bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 276: Mir-139 regulates autophagy in prostate cancer cells through Beclin-1 and mTOR signaling proteins
Authors: Benatar Tania C. , Nam Robert , Sherman Christopher , Amemiya Yutaka , Seth Arun 
Published: September 2020

Abstract
Background: Prostate cancer represents the second leading cause of cancer death in men and develops as a result of the accumulation of genetic and epigenetic alterations. We previously identified a panel of five miRNAs associated with biochemical recurrence and metastasis following prostatectomy from prostate cancer patients using NGS-based whole miRNome sequencing and qPCR-based validation analysis. We have shown that expression of miR-139, one of the downregulated miRNAs from this panel, was associated with improved prognosis in patients with localized prostate cancer. Furthermore, miR-139 overexpression inhibited prostate cancer cell growth through signaling pathways mediated in part through downregulation of AXL and IGF1R. Here, we further investigated the molecular mechanisms by which miR-139 could inhibit prostate cancer cell growth.
Methods: We examined miR-139 transfected PC3, DU145 and LNCaP cells by morphology as well as by cell-based assays, confocal microscopy and immunoblotting.
Results and Discussion: We found that treatment of prostate cancer cells with miR-139 resulted in phenotypic changes characteristic of autophagic cells. Conversion of the microtubule-associated protein light chain 3 (LC3-I to LC3-II) was used to monitor autophagy. We observed that the treatment of PC3 and LNCaP cells with miR-139 increased the conversion of LC3-I to LC3-II protein that was specifically inhibited by addition of miR-139 antagomir. The upregulation of LC3 II was further confirmed by confocal microscopy of PC3 cells treated with miR-139. Mammalian target of rapamycin (mTOR) and Beclin1 are two important autophagy-related molecules playing significant roles in different stages. mTOR negatively regulates autophagy machinery. Treatment of cells with miR-139 inhibited both activation and expression of mTOR. We also observed increased levels of phospho-Beclin 1 in PC3 cells treated with mir-139. Beclin 1 is a scaffold protein that assembles components for promoting or inhibiting autophagy and its phosphorylation controls autophagy. The cargo adaptor protein, p62/SQSTM1, interacts with autophagic substrates, delivering them to the autophagosome for degradation, and is degraded during autophagy. We found that during early treatment of cells with miR-139, p62 expression is inhibited, while at later stages, p62 expression accumulates, suggesting that autophagic flux may be blocked. These results suggest that miR-139 is regulating autophagy in prostate cancer cells at least in part through the mTOR and Beclin-1 proteins.
Citation Format: Tania C. Benatar, Robert Nam, Christopher Sherman, Yutaka Amemiya, Arun Seth. Mir-139 regulates autophagy in prostate cancer cells through Beclin-1 and mTOR signaling proteins [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 276.
Abstract 2778: Mapping the extracellular pH gradient of colorectal cancer after immuno-thermal ablation
Authors: Slovak Ryan J. , Coman Daniel , Ludwig Johannes M. , Kim Hyun S. 
Published: September 2020

