bluuebunny/crossref_metadata_2025 huggingface dataset:
Abstract 2650: Angiogenic inhibitors reduce mycosis fungoides cell survival and enhance cell death
Authors: Rendon-Serna Natalia , Dufies Maeva , Correa-Londono Luis Alfonso , Bermudez-Munoz Olga Maria , Velasquez-Lopera Margarita Maria , Pages Gilles
Published: July 2021

Cutaneous T-cell lymphomas (CTCL) account for approximately 80% of cases of cutaneous lymphomas. Among CTCL, mycosis fungoides (MF) is the most common and most studied type comprising up to 55% of cases. Three different MF stages are recognized in the following order of pathology evolution: patches, plaques and tumor; MF can progress to tumor stage with involvement of other organs, compromising life due to systemic malignant invasion and immunosuppression. Currently, there are not long-lasting treatment for MF. We aimed to understand if angiogenesis, involved in solid tumor progression, could be involved in MF course. For this, we studied both, MF cell lines (HH, Myla) and skin tissues from MF patients at its different stages including healthy skin, according to ethical approval statements. We evaluated the density of blood vessels in skin from MF patients using staining for CD31 by immunohistochemistry. In HH and Myla cell lines, we have studied the presence of angiogenic receptors by western blot and then we have analyzed the effect of 3 angiogenic inhibitors, axitinib, cabozantinib and sunitinib, on cell survival and cell death. We found that there is an increased number of blood vessels in tumor stage compared with patch stage and healthy skin, and in plaque stage compared with patch stage. In HH and Myla cells that express VEGR2 and c-MET, we noticed that although they are sensitive to the angiogenic inhibitors used, axitinib and specially sunitinib reduced significantly cell survival and enhanced cell death at doses between 5-10 micromolar, at 72 hours. We conclude that the density of blood vessels increases with MF progression and thus that angiogenesis takes place during MF pathophysiology. Furthermore, the fact that angiogenic inhibitors already used in tumor treatment provoke MF cell death and affect MF cell survival, suggests that the use of angiogenic inhibitors could be of great interest for controlling MF progression. Future animal model experiments and clinical trials should reveal the potential of angiogenesis inhibition as a new therapeutical option for MF treatment.

Citation Format: Natalia Rendon-Serna, Maeva Dufies, Luis Alfonso Correa-Londono, Olga Maria Bermudez-Munoz, Margarita Maria Velasquez-Lopera, Gilles Pages. Angiogenic inhibitors reduce mycosis fungoides cell survival and enhance cell death [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2650.
Abstract 2656: Muscle wasting in head and neck carcinomas: Investigations on the underlying transcriptional reprogrammation
Authors: Hachicha Héla , Pirozhkova Iryna , Gelin Aurore , Bidault François , Rouffiac Valérie , Dayris Thibault , Ragot Thierry , Breuskin Ingrid , Even Caroline , Gorphe Philippe , Busson Pierre
Published: July 2021

Introduction:

Cancer cachexia is a multifactorial syndrome associated with a skeletal muscle atrophy that affects at least 55% of patients bearing Head and Neck Squamous Cell Carcinomas (HNSCCs). To our knowledge, molecular mechanisms of muscle alterations have never been investigated on clinical muscle specimens from HNSCC patients. The aim of this study was to perform transcriptome profiling by bulk RNAseq on muscle fragments from HNSCC patients with and without cachexia and from nude mice bearing human HNSCC xenografts.

Methods:

Our study was made in three parts: 1) In Vitro investigations on human myoblasts co-cultivated with HNSCC cells. 2) In Situ investigations on muscles collected from nude mice xenografted with HNSSCs cells (FADU). 3) Exploration of clinical muscle samples collected from HNSCC patients. RNA sequencing was used to assess gene expression changes and gene function enrichment related to cachexia, in myoblasts co-cultivated with malignant cells as well as skeletal muscle samples collected from mice xenografted with HNSCC cells (FADU) or HNSCC patients.

Results:

In vitro experimentations showed significant impact of malignant cells on myoblast proliferation and differentiation. In nude mice, FADU xenografts resulted in distant loss of muscle mass at various anatomic locations. RNA sequencing of muscle samples from cachectic patients showed significant differences in gene expression compared with control muscles samples from tumor-free donors. Commonly modified pathways included immune and inflammatory response, mitochondrial metabolism and striated muscle differentiation. The most remarkable finding was the up-regulation of the BIRC3 transcript which encodes the anti-apoptotic protein c-IAP2. This alteration was observed both in human cachectic muscles and murine muscle fragments from tumor-bearing mice.

Conclusions:

Although the mechanism of BIRC3 up-regulation in cachectic muscle cells is not well understood, this finding is likely to be important for patient management in a context of promising therapeutic results obtained using Smac mimetics. Because these compounds are inhibitors of the c-IAP protein family, it will be important in the future to monitor their positive or negative effects on muscle alterations in cachectic HNSCC patients.

