bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1179: Combined genetic and genealogic studies uncover a large BAP1 cancer syndrome kindred, tracing back nine generations to a common ancestor from the 1700s
Authors: Carbone Michele , Flores Erin G. , Emi Mitsuru , Gaudino Giovanni , Pastorino Sandra , Yang Haining , Johnson Todd , Tsunoda Tatsuhiko , Hesdorffer Mary , Pass Harvey I.
Published: July 2016

Germline BAP1 mutations cause a cancer syndrome characterized by high incidence of mesothelioma (MM), uveal melanoma and other cancers, and by very high penetrance, as all individuals carrying BAP1 mutations developed at least one, and usually several, malignancies throughout their lives. Through screening MM patients with histories of multiple cancers, we found four supposedly unrelated patients that shared an identical germline BAP1 mutation. We investigated whether this BAP1 mutation occurred in a ‘hot-spot’ for “de novo” mutations or whether these four MM patients shared a common ancestor. Using molecular genomics analyses we found that they are related. By genealogic studies we traced their ancestor to a couple that emigrated from Germany to North America in the early 1700’s; we traced the subsequent migration of their descendants, who are now living in at least three different US States. Our findings demonstrate that BAP1 mutations are transmitted among subsequent generations over the course of centuries. This knowledge and methodology is being used to identify additional branches of the family carrying BAP1 mutations. Our study shows that the application of modern genomic analyses, coupled with “classical” family histories collected by the treating physician, and with genealogical searches, offer a powerful strategy to identify high-risk germline BAP1 mutation carriers that will benefit from genetic counseling and early detection cancer screening.

Citation Format: Michele Carbone, Erin G. Flores, Mitsuru Emi, Giovanni Gaudino, Sandra Pastorino, Haining Yang, Todd Johnson, Tatsuhiko Tsunoda, Mary Hesdorffer, Harvey I. Pass. Combined genetic and genealogic studies uncover a large BAP1 cancer syndrome kindred, tracing back nine generations to a common ancestor from the 1700s. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1179.
Abstract 1219: Sym015, a novel antibody mixture targeting non-overlapping epitopes of MET, effectively inhibits growth of MET dependent tumors and overcomes resistance to a single monoclonal antibody
Authors: Poulsen Thomas T. , Grandal Michael M. , Jacobsen Helle J. , Hansen Dorte S. , Lindsted Trine , Pedersen Mikkel W. , Horak Ivan D. , Kragh Michael , Lantto Johan
Published: July 2016

The tyrosine kinase receptor MET is involved in progression of a variety of human cancers and constitutes a promising therapeutic target. Particularly, subsets of tumors originating from lung or gastric tissues appear to be truly MET dependent. MET dependency is driven by alterations, such as MET-gene amplification, MET-exon 14 deletion, kinase activating mutations, or autocrine HGF production. Furthermore, MET-amplification has been reported as a key mechanism of de novo resistance to EGFR targeting agents in lung and colorectal cancers.

Sym015, a novel antibody mixture comprising two monoclonal antibodies targeting non-overlapping epitopes on the SEMA domain of MET, was shown to effectively inhibit cell growth in vitro through effective MET degradation. In the present study, we screened a large panel of highly annotated human cancer cell lines for sensitivity to Sym015 in order to identify potential markers of response. Sym015 effectively inhibited growth of cell lines with MET-amplification, MET-exon 14 deletion, and autocrine HGF production, including MET-amplified cell lines with acquired resistance to EGFR targeting agents.

To validate the in vitro findings, a range of cell line- and patient-derived xenograft models with MET amplification or Exon 14 deletion were tested for sensitivity to Sym015 and an analogue of the clinical stage anti-MET monoclonal antibody emibetuzumab (LY2875358). Sym015 effectively inhibited growth of tumors with autocrine HGF production, MET-amplification, and/or Exon 14 deletion, and had superior activity compared to the emibetuzumab analogue in many of the models. Importantly, tumors with a partial response to the emibetuzumab analogue were strongly inhibited by subsequent treatment with Sym015 in two MET-amplified models, one of which also harbors a MET-exon 14 deletion.

In summary, our findings demonstrate a potent antitumor effect of Sym015 in MET-dependent models. The data thus strongly support initiation of clinical trials for patients with MET-amplification and Exon 14 deletions.

Citation Format: Thomas T. Poulsen, Michael M. Grandal, Helle J. Jacobsen, Dorte S. Hansen, Trine Lindsted, Mikkel W. Pedersen, Ivan D. Horak, Michael Kragh, Johan Lantto. Sym015, a novel antibody mixture targeting non-overlapping epitopes of MET, effectively inhibits growth of MET dependent tumors and overcomes resistance to a single monoclonal antibody. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1219.
Abstract 1225: NVP-HDM201: cellular and in vivo profile of a novel highly potent and selective PPI inhibitor of p53-Mdm2
Authors: Jeay Sébastien , Chène Patrick , Ferretti Stéphane , Furet Pascal , Gruenenfelder Bjoern , Guagnano Vito , Guerreiro Nelson , Halilovic Ensar , Hofmann Francesco , Kallen Joerg , Léonard Michelle , Mah Robert , Masuya Keiichi , Ramos Rita , Rynn Caroline , Ruetz Stephan , Stachyra-Valat Thérèse , Stutz Stefan , Vaupel Andrea , Wuerthner Jens , Holzer Philipp
Published: July 2016

