bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 1049A: Deregulation of MITF in the context of inactive MC1R leads to melanocyte transformation
Authors: Alver Tine Norman , Lavelle Timothy J. , Heintz Karen-Marie , Wernhoff Patrik , Nygaard Vegard , Øy Geir F. , Bøe Sigurd L. , Urbanucci Alfonso , Hovig Eivind J.
Published: August 2019

MC1R, the cAMP pathway and the response to solar UV are all elements involved in the growth, differentiation and survival of melanocytes and in malignant melanoma. Here we present a model of melanomagenesis based on the forced expression of MITF-M in the context of MC1R-inactivating variants in a way not dependent on sun exposure. We demonstrate that lentiviral transduction of MITF-M alone leads to consistent transformation of the Hermes4C cell line with inactive MC1R, but not the Hermes3C cell line having a wild type background. Hermes 4c cells transduced with MITF-M are able to form tumors in mice. We mapped MITF binding to chromatin and found that 4C cells displayed enhanced MITF chromatin binding compared to 3C. Expression analysis revealed that the altered binding pattern leads to enhanced transcription of epithelial to mesenchymal transition genes and consequent repression of key melanocyte-specific genes e.g. involved in pigmentation in the background of inactive MC1R. We observed a marked deregulation of AXL and EGFR, which is accompanied by downregulation of PTEN and increased phosphorylation of ERK and PI3K-AKT pathway. In conclusion, we show that MITF-M is a molecular switch in the context of MC1R-inactivating variant which leads to cell reprogramming and melanomagenesis.

Citation Format: Tine Norman Alver, Timothy J. Lavelle, Karen-Marie Heintz, Patrik Wernhoff, Vegard Nygaard, Geir F. Øy, Sigurd L. Bøe, Alfonso Urbanucci, Eivind J. Hovig. Deregulation of MITF in the context of inactive MC1R leads to melanocyte transformation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1049A.
Abstract 1061: Animal models of prostate cancer
Authors: Ponthan Frida , Brown Matthew , Playle Emma , Booth Catherine
Published: August 2019

Background: Prostate cancer is the most common male malignancy and the second most common cause of cancer-related deaths in the United Kingdom and the United States. It’s a heterogeneous disease caused by different genetic mutations where the majority of prostate cancers are indolent and slow growing. However, some tumours are aggressive with the ability to invade local structures and metastasise leading to poor clinical outcome. One of the most important aspects in anti-cancer drug development is the availability of robust preclinical models which allow the efficacy of novel therapies to be examined. We have developed a reproducible, well growing and robust subcutaneous model of LNCaP with an effective response to both conventional standard of care (SOC) chemotherapy and androgen receptor inhibitors.

Materials and Methods: Cells were subcutaneously implanted on the flank of NSG mice and treated with an anti-androgen with or without conventional chemotherapy starting after 15-20 days when the mean tumour volume was 150-200mm3. Animal weight and tumour volume was measured 3x weekly. Androgen receptor (AR) expression was analysed with immunohistochemistry and gene expression analysis before and after treatment.

Results: The LNCaP cell line grew well in vivo with 100% take rate. Vehicle treated tumours doubled in size over the first 6 days of treatment and were 4.5x the mean starting volume by day 17 post-treatment start, when the group was removed from the study. Significant tumour growth inhibition was observed in response to both conventional and anti-androgen receptor treatment consistent with the maintained AR expression confirmed by gene expression analysis.

Conclusion: This high take rate responsive model represents a significant improvement in subcutaneous xenograft models of prostate cancer, and allows the comparison of AR responsive therapies, alone or in combination with conventional SOC or irradiation. Current protocols employing luciferase transfected cells has also enabled evaluation of metastatic disease.

Citation Format: Frida Ponthan, Matthew Brown, Emma Playle, Catherine Booth. Animal models of prostate cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1061.
Abstract 1064: PDX Like Me: A molecular profile-based search tool
Authors: Begley Dale A. , Krupke Debra M. , Neuhauser Steven B. , Richardson Joel E. , Sundberg John P. , Bult Carol J.
Published: August 2019

Patient Derived Xenografts (PDXs) are generated through the engraftment of human tumor tissue into a specialized (usually, immunodeficient) mouse host strain. PDXs are a useful pre-clinical platform for evaluating the efficacy of single agent and combination therapies that are targeted to specific genomic characteristics of a patient’s tumor. We have implemented the PDX Like Me search tool to assist researchers in identifying PDX models whose tumors match - or are similar to - molecular profiles comprised of one or more genes and one or more genomic characteristics (e.g., copy number status, mutation, and expression). The PDX Like Me interface uses a search syntax similar to the Onco Query Language used by the popular cBioPortal resource.

PDX Like Me is one of several search interfaces supported by the Mouse Models of Human Cancer database (formerly, the Mouse Tumor Biology database). MMHCdb (http://tumor.informatics.jax.org) is a comprehensive resource of information on both genetically engineered mouse models (GEMMs) and PDX models of human cancer that has been expertly curated from peer-reviewed scientific publications and direct data submissions from individual investigators and large-scale programs. MMHCdb provides an easy-to-use search interface as well as tools for visualizing associated data from these models. Information in the database is standardized using controlled vocabularies and official gene and mouse strain nomenclature. MMHCdb contains data from spontaneous or endogenously induced tumors from genetically defined mice. MMHCdb holds data on over 7,500 different strains including over 93,000 tumor frequencies and over 2,250 pathology reports with over 6,300 images from over 4,600 references. MMHCdb provides access to clinical, pathology, dosing study results, and genomics data from over 450 PDX models distributed by The Jackson Laboratory. MMHCdb in collaboration with EMBL-EBI has also co-developed the PDX Finder resource to provide a comprehensive global catalog of PDX models available for researchers.