Citation Format: Héla Hachicha, Iryna Pirozhkova, Aurore Gelin, François Bidault, Valérie Rouffiac, Thibault Dayris, Thierry Ragot, Ingrid Breuskin, Caroline Even, Philippe Gorphe, Pierre Busson. Muscle wasting in head and neck carcinomas: Investigations on the underlying transcriptional reprogrammation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2656.
Abstract 2694: An anatomic proteomic atlas of human glioblastoma
Authors: Lam K.H. Brian , Djuric Ugljesa , Batruch Ihor , Richer Maxime , Diamandis Phedias
Published: July 2021

Glioblastoma (GBM) is an aggressive brain tumor with an expected survival of under 15 months despite spirited multimodal therapy. This grim outlook has remained virtually unchanged over the last 40 years and thus necessitates alternative research and therapeutic development strategies. Historically, traditional models of cancer biology have largely considered GBM tissue to be a homogenous mass. Emerging studies however now suggests that the interaction of tumor cells with various normal components of the brain and immune system play an important role in helping cancer grow and develop resistance to therapies. Better understanding of these hallmark features of GBM, collectively known as the “tumor microenvironment” (TME), could lead to the development of new and more effective therapies. In light of this, there is a renewed interest in revisiting our theories of cancer and preserving the microscopic anatomy of GBM in our molecular profiling efforts. Here we leverage laser capture microdissection (LCM) and mass spectrometry-based proteomics, in order to generate a detailed map of the distribution of proteins within tissues across a large number of patients. Specifically, we will isolate, and profile well-understood microscopic components of GBM: cellular tumor (CT), microvascular proliferation (MVP), infiltrating tumor (IT), palisading cells around necrosis (PAN) and normal brain tissue (LE). All of this data will be provided in an online-based GBM atlas as a publicly available resource. This atlas and the associated database for clinical and genomic data will serve as a useful platform for developing therapeutics and testing novel hypotheses related to GBM biology.

Citation Format: K.H. Brian Lam, Ugljesa Djuric, Ihor Batruch, Maxime Richer, Phedias Diamandis. An anatomic proteomic atlas of human glioblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2694.
Abstract 2698: Spatial gene expression profiling in breast cancer
Authors: Spears Melanie , Talebian Vida , Liao Linda , Hopkins Megan , Jain Drashti , Quintayo Mary Anne , Bayani Jane , Cheung Alison , Yaffe Martin , Bartlett John M.
Published: July 2021

Over the last decade breast cancer survival has improved, largely due to the therapies offered to patients with the disease. However, despite the advances in diagnosis and treatment of breast cancer, it is still remains the second leading cause of death from cancer in women. Breast cancer is a heterogeneous disease, this in part, explains why the majority of current therapeutic approaches for cancer work best when multiple agents are combined. The interaction between immune and tumor cells is critical in the development and progression of breast cancer. Here we present in situ transcriptomic profiling, using the NanoString Digital Spatial Profiler (DSP) cancer transcriptomic atlas (CTA) assay, of a cohort of breast cancer lumpectomies to reveal the extent of heterogeneity in pathologically defined unifocal and multifocal cancers. In this study, lumpectomies were processed as whole mounts with serial blocks reviewed. Tissue cores were taken from at least three different regions through the lumpectomy for tissue microarray (TMA) construction, focusing on morphologic/histological differences in addition to the spatial orientation of the sampled region within the lumpectomy. In situ quantification of 1800 tumor and immune genes across 60 patients revealed heterogeneity of tumor and immune genes in most patients. Expression of genes such as HER2, ER and AKT were enriched in the tumor compartment. Whereas, genes such as COLA1, CD68 and CD3 were enriched in the immune compartment. Using a SpatialDecon algorithm for mixed cell deconvolution on the immune areas heterogeneity of the tumor infiltrate at a cell type levels was observed. Fibroblasts and macrophages were prevalent in all samples while immune dense areas also contained B-cells and T-cells. While there are a number of clinically validated transcriptional assays available for breast cancer, we have demonstrated that the immune microenvironment needs to be considered to develop rational stratification of patients to currently available targeted therapies.

Citation Format: Melanie Spears, Vida Talebian, Linda Liao, Megan Hopkins, Drashti Jain, Mary Anne Quintayo, Jane Bayani, Alison Cheung, Martin Yaffe, John M. Bartlett. Spatial gene expression profiling in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2698.
Abstract 2699: Transcriptome analysis identifies molecular markers of tumor-associated high endothelial venules that predict breast cancer survival
Authors: Sawada Junko , Hiraoka Nobuyoshi , Fournier-Goss Ashley , Komatsu Masanobu
Published: July 2021

High endothelial venules (HEVs) are specialized post-capillary venules that recruit naïve lymphocytes to the lymph nodes and are essential for the development of adaptive immunity. HEVs can develop in tumors, and these specialized tumor blood vessels are thought to be important for recruiting naïve T cells and B cells into tumors and locally enhance anti-tumor immunity by promoting tertiary lymphoid structure formation. At present, the genes involved in the development and function of tumor-associated HEVs remain largely unknown. By a comparative transcriptome analysis in human breast cancer, we identified the expression of genes that represents the molecular signature of tumor-associated HEVs distinct from the rest of the tumor vasculature. Tumor vessels highly expressing these genes were associated with the accumulation of T and B lymphocytes and the occurrence of tertiary lymphoid structures, which is known to predict therapeutic response to immune checkpoint inhibitor therapies. Moreover, the Cancer Genome Atlas (TCGA) data show that the high expression of these genes is associated with a significant survival benefit in advanced breast cancer. The molecular signature of HEVs identified here may be useful for guiding immunotherapies and provide a new direction for investigating tumor-associated HEVs and their clinical significance.

