tbuckley/cabot-search huggingface dataset:
Abstract 659: Multiplex cocktails for immunotherapy targets: PD-L1 with tumor specific transcription factors
Authors: David Altree-Tacha, Wei Yuan, George Yang
Journal: Cancer Research (2017)

Abstract Introduction Blocking the interaction between the programmed cell death (PD)-1 protein and one of its ligands, PD-L1, has been reported to generate impressive antitumor responses. Therapeutics targeting PD-L1 in several cancers are currently in clinical trials, and the U.S. FDA has granted accelerated approval for Keytruda (pembrolizumab) to treat patients with advanced (metastatic) non-small cell lung cancer (NSCLC). Various pathology scoring and interpretation methods have been put forth to assay PD-L1 expression, and variability of non-PD-L1 staining across tumors may have important prognostic aspects. PD-L1 expression on tumor cells can be upregulated via activation of CD8+ cytotoxic T lymphocytes. PD-L1 expression is also associated with certain subtypes of tumor-associated lymphocytes, macrophages and dendritic cells. In certain cancers, the cross reactivity of PD-L1 can make interpretation and scoring difficult. Thus, a strategy using IHC multiplex stains could help resolve challenging cases by cocktailing PD-L1 with target tissue transcription factor antibodies along with other potential markers. Method and Materials Formalin-fixed paraffin-embedded tissues and tissue microarrays for various tumor types including lung, bladder and melanoma were processed and cut at 4-5 microns. PD-L1 rabbit monoclonal antibody was cocktailed with the following antibodies: TTF-1 (lung cancer); CD163 (lung cancer); p40 + GATA3 (bladder cancer); and SOX10 (melanoma). Cocktails were detected with double stain detection systems using brown, red and blue chromogens for visualization. Sections were counterstained with a standard hematoxylin or Wiegert’s iron hematoxylin. Results PD-L1 and TTF-1 with or without CD163 (macrophage) and PD-L1 + CD163 were used in lung cancers. The nuclear staining of TTF-1 in lung adenocarcinoma (blue chromogen) helped define tumor positive PD-L1 (DAB) positive cells and CD163 (Fast Red) marked macrophages. PD-L1 combined with p40 and GATA3 was deployed for bladder cancers. PD-L1 membranous expression in tumor cells and co-expression of p40 and GATA3 gave robust nuclear staining in bladder tumor cells. Finally, SOX10 nuclear staining was observed in most melanoma cells and could be easily separated from PD-L1 membrane staining. Conclusion PD-L1 cocktailed with strategic nuclear or cytoplasmic antibodies can help discriminate tumor cells from non-tumor cells, and may facilitate quantitation or scoring methods for more accurate assessment of this key immunotherapy marker. Citation Format: David Altree-Tacha, Wei Yuan, George Yang. Multiplex cocktails for immunotherapy targets: PD-L1 with tumor specific transcription factors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 659. doi:10.1158/1538-7445.AM2017-659
Abstract 4140: Identification of a selective MKLP2/KIF20A inhibitor with highin-vivoantitumor activity
Authors: Yves Collette, Stephanie Miserey-Lenkei, Catherine Guillou, Denis Carniato, Bernard Pau, Bruno Goud, Norbert Vey, Cecile E. Bougeret
Journal: Cancer Research (2017)