MTB is supported by NCI grant CA089713.

Citation Format: Dale A. Begley, Debra M. Krupke, Steven B. Neuhauser, Joel E. Richardson, John P. Sundberg, Carol J. Bult. PDX Like Me: A molecular profile-based search tool [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1064.
Abstract 1066: Drug resistant PDX models for testing targeted drug sensitivity
Authors: Cheng Jijun , Zhang Feifei , Zhou Zhen , Long Yuan , Xu Wenhua , Zhao Shizhu , Chen Hongkui , Lu Shun , Wen Danyi
Published: August 2019

Advanced lung cancer of late stage patients is selectively sensitive to available targeted therapeutics and often develops resistance to early sensitive drug treatment, thus, the patient-derived xenograft (PDX) models are critical to evaluate drug sensitivity and resistance. We established PDX models from routine surgery tissues, as well as from tumor cells in pleural effusion specimens and from biopsies of advanced cancer patients who are refractory to treatments. Genetic profiling of the PDX lung models revealed a group of them with TKI drug-associated EGFR mutations (L858R, T790M, Exon19del, C797S). Among them, are an AZD9291-resistant sub-group with EGFR double mutations (Exon19Δ, T790M) in the clinic before AZD9291 treatment, and yet, acquired resistance to AZD9291 either through emergence C797S mutation or T790M loss in EGFR or through mutations in other pathways, providing insights into potential combo therapies. Indeed, combinations of AZD9291 with Cetuximab as well as with inhibitors targeting other signaling pathways such as MEK, are efficacious against an AZD9291-resistant lung PDX model with EGFR triple mutations (Exon19Δ, T790M, C797S). We also establish valuable lung models resistant to ALK inhibitors, Avastin, PD-1 antibody drugs, and resistant hematopoietic models. In summary, functional drug sensitivity profiling and genetic profiling with drug resistant PDX models provide insights into resistance mechanisms as well as precise therapeutic options against resistance emergence.

Citation Format: Jijun Cheng, Feifei Zhang, Zhen Zhou, Yuan Long, Wenhua Xu, Shizhu Zhao, Hongkui Chen, Shun Lu, Danyi Wen. Drug resistant PDX models for testing targeted drug sensitivity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1066.
Abstract 107: Extracellular vesicles from cancer associated fibroblasts induce drug resistance via integrinβ1/FAK signaling in gastric cancer cells
Authors: Uchihara Tomoyuki , Yonemura Atsuko , Miyake Keisuke , Yasuda Tadahito , Itoyama Rumi , Iwatsuki Masaaki , Baba Yoshifumi , Yoshida Naoya , Baba Hideo , Ishimoto Takatsugu
Published: August 2019

Background: Cancer-associated fibroblasts (CAFs) enhance tumor progression through secretion of soluble factors. Although we demonstrated that direct-interaction between CAFs and cancer cells confer on gastric cancer (GC) cells the ability to invade extracellular matrix (ECM), the molecular mechanism underlying the drug resistance by soluble factors from CAFs has not been determined.

Methods: We have established primary fibroblasts from more than 100 GC patients and collected conditioned medium (CM) of CAFs. Human GC cell lines cultured in normal medium or CAF-CM were examined the drug resistance with or without extracellular matrix (ECM). We isolated extracellular vesicles (EVs) from CAF-CM by ultracentrifugal separation. We performed RNA sequencing of GC cell lines and mass analysis of GC-EVs and CAF-EVs.

Results: GC cells treated with CAF-CM or CAF-EVs did not show morphological change on normal coated plates. Notably, we found that GC cells with CAF-CM or CAF-EVs on ECM-coated plates showed unique network formation and cisplatin resistance. Given that the morphological change appeared within 1-3 hours, we predicted that the stimulation by CAF-EV had an impact on the expression pattern of membrane proteins in GC cells. Mass analysis using isolated CAF-EVs was conducted and revealed that CAF-EVs contained high amount of annexin family protein members. Moreover, we showed that annexin in CAF-EVs is critical for network formation and drug resistance of GC cells. Finally, we identified integrinβ1/FAK signaling was activated in GC cells treated with CAF-EVs during network formation.

Conclusions: These findings highlight the underlying mechanism whereby CAF-EVs in tumor-microenvironment confer GC cells on drug resistance.