Stabilization of p53 protein by preventing its interaction with the negative regulator Mdm2 leads to selective induction of the p53 pathway, thus offering a promising cancer therapeutic strategy in p53 wild-type tumors. In the present study, we show the identification of NVP-HDM201, a novel, highly optimized, and selective inhibitor of the p53-Mdm2 interaction. NVP-HDM201 activates p53 in human cells and induces robust p53-dependent cell cycle arrest and apoptosis, selectively in p53 wild-type tumor cells. Its activity and selectivity has been tested and confirmed across a large panel of cancer cell lines from the Cancer Cell Line Encyclopedia. In vivo, NVP-HDM201 shows a dose-proportional pharmacokinetic (PK) profile and a clear PK/PD relationship, resulting in tumor growth inhibition and regression in SJSA-1 tumor-bearing rats at well-tolerated oral (p.o.) doses. The validation and understanding of its mechanism of action, the overall favorable drug-like properties and the characterization of its on-target toxicological profile in preclinical species strongly supported the initiation of Phase I clinical trials with NVP-HDM201 in pre-selected patients with p53 wild-type tumors.

Citation Format: Sébastien Jeay, Patrick Chène, Stéphane Ferretti, Pascal Furet, Bjoern Gruenenfelder, Vito Guagnano, Nelson Guerreiro, Ensar Halilovic, Francesco Hofmann, Joerg Kallen, Michelle Léonard, Robert Mah, Keiichi Masuya, Rita Ramos, Caroline Rynn, Stephan Ruetz, Thérèse Stachyra-Valat, Stefan Stutz, Andrea Vaupel, Jens Wuerthner, Philipp Holzer. NVP-HDM201: cellular and in vivo profile of a novel highly potent and selective PPI inhibitor of p53-Mdm2. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1225.
Abstract 1265: Dysregulated tyrosine kinase Tyro3 signaling in acute myeloid leukemia
Authors: Eryildiz Fatma , Tyner Jeffrey W.
Published: July 2016

The undesirable outcomes of classical chemotherapy approaches in the treatment of cancer pathogenesis has directed scientists to find more targeted and less toxic therapies. Numerous studies of targeted therapies in cancer have demonstrated that finding target genes that are responsible for cancer pathogenesis can lead to novel targeted cancer therapies. Among these studies, tyrosine kinases have been prominent as effective gene targets. We have employed this approach to study the hematologic malignancy, Acute Myeloid Leukemia (AML).

In this study, we aimed to understand how human tyrosine kinase members, as functional drivers of leukemogenesis, promote tumor progression by activating downstream molecules. For this purpose, an RNAi assisted silencing assay was designed in which every individual tyrosine kinase member was silenced and impact on primary AML cell viability was assessed. Bioinformatic interpretation of the data revealed that TYRO3 is one of the genes showing significant sensitivity to silencing in 7.64% of AML cases tested (n = 484). This data is compatible with previous studies showing aberrant expression of TAM family member tyrosine kinases in several human cancers. In addition, immunoblotting was performed to assess TYRO3 protein expression in several AML cell lines. Several AML lines demonstrated higher expression levels of TYRO3. In a strong correlation with this finding, TYRO3 siRNA was able to partially silence the TYRO3 expression especially in those cell lines that have higher expression levels of TYRO3 and due to this silencing we observed decreased cell viability relative to non-specific siRNA suggesting that high expression of TYRO3 provides a cell survival advantage in AML.

Even though signaling pathways of other TAM family members and their interactions are described in detail, TYRO3 downstream signaling pathway still remains quite undetermined which makes it an intriguing player in cell survival. There are not many small molecules designed to inhibit TAM tyrosine kinases, but our study shows that successful targeted inhibition of this family of receptor tyrosine kinases may represent a promising avenue to provide opportunities to improve therapeutic approaches for patients with AML.

[1] Druker, Brian J. “Imatinib as a paradigm of targeted therapies.” Advances in cancer research 91 (2004): 1-30.

Citation Format: Fatma Eryildiz, Jeffrey W. Tyner. Dysregulated tyrosine kinase Tyro3 signaling in acute myeloid leukemia. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1265.
Abstract 1301: Anti-invasive and antimetastatic effects of ω3-polyunsaturated fatty acids through inhibition of NF-kB/matrix metalloproteinases in ovarian cancer cells
Authors: Jeong Soyeon , Jing Kaipeng , Shin Soyeon , Kim Soyeon , Jeon Young-Joo , Heo Jun-Young , Kweon Gi-Ryang , Park Seung-Kiel , Park Jong-Il , Lim Kyu
Published: July 2016

Ovarian cancer is leading cause of gene cological cancer death in the United States. It is reported that about 14,000 patients will die to ovarian cancer in United States, 2015. ω3- and ω6-PUFAs are considered as essential fatty acids because they cannot be synthesized in mammals. The ω3-desaturase (fat-1) converts ω6-PUFAs to ω3-PUFAs. Cancer cells and transgenic mice stably expressing fat-1 geneare useful models to study the anti-cancer effects of ω3-PUFAs. Here, we show anti-invasive and anti-metastatic action of ω3-PUFA in ovarian cancer.DHA significantly inhibited cell invasion and wound healing activity of human ovarian PA-1 cells. DHA also abolishedthe induction of matrix-metalloproteinases (MMPs) and cyclooxygenase (COX2) as well as the transactivity of NF-κB in PA-1 cells. When ID8 cells were subcutaneously implanted into fat-1 transgenic mice, the tumor size and volume were smallin comparison to wild-type mice. The growth inhibition of tumor was also confirmed with the increase in the level of TUNEL-positive cells. Additionally, when ID8 cells were injected via tail vein of fat1 mice, the lung metastasis was remarkably inhibited in fat-1 transgenic mice. Furthermore, the proliferation of fat-1-stably expressing PA-1 (f-PA-1) cells was more attenuated than that of cells expressing control vectors. The invasion and NF-κB/MMPs promoter activities were also decreased in f-PA-1 cells. These results suggest that ω3-PUFAs inhibit invasion and metastasis of ovarian cancer cells through inhibition of NF-κB/MMPs. Therefore, ω3-PUFAs may contribute an effective and safe therapeutic approach for the chemoprevention and treatment of human ovarian cancer. [This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (MEST) (2007-0054932)]

