citation Format: Yu Chun Kao, Sheng Hsiung Lin, Chun Chieh Yu, Ruei Nian Li, Ming Tsang Wu. Aberrant promoter methylation of HOPX-β, MUC1 genes and global methylation in esophageal squamous cell carcinoma. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C12.
Abstract C65: Membrane type-1 matrix metalloproteinase (MT1-MMP) is a potent regulator of cancer malignant progression through the heparin-binding EGF-like growth factor activation
Authors: Koshikawa Naohiko , Seiki Motoharu
Published: August 2014

MT1-MMP is a potent proinvasive and growth promoting membrane protease in cancer. The roles of MT1-MMP are mediated by its pericellular proteolysis activity. Major substrates of MT1-MMP are collagen I and other extracellular matrices. MT1-MMP also cleaves membrane proteins and modulates their biological activities in cancer progression. However, little is known about the cell surface substrates of MT1-MMP and identification of the substrates should enable a better understanding of its multiple biological functions. Therefore to identify new substrates of MT1-MMP, we purified proteins associating with MT1-MMP in human carcinoma cells and analyzed them by mass spectrometry. As results, we identified over 40 membrane proteins, and found HB-EGF is a novel component of MT1-MMP complex. MT1-MMP cleaves HB-EGF and removes the N-terminal region that is important for binding heparin. Consequently the HB-EGF processing by MT1-MMP converts it into a heparin-independent growth factor with enhanced mitogenic activity and thereby expression of both proteins co-stimulates tumor cell growth in vitro and in vivo. Furthermore ovarian ascites carcinoma cells expressed both MT1-MMP and HB-EGF at high levels and produced MT1-MMP-cleaved HB-EGF in the ascitic fluids. These data suggest that MT1-MMP acts as a regulator of cancer progression on the cell surface.

Citation Format: Naohiko Koshikawa, Motoharu Seiki. Membrane type-1 matrix metalloproteinase (MT1-MMP) is a potent regulator of cancer malignant progression through the heparin-binding EGF-like growth factor activation. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Invasion and Metastasis; Jan 20-23, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;73(3 Suppl):Abstract nr C65.
Abstract A28: Generation and characterization of anti-idiotype antibody ganglidiomab as GD2 surrogate for immunotherapy of neuroblastoma.
Authors: Lode Holger N. , Schmidt Manuela , Seidel Diana , Huebener Nicole , Brackrock Diana , Bleeke Matthias , Reker Daniel , Brandt Sven , Mueller Hans-Peter , Helm Christiane , Siebert Nikolai
Published: August 2014

Purpose: Immunotherapy targeting disialoganglioside GD2 emerges as an important treatment option for neuroblastoma, a pediatric malignancy characterized by poor outcome. Here, we report the generation and characterization of ganglidiomab, a new anti-idiotype antibody to anti-GD2 antibodies of the 14.18 family for monitoring of clinical trials and the development of neuroblastoma vaccines.

Experimental Design and Results: Balb/c mice were immunized with 14G2a and splenocytes harvested to generate hybridoma cells. Clones were screened by ELISA for mouse antibody binding to hu14.18. One positive clone was selected to purify and characterize the secreted IgG protein (κ, IgG1). This antibody bound to anti-GD2 antibodies 14G2a, ch14.18/CHO, hu14.18 and to immunocytokines ch14.18-IL2 and hu14.18-IL2 as well as to NK-92 cells expressing scFv(ch14.18)-zeta receptor. Binding of these anti-GD2 antibodies to the nominal antigen GD2 as well as GD2 specific lysis of neuroblastoma cells by NK-92-scFv(ch14.18)-zeta cells was competitively inhibited by ganglidiomab, proving GD2 surrogate function and anti-idiotype characteristics. The dissociation constants of ganglidiomab from anti-GD2 antibodies ranged from 10.8 ± 5.01 to 53.5 ± 1.92 nM as determined by Biacore analyses using “steady state” analysis. The sequences of framework- (FRs) and complementarity determining -regions (CDRs) of ganglidiomab were identified. Finally, we demonstrate induction of a GD2 specific humoral immune response after vaccination of mice with ganglidiomab effective in mediating GD2 specific killing of neuroblastoma cells.