Citation Format: Tomoyuki Uchihara, Atsuko Yonemura, Keisuke Miyake, Tadahito Yasuda, Rumi Itoyama, Masaaki Iwatsuki, Yoshifumi Baba, Naoya Yoshida, Hideo Baba, Takatsugu Ishimoto. Extracellular vesicles from cancer associated fibroblasts induce drug resistance via integrinβ1/FAK signaling in gastric cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 107.
Abstract 1081: Smad3 silences neutrophil anticancer activity in the tumor microenvironment
Authors: Tang Patrick Ming-Kuen , Tang Philip Chiu-Tsun , Chung Jeff Yat-Fai , Huang Xiao-Ru , To Ka-Fai , Lan Hui-Yao
Published: August 2019

Tumor microenvironment (TME) is a new therapeutic target for cancer, however it is highly heterogeneous and still largely unknown. Recently, we revealed that Smad3 is essential for TME-driven cancer progression, better understanding of the underlying mechanism may uncover novel therapeutic targets for cancer. Indeed, Smad3-dependent TME (Smad3-TME) is originated from bone marrow, where neutrophils are the most abundant leukocytes but their role in cancer is still controversial. Here, we are the first study to reveal the regulatory role of Smad3 in tumor-associated neutrophils (TAN). Interestingly, we observed that knockout of Smad3 (Smad3-KO) dramatically increased TAN population in mice bearing syngeneic melanoma B16F10 and lung carcinoma LLC, where the anticancer N1 phenotypes were significantly induced in the SMad3-KO TME-isolated TAN. Unexpectedly, adoptive transfer of Smad3-KO bone marrow-derived neutrophils markedly suppressed the progression of LLC-tumor on Smad3-WT mice in vivo, and their enhanced cancer-killing activity was confirmed by co-culture assay in vitro. Furthermore, microarray analysis identified a total of 4416 differentially expressed genes associated with Smad3-KO neutrophils under cancer condition in vitro, but the presence of Smad3 largely decreased the transcriptome diversity; especially on genes associated with virus defense response including an anticancer cytokine IL-28. Silencing of IL-28 significantly reduced the anticancer efficiency of Smad3-KO TAN on LLC-bearing mice in vivo. Mechanistically, we uncovered that IL-28A serves as a direct Smad3 target gene and specifically down-regulated in neutrophils under TGF-β1 or LLC-secretome stimulations; showing by ChIP and dual-luciferase reporter assays in vitro. Thus, targeting Smad3 precisely on TAN may represent as a novel and effective immunotherapy for cancer. Acknowledgments: This study was supported by Lui Che Woo Institute of Innovative Medicine (CARE program), Research Grants Council of Hong Kong (GRF 14117815, 14121816, 14163317, C7018-16G, TRS T12-402/13N), Health and Medical Research Fund (03140486, 14152321), Innovation and Technology Fund of Hong Kong (ITS/068/18), Direct Grant for Research CUHK (2017.002).

Note: This abstract was not presented at the meeting.

Citation Format: Patrick Ming-Kuen Tang, Philip Chiu-Tsun Tang, Jeff Yat-Fai Chung, Xiao-Ru Huang, Ka-Fai To, Hui-Yao Lan. Smad3 silences neutrophil anticancer activity in the tumor microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1081.
Abstract 1085: Neuropilin-1 deficient macrophages as a treatment strategy for glioblastoma
Authors: Caponegro Michael D. , Tsirka Stella E.
Published: August 2019

Post-diagnosis survival of grade IV glioblastoma (GB) is a dismal 14 months in patients receiving standard of care therapy. With the development of immunotherapeutic strategies targeting non-cancerous, stromal cells of the tumor microenvironment (TME), new avenues are being pursued in clinical trials and pre-clinical animal models of advanced GB. Glioma-associated microglia/macrophages (GAMs) of the TME promote chemoattraction, immune suppression, neoangiogenesis, and tumor cell survival, which parallels tumor-infiltrating macrophage behavior of other solid malignancies. GAMs are now recognized as a potentially practical immunotherapeutic target in GB. Previously in our lab, we have shown that GAM-specific deletion of the cell surface co-receptor Neuropilin-1 (NRP1) drastically reduces pro-tumorigenic behavior of infiltrating microglia and/or macrophages in murine glioma by reducing immunosuppressive tendencies and neoangiogenesis. As an extension to our genetically-ablated mouse model, we present here a platform by which these NRP1-deficient monocytic cells can be harvested ex vivo and locally delivered into the TME of the bulk glioma lesion, to exert anti-tumorigenic effects. To our knowledge, this is the first attempt to deliver cells of monocytic origin intratumorally as a therapeutic option.

Citation Format: Michael D. Caponegro, Stella E. Tsirka. Neuropilin-1 deficient macrophages as a treatment strategy for glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1085.
Abstract 1086: Immune checkpoint inhibitors CD300A and LAIR1 are associated with poor prognosis in clear cell renal cell carcinoma
Authors: Sjolund Jonas , Lindgren David , Nilsson Helén , Johansson Martin E. , Axelson Hakan , Pietras Kristian
Published: August 2019

Immune checkpoint inhibitors targeting the programmed cell death-1 receptor (PD-1) have been shown to improve the survival of patients with clear cell renal cell carcinoma (CCRCC). However, a benefit for CCCRC patients was observed with the FDA-approved PD-1 inhibitor Nivolumab irrespective of PD-1 ligand (PD-L1) expression status indicating there are potentially additional CCRCC checkpoint molecules worth exploring for targeted therapy. We therefore analyzed RNAseq data (n=534, CCRCCs; n=72, normal kidney specimens) from the TCGA project, and found that PD-L1 was not upregulated in CCRCC when compared to normal tissue whereas other checkpoint members i.e. CD300A and LAIR1 were among the most significantly upregulated genes in CCRCCs. In terms of overall survival, high expression of PD-L1 was associated with better outcome in CCRCCs while high expression of CD300A and LAIR1 were significantly associated with worse outcomes. We confirmed these findings in two independent cohorts of CCRCC patients. Furthermore, we performed cell type enrichment analysis, which showed that CD300A and LAIR1 are most likely expressed by tumor-associated macrophages in the CCRCC tumor microenvironment. Our results suggest that PD1/PDL1 might be suboptimal targets in CCRCC and that there are additional unexplored immune checkpoint molecules that warrant our attention.