Citation Format: Soyeon Jeong, Kaipeng Jing, Soyeon Shin, Soyeon Kim, Young-Joo Jeon, Jun-Young Heo, Gi-Ryang Kweon, Seung-Kiel Park, Jong-Il Park, Kyu Lim. Anti-invasive and antimetastatic effects of ω3-polyunsaturated fatty acids through inhibition of NF-kB/matrix metalloproteinases in ovarian cancer cells. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1301.
Abstract 1315: Nanoparticles targeting RAF/ERK-driven cell-autonomous resistance to sorafenib for effective treatment of hepatocellular carcinoma
Authors: Chen Yunching , Liu Ya-Chi , Lin Ts-Ting , Ramjiawan Rakesh , Duda Dan
Published: July 2016

Sorafenib is the only systemic therapy approved for advanced hepatocellular carcinoma (HCC). Sorafenib's efficacy has been attributed in part to inhibition of cancer cell proliferation due to B- and CRAF targeting in HCC cells. However, cell autonomous mechanisms promote evasion from sorafenib treatment, leading to moderate survival benefit. Herein, we demonstrated that the effects of sorafenib on HCC cell viability were initially independent of RAF kinase inhibition and were mediated in part by p38MAPK inhibition. Moreover, sorafenib treatment led to RAF heterodimerization (BRAF/CRAF) and subsequent ERK activation and thus converted HCC cell dependence for survival signaling to the RAF/ERK pathway. Inhibition of ERK pathway by the MEK inhibitor AZD6244 significantly enhanced the therapeutic efficacy of sorafenib efficacy on HCC in vitro and in vivo. Furthermore, we developed HCC-targeted nanoparticles (NPs) to efficiently co-deliver both sorafenib and AZD6244 into HCC, downregulate RAF/ERK pathway, suppress angiogenesis and enhance anti-cancer effect in the orthotopic HCC model. In conclusion, sorafenib treatment leads to rapid MAPK activation due to BRAF heterodimerization as a cell-autonomous mechanism of treatment resistance. Co-delivery of sorafenib and MEK inhibitor AZD6244 using HCC targeted NPs downregulates the compensatory activated RAF/ERK pathway and overcomes resistance to sorafenib in HCC.

Citation Format: Yunching Chen, Ya-Chi Liu, Ts-Ting Lin, Rakesh Ramjiawan, Dan Duda. Nanoparticles targeting RAF/ERK-driven cell-autonomous resistance to sorafenib for effective treatment of hepatocellular carcinoma. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1315.
Abstract 1327: Gold nanorods based hypoxia-targeted photothermal cancer therapy
Authors: Xie Huan , Krishnan Sunil
Published: July 2016

PURPOSE: Tumor hypoxia is a well-recognized driver of resistance to traditional cancer therapies such as chemotherapy and radiation therapy. Gold nanorods (GNR), with unique dimension and optical properties, have shown superior capability for tumor photothermal ablation. This study aimed to develop a new nanoconstruct composed of gold nanorods (GNRs) conjugate to anti-carbonic anhydrase IX (CAIX) antibody that specifically binds to CAIX, a biomarker of hypoxia, to facilitate targeting tumor hypoxic areas for focused photothermal ablation. METHODS: GNR were synthesized according to the seed-mediated growth method with modification. Transmission electron microscopy (TEM) were used to characterize the size and shape of the particles. The GNR/anti-CAIX conjugates were prepared through a bifunctional polyethylene glycol (PEG). The conjugates were characterized by UV-Vis spectra, particle sizes and zeta potentials. The binding affinity and specificity of the GNR/anti-CAIX were evaluated with ELISA, cell adhesion assay and cell photothermal treatment. HT29 colon cancer cell line (overexpressing CAIX) and NIH3T3 cell line (no expressing CAIX) were used in our study. Near-infrared (NIR) ablation was performed on HT29 tumors in mice that treated with PBS, GNR-PEG and GNR/anti-CAIX. Biodistribution were conducted on the latter two groups. RESULTS: TEM images showed that the synthesized GNR had an aspect ratio of about 3 (8 × 24 nm). The changes of UV-vis spectra, particle sizes and zeta potentials confirmed the success of the conjugation. ELISA suggested that the number of anti-CAIX per GNR was 4.8 ± 0.3. In the cell binding assay, GNR/anti-CAIX only showed specific and high degree binding to HT29 cells but not to NIH3T3 cells. NIR laser treatment of HT29 cells resulted in significantly higher killing of GNR/anti-CAIX-targeted cells than of controls. NIR ablation of HT29 tumors in mice showed no tumor regression in the sham-treated group, regression but recurrence in the non-targeted-GNR group, and complete tumor regression in the targeted-GNR group. Biodistribution study showed significant higher accumulation in tumor in the targeted-GNR group. CONCLUSIONS: GNR/anti-CAIX nanoconstructs show promise as highly efficient targeting and photothermal ablation agents for cancer treatment.