Citation Format: Junko Sawada, Nobuyoshi Hiraoka, Ashley Fournier-Goss, Masanobu Komatsu. Transcriptome analysis identifies molecular markers of tumor-associated high endothelial venules that predict breast cancer survival [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2699.
Abstract 274: Comparison of proteomics identification pipelines for lymphocyte characterization
Authors: McCown Michaela A. , Allen Carolyn , Machado Daniel D. , Payne Samuel H.
Published: July 2021

Proteomics analyses can be adapted to samples as small as a single cell, however such sample reduction severely limits the number proteins identified and quantified. Multiple software pipelines exist for processing the raw data to quantitative protein tables. For very small sample amounts, the sensitivity of software algorithms can have a significant impact on the final results. We collected whole blood samples for two subjects by venous blood draws. Using antibody guided flow cytometry cell sorting, we separated B and T cell lymphocytes. For two subjects and two different blood collections, five replicates of 145 B cells or T cells were prepared for mass spectrometry for proteomics using the AutoPOTS workflow. Using the raw files from these forty runs, we perform protein quantification with a selection of common tools such as MaxQuant, ProteomeDiscoverer, and FragPipe and compare the number of identified and quantified proteins. In small sample proteomics, the primary objective is obtaining the most quantified proteins. Although each tool has advantages in terms of usability, speed, and sensitivity, our primary concern is sensitivity. At such a low sample input, we observe that the number of quantified proteins increases by 30% using the most sensitive algorithm. For identifying and quantifying proteins from small samples, the sensitivity of the software is the most important factor to consider. In addition to the identification sensitivity of each algorithm, we are exploring optimal parameters for increased protein quantification. Differences between samples appear regardless of the processing tool, showing that using a less sensitive tool will still characterize the trends but with a significant cost to the number of proteins. Using the most sensitive algorithm for small sample proteomics will greatly improve the number of proteins identified and quantified.

Citation Format: Michaela A. McCown, Carolyn Allen, Daniel D. Machado, Samuel H. Payne. Comparison of proteomics identification pipelines for lymphocyte characterization [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 274.
Abstract 2765: The indoleamine 2,3 dioxygenase-1 pathway drives intratumoral B cell maintenance
Authors: Johnson Burles Avner , Aragaki Adam K. , Williams Donna M. , Rogers Ophelia , Luo Li , Xian Lingling , Chia Lionel , Hahn Noah M. , Desiderio Stephen , Johnson Theodore S. , McConkey David J. , Resar Linda M.S.
Published: July 2021

Immunotherapy has revolutionized cancer treatment by improving survival in many cancer subtypes. While tumor infiltrating B (TIL-B) cells correlate with response to immunotherapy in selected solid tumors, they portend resistance to BRAF inhibitors in BRAF mutant melanoma. Thus, the mechanisms underlying TIL-B cell function within the tumor microenvironment have remained elusive. Here, we discovered that inhibition of the indoleamine 2,3 dioxygenase-1 (IDO1) pathway with D-1-methyl-tryptophan (D-1MT) markedly decreases TIL-B cells in a preclinical model of melanoma. Single cell RNA sequencing (scRNAseq) of murine melanoma demonstrate TIL-B cells are heterogeneous: while some B cells express markers consistent with an immune stimulatory phenotype, others express markers consistent with immune inhibitory function. D-1MT decreased splenic B cells and bone marrow derived B cell precursors in tumor bearing mice, suggesting that IDO1 pathway inhibition impedes B cell maturation. In four distinct human cancer subtypes, intratumoral IDO1 expression consistently correlates with high expression of a pan-B cell gene signature. Further, analysis of scRNAseq data from a cohort of patients with melanoma revealed that most TIL-B cells express IDO1. In mice, D-1MT also decreases intratumoral myeloid derived suppressor cells (MDSCs), which are essential for maintenance of TIL-B cells. Surprisingly, we found that D-1MT enhanced anti-tumor effects of the BRAF inhibitor, vemurafenib, in a preclinical model of BRAF mutant melanoma, suggesting that targeting B cells in this setting may be beneficial. Together, our data reveal a novel paradigm for the IDO1 pathway in regulating TIL-B cells, and uncovered IDO1 as a potential targetable mechanism of resistance to BRAF inhibition in melanoma.