Note: This abstract was not presented at the meeting.

Citation Format: Jonas Sjolund, David Lindgren, Helén Nilsson, Martin E. Johansson, Hakan Axelson, Kristian Pietras. Immune checkpoint inhibitors CD300A and LAIR1 are associated with poor prognosis in clear cell renal cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1086.
Abstract 1097: Establishment of murine immune and stroma reference bench for TME study on in vivo models
Authors: Xue Jia , An Xiaoyu , Chen Xiaobo , Qian Wubin , Li Henry Q. , Guo Sheng
Published: August 2019

Cancer is a heterogeneous disease and highly related to immunological system. Experimental animal models such as patient derived xenograft (PDX) are excellent system to understand tumor microenvironment (TME), especially for immunotherapy. Deconvolution analysis on bulk tissue genomics of these in vivo tumor models is one of cost-effective ways to investigate TME. Several groups have released robust deconvolution algorithms with human immune cell signature. Our previous research on ~1600 transcriptomes of PDX collection identified various human immune and stromal contents in 24 different cancer types. However, no appropriate mouse immune signature generated from single standard platform could be used so far to study murine immune and stromal contents. To this end, we established a reference dataset by performing transcriptome sequencing of over 10 types of mouse immune and stromal cells, including different subtypes of B cells, T cells, monocytes, macrophages, dendritic cells, neutrophils, natural killer cells, cancer associated fibroblasts, glial cell and epithelial cell, etc. These cells were isolated from 3 different tissues (bone marrow, lymph node and spleen) on tumor-bearing and non-tumor-bearing mice covering a variety of mouse strains. We identified the transcriptomic landscape of murine immune system as well as difference across source tissues/organs and mouse strains. Based on this reference set, which is generated from single standard protocol, we defined precision gene signatures for studied cells and refined deconvolution algorithms for mouse TME studies. We further evaluated multiple algorithms by flow cytometry. In conclusion, our murine immune and stroma reference dataset will be essential basis for model selection in immunotherapy study in future.

Note: This abstract was not presented at the meeting.

Citation Format: Jia Xue, Xiaoyu An, Xiaobo Chen, Wubin Qian, Henry Q. Li, Sheng Guo. Establishment of murine immune and stroma reference bench for TME study on in vivo models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1097.
Abstract 1099: Organotypic tumor tissue slices provide versatile platform for immuno-oncology
Authors: Sivakumar Ramya , Chan Marina , Yeung Raymond , Gujral Taranjit S.
Published: August 2019

Organotypic tissue slices represent one of the most physiologically-relevant culture systems for studying the tumor microenvironment. The thin slices of tumor cut out using automated vibratome retain the original architecture and cellular composition of the tumor from several days up to weeks depending on the investigated tumor model. The slice culture has been emerging as a versatile platform for studying cancer cell-host cell interactions. However, a thorough characterization of the multicellular organotypic culture system, particularly its immune component, is still unresolved to allow a better understanding of its potential applications. Here, we carried out a comprehensive characterization of immune cell composition in organotypic tumor slices prepared from several different syngeneic mouse tumor models. We generated several syngeneic mouse tumor models and harvested the primary tumors to make tumor slices. We then immunophenotyped the major tumor infiltrating leukocytes (TILs) by flow cytometry to understand the immune makeup of these slices immediately after the harvest, over a period of 7 days and upon treatment of various immuno-modulators. We found that the immune compositions of tumor slices do not deviate from that of the tumor cores, immediately after harvest, across all the syngeneic tumor models. During the 7-day follow-up, despite an initial decline in the overall cell viability which stabilized on day 3, we observed a consistent percentage of TILs out of total viable cells, suggesting that the relative composition of tumor and immune populations can be maintained in the tumor slice over time. Among the TILs, macrophages were the predominant immune cell type detected, similar to that observed in many human solid tumors. Macrophages remained viable till day 7 while other cell types were mostly viable till day 5, providing a window of time sufficient to support many short-term perturbation studies. Further, we found that these immune cell populations were responsive to the treatment with immunomodulators. For example, we observed an upregulation of co-stimulatory markers, CD86 and MHC-II, on macrophages and DCs upon stimulation with the pro-inflammatory cytokine IFN γ, further supporting the application of tumor slice in the study of immuno-oncology. Overall, these data support the approach of utilizing tumor slice culture as a platform for studying tumor microenvironment and evaluating efficacy of immune modulatory drugs.