GRANT SUPPORT: This work was performed under funding from NIH/NIGMS grant (1SC3GM102018-01) and NIH/NIMHD/RCMI grant (5G12RR003045-21).

Citation Format: Huan Xie, Sunil Krishnan. Gold nanorods based hypoxia-targeted photothermal cancer therapy. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1327.
Abstract 1408: Tumor-targeting Salmonella typhimurium A1-R promotes CD8+ T cell infiltration in pancreatic cancer in an orthotopic syngeneic mouse model
Authors: Murakami Takashi , Hiroshima Yukihiko , Zhang Yong , Zhao Ming , Kiyuna Tasuku , Matsuyama Ryusei , Chishima Takashi , Tanaka Kuniya , Bouvet Michael , Endo Itaru , Hoffman Robert M.
Published: July 2016

The effect of tumor-targeting Salmonella typhimurium A1-R on CD8+ cell infiltration was determined on Pan02 murine pancreatic adenocarcinoma cells implanted orthotopically on the pancreatic tail of C57BL/6 mice. Three weeks after orthotopic implantation, mice were randomized into G1: untreated control group (n = 8); and G2: Salmonella typhimurium A1-R treatment group (n = 8, 1 × 107 CFU/body, iv, weekly, 3 weeks). On the 22nd day from initial treatment, all mice were sacrificed and tumors were harvested. The tumor volume ratio was defined as ratio of tumor volume at the 1st day to the 22nd day. H&E staining was performed to evaluate tumor destruction which was classified into 4 categories according to the Evans grading system: Grade I; 0-9% destruction, Grade IIA; 10-50% destruction, Grade IIB; 51-90% destruction, Grade III; few (<10%) viable cancer cells. Immunohistochemical staining using anti-mouse CD8+ antibody was performed to count tumor infiltrating CD8+ cells. The tumor volume ratio was significantly lower in G2 (3.0 ± 2.8) than G1 (39.9 ± 30.7, p < 0.01). Six mice in G1 had peritoneal dissemination whereas no mice showed peritoneal dissemination in G2 (p < 0.01). Significant tumor destruction was observed in G2 (Grade I: 0, Grade IIA: 1, Grade IIB: 6, Grade III: 1) compared to G1 (Grade I: 5, Grade IIA: 3, Grade IIB: 0, Grade III: 0, p < 0.01). Tumor infiltrating CD8+ cells per high-magnification field were significantly higher in G2 (133.5 ± 32.2) than G1 (45.1 ± 19.4, p < 0.001). Strong tumor destruction correlated with higher numbers of tumor-infiltrating CD8+ cells per high-magnification field (Grade I: 35.5 ± 18.3, Grade IIA: 77.3 ± 32.8 Grade IIB and III: 134.7 ± 34.8). The present study demonstrates that Salmonella typhimurium A1-R promotes CD8+ T cell infiltration in a syngeneic pancreatic cancer mouse model.

Citation Format: Takashi Murakami, Yukihiko Hiroshima, Yong Zhang, Ming Zhao, Tasuku Kiyuna, Ryusei Matsuyama, Takashi Chishima, Kuniya Tanaka, Michael Bouvet, Itaru Endo, Robert M. Hoffman. Tumor-targeting Salmonella typhimurium A1-R promotes CD8+ T cell infiltration in pancreatic cancer in an orthotopic syngeneic mouse model. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1408.
Abstract 1414: Treatment outcome under cetuximab-based therapy and individual ADCC capability in head and neck cancer
Authors: Lo Nigro Cristiana , Lattanzio Laura , Vivenza Daniela , Monteverde Martino , Tonissi Federica , Strola Giuliana , Merlano Marco , Milano Gerard
Published: July 2016

Introduction

Cetuximab is a IgG1 monoclonal antibody against epidermal growth factor receptor (EGFR) used for head and neck squamous cell carcinoma (HNSCC) treatment. In addition to EGFR targeting, cetuximab is able to develop an antibody-dependent cell-mediated cytotoxicity (ADCC), triggered by Fc receptors (Fcγ-R) on Natural Killer cells, macrophages and polymorphonucleated leukocytes.

This prospective study examined the possible prognostic value of cetuximab-mediated ADCC response in a cohort of HNSCC patients.

Patients and Methods

There were 63 patients treated with curative intent with chemo-radiotherapy (CT-RT), associated (N = 39) or not (N = 24) with cetuximab. Peripheral blood samples were collected at start of therapy. Intrinsic ADCC was evaluated from ex vivo NK-dependent activity measuring LDH release through the Cytotox 96® non radioactive cytotoxicity assay, previously standardized in our laboratory (Crit Rev Oncol Hematol, 2015). EGFR tumoral expression was analyzed by immunohistochemistry.

Results

Median ADCC response at treatment start for all patients was 56.1% (range 5-99%). Analysing the whole population, patients with ADCC above the median value (high) showed a better OS as compared to patients with ADCC below the median (low), HR = 0.3827 (95% CI, 0.1464 to 1.000), p = 0.05 (Long-rank Mantel-Cox Test). A sub-group analysis confirmed a possible link between high ADCC and a better OS (CT-RT group, p = 0.223 and CT-RT+cetuximab group, p = 0.244).