Abstract Mitotic kinesins are essential regulators of cancer cell replication and migration. The mitotic kinesin MKLP2/KIF20A, a member of the kinesin-6 family, plays an essential role during cytokinesis and was identified as a potential new target for cancer chemotherapy1. We have previously identified Paprotrain, a new synthetic compound, as the first selective MKLP2 inhibitor2. Recently, we obtained Paprotrain analogues with higher potency on MKLP23. Herein we describe the identification and characterization of BKS0349, a new potent analogue of Paprotrain. BKS0349 compound is 10 times more potent than paprotain on MKLP2 inhibition and has shown an even more restricted specificity profile when tested on a large set of kinases. In-vitro this compound is highly cytotoxic on a wide panel of human cancer cell lines (IC50 ranged 10-70 nM, which corresponds to 1000 fold improvement of paprotrain potency) while no toxicity is observed on human normal cells such as peripheral blood mononuclear cells (PBMC) and primary hepatocytes (IC50 &amp;gt;50µM). BKS0349 compound is well tolerated in mice using repeated intravenous administrations (200 mg/kg, twice a week for 4 weeks). In xenografted nude mice, in-vivo treatment with BKS0349 compound demonstrates a high antitumor activity against various human cancer cell models, either sensitive or resistant to some standard-of-care treatments. In addition, human cancer cells treated in-vitro as well as in-vivo with BSK0349 compound display Golgi scattering and a mitotic arrest leading to cell death, as hallmarks of BKS0349’ mode of action. These findings show that MKLP2 is a potential new target for cancer chemotherapy and BKS0349 a good candidate to be developed for cancer treatment. 1Paclitaxel targets FOXM1 to regulate KIF20A in mitotic catastrophe and breast cancer paclitaxel resistance. P. Khongkow et al. Oncogene, 2015, 1-13. 2Relocation of Aurora B and Survivin from centromeres to the central spindle impaired by a kinesin-specific MKLP-2 inhibitor. S. Tcherniuk et al. Angew. Chem. Int. Ed., 2010, 49:8228-8231. 3New MKLP-2 inhibitors in the paprotrain series: design, synthesis and biological evaluations. C. Labrière et al. Bioorg and Med Chem, 2016, 24:731-734. Note: This abstract was not presented at the meeting. Citation Format: Yves Collette, Stephanie Miserey-Lenkei, Catherine Guillou, Denis Carniato, Bernard Pau, Bruno Goud, Norbert Vey, Cecile E. Bougeret. Identification of a selective MKLP2/KIF20A inhibitor with high in-vivo antitumor activity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4140. doi:10.1158/1538-7445.AM2017-4140
Abstract 680: Prognostic impact of OPN and DKK1 in patient of hepatocellular carcinoma after hepatectomy
Authors: YunSung Seo, Hye Rim Byeon, In Hae Park, Seung Duk Lee
Journal: Cancer Research (2017)

Abstract The new biomarkers are essential for improving the survival and prognosis of hepatocellular carcinoma(HCC) patients. Alpha-fetoprotein(AFP) is the most widely used biomarker. But the low sensitivity and specificity limits its clinical application. Recent study validated the diagnostic capability of osteopontin(OPN) and dickkopf-1(DKK1) and assessed the combination of AFP, DKK1, and OPN as a panel for the diagnosis of HCC. Based on these previous studies, we hypothesized that combination of OPN and DKK1 can be used to as a marker for prognosis to patient of HCC after hepatectomy. From January 2006 to December 2008, patients undergoing hepatectomy for hepatocellular carcinoma were screened serum that has been stored in the Bank of tumor. To target the selected patient, it was confirmed that the remaining tissue specimen is stored after diagnosis. Serum of the patients was used to investigate the OPN and DKK1 by ELISA. In the paraffin block were prepared unstained slide and OPN and DKK1 level checked by IHC. It examined the correlation between prognosis and biomarkers through statistical analysis. AFP, OPN(serum level) and DKK1(serum level) are an independent prognostic factor for overall survival(OS) in HCC after hepatectomy(n=60, P=0.0204, 0.0167 and 0.0455 respectively). New biomarkers combinations based on the AFP existing biomarker are showed a falling curve of the overall survival(OS) and disease-free survival(DFS) in Kaplan Meier curve. In conclusion, combination of OPN, DKK1 and AFP as a biomarker could support the correct diagnosis for HCC after hepatectomy. Note: This abstract was not presented at the meeting. Citation Format: YunSung Seo, Hye Rim Byeon, In Hae Park, Seung Duk Lee. Prognostic impact of OPN and DKK1 in patient of hepatocellular carcinoma after hepatectomy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 680. doi:10.1158/1538-7445.AM2017-680
Breath Measurements as Volatile Organic Compound Biomarkers
Authors: Lance Wallace, Timothy Buckley, Edo Pellizzari, Sydney Gordon
Journal: Environmental Health Perspectives (1996)