Citation Format: Ramya Sivakumar, Marina Chan, Raymond Yeung, Taranjit S. Gujral. Organotypic tumor tissue slices provide versatile platform for immuno-oncology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1099.
Abstract 1101: Characterization of spatial distribution of tumor-infiltrating CD8+ T cells refines their prognostic impact in pancreatic cancer
Authors: Masugi Yohei , Abe Tokiya , Kitago Minoru , Shinoda Masahiro , Sakamoto Michiie
Published: August 2019

Background: The abundance of cytotoxic T cell infiltrates has important implications for patient outcome and therapeutic design in pancreatic ductal adenocarcinoma (PDAC), although intratumoral heterogeneity remains a major challenge for the better understanding of complicated immune microenvironment of PDAC. We aimed to characterize spatial CD8+ cell distribution within PDAC tissues, which might refine the prognostic role of tumor-infiltrating CD8+ lymphocytes.

Methods: We measured CD8+ cell density in tumor center, tumor margin, or whole tumor area, as well as CD8+ cell proximity to carcinoma cells, using fluorescent multiplex immunohistochemistry-based tissue computation on whole tissue sections of 214 surgically resected PDACs. Multivariable Cox proportional hazards regression analysis was performed to assess the association of CD8+ cell density with pancreatic cancer-specific survival, adjusting for major clinicopathologic and immune-related features, including the programmed cell death 1 ligand 1 (CD274, PD-L1) expression and the extent of tertiary lymphoid structures.

Results: We found a substantial heterogeneity in CD8+ cell density with, on average, a 2-fold-lower density in tumor center than in tumor margin. In multivariable model, the trend of prolonged survival with higher CD8+ cell density was stronger for tumor center, compared to tumor margin or whole tumor area (P for trend = 0.002, 0.07, or 0.009, respectively). Tumor CD274 expression and extensive tertiary lymphoid structures were appeared to be associated with higher CD8+ cell density in tumor margin, but not with CD8+ cell density in tumor center. The mean distance from tumor cells to the adjacent CD8+ cells correlated inversely with CD8+ cell density in tumor center (Spearman correlation efficient, -0.89), although the density of CD8+ cells proximate to cancer cells did not show any prognostic correlation.

Conclusion: Our data have shown a prognostic significance of CD8+ cell density in tumor center of PDAC, where CD8+ cell infiltration has been severely limited, suggesting a major role of pro-tumorigenic effect of immunosuppressive microenvironment in pancreatic cancer.

Citation Format: Yohei Masugi, Tokiya Abe, Minoru Kitago, Masahiro Shinoda, Michiie Sakamoto. Characterization of spatial distribution of tumor-infiltrating CD8+ T cells refines their prognostic impact in pancreatic cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1101.
Abstract 1102: Time and spatial distribution of monocytic lineages in the process of carcinogen-induced oral cancer
Authors: Chen Chi Jui
Published: August 2019

Head and neck cancer (HNC) is the sixth among the top ten cancers in the world. Among this group of disease, oral cancer has been one of the top five killers in males for many years in Taiwan. Despite combined treatment, the prognosis for HNC patients is, in general, poor, which has strongly stimulated research on finding new strategies specific for HNC. To find a new treatment protocol for HNC, we examined the tumor microenvironment of a carcinogen (4-Nitroquinolone-1-oxide,4-NQO)-induced oral cancer. The immunohistochemical staining (IHC) results showed that this tumor has relative low density of vessels and high percentage of hypoxia around tumor edge. In addition, we found a layer of α-SMA positive cancer-associated fibroblast (CAF) in the junction of tumor and normal tissues. Further, we also found that a cluster of high density of CD68+ macrophages and Gr-1+ cells in this junction. To our surprise, very few tumor-associated macrophages (TAMs) could be identified in other tumor regions except this junction. In addition, the analysis of blood sample during tumor development found that the population of monocytic and granulocytic myeloid-derived suppressor cells (M-MDSCs and G-MDSCs) in the whole blood increases with the growth of the tumor, which has good correlation with the decrease of CD8+ and CD4+ T cells. The unique spatial distribution of myeloid cells in tumor tissue and consistent dynamic change of MDSC and T cells in the blood provides us an ideal tumor model for designing new strategies specific for HNC. We are currently using this tumor model to study the potential of combining immunotherapy with boron neutron capture therapy (BNCT) for HNC.

Citation Format: Chi Jui Chen. Time and spatial distribution of monocytic lineages in the process of carcinogen-induced oral cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1102.
Abstract 1105: Cisplatin treatment alters the preference of melanoma metastasis to liver associated with Ly6G positive cells
Authors: Chun-Hsiang Chang
Published: August 2019

This study examined the influence of cisplatin on host stroma and its consequence on metastasis using a metastatic melanoma tumor model. This preliminary result indicates that Gr-1+ cells could play important role on cisplatin treatment-directed liver metastasis. To further examine the role of Gr-1+ cells on this phenomena, Gr-1Ab was administered at day 16 after tumor inoculation. Results show that Gr-1Ab treatment could effectively decrease the Ly6G+ cells in both lung and liver and without significant reduced in lung-metastasis but, increased the metastasis in lung. To identify the effect of cisplatin treatment on B16-F0 melanoma metastasis, normal mice were treated with cisplatin prior to iv inject of B16-F0 melanoma cells. Cisplatin prevented treatment, similar to Gr-1 Ab therapy, decreased Ly6G+ cell in lung and liver tissue. And, it is minor difference of lung metastasis compare to Gr-1 Ab treatment, but more metastatic site than it. It demonstrates that cisplatin therapy induced liver-metastasis could associate with Ly6G+ cells. This study concludes that the cisplatin treatment could alter the preference of tumor metastasis, which is associated with the reduction of Ly6G+ cells It could be a potential target to enhance therapy efficiency.