Citation Format: Burles Avner Johnson, Adam K. Aragaki, Donna M. Williams, Ophelia Rogers, Li Luo, Lingling Xian, Lionel Chia, Noah M. Hahn, Stephen Desiderio, Theodore S. Johnson, David J. McConkey, Linda M.S. Resar. The indoleamine 2,3 dioxygenase-1 pathway drives intratumoral B cell maintenance [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2765.
Abstract 2768: A novel non-terminal tumor sampling procedure using fine needle aspiration supports biomarker discovery
Authors: Sitnikova Suzanne I. , Munnings-Tomes Sophie , Kentner Stacy R. , Mulgrew Kathy , Espana-Agusti Judit , Zhang Tianhui , Ilieva Kristina M. , Ghadially Hormas M. , Robinson Matthew J. , Wilkinson Robert W. , Dovedi Simon J.
Published: July 2021

Immunotherapy is part of the standard of care for oncology however, durable objective responses remain limited to a subset of patients. As such there is a crucial need to identify biomarkers that can predict/enrich for treatment response. So far, the majority of proposed biomarkers consist of features of the tumor microenvironment (TME). However, in preclinical mouse models, the collection of tumor tissue for this type of analysis is a terminal procedure obviating the ability to directly link potential biomarkers to response/outcome following treatment. Therefore, we have developed and validated a novel non-terminal tumor sampling method to biopsy the TME in mouse models based on fine needle aspiration. We show that this technique enables repeated in-life sampling of subcutaneous flank tumors without impacting tumor growth or animal welfare. This method yields sufficient sample for flow cytometric analysis of tumor-infiltrating immune cells and we demonstrate that it is able to recapitulate results obtained with the current methodology of whole tumor analysis. We propose that this method represents a simple, fast, minimally invasive technique to enable analysis of the TME, reduction in the number of animals used for preclinical oncology studies and refinement of pharmacodynamic analysis. Additionally, its unique ability to provide longitudinal TME sampling can support investigation of biomarkers of response to treatment.

Citation Format: Suzanne I. Sitnikova, Sophie Munnings-Tomes, Stacy R. Kentner, Kathy Mulgrew, Judit Espana-Agusti, Tianhui Zhang, Kristina M. Ilieva, Hormas M. Ghadially, Matthew J. Robinson, Robert W. Wilkinson, Simon J. Dovedi. A novel non-terminal tumor sampling procedure using fine needle aspiration supports biomarker discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2768.
Abstract 2795: Estimation of cell lineage and inter-cell interactions from spatial transcriptomics data
Authors: Ru Beibei , Huang Jinlin , Zhang Yu , Aldape Kenneth , Jiang Peng
Published: July 2021

The spatial transcriptomics (ST) technology has enabled geographical profiling of tumor gene expression. However, each ST spot may detect mixture signals from diverse immune or malignant cells of unknown lineages, and local tissue densities may vary significantly across regions. Therefore, the decomposition of ST cell lineages remains a challenge that cannot be resolved by previous decomposition methods for fixed cell types in bulk tumors. We developed the Spatial Cell Estimator (SpaCE) to infer the cell identities and intercellular interactions for tumor ST data. SpaCE first estimates the cancer cell clonal abundance through modeling the copy number segmental variations across ST spots. Then, we utilized a constrained regression to determine the hierarchy of immune cell lineages and local tissue densities. The prediction accuracy was validated through both simulation and double-blinded pathology annotations on hematoxylin and eosin images. Based on reliable cell lineage inference, SpaCE can further reveal how intercellular interactions affect the pathway and gene activities in distinct regions to modulate the cancer progression.

Citation Format: Beibei Ru, Jinlin Huang, Yu Zhang, Kenneth Aldape, Peng Jiang. Estimation of cell lineage and inter-cell interactions from spatial transcriptomics data [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2795.
Abstract 2796: Pan-cancer characterization of tumor immune microenvironment
Authors: Chu Tianqing , Bei Zhang
Published: July 2021

Background: Anti-PD1/PDL1 therapy has been established as standard care in multiple tumor types, however, many patients will not benefit from this treatment as lack of infiltrating immune cells. Our aim was to explore the pan-cancer immune microenvironment and whether they impacted by ICI biomarkers.

Method: 7-plex mIHC was performed to assessed immune cell infiltrations were assessed. Positive PD-L1 expression was defined as ≥1% of tumor cells with membranous staining. Statistical analyses were performed using R version 3.4.4.

Result: Among 520 samples, CD8+T cell infiltration in tumor was assessed in Esophagus, Liver, Biliary Tract, Lung, Colorectum, Stomach tumors, ordered by descending median density at the center of the tumor. The median density at the periphery was almost double than the median density at the center. In addition, we have observed high TAM.M1 density in Bladder urinary Tract, Biliary Tract, Liver, with a higher or equivalent level of infiltration in stroma than in tumor. While in contrast, the TAM.M2 cell density was higher in stroma than in tumor.

Regarding to PDL1 status in colorectum, there was little difference in the degree of CD8+ T cell infiltration between stroma and tumor. In the term of MSI status, CD8+T cell infiltration was significantly higher in MSIH colorectum tumor.

We compared the proportion of above immune cells. In stroma, except lung tumor, CD8+T cells made up the largest share of immune cells, following by NK cells. In tumor, except for lung cancer, NK cells accounted for the highest proportion. Cancer with the highest proportion of NK cells is bladder/urinary tract cancer. Although the proportion of NK cells in lung cancer is also high, macrophages proportion is the highest, especially TAM.M2.

Conclusions: Together, these data outlined the immune infiltration inter-, extra-tumor, and associations with TMB, MSI status. Tumor immune microenvironment might serve as promising predictive factors for immunotherapy.