Abstract
Background: Biosensor imaging of redundant deviation in shifts (BIRDS) is a non-invasive MR imaging modality that maps extracellular pH (pHe) by directly detecting paramagenetically shifted non-exchangeable protons of exogeneous agents. Previously it has been used to characterize various tumors in the rat brain and to monitor the response to loco-regional treatment of liver cancer. The purpose of this study was to investigate the feasibility of utilizing pHe mapping with BIRDS to differentiate ablated and non-ablated tumors in the setting of systemic immunotherapy of murine colorectal cancer (CRC).
Materials and Methods: Eight BALB/c mice (Charles River Laboratories, Wilmington, MA, USA) were implanted with bilateral flank injections of 0.5 × 106 CT26.WT colorectal cancer cells (CRL- 2638; ATCC, Manassas, VA). Mice were randomly assigned into 3 treatment groups; group 1 received injections of sham antibodies (InVivoMAb IgG controls), group 2 was treated with dual anti-PD-1 and anti-CTLA-4 antibodies (Clone J43 & Clone 9D9; BioXcell, West Lebanon, NH, USA), III). Group 3 received cryoablation in combination with systemic injections of dual checkpoint inhibitors. Mapping of pHe was performed between 3-10 days after the initiation of treatment and was immediately followed by sacrifice.
Results: The mean pHe of untreated, control tumors was significantly lower than the average pHe of tumors treated with immuno-thermal ablations (6.6 vs 6.96; p&lt;0.05). Among the mice in the ablation groups, the targeted tumors demonstrated significantly less acidity on average than the distal, off-target tumors (6.96 vs 6.63; p&lt;0.05). The average pHe of tumors in the DICB group was not significantly different than any other group (6.75 vs 6.6, 6.96 & 6.63; p&gt;0.05).
Conclusions: BIRDS MR quantification was feasible for imaging murine CRC model. BIRDS quantification of pHe could serve as an imaging non-invasive biomarker for metabolic changes of the tumor microenvironment in a murine CRC model with Immuno-Thermal Ablation Therapy.
Significance at a GlanceTreatment GroupMean Extracellular pHSignificant DifferencesControl6.6Vs Target Ablated Tumors (p&lt;0.05)Dual Immune Checkpoint Blockade (DICB)6.75NoneTargeted tumors in Ablated group6.96Vs Control (p&lt;0.05) Vs Off-target Tumors (p&lt;0.05)Off-target tumors in Ablated Group6.63Vs Ablated Tumors (p&lt;0.05)
Citation Format: Ryan J. Slovak, Daniel Coman, Johannes M. Ludwig, Hyun S. Kim. Mapping the extracellular pH gradient of colorectal cancer after immuno-thermal ablation [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2778.
Abstract 2780: In vivo quantitative molecular imaging for receptor-specific contrast and image guided surgery of glioblastoma
Authors: Bano Shazia 
Published: September 2020

Abstract
Quantitative, noninvasive imaging utilizing the full potential of antibody targeted nanomedicines has recently emerged as a new paradigm in cancer therapeutics. However, the specificity of these nanomedicines to the molecular targets in in vivo is much controversial.
We have leveraged a molecular-specific approach for the simultaneous imaging and quantitation of in vivo EGFR expression levels, using NIR-activable cetuximab targeted nanomedicine. Using quantitative molecular imaging, the EGFR specificity of these targeted nanomedicines is measured, following i.v. administration in glioblastoma tumors bearing mice.
For the first time we demonstrate, that in vivo specificity of ligand targeted nanomedicines correlates positively with cellular expression levels of EGFR established by flow cytometry (Pearson's r = 0.93, p &lt; 0.0003).Importantly, we show that the EGFR concentrations in glioblastoma (with varying expressions of EGFR) closely match with those previously derived from dual-reporter imaging using fluorescent conjugates.
This first demonstration of non-invasive quantitation of targeted nanomedicine binding introduces an unexplored approach to leverage targeted nanomedicines as a critical component in the design and validation process of cancer therapeutics. Such an informed methodology promises to expedite translation and minimize failures that often occur after substantial investment.
Citation Format: Shazia Bano. In vivo quantitative molecular imaging for receptor-specific contrast and image guided surgery of glioblastoma [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2780.
Abstract 2784: Intratracheally delivered perflurocarbon nanoparticles to enhance radiotherapy against lung cancer
Authors: Wu Lina , Xie Jin 
Published: September 2020