A brief review of the uses of breath analysis in studies of environmental exposure to volatile organic compounds (VOCs) is provided.The U.S. Environmental Protection Agency's large-scale Total Exposure Assessment Methodology Studies have measured concentrations of 32 target VOCs in the exhaled breath of about 800 residents of various U.S. cities.Since the previous 12-hr integrated personal air exposures to the same chemicals were also measured, the relation between exposure and body burden is illuminated.Another major use of the breath measurements has been to detect unmeasured pathways of exposure; the major impact of active smoking on exposure to benzene and styrene was detected in this way.Following the earlier field studies, a series of chamber studies have provided estimates of several important physiological parameters.Among these are the fraction, f, of the inhaled chemical that is exhaled under steady-state conditions and the residence times, t, in several body compartments, which may be associated with the blood (or liver), organs, muscle, and fat.Most of the targeted VOCs appear to have similar residence times of a few minutes, 30 min, several hours, and several days in the respective tissue groups.Knowledge of these parameters can be helpful in estimating body burden from exposure or vice versa and in planning environmental studies, particularly in setting times to monitor breath in studies of the variation with time of body burden.Improvements in breath methods have made it possible to study short-term peak exposure situations such as filling a gas tank or taking a shower in contaminated water.
Abstract 570: Transparent tumor tomography (T3): 3D spatial immunoanalysis for PD-L1 immune checkpoint blockade therapy
Authors: Steve Seung-Young Lee, Vytautas P. Bindokas, Stephen J. Kron
Journal: Cancer Research (2017)

Abstract Untangling the complexity of programmed death-ligand 1 (PD-L1) expression within a heterogeneous tumor microenvironment is an urgent challenge in PD-1/PD-L1 immune checkpoint blockade therapy. Here, we address this challenge with a method, termed transparent tumor tomography (T3), facilitating three-dimensional (3D) visualization and spatial analysis of distributions of multiple biomarkers regarding to cancer cells, vasculature, and immune cells in context in the tumor microenvironment. With T3 analysis of transgenic mouse mammary tumors immunostained against Her2, CD45, Ki-67, CD31, and PD-L1, we reveal that PD-L1 expression within the tumor microenvironment is highly adaptable for efficiently preventing immune cell infiltration into the tumor. Stronger correlation of Her2 and PD-L1 expression in the tumor periphery where has a high CD45+ immune infiltrate density is determined by tumor-wide analysis. Also, tomographic analysis shows blood vessels expressing PD-L1 in the tumor core, where PD-L1 expression is lower. Furthermore, high-resolution T3 image localizes PD-L1 expression to a region between the endothelium and the surrounding smooth muscle cells in blood vessels. We investigate spatial pharmacokinetics of anti-PD-L1 antibody in the whole mouse mammary tumor in the context of hypoxia, CD31+ blood vessels, and target PD-L1+ cells. The 3D anti-PD-L1 antibody distribution is fit to a two-compartment pharmacokinetic model, yielding estimated distribution half-life of 4.7 min and terminal half-life of 2.5 days. We also evaluate anti-tumor immune responses after PD-L1 blockade therapy using T3. We observe broad distribution of tumor infiltrating CD3+CD8+ cytotoxic T cells in 3D tumor section following combination therapy of radiation and anti-PD-L1 antibody compared to PBS, anti-PD-L1 antibody alone, or radiation alone treatment group. Moreover, we apply T3 for immunoanalysis of whole core needle biopsy. We spatially map PD-L1 expression and granzymeB-producing CD3+CD8+ cytotoxic T cells in pre-treatment and in-treatment core needle biopsies at cellular resolution and in three dimensions. Meanwhile, T3 analysis is nondestructive, allowing secondary analysis by IHC and/or IF. We anticipate that T3 can be applied broadly to facilitate preclinical studies of immunotherapy and also find use in spatial, multiparameter analysis of patient biopsies, particularly to improve predictive testing and analysis of immune responses to tumor immunotherapy. Citation Format: Steve Seung-Young Lee, Vytautas P. Bindokas, Stephen J. Kron. Transparent tumor tomography (T3): 3D spatial immunoanalysis for PD-L1 immune checkpoint blockade therapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 570. doi:10.1158/1538-7445.AM2017-570
Abstract 1019: <i>In vivo</i> CRISPR screening identifies Ptpn2 as a target for cancer immunotherapy
Authors: Robert T. Manguso, Hans W. Pope, Margaret D. Zimmer, Flavian D. Brown, Kathleen B. Yates, Brian C. Miller, Natalie B. Collins, Kevin Bi, Martin W. Lafleur, Vikram R. Juneja, Sarah A. Weiss, David E. Fisher, David E. Root, Arlene H. Sharpe, John G. Doench, W Nicholas Haining
Journal: Cancer Research (2017)