Citation Format: Tianqing Chu, Zhang Bei. Pan-cancer characterization of tumor immune microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2796.
Abstract 2803: Classification of lung cancer histology images using deep learning
Authors: YANG Yan , ZHEN Tiantian , SHI Huijuan , XIE Weidong , HAN Anjia , WANG Xunzhang
Published: July 2021

The pathologist's diagnosis on glass slides is the basis for clinical and pharmaceutical research and, more importantly, for the decision on how to treat the patient. Diagnosing pathology slides at microscopy-level is a complicated task that requires years of pathologist training. Only in recent years, computational pathology has emerged to facilitate computer-assisted diagnostics and to enable a digital workflow for pathologists. Meanwhile, deep learning architectures have great potential in pathology imaging, since the enormous success and advancement had been achieved in solving image classification tasks, which would reform the digital workflow with higher efficiency, greater capacity. In this study, we utilize various computational methods based on deep learning framework and build a stand-alone diagnostic tool to effectively classify histopathology images. The dataset of tissue slides consists of 135,000 samples from eligible patients with lung cancer in the First Affiliated Hospital of Sun Yat-sen University. The slide images were intercepted in non-overlapping 1500 × 1000 pixel windows at a magnification of ×40. These tiles above were labeled, and split into a training set, for learning the CNN parameters, and validation set, for validating the model, and test set, for evaluating its performance. Then, features were extracted from the heat map to train a slide aggregation model. The malignant classification of lung cancer by the decision support system include the subtype of adenocarcinoma and squamous carcinoma. An accuracy of 86% for non-cancer (normal) vs. malignant (i.e. adenocarcinoma or squamous carcinoma) on slide tile model, as well as an accuracy of 99% for the multi-label classification on slide aggregation model was obtained in the test set. We believe that our method provides an innovative and reliable means for making diagnostic suggestions, and could be deploy as next-generation, artificial intelligence-enhanced CAD technology in diagnostic pathology,which would empower pathologists' efficiency and accuracy to ultimately provide better patient care.

Citation Format: Yan YANG, Tiantian ZHEN, Huijuan SHI, Weidong XIE, Anjia HAN, Xunzhang WANG. Classification of lung cancer histology images using deep learning [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2803.
Abstract 2807: Variations in glycolytic activity of HCC subtypes as monitored by in vivo h 13C MRI
Authors: Dou Qianhui , Grant Aaron K. , Callahan Cody , Ahmed Muneeb , Tsai Leo L.
Published: July 2021

Hepatocellular carcinoma (HCC) is the 4th leading cause of cancer-related death worldwide. Despite therapeutic advancements, a significant proportion of patients progress, highlighting the need to improve diagnostic approaches to monitor therapy. Radiofrequency ablation (RFA), an accepted locoregional therapy for early HCC, can stimulate more aggressive tumor biology within under-treated or distant tumors. Increased glycolytic activity is a hallmark feature of cancer, but variations in metabolism and its link to tumor aggression can impact the sensitivity of HCC to therapies affecting these pathways. Tumor metabolic phenotypes can be characterized by hyperpolarized 13C MRI (h13C MRI), which can also be used to assess treatment response. In this study, we used orthotopic rat HCC models with two different cell lines N1S1 and MCA-RH7777, and analyzed their mRNA expression of glycolysis-related genes and in vivo glycolytic activity with h13C MRI. We then partially-treated tumors with radiofrequency ablation (RFA) and used h13C MRI to measure in vivo tumor metabolism before and 3 days after RFA to compare them to histologic and phenotypical measures of disease progression.

N1S1 tumors grew only as single lesions, while MCA-RH7777 rapidly produced intra-hepatic and distal metastases. Glycolysis-related genes including HK2, HK3, PFKFB3, and MCT4 were highly expressed in N1S1, but not in MCA-RH7777, while GPT1 was highly expressed in MCA-RH7777 but not in N1S1. In vivo h13C MRI demonstrated significantly increased 13C lactate flux in N1S1 tumor compared to the background normal liver, while 13C lactate flux in MCA-RH7777 rat model did not, consistent with mRNA expression patterns.

Tumor 13C lactate flux was significantly increased and 13C alanine flux was significantly decreased following partial RFA treatment to N1S1 tumors when compared to untreated tumors, which was associated with an increase in tumor Ki67 expression. However, no change was seen on h13C MRI between the RFA and control arms in MCA-RH7777 tumors, even though growth and total tumor burden progressed significantly more following RFA. In contrast to N1S1, there was also no difference between 13C lactate flux and 13C alanine flux within MCA-RH7777 tumors and the neighboring normal hepatic parenchyma, suggesting that glycolytic activity in these tumors did not drive RFA-stimulated progression. Targeting of glycolytic pathways as a potential therapy for HCC may therefore be limited to certain subtypes, and h13C MRI may be used to select for sensitivity.