Abstract
Radiotherapy (RT) is a major treatment modality for lung cancer. Despite of technological advances, the five-year survival rate remains low. There is an urgent need of new and effective approaches that can selectively sensitize lung cancer cells to radiation. In this proof-of-concept study, we tested whether intratracheally(IT) delivered perfluorocarbon nanoparticles (PFC NPs) can enhance radiotherapy against lung cancer. We hypothesize that the approach can capitalize on the oxygen-loading capacity of PFC NPs and solve the unfavorable delivery problem associated with systemically injected PFC NPs. Therapy studies were performed in mice bearing subcutaneously inoculated H460 tumors and in orthotopicVX2 tumor models established on New Zealand white rabbits. PET/CT, 19F-MRI, and DCE-MRI were employed to assess PFC NPs delivery and treatment outcomes. 1H-MRS was performed on tumors from different treatment groups and the results were compared with in vivo imaging. For both tumor models, animals were divided into four groups (n=3 for rabbits and 5 for mice): (1) control; (2) PFC NPs only; (3) radiotherapy only; and (4) RT+PFC NPs group. Tumor tissues were subjected to pathology analysis and the results were compared with imaging data. Urine samples were collected and analyzed by UPLC-Q-TOF-MS/MS to assess metastasis. Our results showed that combining IT delivered PFC NPs and radiotherapy led to significantly enhanced tumor growth inhibition. Specifically, 19F-MRI confirmed that PFC NPs can deliver O2 to tumors and increase the mean pO2. DCE-MRI showed distinct tumor blood perfusion behaviors in the RT+PFC NPs group (P&lt;0.05).We also compared the 18F-FDG accumulation in lesions in the early dynamic perfusion phase with the standard static metabolic phase by analyzing the peak value of the SUV mean, time to peak value, and the rate of the SUVmean increase. We found significant difference between the two phases in the RT+PFCNPs group. 1H-MRS found a decreased Cho/Cr ratio in animals treated by RT+PFC, suggesting that the combination treatment improved tumor suppression. Meanwhile, urine glutamine content is elevated in the RT+PFCNPs group, indicating a tendency to return to normal after treatment. Overall, our studies suggest the potential of IT delivered PFCNPs to enhance lung cancer radiotherapy. The delivery of the particles can be image-guided for efficient and site-specific radiosensitization. The approach holds great promise in clinical translation.
Citation Format: Lina Wu, Jin Xie. Intratracheally delivered perflurocarbon nanoparticles to enhance radiotherapy against lung cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2784.
Abstract 2814: Establishment and optimization of a drug-resistant PDX-based platform using in vivo and in vitro approaches
Authors: Zhuang Xiaoying , Xie Nan , Jin Lei , Zeng Wengang , Zheng Yuhui , Zhang Chunling , Cai Liping , Sun Hanmei , Xu Jie , Gao Wenjie , Ji Qunsheng 
Published: September 2020

Abstract
Induced resistance after anticancer agent treatment is the main reason for cancer relapse and short median survival of patients. Here we employed in vivo and in vitro approaches to establish and optimize a drug-resistant platform based on patient-derived xenografts (PDXs). PDXs derived from lung cancer patients who acquired drug resistance reproduced the insensitivity to standard target therapies in mice. Alternatively, resistance was induced in established PDX by continuously exposing with dosage increasing, which demonstrated the induced response declining after the treatment of one second-line therapy drug (Irinotecan). These drug resistant models were additionally confirmed by the in vitro 3D cell based assay, which mimicked the precise histologic structure and cellular composition of the corresponding patient samples. These results from in vivo and in vitro studies displayed good correlation, suggesting that this platform of PDXs and 3D cell based assay can be applied to preclinical development, mechanism research and new drug screen to overcome treatment resistance and have the potential for application to personalized medicine programs.
Citation Format: Xiaoying Zhuang, Nan Xie, Lei Jin, Wengang Zeng, Yuhui Zheng, Chunling Zhang, Liping Cai, Hanmei Sun, Jie Xu, Wenjie Gao, Qunsheng Ji. Establishment and optimization of a drug-resistant PDX-based platform using in vivo and in vitro approaches [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2814.
Abstract 2835: Effector T-cell shape is a potential digital biomarker for assessing CD8 lymphocyte activation in colorectal cancer (CRC) tumors
Authors: Zwing Natalie , Li Xiao , Hibar Derrek , Failmezger Henrik , Nie Yao , Gaire Fabien , Korski Konstanty 
Published: September 2020