Citation Format: Chang Chun-Hsiang. Cisplatin treatment alters the preference of melanoma metastasis to liver associated with Ly6G positive cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1105.
Abstract 1109: CPAP promotes HCC angiogenesis and metastasis via interacting with and enhancing STAT3 signaling
Authors: Chen Ruo Yu , Hung Liang-Yi
Published: August 2019

Centrosomal P4.1-associated protein (CPAP) is a centrosomal protein and can as a transcriptional coactivator of STAT5 and NF-κB in cancer. Our previous studies indicated that CPAP is overexpressed in tumor tissue and can increase TNFα-mediated NF-κB activation in HCC. Here, we demonstrated that overexpressed CPAP increases tumor growth, angiogenesis, as well as metastasis ex vivo and in vivo. We found that CPAP increases these malignant abilities of tumor cells are through IL-6/STAT3 signaling. We demonstrated that CPAP directly interacts with C-terminal domain of STAT3 to increase STAT3 activity. Overexpression of CPAP enhances IL-6/STAT3/IL-8-mediated angiogenesis and other metastatic genes expression such as CD44 and MCAM. These results indicated that CPAP leads to HCC malignancy and metastasis by increasing STAT3 activity through directly interaction. Clinically, CPAP positively correlates with IL-8 in HCC with vascular invasion; and also positively correlates with CD44 and MCAM in HCC tissues. In summary, our findings shed light on the importance of CPAP to act as a potential therapeutic target for inhibiting the IL-6/STAT3-mediated angiogenesis pathway and treating metastatic HCC.

Citation Format: Ruo Yu Chen, Liang-Yi Hung. CPAP promotes HCC angiogenesis and metastasis via interacting with and enhancing STAT3 signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1109.
Abstract 1118: FOXQ1 stimulates WNT-signaling through transcriptional activation of WNT5B to promote the epithelial to mesenchymal transition
Authors: Mitchell Allison V. , Wu Guojun
Published: August 2019

Aberrant induction of the epithelial to mesenchymal transition (EMT) is an acknowledged mechanism of acquired metastatic competence and drug resistance in many epithelial cancer types. Both physiological and pathological forms of EMT are thought to occur through a network of transcription factors (TFs) that drive cell plasticity and the acquisition of stem-like traits. The forkhead box transcription factor, FOXQ1, is a potent driver of EMT. FOXQ1 has been shown to be specifically upregulated in the triple negative subtype of breast cancer (TNBC) where is it associated with worse clinical prognosis. However, the role of FOXQ1 within the EMT transcriptional network has not been extensively characterized. In this study, we employed chromatin immunoprecipitation and RNA sequencing in a human mammary luminal epithelial cell line with ectopic FOXQ1 (HMLE/FOXQ1), as a model of breast EMT. We identified FOXQ1 localization within the promoter regions of prominent EMT-TFs, including TWIST1, ZEB1, FOXC2 and SIX2. Further, FOXQ1 direct gene targets are enriched in signaling pathways previously associated with EMT, including the WNT-signaling pathway. Within the WNT pathway, the WNT5B signaling ligand was a direct target for transcriptional activation by FOXQ1. Gene expression analysis in a panel of breast cancer cell lines revealed both FOXQ1 and WNT5B are enriched in the basal B subtype of TNBC. FOXQ1 and WNT5B expression were also found to be upregulated and correlated downstream of TGFβ induced EMT. Disruption of WNT5B expression attenuated EMT-associated stem-like traits, including decreases in mammosphere formation and in the CD44+/CD24- population, and significantly reduced cell invasion. Analysis of breast cancer gene expression from the TCGA revealed that FOXQ1 and WNT5B expression are correlated across all breast samples (r=0.46) and differentially upregulated in ER- breast cancer. Additionally, both FOXQ1 and WNT5B expression were associated with worse recurrence-free survival. Our data supports that WNT5B is a critical FOXQ1 target for mediating EMT features and suggests that disrupting the FOXQ1-WNT5B axis could be a potential strategy to target the EMT cell population in TNBC patients.