Citation Format: Qianhui Dou, Aaron K. Grant, Cody Callahan, Muneeb Ahmed, Leo L. Tsai. Variations in glycolytic activity of HCC subtypes as monitored by in vivo h 13C MRI [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2807.
Abstract 2817: Metabolic heterogeneity in breast cancer integrative clusters to screen metabolic vulnerabilities for diagnosis and treatment response
Authors: Ros Susana , Wright Alan J. , Bulat Flaviu , Greenwood Wendy , Hu De-en , Callari Maurizio , Chia Ming Li , Serra Violeta , Rueda Oscar , Bruna Alejandra , Caldas Carlos , Brindle Kevin M.
Published: July 2021

Introduction: Metabolic adaptation occurs in transformed cells and can be exploited clinically for diagnosis and disease monitoring. Metabolic profiles can be influenced by genetic heterogeneity, both within and between tumors. Breast tumors show considerable intra and inter tumor genomic heterogeneity, which partly accounts for the variable clinical course of the disease and response to treatment. Imaging tumor metabolism may provide a metabolic signature that can be used to stratify patients.

Methods: We have investigated eight patient-derived xenograft (PDX) models of breast cancer to determine whether metabolic imaging can be linked to different “integrative clusters” (Int-Cluster), which are defined by their distinctive genomic profiles and clinical outcomes. Metabolic imaging was performed using PET measurements of the uptake, phosphorylation, and trapping of the glucose analog, 2-deoxy-2-[18F]fluorodeoxyglucose ([18F]FDG), catalyzed by hexokinase II, and 13C magnetic resonance spectroscopy (MRS) measurements of hyperpolarized 13C label exchange between [1-13C]pyruvate and the endogenous tumor lactate pool, catalyzed by lactate dehydrogenase A.

Results: Lactate labelling was higher in Int-Clusters dominated by ER- basal-like PDXs from patients with a poor short-term prognosis. Lower levels of lactate labelling were observed in Int-Clusters dominated by estrogen receptor positive (ER+) PDXs and the Pam50 subtypes, Luminal A and B, from patients which have an intermediate to good prognosis. More importantly, tumors that had a similar genomic profile nevertheless showed differences in lactate labeling, that was not observed in the [18F]FDGPET measurements. Although lactate labelling was low in the ER+ tumor models, decreases in lactate labeling could be used to detect treatment response, for example to drugs targeted at the most frequently mutated oncogene in this tumor subtype, PI3KCA.

Discussion: These results illustrate the potential importance of metabolism-based stratification of breast cancer and demonstrate that metabolic imaging can provide diagnostic information that could be important for non-invasive diagnosis and for designing and monitoring therapeutic strategies.

Citation Format: Susana Ros, Alan J. Wright, Flaviu Bulat, Wendy Greenwood, De-en Hu, Maurizio Callari, Ming Li Chia, Violeta Serra, Oscar Rueda, Alejandra Bruna, Carlos Caldas, Kevin M. Brindle. Metabolic heterogeneity in breast cancer integrative clusters to screen metabolic vulnerabilities for diagnosis and treatment response [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2817.
Abstract 2820: A human Claudin-3 monoclonal antibody as a potential multimodal theranostic probe in ovarian cancer
Authors: Oh Sera , Yang Hobin , Oh Ho Rim , Lee Chul-Hee , Kim Young-Hwa , Cheon Gi Jeong , Kang Keon Wook , Shin Young Kee , Youn Hyewon
Published: July 2021

Introduction: Claudin-3 (CLDN3), a tight junction protein, regulates cell-to-cell interactions in epithelial or endothelial cell sheets. During tumorigenesis, epithelial cells are transformed and tumor cells proliferate through out-of-plane division, resulting in external exposure of CLDN3. This alteration of CLDN3 expression is associated with cancer progression, correlating with malignancy in various carcinomas. Since CLDN3 is particularly overexpressed in most ovarian cancers and used as an effective diagnostic marker, we tested the possibility of using a CLDN3-specific antibody as a novel imaging probe.

Materials and Methods: After reducing the CLDN3 specific antibody to expose the -SH group, click chemistry was used to conjugate radioactive isotope 111In or fluorescent protein FNR648. Human ovarian cancer OVCAR-3 cells and human glioblastoma (U87MG) cells were used as CLDN3 positive and negative cells, respectively. Flow cytometry was used to measure the binding of CLDN3 IgG1 monoclonal antibody to theses cell lines. To establish xenograft model, OVCAR-3 cells were injected subcutaneously into the mice. 111In-labeled CLDN3 antibody (370 kBq/50 μl) was administered intravenously to mice with. After 24 hours, organs including tumor were excised and measured with a γ-counter. Images were acquired with IVIS and SPECT/CT.

Results: The labeling efficiency of NOTA-111In or antibody-NOTA-111In was 98.52% or 100%, respectively. FNR648 labeled CLDN3 antibody was bound to the cell surface of OVCAR-3 at 83.4% and to U87MG at 5.7%, respectively. In OVCAR-3 tumor xenografted mice, mice injected with CLDN3 antibody showed 2.5-fold higher tumor uptake (20.4 ± 7.4% ID/g) than mice injected with human IgG (8.8 ± 2.6% ID/g) at 24 hour p.i. The fluorescence signal of CLDN3 antibody peaked at 24 hour p.i.