Abstract
With the recent advances in cancer immunotherapy it is critical to identify innovative biomarkers, reflecting the interaction between the tumor and its immune microenvironment. As a model system, CRC tumors are considered as non-immunogenic cancers with minor exception of microsatellite instable (MSI) cases that, in contrast to microsatellite stable (MSS) tumors, are characterized by a high tumor mutational burden (TMB) due to defects in mismatch repair (MMR) genes. These characteristics make MSI tumors more visible for the immune system, causing higher lymphocytic influx. When analyzing T cell kinetics, live cell imaging studies shown that T cells elongate while trafficking and are rounded while interacting or resting in the tissue. Based on these initial observations, we hypothesized that by assessing the density, distribution and shape of CD8 cytotoxic T cells in tumor areas, we could find a surrogate combined digital image-derived biomarker reflecting tumor immune microenvironment dynamics. To visualize this fundamental correlation, multiplexed immunohistochemistry was performed on 74 selected cases of primary CRC from our tissue bank collection with subsequent whole slide image analysis trained by machine learning. The CD8 T cell shape was assessed by nuclear eccentricity, along with PD1 and Ki67 activation status of the detected cell. Spatial plots revealed a haphazard T cell distribution in the overall tumor area, locating round and elongated CD8 T cells in both, tumor epithelium and tumor stroma compartments. Further discrimination of CD8 T cells according to PD1 and Ki67 positivity, showed higher activation / exhaustion in the round shaped T lymphocytes. In addition, we correlated the T cell shape with the MMR status and TMB assigned to the patient. Finally, integration of differential gene expression analysis provided insights into the role of the T cell shape in distinct tumor immune microenvironment conditions. With this novel approach, we discovered a new potential digital biomarker associated with the T effector cell function. Our results warrant further validation on bigger CRC patient cohorts, other tumor indications and through expanding the image analysis scope.
Citation Format: Natalie Zwing, Xiao Li, Derrek Hibar, Henrik Failmezger, Yao Nie, Fabien Gaire, Konstanty Korski. Effector T-cell shape is a potential digital biomarker for assessing CD8 lymphocyte activation in colorectal cancer (CRC) tumors [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2835.
Abstract 2844: New breast cancer protein as a potential biomarker in liquid biopsies
Authors: Ibrahim Safaa A. , riehl Valerie , Beaman Kenneth D. 
Published: September 2020

Abstract
The decline in breast cancer mortality over the past 2 decades is primarily the result of advances in early detection and treatment. However, the five year relative survival rate of metastatic breast cancer (MBC) patients is only 27%. This requires the need of new biomarkers for early detection of MBC. Tumors secrete microvesicles (MV) in large quantities to blood circulation in order to communicate with distant cells such as immune cells and prepare the metastatic niche. Our recent studies showed that MV from metastatic breast cancer cells contain a specific peptide from the a2 isoform vacuolar ATPase (a2NTD). This a2NTD is undetectable in mammary epithelial cells MV but expressed abundantly by MV produced by breast tumor cells. We have previously shown that the MV containing a2NTD modulate immune cells, promotes tumor growth, metastasis and positively correlates with the invasiveness of breast cancer cells. Identifying the role of a2NTD in regulating immune cells in liquid biopsy will provide a new biomarker for breast cancer. Treatment of whole blood from healthy volunteers with MDA MB-231 MV or recombinant a2NTD drives the neutrophil transition from High density neutrophils (HDN) to Low density neutrophils (LDN). Flow cytometrical analysis showed that LDN possess the PMN-MDSCs markers CD11b+ CD15+ CD14- CD33- HLA-DR-. In addition a2NTD treatment was associated with decreased internal staining of Arg-1 in neutrophils that suggests the release of Arg-1. This data suggest that a2NTD can reprogram neutrophils to exert immunosuppressive phenotype. Further characterization of surface markers as well as functional assays demonstrate the exact phenotype of neutrophils. Together studying the impact of a2NTD present in MV on immune alteration during metastasis can be potential predictors of the growth and metastasis of breast cancer.
Citation Format: Safaa A. Ibrahim, Valerie riehl, Kenneth D. Beaman. New breast cancer protein as a potential biomarker in liquid biopsies [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2844.
Abstract 2847: Multiplex bead array approach for quantitative profiling of 95 cancer-immunity biomarkers in breast cancer
Authors: Lie Wen-Rong , Davies Alexander , Pepin Danielle , Godeny Michael 
Published: September 2020