Conclusion: We generated and characterized a novel anti-idiotype antibody ganglidiomab for immune monitoring of clinical trials with anti-GD2 antibodies and provide an important baseline for the development of anti-idiotype vaccines against malignancies expressing GD2.

Citation Format: Holger N. Lode, Manuela Schmidt, Diana Seidel, Nicole Huebener, Diana Brackrock, Matthias Bleeke, Daniel Reker, Sven Brandt, Hans-Peter Mueller, Christiane Helm, Nikolai Siebert. Generation and characterization of anti-idiotype antibody ganglidiomab as GD2 surrogate for immunotherapy of neuroblastoma. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A28.
Abstract A53: Alpha particles and induction of an antitumor immune response.
Authors: Gorin Jean-Baptiste , Gouard Sébastien , Chérel Michel , Faivre-Chauvet Alain , Ménager Jérémie , Morgenstern Alfred , Bruchertseifer Frank , Davodeau François , Gaschet Joëlle
Published: August 2014

Objectives: Radioimmunotherapy (RIT) is an approach allowing targeting of radioelements to tumours, usually through the use of antibodies specific for tumour antigens. The radiations emitted by the radioelements then induce direct killing of the targeted cells as well as indirect killing through bystander effect. Interestingly, it has been shown that ionizing radiation, in some settings of external radiotherapy, can foster an immune response directed against tumour cells. Our research team is dedicated to the development of alpha RIT, i.e RIT using alpha particle emitters, we therefore decided to study the effects of such particles on tumour cells in regards to their immunogenicity.

Material and Methods: We studied the effects of bismuth-213, an alpha emitter, on cellular death and autophagy in a mouse (5T33) and a human (LP-1) myeloma cell line. Using flow cytometry analysis, we measured the expression of MHC molecules and several DAMPs at the cell surface to determine whether irradiation with bismuth-213 could cause the tumour cells to be recognized by the immune system. We also tested the irradiated cell culture media by ELISA to check whether DAMPs were secreted after treatment with bismuth-213. Finally, a co-culture of dendritic cells with irradiated tumour cells or their culture media was performed to test whether it would induce dendritic cell activation.

Results: Only slight levels of apoptosis and necrosis were detected within the 48 hours following irradiation in the studied cell lines, however they both exhibited signs of autophagy. No increase in membrane or extracellular expression of DAMPs was observed after treatment with bismuth-213, or in the expression of MHC class I and II. However, the co-culture experiments indicated that soluble factors are released in the culture media after irradiation with bismuth-213, which are able to induce dendritic cell activation.

Conclusion: These data suggest that alpha particles would be of interest in rising an immune response associated to RIT.

Citation Format: Jean-Baptiste Gorin, Sébastien Gouard, Michel Chérel, Alain Faivre-Chauvet, Jérémie Ménager, Alfred Morgenstern, Frank Bruchertseifer, François Davodeau, Joëlle Gaschet. Alpha particles and induction of an antitumor immune response. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A53.
Abstract A56: The number of MDSC (Gr1+/CD11b+) and T lymphocytes in blood of canine patients during mammary and skin tumors development.
Authors: Mucha Joanna , Homa Agata , Majchrzak Kinga , Motyl Tomasz , Król Magdalena
Published: August 2014

The very recent studies have indicated an important role of myeloid derived suppressor cells (MDSC) in the tumor progression and metastasis. They are thought to suppress the immune system and promote angiogenesis via STAT3 activation.