Citation Format: Allison V. Mitchell, Guojun Wu. FOXQ1 stimulates WNT-signaling through transcriptional activation of WNT5B to promote the epithelial to mesenchymal transition [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1118.
Abstract 112: Patient derived models reveal impact of the tumor microenvironment on therapeutic response
Authors: Shafi Ayesha A. , Schiewer Matthew J. , Leeuw Renée de , Dylgjeri Emanuela , McCue Peter A. , Shah Neelima , Gomella Leonard , Lallas Costas , Trabulsi Edouard , Centerera Margaret , Hickey Theresa , Butler Lisa , Raj Ganesh , Tilley Wayne , Cukierman Edna , Knudsen Karen E.
Published: August 2019

Prostate cancer (PCa) is the most common non-cutanenous cancer and the second leading cause of cancer-related death in American men. Androgen receptor (AR) is a hormone-activated transcription factor that plays an important role in both the development and progression of PCa. Androgen deprivation therapy is a common first-line therapy for disseminated disease. However, virtually all tumors become resistant to such therapy and the tumor recurs. Thus, there is a vital need for the development of novel, more effective drugs. One major hurdle in this aspect is the lack of adequate preclinical models. Current models do not effectively recapitulate the heterogeneity and the microenvironment of human PCa tumors, significantly hindering the ability to accurately predict therapeutic response. Our collaborative group has utilized and characterized a method to culture patient tumors ex vivo, termed Patient Derived Explant (PDE). This approach maintains the integrity of the native tumor microenvironment, tumor tissue morphology, and molecular signaling. Importantly, our PDE model can be manipulated both chemically (drugs/compounds) and genetically (shRNA) in order to determine specific reactions and mechanisms of response on individual tumor growth. Furthermore, with this model we can quantitatively assess drug efficacy on numerous parameters (i.e. AR levels, Ki67 staining, apoptosis screening, and desmoplasmic indices). Data to be discussed will assess the variances in response to AR-directed therapeutics and underlying mechanisms of action, while also utilizing TME characteristics as a means to predict response to therapy. In addition, we can potentially identify clinically relevant subpopulations of patients and molecularly profile their cultured tissue to uncover new pathways for therapeutic intervention. Thus, the PDE model allows for a comprehensive evaluation of individual tumors in their native microenvironment to ultimately develop more effective therapies. This study will have transformative clinical impact discerning novel metrics for the inclusion of precision medicine for advanced PCa.

Citation Format: Ayesha A. Shafi, Matthew J. Schiewer, Renée de Leeuw, Emanuela Dylgjeri, Peter A. McCue, Neelima Shah, Leonard Gomella, Costas Lallas, Edouard Trabulsi, Margaret Centerera, Theresa Hickey, Lisa Butler, Ganesh Raj, Wayne Tilley, Edna Cukierman, Karen E. Knudsen. Patient derived models reveal impact of the tumor microenvironment on therapeutic response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 112.
Abstract 1122: CD44 as potential therapeutic target for medulloblastoma metastasis
Authors: Qi Dan , Wang Fengfei , Fonkem Ekokobe , Huang Jason H. , Wu Erxi
Published: August 2019

Medulloblastoma (MB) is a commonly diagnosed pediatric brain tumor that shows highly variable rates of morbidity and mortality. Metastasis is the leading cause of death in patients with MB, approximately 30% of MB patients have metastatic tumors right at diagnosis. Therefore, there is a critical need to dissect the molecular mechanisms that govern MB progression and metastasis. In our previous work, we found that DHA and VP16 synergistically inhibit MB cell proliferation and induce cell death accompanied with a marked reduction in the expression of CD44 standard form (CD44s). In this study, we show that MB cells express CD44s mainly and the elevated level of CD44s enhances MB cell invasion, suggesting that CD44s may play a crucial role in MB metastasis. In addition, we observed CD44s expression only in MB patients with metastasis using a small cohort of samples. Large database analysis show that MB patients with elevated levels of CD44 have poorer outcomes. Moreover, CD44 knockdown markedly reduces the expressions of transcription factor c-Myc and the platelet derived growth factor receptor (PDGFR) β, both are therapeutic targets for MB. Interestingly, PDGFRβ knockdown abolishes CD44s expression and downregulates the level of c-Myc. However, there is no expression change of CD44s and PDGFR β after the knockdown of c-Myc. These results indicate that the expression and function of CD44s could be critical to MB progression and metastasis, highlighting the potential therapeutic targets for MBs with metastasis.

Citation Format: Dan Qi, Fengfei Wang, Ekokobe Fonkem, Jason H. Huang, Erxi Wu. CD44 as potential therapeutic target for medulloblastoma metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1122.
Abstract 1124: S100A14 suppresses metastasis of nasopharyngeal carcinoma via inhibition of NF-kB signaling
Authors: Meng Dong-Fang , Qian Chao-Nan
Published: August 2019

Nasopharyngeal carcinoma (NPC) has a high incidence rate in southern China and Southeast Asia. Distant metastasis is the main reason for treatment failure. By analyzing our previously published whole genomic expression profiling data comparing low- versus high-metastasis NPC cells, we found that S100A14 expression was dramatically suppressed in high-metastasis cells. The reduced expression of S100A14 protein in the primary NPC collated with poorer overall survival of the NPC patients. Our preliminary experiments showed that S100A14 could inhibit the migratory and invasive abilities, as well as the epithelial-mesenchymal translation of NPC cells. RNA sequencing analyses found that NF-kB signaling pathway was the most dramatically influenced pathway by S100A14 with a negative relationship found between S100A14 and NF-kB signaling. We therefore propose a hypothesis that S100A14 inhibits NPC metastasis through inhibition of NF-kB signaling. In the proposed study, we will use a series of in vitro and in vivo approaches to clarify the inhibitory effects of S100A14 in terms of nuclear localization of P50 and P65, expression of IL-1beta, phosphorylation activation of the key molecules in NF-kB signaling pathway, as well as the in vivo metastasis of NPC cells. The accomplishment of the proposed study will reveal a novel mechanism of inhibiting NPC metastasis via inhibitory effects of S100A14 upon NF-kB signaling, which should be composed of drugable targets for metastasis prevention and treatment.