Conclusion: We successfully conjugated radioisotope and fluorescent protein using the CLDN3 specific antibody. Since the specific binding of CLDN3 antibodies to OVCAR-3 tumors has been validated in a mouse model and diagnostic radionuclides can be replaced with therapeutic radionuclides, this human monoclonal antibody could be used as a useful theranostic probe.

Citation Format: Sera Oh, Hobin Yang, Ho Rim Oh, Chul-Hee Lee, Young-Hwa Kim, Gi Jeong Cheon, Keon Wook Kang, Young Kee Shin, Hyewon Youn. A human Claudin-3 monoclonal antibody as a potential multimodal theranostic probe in ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2820.
Abstract 2833: Mesothelin expression in human tumor types: a tissue microarray study on more than 13,000 tumor samples
Authors: Weidemann Sören , Gorbokon Natalia , Höflmayer Doris , Möller Katharina , Dum David , Luebke Andreas , Kluth Martina , Hube-Magg Claudia , Blessin Niclas , Sauter Guido , Izbicki Jakob , Simon Ronald , Steurer Stefan , Marx Andreas , Krech Till , Jansen Kristina , Lennartz Maximilian
Published: July 2021

Mesothelin is a glycoprotein which is normally expressed in only few non-vital tissues. Due to its membranous location and expression in various cancer types, it represents an attractive molecule for target-specific cancer therapies. To examine, what tumor types might benefit most from anti-mesothelin therapies, a set of tissue microarrays containing samples from 13,298 tumors from 122 different tumor types and 608 samples from 76 different normal tissue categories were analyzed for mesothelin expression by immunohistochemistry. Sixty-six (54%) of these 122 tumor types and subtypes showed at least occasional weak staining, including 49 tumor types with at least one strongly positive sample. Tumor types with highest prevalence of mesothelin positivity included ovarian cancers (serous 97%, clear cell 83%, endometroid 77%, mucinous 71%, Müllerian mixed tumors 65%), pancreatic adenocarcinoma (81%), endometrial cancers (clear cell 71%, serous 57%, Müllerian mixed tumors 50%, endometroid 45%), malignant mesothelioma (71%), and lung adenocarcinoma (55%). Mesothelin positivity was particularly rare or absent in urothelial carcinoma (10% positive out of 925), breast cancer (6% of 1,285), renal cell cancer (7% of 1,049), adenocarcinoma of the prostate (0 of 498), thyroid carcinomas (0 of 238), and soft tissue tumors (0 of 920). Significant associations with patient outcome or parameters of malignancy were not observed in 1,072 breast cancers of no special type, 925 urinary bladder cancers, 386 serous ovarian cancers, 266 lung cancers or 373 stomach cancers. Taken together, our systematic and highly standardized analysis identified numerous clinically important cancer types with frequent and high level mesothelin expression. These tumor types might benefit most from future anti-mesothelin therapies. The prognostic relevance of mesothelin appears to be low in the analyzed tumor types.

Citation Format: Sören Weidemann, Natalia Gorbokon, Doris Höflmayer, Katharina Möller, David Dum, Andreas Luebke, Martina Kluth, Claudia Hube-Magg, Niclas Blessin, Guido Sauter, Jakob Izbicki, Ronald Simon, Stefan Steurer, Andreas Marx, Till Krech, Kristina Jansen, Maximilian Lennartz. Mesothelin expression in human tumor types: a tissue microarray study on more than 13,000 tumor samples [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2833.
Abstract 2857: Asparagine synthetase regulate β-catenin dependent lung cancer metastasis
Authors: Cai Dong-Jing , Hu Cheng-Ping , li Li , Zhang Zi-Yu , Li Yuan-Mao , Li Min
Published: July 2021

Objective: Lung cancer was a malignant tumor with high morbidity and mortality. The lead cause of lung cancer-related death was metastasis. Asparagine synthetase (ASNS) that was a key enzyme to convert glutamine to asparagine played important role in cell metabolism. Based on previous metabolomics data, the high level of asparagine synthase was significantly correlated with tumor metastasis. The purpose of this study was to investigate the role of asparagine synthase in lung cancer metastasis, furthermore for anti-cancer target therapy.

Method: For pursuing high ASNS correlated to metastasis, a cohort were collected from Xiangya hospital. And selected paired lung cancer cell lines were used for discovering potential molecular interactions. Importantly, some key molecules or pathways were found by cell biology study and bioinformatics analysis. In the part of cellular and molecule study, Transwell, scratch test, Western blot, immunofluorescence, immunohistochemistry and co-immunoprecipitation experiments were used. The results of the inquiry were validated by using a mouse metastasis model.

Results: With provided from Xiangya hospital and TCGA cohort, we could validate high ASNS had relations to patient's metastasis and survival. In part of cell biology study, high wild-type ASNS could promote lung cancer cell metastasis and proliferation in vivo and vitro. Overexpression of amidotransferase-dysfunction ASNS had partially potential to drive metastasis but have nothing to increase cell proliferation. Furthermore, Wnt/β-catenin cascades were responsible for tumor mestasis among wild-type and amidotransferase-mutated type groups. The accumulation and nuclear location of β-catenin definitely depend on high ASNS expression even with amidotransferase mutation. With processing, ASNS possible interacted with key proteins located in Wnt/β-catenin signaling pathway to mediate signal conduction. But the activation of ASNS amidotransferase was necessary to promoted tumor proliferation.