The aim of present study was two-fold: 1) to assess the percentage of MDSCs (CD11b+/Gr1+) in canine blood leukocytes and percentage of MDSCs that express p-STAT3 at high level; and 2) to assess the percentage of CD3+, CD4+, CD8+ and Treg (CD4+/FoxP3+) in canine blood lymphocytes collected from patients with various clinical stage of mammary and skin tumors comparing to control healthy dogs. The samples were analyzed using BD FACS AriaII.

The results of our study indicate that percentage of Gr1-positive cells is significantly increased (p<0.05) in patients with stage IV tumor (4.6%, compared to control and stage I, II and III tumor: 2.38%, 0.8%, 0.65%, 1.2%, respectively). The number of Treg cells was significantly decreased (p<0.001) in patients with stage III and IV tumor (1.06% and 1.03%, respectively) compared to control (4.82%), stage I and II tumor (6.5% and 3.95%, respectively). Similarly, the number of CD4+ cells was significantly decreased (p<0.01 and p<0.001) in stage III and IV tumors (14.8% and 2.27%, respectively) compared to control (29.84%), and stage I and II tumors (26.20% and 18.75%, respectively). Thus, the ratio of CD4/CD8 was also significantly decreased (p<0.01 and p<0.001) in stage III and IV tumors (respectively). The number of CD3+ cells was also significantly (p<0.01) decreased in stage III and IV tumors (5.23% and 4.46%, respectively), comparing to control (11.66%), and stage I and II tumors (13.57% and 7.4%).

Our results indicate significant immunosuppression in patients with advanced tumors that can be related to high MDSCs level. High level of these cells may be also related with metastasis formation in patients with advanced tumors.

This work was supported by grant no IP2011 027171 from the Ministry of Sciences and Higher Education.

Citation Format: Joanna Mucha, Agata Homa, Kinga Majchrzak, Tomasz Motyl, Magdalena Król. The number of MDSC (Gr1+/CD11b+) and T lymphocytes in blood of canine patients during mammary and skin tumors development. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A56.
Abstract A74: The chemoattractant chemerin suppresses melanoma by recruiting natural killer cell antitumor defenses.
Authors: Pachynski Russell , Zabel Brain , Monnier Justin , Gentles Andrew , Swanson Christina , Kohrt Holbrook , Hadeiba Husein , Alizadeh Ash , Butcher Eugene
Published: August 2014

Infiltration of specialized immune cells regulates the growth and survival of neoplasia. Here, in a survey of public whole genome expression datasets we found that the gene for chemerin, a widely expressed endogenous chemoattractant protein, is down-regulated in melanoma as well as other human tumors. Moreover, high chemerin messenger RNA expression in tumors correlated with improved outcome in human melanoma. In experiments using the B16 transplantable mouse melanoma, tumor-expressed chemerin inhibited in vivo tumor growth without altering in vitro proliferation. Growth inhibition was associated with an altered profile of tumor-infiltrating cells with an increase in natural killer (NK) cells and a relative reduction in myeloid-derived suppressor cells and putative immune inhibitory plasmacytoid dendritic cells. Tumor inhibition required host expression of CMKLR1 (chemokine- like receptor 1), the chemoattractant receptor for chemerin, and was abrogated by NK cell depletion. Intratumoral injection of chemerin also inhibited tumor growth, suggesting the potential for therapeutic application. Preliminary data in other adenocarcinoma tumor types (eg breast) show similar results, supporting broad applicability of such an approach. These results show that chemerin, whether expressed by tumor cells or within the tumor environment, can recruit host immune defenses that inhibit tumorigenesis and suggest that down-regulation of chemerin may be an important mechanism of tumor immune evasion.