Importantly, considering CT-RT+cetuximab treatment and EGFR tumoral expression (N = 21), patients with EGFR 3+ status and high ADCC showed an improved OS as compared to patients with ADCC below median value, HR = 0.09031 (95% CI, 0.009983 to 0.8170), p = 0.032 (Long-rank Mantel-Cox Test). Median OS values for patients with high EGFR expression were 34.8 months (range 9.1-53.7) and 13.1 months (range 2.1-39.5) for patients with respectively high ADCC and low ADCC.

Conclusion

ADCC is shown to be a strong driver of cetuximab-based therapy outcome in HNSCC. High tumoral EGFR expression and high ADCC confer a particularly long overall survival. These data suggest an ADCC-based and EGFR expression-based precision therapy under cetuximab in HNSCC and open perspectives for NK boosting.

Citation Format: Cristiana Lo Nigro, Laura Lattanzio, Daniela Vivenza, Martino Monteverde, Federica Tonissi, Giuliana Strola, Marco Merlano, Gerard Milano. Treatment outcome under cetuximab-based therapy and individual ADCC capability in head and neck cancer. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1414.
Abstract 1419: Characterization of the evolution of immune response in lung squamous carcinogenesis
Authors: Mascaux Celine , Angelova Mihaela , Beane Jennifer , Hijazi Kahkeshan , Antoine Geraldine , Willard Gallo Karen , Ninane Vincent , Burny Arsène , Sculier Jean-Paul , Spira Avrum , Galon Jérôme
Published: July 2016

Molecular profiling of from pre-invasive bronchial lesions, and particularly the evolution of immune response, may identify promising new biomarkers for early detection and targets for chemoprevention/treatment of lung cancer.

mRNA expression was analyzed (Agilent microarrays) from fresh frozen human bronchial biopsies (N = 122, 77 patients) at successive morphological stages of lung squamous carcinogenesis. Modules of co-expressed genes were identified among the stage associated genes, selected using a linear mixed-effects model adjusting for smoking status, gender, and history of cancer as fixed effects and patient as a random effect. Cell type specific signatures were applied to the genes co-expressed in these modules in order to identify the immune response signal through the stages of lung carcinogenesis.

Genes expression alterations were grouped into 4 successive molecular steps. Based on their behavior across the 4 steps, 8 modules of genes with different patterns were observed: gene modules primarily up-regulated among the early or late steps, up-regulated in a linearly across the 4 steps, as well as down- then up-regulated (biphasic). Genes signing the presence of activating CD8 and CD4 T cells, as well as some memory B cells were progressively and linearly up-regulated from metaplasia through all stages of carcinogenesis. At the later stages, the transition to high-grade dysplasia, the most significant gene expression changes included immune/inflammatory response genes. At this stage a very strong adaptive immune response is noted involving T cells - mainly Th1 and T regulators - and dendritic cells. In addition, immunosuppressive signals, corresponding to negative co-inhibitory molecules, also significantly appears at the high-grade dysplasia stage.

The adaptive CD4/CD8 related immune response starts from the earlier stages (metaplasia) of lung carcinogenesis. The significant modification of inflammatory/immune response genes in association with high-grade lesions suggests, at this critical stage of carcinogenesis, a major role of the surrounding microenvironment with an adaptive immune response, driven by Th1 and Treg cells, and but also, before tumor invasion across the basal membrane, a tumor versus host immunosuppressive response.

Citation Format: Celine Mascaux, Mihaela Angelova, Jennifer Beane, Kahkeshan Hijazi, Geraldine Antoine, Karen Willard Gallo, Vincent Ninane, Arsène Burny, Jean-Paul Sculier, Avrum Spira, Jérôme Galon. Characterization of the evolution of immune response in lung squamous carcinogenesis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1419.
Abstract 1422: Dietary 2-deoxy-D-glucose (2-DG) reduces TLR-4 induced acute inflammation and prevents chronic inflammation driven carcinogenesis
Authors: Pandey Sanjay , Singh Saurabh , Anang Vandana , Shah Dhananjay K. , Seth Saurabh , Bhatt Anant N. , Manda Kailash , Roy Bal G. , Natarajan K , Dwarakanath Bilikere S.
Published: July 2016

Inflammation is a critical component and contributor of carcinogenesis. Toll like receptors (TLRs) are key regulators of innate immunity and inflammatory signaling. Besides various microbe-associated molecular patterns, TLRs can also bind to endogenously generated damage associated molecular patterns (DAMPS) ligands such as heat shock proteins, surfactant protein A18 and homobility group box-1(HMGB1). While HMGB1, a TLR-4 ligand is over-expressed in several human neoplasms, preclinical studies in mouse tumor models have strengthened the role of TLR-4-HMGB1 axis in promoting carcinogenesis. TLR-4 dependent inflammation and stimulation is accompanied with a switch from respiratory to glycolysis dominated metabolism, which is critical for the differentiation and maturation of myeloid cells thus affecting the outcome of an inflammatory disease. In the present studies we investigated the potential of the glycolytic inhibitor 2-deoxy-D-glucose (2-DG) as an energy restriction mimetic agent for countering the acute and chronic inflammation which can be employed as a cancer preventive strategy. Oral administration of 2-DG in drinking water (0.4% w/v) for 7 days reduced the diethyl nitrosamine (DEN) induced acute inflammation reflected by reduced the serum levels of TNF(∼4 fold) and IL-6(∼7 fold). when administered chronically, 2-DG reduced the incidence (∼50%) and the number of papillomas (∼84%) in DMBA-TPA skin carcinogenesis model, which correlated with the reduced levels of TNF. In an acute inflammation model established using LPS (a TLR-4 ligand), we observed that 2-DG reduced the systemic and macrophage specific inflammatory response in LPS injected mice. 2-DG (1mM) significantly reduced the levels of co-stimulatory surface markers (CD80 and CD86) and associated ROS and NOS effecter functions in mouse macrophagic cell line (RAW 264.7). These studies suggest that dietary administration of 2-DG can be used as a strategy for preventing acute inflammation and associated carcinogenesis.