Citation Format: Dong-Fang Meng, Chao-Nan Qian. S100A14 suppresses metastasis of nasopharyngeal carcinoma via inhibition of NF-kB signaling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1124.
Abstract 1142: An activity-dependent proximity ligation platform for spatially resolved and multiplexed quantifications of active enzymes in single cells
Authors: Li Gang , Moellering Raymond E.
Published: August 2019

Integration of chemical probes into proteomic workflows enables the interrogation of protein activity, rather than abundance. Current methods limit the biological contexts that can be addressed due to sample homogenization, signal-averaging, and bias toward abundant proteins. To expand the application scope of chemical proteomics and explore the cellular heterogeneity on enzyme activities, we developed a new platform that integrates family-wide chemical probes with proximity-dependent oligonucleotide amplification and imaging to quantify enzyme activity in native contexts with high spatial and single cell resolution. Application of this method, activity-dependent proximity ligation (ADPL), to serine hydrolase and cysteine protease enzymes enables quantification of differential enzyme activity resulting from endogenous changes in localization and expression. In a competitive format, small molecule target engagement with endogenous proteins in live cells can be quantified. Retention of sample architecture enables interrogation of complex environments such as cellular co-culture and patient samples. We also successfully applied ADPL in xenograft tissue sample, exhibiting the application prospect in primary patient tissue. Additionally, implementation of barcoded antibody-oligo direct conjugation enabled multiplexed readout of active enzymes within and between enzyme families, providing the possibility of simultaneous biomarker detection. Together, this work supports ADPL should be amenable to diverse and multiplexed protein families to detect active enzymes at scale and resolution out of reach with current methods.

Citation Format: Gang Li, Raymond E. Moellering. An activity-dependent proximity ligation platform for spatially resolved and multiplexed quantifications of active enzymes in single cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1142.
Abstract 1144: Imaging radiotherapy induced pulmonary fibrogenic changes with integrin-PET
Authors: Saleem Azeem , Helo Yusuf , Searle Graham , Dekaj Fatjon , Cook Jo , Win Zarni , Gunn Roger , Wells Paula
Published: August 2019

Integrin αvβ6 plays a key role in the fibrotic pathway by activation of Transforming Growth Factor β. Expression of αvβ6 integrin in alveolar epithelium increases after radiotherapy (RT) before onset of fibrosis and anti- αvβ6 therapy prevents fibrogenesis. Post-RT lung fibrosis, a major barrier to improved cure rate presents as progressive pulmonary injury. The utility of PET imaging with a αvβ6 integrin ligand was evaluated in patients with non-small cell lung cancer following pulmonary RT.

Subjects who had received pulmonary RT within 6 months were recruited. PET imaging of the lungs over 60 minutes was performed after administration of a fluorine-18 radiolabelled 20 amino acid αvβ6-specific peptide ligand, [18F]-FBAA20FMDV2 (IMAFIB; Cancer Research Technologies (CRT) patented). PET-CT scans were co-registered with the RT planning (RTP) scans and segmented to regions corresponding to RT doses of > 40 Gy (excluding tumour), 25-40 Gy, 15-25 Gy, 8-15 Gy and < 8 Gy. Time activity curves and the standardised uptake values (SUVs) were calculated and comparisons made between RT dose and PET uptake (SUV). Linear regression between SUV and RT dose was performed for each subject and normalised to an intercept of 1.

6 subjects (3M; 3 F) aged 51-75 years underwent an IMAFIB-PET scan between 6 -22 weeks after RT completion and mean IMAFIB radioactivity of 45.4 (range 8.9 – 122) MBq administered. 5 subjects received conformal external beam RT, while one subject received stereotactic RT (SBRT). A correlation between a higher PET uptake (SUV) and higher RT doses received by the lung was observed in all subjects with a significant relationship (p < 0.05) in 3 of the 6 subjects (table). The averaged normalised SUV across all subjects increased monotonically with RT dose, between 1.05 ± 0.14 at 4 Gy and 1.58 ± 0.31 at 52 Gy.

Subject No.111216172021RT dose and fractionation55Gy in 5 fractions. SBRT64 Gy in 32 fractions45 Gy in 15 fractions50 Gy in 20 fractions64 Gy in 32 fractions64 Gy in 32 fractionsTime between completion of RT and PET scan (weeks)822136714Gradient of relationship between PET uptake and RT dose0.0290.0050.0100.0060.0030.008Intercept1.3050.5040.8370.3910.6910.469r20.8410.5390.8360.8790.3100.672p value0.0280.1580.0300.0180.1630.089

A relationship between IMAFIB PET uptake and RT dose is consistent with RT dose-related activation of αvβ6 integrin in the irradiated lung and to the induction of lung fibrosis in subjects receiving pulmonary RT.

Citation Format: Azeem Saleem, Yusuf Helo, Graham Searle, Fatjon Dekaj, Jo Cook, Zarni Win, Roger Gunn, Paula Wells. Imaging radiotherapy induced pulmonary fibrogenic changes with integrin-PET [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1144.