Citation Format: Russell Pachynski, Brain Zabel, Justin Monnier, Andrew Gentles, Christina Swanson, Holbrook Kohrt, Husein Hadeiba, Ash Alizadeh, Eugene Butcher. The chemoattractant chemerin suppresses melanoma by recruiting natural killer cell antitumor defenses. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A74.
Abstract A76: Comparative mRNA and microRNA expression profiling of canine mammary carcinoma cell lines and macrophages grown as a co-culture in vitro.
Authors: Król Magdalena , Pawlowski Karol , Majchrzak Kinga , Majewska Alicja , Motyl Tomasz
Published: August 2014

Solid tumors comprise of various cells, including cancer cells, resident stromal cells, migratory haemopoietic cells and other. These cells regulate tumor growth and metastasis. Macrophages constitute probably the most important element of all interactions within the tumor microenvironment. However, the molecular mechanism, that guides tumor environment, still remains unknown. Exploring the underlying molecular mechanisms that orchestrate these phenomena has been the aim of our study.

The co-cultures of five various canine mammary carcinoma cell lines and macrophages were established and maintained for 72 hrs. After that, having sorted the cells, an integrated analysis of genome-wide mRNA and microRNA expression profiles have been assessed.

The analysis showed that the up-regulated genes in the cancer cell lines grown as co-culture with macrophages are involved mainly in: macrophages activation, cell motion, mammary gland development, cell-cell adhesion, and angiogenesis. Basides, the presence of macrophages in the cancer environment induces acquisition of the macrophage antigens and phenotype in cancer cells.

The up-regulated genes in macrophages grown as co-cultre with cancer cells were involved mainly in cell-cell adhesion and cell-matrix adhesion.

We found that co-culture of cancer cells and macrophages regulates expression of several important miRNAs.

Taken together, this integrated comparative study generated a scheme of the molecular changes that occur during interactions between macrophage and cancer. Because as far as we realize this is the first analysis of changes in miRNA expression in cancer cells due to a presence of macrophages, our study can serve as a valuable source for future studies on a tumor microenvironment, and some of the highlighted genes, miRNAs, pathways or processes may be useful for diagnostic or therapeutic purposes.

This work was supported by grant no N N308012939 from the Ministry of Sciences and Higher Education.

Citation Format: Magdalena Król, Karol Pawlowski, Kinga Majchrzak, Alicja Majewska, Tomasz Motyl. Comparative mRNA and microRNA expression profiling of canine mammary carcinoma cell lines and macrophages grown as a co-culture in vitro. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A76.
Abstract A94: Imatinib modulates CD4+ T cells in gastrointestinal stromal tumor (GIST).
Authors: Balachandran Vinod P. , Cavnar Michael , Zeng Shan , Bamboat Zubin , Ocuin Lee , Obaid Hebroon , Sorenson Eric , Kim Teresa , Popow Rachel , DeMatteo Ronald P.
Published: August 2014

Introduction: Imatinib mesylate partially exerts its antitumor effects in GIST by inhibiting tumor production of the immunosuppressive enzyme indoleamine 2,3-dioxygenase (Ido), thereby activating antitumor CD8+ T cells. The role of conventional CD4+ T cells (CD4+FoxP3-) in imatinib therapy is unknown. We examined the effects of imatinib on CD4+ T cells in GIST.

Methods: We injected imatinib or saline intraperitoneally for one week to KitV558-/+ mice that develop spontaneous GISTs. 1-methyl-D-tryptophan (D-1MT), an Ido inhibitor was given via oral gavage. We assessed treatment response by tumor weight and immune cells by flow cytometry. Regulatory T cells (Treg) were defined as CD4+FoxP3+. Matched blood and tumor specimens from GIST patients were freshly analyzed for immune composition. Tumors were classified as untreated, sensitive, or resistant to imatinib based on radiologic assessment just before the time of surgery.