Citation Format: Sanjay Pandey, Saurabh Singh, Vandana Anang, Dhananjay K. Shah, Saurabh Seth, Anant N. Bhatt, Kailash Manda, Bal G. Roy, K Natarajan, Bilikere S. Dwarakanath. Dietary 2-deoxy-D-glucose (2-DG) reduces TLR-4 induced acute inflammation and prevents chronic inflammation driven carcinogenesis. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1422.
Abstract 1433: A mobile app for health related quality of life and symptom assessment in patients with primary brain tumors in an outpatient oncology clinic
Authors: Rowe Lindsay , Dong Tuo , Armstrong Terri , Mackey Megan , Gilbert Mark , Krauze Andra , Camphausen Kevin
Published: July 2016

Purpose: Primary brain tumors, and their treatments, can have a significant impact on patient quality of life due to altered mentation, mood changes, memory loss, and neurologic deficits. It has therefore been suggested that patient related outcomes need to be included as endpoints for treatment efficacy. The objective of this project was to develop and evaluate a mobile application (GlioNCI), in reporting health-related quality of life measures and symptom scoring in patients with primary brain tumors treated and followed in an outpatient oncology clinic.

Experimental Procedures and Results: Apple's Xcode Integrated Development Environment (IDE) was used to develop GlioNCI using the Swift programming language. The parameters and scores chosen had previously been validated in primary CNS tumors. Medication and steroid use is captured at each encounter, as are seizure characteristics, frequency and management. Health related quality of life questionnaires included portions of the EORTC QLQ-C30, and EORTC QLQ-BN20, and MD Anderson Symptom Inventory Brain Tumor Module, which have been validated in patients with cancer and primary CNS tumors respectively. Patients’ symptoms and effects on function are collected with the Activities of Daily Living scale, and the Instrumental Activities of Daily Living Scale. Mood and mental state were addressed using the Hospital Anxiety and Depression Scale. Separate questionnaires for different encounters and time points in outpatient treatment were included to assess care provider, as well as patient related measures, allowing for flexibility of the app to the needs of both. A mobile app was developed to collect, and archive in a database, input and feedback from both patients and care providers in oncology clinics.

Conclusion: The creation of a patient centered quality of life and symptom assessment mobile app, following patients with primary brain tumors over their course of treatment and in follow-up is feasible. Pilot testing and patient evaluation of the application in an oncology clinic will be used to validate GlioNCI as a tool in outpatient clinics.

Citation Format: Lindsay Rowe, Tuo Dong, Terri Armstrong, Megan Mackey, Mark Gilbert, Andra Krauze, Kevin Camphausen. A mobile app for health related quality of life and symptom assessment in patients with primary brain tumors in an outpatient oncology clinic. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1433.
Abstract 1438: Preserving genital nerve function in prostate cancer: imaging with a molecular imaging agent
Authors: Le Roux Lucia , Ramos Daniela , Flores Leo , Schellingerhout Dawid
Published: July 2016

Purpose: To establish proof-of-concept for the use of a molecular nerve imaging agent in demonstrating nerve tissue at risk during prostate surgery, and thus preventing iatrogenic sexual dysfunction. Erectile Dysfunction (ED) is a common complication of all forms of prostate cancer treatment, with incidence varying from 13-97% depending on the series quoted. There is broad agreement that treatment related damage to the neurovascular bundles is a root cause of the condition, with strong indications from many studies that preservation of the neurovascular bundles avoids post-treatment ED. The goal of this study is to use a novel neural imaging agent based on the non-toxic Tetanus Toxin C-fragment (TTc) to allow the visualization of the anatomy of the genital nerves, thus facilitating nerve-sparing therapies.

Materials and methods: C57 Black male mice were anaesthetized (1-2% isofluorane/oxygen) and injected with 40 μg/10 μl of fluorescently labeled TTc790 in the tunica albuginea of the left corpus cavernosum. Animals were imaged live over a period of 3 hours post injection with a Xenogen IVIS 200 small animal imager. Animals were then euthanized, and the complete male reproductive system was surgically exposed and imaged ex-vivo with a Zeiss AxioZoom16 dissection microscope. The cavernosal nerves, prostatic tissue with intact nerve plexi were dissected and prepared for cryo-sectioning followed by fluorescent immuno-histology using standard methods.

Results: En bloc excision of the prostate with the penis allowed imaging of the genital nerves using the retrograde transported neural marker, TTc790. We noted fluorescent TTc-related signal uptake along the expected course of the nerve tract ipsilateral to the injection site, and were able to clearly demonstrate the pelvic ganglion within the capsule of the prostate. Immuno-histological assessment with post cryo-sectioning confirmed our surgical microscopy findings, including our demonstration of the pelvic ganglion.