Abstract Despite the dramatic clinical success of cancer immunotherapy with PD-1 checkpoint blockade, most patients don’t experience sustained clinical benefit, suggesting that additional therapeutic strategies are needed. Functional genomic screens in cancer cells to discover new therapeutic targets are usually carried out in vitro where interaction with the immune system is absent. Here we report a pooled, loss-of-function genetic screening approach using CRISPR/Cas9 genome editing that is conducted in vivo in mouse transplantable tumors treated with vaccination and PD-1 checkpoint blockade. We tested 2,400 genes expressed by melanoma cells for those that synergize with or cause resistance to checkpoint blockade, and recovered the known immune evasion molecules, PD-L1 and CD47. Loss of function of multiple genes required to sense interferon-y caused resistance to immunotherapy. Deletion of Ptpn2, a pleotropic protein tyrosine phosphatase improved response to immunotherapy. In vivo, Ptpn2 deficient tumors showed increased infiltration of activated CD8+T cells. In vitro, Ptpn2 loss by tumor cells increased antigen presentation to T cells. Biochemical, transcriptional and genetic epistasis experiments demonstrated that loss of function of Ptpn2 sensitizes tumors to immunotherapy by enhancing interferon-y-mediated effects on the tumor cell. Thus, augmenting interferon-y signaling in tumor cells could increase the efficacy of immunotherapy. More generally, in vivo genetic screens in tumor models can identify new immunotherapy targets and rationally prioritize combination therapies. Citation Format: Robert T. Manguso, Hans W. Pope, Margaret D. Zimmer, Flavian D. Brown, Kathleen B. Yates, Brian C. Miller, Natalie B. Collins, Kevin Bi, Martin W. Lafleur, Vikram R. Juneja, Sarah A. Weiss, David E. Fisher, David E. Root, Arlene H. Sharpe, John G. Doench, W Nicholas Haining. In vivo CRISPR screening identifies Ptpn2 as a target for cancer immunotherapy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1019. doi:10.1158/1538-7445.AM2017-1019
Long-Term Cold Adaptation Does Not Require FGF21 or UCP1
Authors: Susanne Keipert, Maria Kutschke, Mario Ost, Thomas Schwarzmayr, Evert M. van Schothorst, Daniel Lamp, Laura Brachthäuser, Isabel Hamp, Sithandiwe E. Mazibuko, Sonja Hartwig, Stefan Lehr, Elisabeth Graf, Oliver Plettenburg, Frauke Neff, Matthias H. Tschöp, Martin Jastroch
Journal: Cell Metabolism (2017)
Abstract 1993: Acidic tumor microenvironment stimulation of GPR4 alters cytoskeletal dynamics and migration of vascular endothelial cells
Authors: Elizabeth A. Krewson, Li V. Yang, Lixue Dong
Journal: Cancer Research (2017)

Abstract The acidic tumor microenvironment results from aberrant vasculature, insufficient oxygen delivery and aerobic glycolysis of cancer cells. As cancer progresses, the acidic microenvironment can lead to structural changes of endothelial cells (ECs) resulting in increased vascular permeability which may increase metastasis. Extracellular acidification can activate cellular signaling molecules, including G-protein coupled receptor 4 (GPR4), which is prominently expressed in ECs. We have previously reported that acidic activation of GPR4 induces the transcription of several pro-inflammatory and ER-stress related genes in ECs. The purpose of the study is to identify the functional response of acidosis-induced GPR4 activity using Human Umbilical Vein Endothelial Cells (HUVECs) as a model system. The expression of GPR4 was overexpressed (HUVEC/GPR4) or knocked down (GPR4 shRNA) using several genetic constructs. HUVECs were treated with media buffered to pH 6.4 or to physiological pH 7.4. Permeability of a HUVEC cell monolayer was assessed by quantifying gap formation as one indicator. Cell migration was assessed through wound-healing assays. Cytoskeletal dynamics were also investigated using Rhodamine Phalloidin to investigate actin stress fiber formation and immunocytochemistry to investigate focal adhesion dynamics. Our results showed acidic pH induced GPR4 activation changed the cytoskeletal phenotype, decreased cellular migration, and proliferation in HUVECs compared to the physiological pH treatment. There was also a further increase in gap area of a HUVEC/GPR4 monolayer and formation of actin stress fibers in response to acidic conditions. Additionally, altering expression of phosphorylated paxillin and focal adhesion kinase were observed under acidosis/GPR4 mediated stress. In conclusion, the proton sensor GPR4 induces cytoskeletal variations, reduces cellular migration, and proliferation in response to an acidic tumor microenvironment. Citation Format: Elizabeth A. Krewson, Li V. Yang, Lixue Dong. Acidic tumor microenvironment stimulation of GPR4 alters cytoskeletal dynamics and migration of vascular endothelial cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1993. doi:10.1158/1538-7445.AM2017-1993
Abstract 5800: Epidermal growth factor-induced ANGPTL4 enhances anoikis resistance and tumor metastasis in head and neck squamous cell carcinoma
Authors: Yu-Han Liaong
Journal: Cancer Research (2017)