Results: Imatinib increased CD4+ number, activation, and degranulation in the draining lymph node (DLN) but not the spleen of GIST mice (p<0.05). In the tumor, imatinib increased CD4+ number, activation, proliferation, and the intratumoral CD4+/Treg ratio (p<0.05). Conversely, Ido inhibition did not alter CD4+ number, activation, proliferation, or the CD4+/Treg ratio in the DLN or tumor. In untreated human GISTs (n=14), CD4+ T cells demonstrated greater activation and memory phenotype compared to autologous blood CD4+ T cells, but secreted Th2 cytokines on in vitro restimulation (p<0.05). Imatinib treatment altered the intratumoral CD4+/Treg ratio, with a higher ratio in sensitive tumors (n=17) compared to resistant tumors (n=12; p<0.05).

Conclusion: In mouse GIST, imatinib activates CD4+ T cells independently of Ido and increases the intratumoral CD4+/Treg ratio, a hallmark of immunologic outcome. Similarly, imatinib sensitive human GISTs have a higher CD4+/Treg ratio than resistant tumors. Hence, CD4+ T cells may contribute to the antitumor effects of imatinib in GIST. Combination immunotherapy with imatinib and CD4+ T cell modulating agents may be a promising therapy for GIST.

Citation Format: Vinod P. Balachandran, Michael Cavnar, Shan Zeng, Zubin Bamboat, Lee Ocuin, Hebroon Obaid, Eric Sorenson, Teresa Kim, Rachel Popow, Ronald P. DeMatteo. Imatinib modulates CD4+ T cells in gastrointestinal stromal tumor (GIST). [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr A94.
Abstract B32: Preclinical testing of three immune adjuvants in vaccination therapy for invasive canine meningioma.
Authors: Andersen Brian M. , Pluhar G Elizabeth , Seiler Charles E. , Xiong Zhengming , Goulart Michelle R. , O'Sullivan Matthew Gerry , Hunt Matthew A. , Schiaffo Charles E. , Ferguson David M. , Ohlfest John R.
Published: August 2014

Malignant and atypical meningiomas are resistant to standard therapies, but therapeutic vaccines have not been tested preclinically or clinically. Pet dogs with naturally occurring meningioma are an underutilized model for aggressive human meningioma and an outstanding opportunity for assessing experimental therapeutic approaches. We treated 11 meningioma-bearing dogs with surgery and vaccine immunotherapy consisting of autologous tumor cell lysate combined with CpG or imiquimod. Interferon gamma-elaborating T cells were detected in the peripheral blood of two of cases, but vaccine-induced tumor-reactive antibody responses were found in the sera of all dogs. Systemic antibody responses were polyclonal, recognizing intracellular and cell surface antigens, and heat shock protein 60 was identified as one common antigen. Tumor-reactive antibodies bound allogeneic canine and human meningiomas, demonstrating common antigens across breed and species. Histological analysis revealed robust infiltration of antibody-secreting plasma cells into the brain around the tumor in treated compared to pre-treatment samples. Tumor-reactive antibodies were capable of inducing antibody dependent cell-mediated cytotoxicity to autologous and allogeneic tumor cells. Moreover, median survival for lysate/CpG and lysate/imiquimod vaccination was 646 days versus 222 days in historic surgery controls (p<0.05). Compared to CpG, a novel imidazoquinoline with TLR7/8 and inflammasome activity, compound 528, was equally potent at activating canine PBMCs. A prospective randomized two-arm trial was recently launched, using lysate—compound 528 vaccines, versus surgery controls. Mean follow-up is 139 days at the time of writing, and recruitment is ongoing. These data demonstrate the feasibility and immunologic efficacy of vaccine immunotherapy for treating a large animal model of human meningioma and warrant further development towards human trials.