Conclusions: We demonstrate the use of TTc790 as a nerve imaging agent for showing the anatomy of the cavernosal nerves and pelvic ganglion complex by intra-operative fluorescent imaging. This suggests that, with further work, image guidance could be provided to physicians and surgeons to spare these nerves during prostate carcinoma treatments, allowing the preservation of genital nerve function and improving quality of life in prostate cancer survivors.

Citation Format: Lucia Le Roux, Daniela Ramos, Leo Flores, Dawid Schellingerhout. Preserving genital nerve function in prostate cancer: imaging with a molecular imaging agent. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1438.
Abstract 1457: Regulatory T cells and its impact on prostate cancer development and clearance
Authors: Kumar Sanjay , Stokes James , Scissum Gunn Karyn , Singh Udai , Ponnazhagan Selvarangan , Mishra Manoj K.
Published: July 2016

Prostate Cancer (PCa) is the most common non-skin malignancy and the most commonly diagnosed cancer in men in United States. The immunotherapeutic role of immune cells in regulation of PCa have been studied but still the specific roles of regulatory T cells needs further investigation. In this study, the role of immune cells, particularly the role of regulatory T cells, in tumor development and progression was analyzed using TRAMP (transgenic adenocarcinoma of mouse prostate) model system. TRAMP cells (TRAMP C1, C2 and C3) were used to induce tumor in C57/B6 mouse. Interestingly, the TRAMP-C3 is non-tumorigenic whileTRAMP-C1 and TRAMP-C2 are tumorigenic. Briefly, tumor induction studies were performed on different groups of mice. Mice were inoculated with these three cell lines, tumors were analyzed at different time points, and the percentage and absolute number of different immune cells (CD4, CD8, NK, NKT, Macrophages, regulatory T cells, and Dendritic cells) were analyzed. Our data demonstrated that the capacity of TRAMP-C1 and TRAMP-C2 cells to form tumors and the inability of TRAMP-C3 cells to induce tumors is mediated by number of regulatory T cells in the tumor microenvironment. Therefore, the data suggest an understanding of function and effect of regulatory T cells during tumor progression and clearance is needed to successfully develop a targeted therapy to modulate the number of immune cells in the tumor microenvironment.

Citation Format: Sanjay Kumar, James Stokes III, Karyn Scissum Gunn, Udai Singh, Selvarangan Ponnazhagan, Manoj K. Mishra. Regulatory T cells and its impact on prostate cancer development and clearance. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1457.
Abstract 1458: Toll-like receptors 1, 2, 4 and 6 expression levels in diffusely infiltrative astrocytomas
Authors: Moretti Isabele F. , Franco Daiane Gil , Silva Roseli , Galatro Thais F. , Oba-Shinjo Sueli M. , Marie. Suely K.N.
Published: July 2016

Toll-like receptors (TLRs) are the first receptors of the immune system that identify disturbances in homeostasis, capable of recognizing molecules associated to cell and tissue damage. TLRs signaling pathway activates inflammatory transcription factors, mainly Nuclear Factor-kappa B (NF-kB) and Interferon Regulatory Factors (IRF3/7). Due to their importance in inflammatory process, TLRs are research hot targets for therapy in several diseases, including cancer. This study aims to analyze TLRs (1, 2, 4 and 6) expressions in human diffusely infiltrative astrocytomas, WHO grades II to IV of malignancy (AGII-AGIV), and correlate their expressions among them and with the clinical outcome of AGIV (glioblastoma, GBM) patients. TLRs mRNA levels were evaluated by qRT-PCR in 143 astrocytoma samples and 22 non-neoplastic (NN) brain specimens from epilepsy surgery. Upregulated mRNA levels of all studied TLRs were observed in astrocytomas compared to NN cases (p<0.05), except for TLR2 in AGII. TLR1 and TLR2 median expression values increased according to astrocytoma malignancy. TLRs mRNA expressions presented positive correlation with significance (p<0.05) in all grades of astrocytoma, except for TLR2 and TLR4 in AGIII. Strong associations (r<0.7) were detected with TLR1-TLR2 mRNA expressions among AGII, AGIII and AGIV, TLR1-TLR4 in AGIII, and TLR6-TLR1 in AGII. In GBM patients, TLR2 upregulation was associated to better outcome with longer overall survival (p<0.007). The same was verified for concomitant upregulation of TLR1, 2 and 4 (p<0.05). Aiming to analyze the distribution of these receptors in different cells present in the tumor, we performed immunofluorescence staining of GBM cell lines (A172 and U87MG). All TLRs were detected in both cell lines, indicating that their expression were not limited to immune cells. The current literature has described controversial roles for the TLRs in cancer, either activating an anti-inflammatory condition leading to tumor progression or a pro-inflammatory state leading to tumor abrogation. TLRs distribution among different cells in the tumor environment might be relevant for the inflammatory process. Our findings of TLRs on tumor cells may suggest a tendency towards an anti-inflammatory state. The comprehension of TLRs role in each cell type, that compose the tumor, may help to determine the signaling pathways involved in diffusely infiltrative astrocytoma progression, and potential new therapeutic strategies.