Abstract Epidermal growth factor (EGF) is important for cancer cell proliferation, angiogenesis and metastasis in many types of cancer. However, the mechanisms involved in EGF-induced head and neck squamous cell carcinoma (HNSCC) metastasis remain largely unknown. In this study, we reveal that angiopoietin-like 4 (ANGPTL4) plays an important role in the regulation of EGF-induced cancer metastasis. We showed that EGF-induced ANGPTL4 expression promoted anoikis resistance and cancer cell migration and invasion in head and neck squamous cell carcinoma (HNSCC). In addition, depletion of ANGPTL4 inhibited EGF-induced cancer cell invasion. Autocrine production of EGF-induced ANGPTL4 regulated the expression of MMPs. The induction of MMP-1 gene expression by ANGPTL4-activated integrin β1 signaling occurred through the AP-1 binding site in the MMP-1 gene promoter. Furthermore, down-regulation of MMP-1 impeded EGF- and recombinant ANGPTL4-enhanced HNSCC cell migration and invasion. Depletion of ANGPTL4 significantly blocked EGF-primed anoikis resistance and extravasation. Moreover, depletion of ANGPTL4 significantly blocked EGF-primed metastatic seeding of tumor cells and MMP-1 expression in lungs. However, no effect of ANGPTL4 on tumor growth was observed. These results suggest that EGF-induced expression and autocrine production of ANGPTL4 enhances HNSCC metastasis via the up-regulation of MMP-1 expression. Inhibition of ANGPTL4 expression may be a potential strategy for the treatment of EGFR-mediated HNSCC metastasis. Citation Format: Yu-Han Liaong. Epidermal growth factor-induced ANGPTL4 enhances anoikis resistance and tumor metastasis in head and neck squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5800. doi:10.1158/1538-7445.AM2017-5800
Abstract 3424: Role of miR-489 in mammary gland development and Her2 mediated tumorigenesis
Authors: Yogin Patel, Mithil Soni, Shou Liu, Hexin Chen
Journal: Cancer Research (2017)

Abstract HER2 overexpression is linked with poor prognosis and outcome in breast cancer. In our previous study, we have found miR-489 was specifically down regulated by HER2 overexpression. Restoration of miR-489 in multiple breast cancer cell lines significantly inhibited cell growth in vitro and decreased tumor growth in xenograft mice. To study role of miR-489 in Her2 mediated tumorigenesis, for the first time we generated MMTV-miR-489 transgenic mice, which overexpress miR-489 specifically in mammary gland. Our qRT-PCR data has confirmed transgenic mice have significantly more miR-489 expression than FVB mice. We used western blot to further validate our model system and found significant downregulation of miR-489 targets DEK and PTPN11. To find out whether miR-489 has any role in mammary gland development, mammary gland whole mount was performed from wild type FVB mice and MMTV-miR-489 mice at different age. Mammary gland from MMTV-miR-489 mice demonstrated reduction in growth at early age. Our immunohistochemistry staining demonstrated significantly reduction in Ki-67 positive cells in MMTV-miR-489 mammary gland at early age, further confirming reduced growth in MMTV-miR-489 mice. However, we found no significant effect on weight of litters of MMTV-miR-489 female since after 8-week mammary gland was able to recover growth. To find out effect of miR-489 overexpression on Her2 mediated tumorigenesis, we generated double transgenic mice MMTV-Her2/miR-489 by crossing MMTV-miR-489 mice with MMTV-Her2 mice. We have observed significant delay in tumor onset and reduced tumor growth in MMTV-Her2/ miR-489 mice compare to MMTV-Her2 mice. We have also observed less number of metastatic site in lung by performing H and E staining of lung. Our IHC data showed reduction in PTPN11 and DEK in miR-489 overexpress mammary tumor. Surprisingly, we found significant reduction of miR-489 expression in mammary tumors of MMTV-Her2-miR-489 when compared with normal mammary gland of same mice. Overall, our results indicated miR-489 overexpression suppresses mammary gland development at early age, reduced mammary tumorigenesis and decrease lung metastasis by targeting PTPN11 and DEK. Citation Format: Yogin Patel, Mithil Soni, Shou Liu, Hexin Chen. Role of miR-489 in mammary gland development and Her2 mediated tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3424. doi:10.1158/1538-7445.AM2017-3424