Conclusion: ASNS promoted metastasis by mediating Wnt/β-catenin signaling conduction even without ASNS amidotransferase activity, while cell proliferation was required for amidotransferase function. Maybe dual inhibition of ASNS and Wnt/β-catenin would be an effective target for anti-tumor metastasis and proliferation.

Key Words: ASNS, metastasis, lung cancer, β-catenin

Citation Format: Dong-Jing Cai, Cheng-Ping Hu, Li li, Zi-Yu Zhang, Yuan-Mao Li, Min Li. Asparagine synthetase regulate β-catenin dependent lung cancer metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2857.
Abstract 2859: Understanding metastatic cancer biology
Authors: Christensen Ditte Sigaard , Ahrenfeldt Johanne , Birkbak Nicolai Juul
Published: July 2021

Introduction: Metastatic disease is responsible for 90% of all cancer death. Understanding the process of how primary tumors achieve metastatic potential is of great importance and paramount to the development of precision medicine that may limit the aggressive distant spread of metastatic cancer. It is hypothesized that during cancer evolution, cells within the primary tumor obtain metastatic potential through acquisition of specific somatic alterations. However, the defining ability that allows metastasis remains unknown. By comparing genomic profiles of primary and metastatic cancers we wish to investigate if potential metastatic gate-keeper mutations exist, defined as alterations to individual genes or pathways that are required to facilitate metastatic dissemination.

Material and methods: Here, we analyzed panel-based DNA sequencing datasets from the GENIE (Genomics Evidence Neoplasia Information Exchange) project. Analyses were performed on 123 shared genes from the 6 most common panels, selected to include as many patients as possible while analyzing as many genes as possible. In total 43,323 patients had mutations in one or more of the 123 shared genes across 37 different cancer types. Using bioinformatic tools, we compared genomic alterations in primary versus metastatic samples.

Results: Out of the 123 genes we found 26 significantly enriched genes and 30 significantly depleted genes in metastatic disease in at least one cancer type. Overall, TP53 was the most mutated gene across all cancer types and the most enriched gene with significant enrichment in metastatic prostate cancer, non-small cell lung cancer and soft tissue sarcoma. Out of the 30 depleted genes we identified CTNBB1 as significantly depleted in four different cancer types and PIK3CA significantly depleted in two. PIK3CA is one of the most commonly mutated cancer genes, and a target of several precision therapies. Finding of the gene being depleted may indicate a less significant role in the development of metastatic disease. Additionally, we found a higher tumor mutation burden and increased levels of chromosomal instability for metastatic samples compared to primary.

Conclusions: With this analysis we demonstrate how the power of large datasets can be utilized to make novel inferences on cancer biology. We observed significant enrichment in overall mutation counts and copy number alterations. Furthermore, we identified both enrichments and depletions of specific alterations in metastatic disease potentially revealing how certain driver gene combinations associate with cancer progression more commonly than others.

Citation Format: Ditte Sigaard Christensen, Johanne Ahrenfeldt, Nicolai Juul Birkbak. Understanding metastatic cancer biology [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2859.
Abstract 2860: NOSTRIN-induced inhibition of pancreatic cancer progression involves CCBE1 in an eNOS-independent manner
Authors: Zhang Lin , Yang Shouhui , Wang Limin , He Peijun , Yuuki Ohara , Valenzuela Paloma , Tang Wei , Gaedcke Jochen , Ghadim B. Michael , Gaida Matthias M. , Bergmann Frank , Alexander H. Richard , Hanna Nader , Ryan Brid M , Hussain S. Perwez
Published: July 2021

NOSTRIN represses disease aggressiveness in pancreatic cancer by suppressing eNOS-mediated NO production. We have further explored the NOSTRIN-mediated eNOS-independent pathways that contribute to its tumor inhibitory function. Gene expression analysis identified CCBE1 as a target of NOSTRIN. We provide experimental and clinical evidence that NOSTRIN functions a negative regulator in pancreatic cancer progression through inhibiting the expression of CCBE1. Furthermore, CCBE1 expression is regulated by NOSTRIN independent of eNOS. NOSTRIN attenuates protein-protein interaction between ABL1 and PKD2 resulting in the suppression of PKD2 and NFkB p50 activation. Inhibition of PKD2 and NFkB p50 activation results in the suppression of CTCF, which is as a transcription factor of CCBE1. CCBE1 expression is decreased as the down-stream gene of CTCF. Taken together, CCBE1 may be a potential therapeutic target in pancreatic cancer.

Citation Format: Lin Zhang, Shouhui Yang, Limin Wang, Peijun He, Ohara Yuuki, Paloma Valenzuela, Wei Tang, Jochen Gaedcke, B. Michael Ghadim, Matthias M. Gaida, Frank Bergmann, H. Richard Alexander, Nader Hanna, Brid M Ryan, S. Perwez Hussain. NOSTRIN-induced inhibition of pancreatic cancer progression involves CCBE1 in an eNOS-independent manner [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2860.