Citation Format: Isabele F. Moretti, Daiane Gil Franco, Roseli Silva, Thais F. Galatro, Sueli M. Oba-Shinjo, Suely K.N. Marie. Toll-like receptors 1, 2, 4 and 6 expression levels in diffusely infiltrative astrocytomas. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1458.
Abstract 151: Modeling the rise of intratumoral heterogeneity in growing, static, and regressing human colorectal polyps
Authors: Sievers Chelsie K. , Zou Luli , Pickhardt Perry J. , Matkowskyj Kristina A. , Albrecht Dawn , Kim David H. , Moawad Fouad J. , Cash Brooks D. , Reichelderfer Mark , Newton Michael A. , Halberg Richard B.
Published: July 2016

Benign adenomatous colon polyps are thought to be transformed into cancers through the stepwise accumulation of mutations. However, not all polyps progress. A significant number remain static in size, regress, or resolve completely. The mechanisms underlying these differential fates are unknown, and currently there are no biological characteristics that can reliably predict which polyps will grow or progress into invasive cancer. To determine the mutational landscape, targeted next generation sequencing was performed on a unique collection of small (6-9mm) colorectal polyps with known growth rates based on interval imaging with CT colonography. To determine spatial location of identified mutations within a polyp, micro-dissection was performed followed by quantitative PCR to validate low frequency mutations. The mutational landscape of small polyps is varied both within and among individual polyps. Polyps carried 0-3 pathogenic mutations with the most frequent being in APC (67%, 32/48), KRAS/NRAS (17%, 8/48), BRAF (17%, 8/48), FBXW7 (10%, 5/48), and TP53 (8%, 4/48). Additionally, 13% (6/48) contained driver mutations at varied mutant allele frequencies, indicating the presence of subclonal populations. In silico modeling of tumor growth was used to determine the likely size at which additional driver mutations arose in order to observe those varied frequencies. This model of colon tumor growth was adapted so that mutations occur with a given probability of 10−5, which may change the fitness positively or negatively, and the lineage from these mutant subpopulations was tracked during tumor growth. In silico polyps were sectioned and mutant allele frequency was recorded and compared to the frequencies observed from the targeted sequencing of human polyps. Contrary to the slow step-wise accumulation of mutations theory, these data indicate small colonic polyps can have multiple pathogenic mutations in crucial driver genes that arise early in a tumor's existence. Understanding the molecular pathway of tumorigenesis and clonal evolution in polyps that are at risk for progressing to invasive cancers will allow us to begin to better predict which polyps may be more likely to progress into adenocarcinomas and which patients are predisposed to developing advanced disease.

Citation Format: Chelsie K. Sievers, Luli Zou, Perry J. Pickhardt, Kristina A. Matkowskyj, Dawn Albrecht, David H. Kim, Fouad J. Moawad, Brooks D. Cash, Mark Reichelderfer, Michael A. Newton, Richard B. Halberg. Modeling the rise of intratumoral heterogeneity in growing, static, and regressing human colorectal polyps. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 151.
Abstract 1527: Urinary miRNAs as predictors of tumor metastasis in localized patients with clear cell renal cell carcinoma: a pilot study
Authors: Shu Xiang , Hildebrandt Michelle AT , Borges dos Reis Rodolfo , Ye Yuanqing , Gu Jian , Wood Christopher G. , Wu Xifeng
Published: July 2016

Introduction: 25-50% of localized renal cell carcinoma (RCC) patients will eventually develop metastasis which has a dramatic adverse effect on prognosis following treatment. Unfortunately, reliable biomarkers predicting RCC metastasis are still lacking. MicroRNAs (miRNAs) are important regulators for vital biological processes in humans. Urine miRNAs(UmiRNAs) present promising potential as biomarkers for diagnosis of urological cancers. However, no study has assessed the predictive value of UmiRNAs for RCC metastasis.

Materials and Methods: We prospectively collected pre- and post-surgery urine samples from 117 patients with localized clear cell RCC (ccRCC,T1-3 N0/x M0). UmiRNAs were extracted and purified from 250 μl urine supernatants and measured using RT-PCR. Relative quantifications (RQ) of 137 selected miRNAs were calculated by 2−ΔΔct method. The ratio of post/pre RQs were tested for the association with distant metastasis using multivariable Cox regression models.

Results: More than half of the patients were diagnosed at pT3 (53%) or grade III-IV (69%). Twenty-two patients (18.8%) had metastasis during a mean follow-up of 73 months. The levels of 34 UmiRNAs were significantly changed after nephrectomy (p<0.05, 29 of 34 significantly reduced). The 50% or more reduced post-nephrectomy levels of five UmiRNAs (miR-191-5p, miR-324-3p, miR-186-5p, miR-93-5p, and miR-30b-5p) independently conferred a 2- to 4-fold increased risk of metastasis with miR-191-5p being the most significant one (hazard ratio [HR] = 4.16, 95% confidence interval [CI] = 1.38-12.58, p = 0.011). Stratified analyses by stage and Fuhrman grade revealed that the associations were most profound in pT3 patients for miR-191-5p, miR-324-3p, and miR-186-5p. Ingenuity pathway analysis showed that the predicted target genes of these five UmiRNAs were enriched in cell survival and proliferation networks.

Conclusion: Our study suggests that urinary miR-191-5p, miR-324-3p, and miR-186-5p are potential biomarkers for the prediction of metastasis in localized, particularly pT3 ccRCC patients after nephrectomy. These findings may help improve individualized patient management in the clinic.

Citation Format: Xiang Shu, Michelle AT Hildebrandt, Rodolfo Borges dos Reis, Yuanqing Ye, Jian Gu, Christopher G. Wood, Xifeng Wu. Urinary miRNAs as predictors of tumor metastasis in localized patients with clear cell renal cell carcinoma: a pilot study. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 1527.