Citation Format: Brian M. Andersen, G Elizabeth Pluhar, Charles E. Seiler, Zhengming Xiong, Michelle R. Goulart, Matthew Gerry O'Sullivan, Matthew A. Hunt, Charles E. Schiaffo, David M. Ferguson, John R. Ohlfest. Preclinical testing of three immune adjuvants in vaccination therapy for invasive canine meningioma. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B32.
Abstract B44: Enabling in vivo visualization of cellular therapies in the clinic: Maturation of αDC1 data sets.
Authors: Helfer Brooke M. , Balducci Anthony , Urban Julie , Butterfield Lisa , Ahrens Eric , Kalinski Pawel , Wesa Amy
Published: August 2014

Cellular therapies are increasingly being used in the clinic to modify a patient's immune function. Human dendritic cells (DC) have great clinical promise as an immunotherapeutic intervention against cancer, but questions concerning the dispersion and persistence of these cells, as well as correlations to therapeutic potency remain. Here we utilize a perfluoropolyether (PFPE) magnetic resonance imaging (MRI) tracer agent that aims to address these questions in the assessment of α-type-1-polarized dendritic cells (αDC1). αDC1 cells, producing high levels of interleukin-12p70 with putative strong lymph node homing capabilities, have been evaluated in support of an IND incorporating post-transfer MR imaging of the therapeutic cells. Important therapeutic functions, such as cytokine production, cellular phenotype, and antigen processing are maintained. Transitioning the protocol from the laboratory into clinical production, taking into account GMP procedural changes, resulted in both maintained cellular properties and detectable signal by 3T MRI.

In vivo results demonstrate the capability of MRI to detect and quantify labeled cells with a high degree of specificity. With nominal exogenous fluorine naturally present in tissue, labeled cells appear with little background. Pairing this signal with conventional proton MRI from the same imaging session, cells are able to be traced to their precise anatomical location. The MRI tracking capabilities, safety profiles, and use of 19F labeling in preclinical data sets demonstrate the ability of 19F to be used in clinical applications to strengthen the understanding of cellular therapeutics.

Citation Format: Brooke M. Helfer, Anthony Balducci, Julie Urban, Lisa Butterfield, Eric Ahrens, Pawel Kalinski, Amy Wesa. Enabling in vivo visualization of cellular therapies in the clinic: Maturation of αDC1 data sets. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B44.
Abstract B58: CD40 combined toll-like receptor 3 ligation restore morphine-induced reducing dendritic cells-mediating immunity and antitumor effect.
Authors: Chang Ming-Cheng , Chen Yu-Li , Sun Wei-Zen , Cheng Wen-Fang
Published: August 2014

Morphine has been regarded as a kind of medications with great effective for the management of postoperative pain in surgery and cancer pain with compromising host immunity. However, the mechanisms of how morphine influences the host immunity are still unclear. Our previous study has revealed that morphine could promote effector T cell apoptosis and suppress anti-tumor effect enhanced by antigen-specific DNA vaccine. In current study, we evaluated how morphine interrupted host immunity especially on dendritic cells (DCs) and the possible mechanisms. Morphine could delay the differentiation and maturation of the bone marrow monocyte (BMM)-derived DCs. Furthermore, morphine could inhibit the secretion of IL-6 and TNF-α; but stimulate the excretion of IL-10 from BMM-derived DCs. Besides, morphine could suppress the maturation processes of BMM-DC via the activation of MAPK and down-regulation of P38. When mice administrated with an adjuvant mixture containing the TLR3 ligand-poly (I:C) and agonistic anti-CD40 antibodies, a significant increase DCs property, antigen presenting activity and MAPK/ERK/p38 protection effects were observed. Finally, the TLR3 ligand-poly(I:C) and anti-CD40 Ab vaccinated mice treated with morphine could maintain the anti-tumor effects. Our findings reveal that morphine treatment suppressed DCs maturation and TLR3 ligand-poly(I:C) with anti-CD40 Ab administration may serve as a useful approach to treat the profound immunosuppressive function and prevent tumorigenesis after long-term morphine treatment.

Citation Format: Ming-Cheng Chang, Yu-Li Chen, Wei-Zen Sun, Wen-Fang Cheng. CD40 combined toll-like receptor 3 ligation restore morphine-induced reducing dendritic cells-mediating immunity and antitumor effect. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B58.