Abstract
Immune regulators, including immune checkpoint proteins and cytokines/chemokines, have an emerging role as both disease/treatment biomarkers and targeted agents in cancer therapies. To contribute to the understanding of the roles of these immune regulators in cancer, we have developed 3 new Luminex-based multiplex immunoassay panels, a 48-plex human cytokine/chemokine panel, a 17-plex human immune checkpoint protein panel, and a 30-plex human immune checkpoint protein panel, to simultaneously quantitate the expression levels of 95 key immune regulator proteins in biofluids or cell/tissue homogenates. Here we report the quantitative profiles of these 95 immune regulators in 4 types of breast cancer samples: cancer versus healthy control serum samples, the lysates from breast cancer tumor biopsies versus the adjacent normal tissues, the conditioned media and lysates from the established breast cancer cell lines, and breast cancer cell-derived exosomes. Analysis of the circulating immune protein signatures generated from this multiplex approach reveals an elevated level of IL-6, IL-27, M-CSF, MDC, MIG, sCD27, sTIM3, sCD40, Galectin-3, Galectin-1, FGL-1, BAFF and low level of EGF and sCD40L in breast cancer serum samples compared to the healthy serum controls. The expression profiling of tumor and adjacent normal tissues from 3 patients with metastatic breast cancer revealed differential expression of multiple protein markers including IL-6, IL-8, MIG, IL-1ra, IL-18, IP-10, MCP-1, MIP-1b, VEGF, BTLA, HVEM, CTLA-4, CD40, TLR-2, Siglec-9, CD25, Granzyme B, APRIL, BAFF, Nectin-2, Nectin-4, E-cadherin, and IDO-1 in the matched lysates. We performed similar analysis using cultured breast tumor cells. In these samples we identified markers with significantly altered expression in conditioned media, cell lysates, and tumor cell-derived exosomes from MCF10CA1d tumor cells versus MCF-10A non-tumorigenic mammary epithelial cells as well as triple negative HMT-3522 T4-2 and MDA-MB-231 breast cancer cells. Altogether, our results suggest Luminex-based profiling allows for sensitive and versatile multiplexed analysis of stimulating and inhibitory immune mediators in circulation, tissues, and cell lines which will assist in the discovery of biomarkers and therapeutic targets for cancer interventions.
Citation Format: Wen-Rong Lie, Alexander Davies, Danielle Pepin, Michael Godeny. Multiplex bead array approach for quantitative profiling of 95 cancer-immunity biomarkers in breast cancer [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2847.
Abstract 2854: Multi-omic considerations: Exploring LBgard blood tubes for proteomic analysis
Authors: Vasquez Jacob M. , Desharnais Joel 
Published: September 2020

Abstract
Expanding the number of -omic (genomic, proteomic, etc.) analyte classes, a single blood draw will serve to increase the scientific value of that sample by affording additional multi-variant analyses. The detection and monitoring of many chronic diseases and cancers requires multi-factorial/multi-analyte measurement. Specialized blood collection tubes (e.g., liquid biopsy tubes) are routinely used in the development of minimally invasive methods to monitor key cancer biomarkers in blood. The suitability of plasma isolated from liquid biopsy blood tubes for quantitative proteomics studies needs further exploration. Herein we performed proteomic analyses of whole blood collected and stored in LBgard Blood Tubes.
In this study, matched blood samples from 3 healthy donors were collected in LBgard Blood Tubes and K3EDTA (EDTA) tubes, which is the standard blood collection tube used for plasma proteomic analyzes. Blood collected in LBgard Blood Tubes was stored for either 5 days or 7 days at ambient temperature. Following high-abundant plasma protein depletion, plasma proteins were analyzed in quantitative discovery proteomics by liquid chromatography tandem-mass spectrometry (TMT-LC-MS/MS). Over 800 unique proteins were identified by mass spectrometry in the plasma samples analyzed. Of those, &gt;90% were found to be of similar abundance between EDTA Day 0 plasma samples and plasma isolated from whole blood collected in LBgard Blood Tubes. After storage of the LBgard Blood Tube samples for up to 7 days at ambient temperature, &gt;85% of the proteins demonstrated similar abundance in both stored LBgard Blood samples and EDTA tube samples processed immediately after collection. Samples were also evaluated via ELISA to quantify the levels of proteins known to be cancer related. LBgard Blood Tubes prevented changes in the plasma concentrations of several protein biomarkers known to be sensitive to preanalytical variability in samples stored for 5 days at ambient temperature as compared to samples stored in EDTA tubes under the same conditions.
Utilizing tandem-mass tag spectrometry and immunoassay methodologies, we have determined the compatibility of LBgard Blood Tube for determining proteomic profiles of blood samples. Additionally, blood collected in LBgard Blood Tubes was shown to have consistent concentrations of protein after storing the collected blood sample for several days, with enhanced protein detection as compared to EDTA tubes.
Citation Format: Jacob M. Vasquez, Joel Desharnais. Multi-omic considerations: Exploring LBgard blood tubes for proteomic analysis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 2854.