bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 274: Multifaceted function of microRNA-299-3p fosters an antitumor environment through modulation of androgen receptor and VEGFA signaling pathways in prostate cancer cells
Authors: Ganapathy Kavya , Staklinski Stephen , Hasan Md Faqrul , Ottman Richard , Andl Thomas , Berglund Anders , Park Jong , Chakrabarti Ratna 
Published: September 2020

Abstract
MicroRNAs (miRNAs) play crucial roles in the regulation of prostate cancer (PCa) through modulation of signaling pathways. Androgen receptor (AR) signaling is central to PCa and PCa therapy. The therapeutic suitability of miRNAs that target AR has not been investigated thoroughly. Here, we illustrate the clinical relevance, functional significance and therapeutic benefit of miR-299-3p, an AR targeting microRNA, in PCa progression. We noted a loss of expression of miR-299-3p in prostate tumors compared to noncancerous prostate tissue in clinical specimens. Studies on the impact of miR-299-3p restoration on AR function, phenotypic changes associated with cancer progression and involvement of the downstream components of the AR signaling pathway were conducted using in vitro and in vivo model systems. Replenishment of miR-299-3p in C4-2B, 22Rv-1, and PC-3 cells contributed to cell cycle arrest, reduced cell proliferation, migration and increased expression of apoptotic markers. Additionally, expression of miR-299-3p reduced AR, PSA and VEGFA expression. AGO-RNA pulldown experiment showed enrichment of AR, VEGFA and miR-299-3p in the RISC in the extracts of C4-2B cells overexpressing miR-299-3p. Overexpression of miR-299-3p inhibited epithelial-mesenchymal transition signaling, through inhibition of expression of Slug, TGF-Β3, phospho-AKT, and phospho-PRAS40 and up-regulation of E-cadherin. Replenishment of miR-299-3p showed reduced tumor growth in xenograft mice models. Furthermore, we also tested the benefit of miR-299-3p restoration in combination treatment with enzalutamide and docetaxel which showed a synergistic effect on drug sensitivity. In conclusion, this study has identified novel mechanisms of antitumor and antimigration functions of miR-299-3p through modulation of AR and VEGFA signaling pathways which leads to improved drug sensitivity of PCa.
Citation Format: Kavya Ganapathy, Stephen Staklinski, Md Faqrul Hasan, Richard Ottman, Thomas Andl, Anders Berglund, Jong Park, Ratna Chakrabarti. Multifaceted function of microRNA-299-3p fosters an antitumor environment through modulation of androgen receptor and VEGFA signaling pathways in prostate cancer cells [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 274.
Abstract 2750: An efficient multiplexing approach to avoid batch effects in single cell RNA- and Ab-seq studies demonstrated in a mouse model of chronic inflammation and obesity
Authors: Song Hye-Won , Baracho Gisele , Taylor Ian , Widmann Stephanie , Mortimer Stefanie 
Published: September 2020

Abstract
Obesity can weaken the body's immune system and reduce its ability to fight off infections. Current analysis on immune cells from obese mouse models suggests that the immune cells suffer from chronic inflammation. To understand the root cause of this problem, we used single cell sequencing to examine thousands of immune cells isolated from primary and secondary lymphoid organs as well as adipose tissue and compared a diet-induced obesity mouse model with control mice. Cells were stained with 30 BD® AbSeq DNA-barcoded antibodies to enable multiomic analysis, i.e. examining protein alongside mRNA expression in tandem. We also utilized DNA-barcoded universal antibodies from the BD® Single-Cell Multiplexing Kit, which allowed us to combine 8 samples from different mice and tissue types into a single pooled sample, significantly reducing experimental scale and cost while eliminating potential batch effects. The pooled samples were loaded on the BD Rhapsody™ system to perform cell lysis and individual mRNA and cell barcoding, allowing measurement of ~400 immune-related mRNAs and 30 proteins at the single cell level. We were able to efficiently de-multiplex the pooled samples after sequencing. The targeted mRNA and AbSeq panel provided robust clustering of immune cell types and showed that genes related to critical immune responses, including inflammation and lymphocyte activation, are differentially regulated in specific immune-cell subsets in the obese mouse model. Using this multiomic analysis of genes differentially regulated in immune cells from different tissues, we propose a model to explain the immuno-phenotype we observed in obese mouse.
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Citation Format: Hye-Won Song, Gisele Baracho, Ian Taylor, Stephanie Widmann, Stefanie Mortimer. An efficient multiplexing approach to avoid batch effects in single cell RNA- and Ab-seq studies demonstrated in a mouse model of chronic inflammation and obesity [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 2750.
Abstract 2759: Immune cell killing assay for analyzing the effects of immunomodulating agents in vitro
Authors: Mäki-Jouppila Jenni H. , Halleen Jussi M. , Fagerlund Katja M. 
Published: September 2020

Abstract
Immuno-oncology has emerged as one of the most promising new approaches in cancer drug development. Greater cancer cell specificity of treatments can be achieved when the immune cells of a patient are used against tumors. Therefore, it is important to study the effects of immune cells on cancer cells using different models in vitro and in vivo. Our aim was to validate an in vitro T cell killing assay where activated peripheral blood mononuclear cells (PBMCs) target different cancer cells and increase their apoptosis.
MCF-7 human breast carcinoma cells (ATCC) were used as target cells and were seeded on 96-well plates. 24 hours later, human PBMCs (Lonza) were added to the culture as effector cells. Simultaneously, the PBMCs were activated with 100 ng/ml anti-CD3 and 10 ng/ml IL2. Apoptosis was detected using IncuCyte Annexin V reagent, which was introduced to the cells at the same time with activators. Cell growth, proliferation and apoptosis were monitored with a live cell imager IncuCyte S3 (Sartorius) for three days. Cell viability was determined by CellTiter Glo (Promega) at the end of the study.
The target-to-effector ratio of 1:5 and 1:10 both showed an increase in apoptosis of MCF-7 cells in the wells with activated PBMCs. This was detected with a confluence parameter (percentage) of IncuCyte S3.
The T cell killing assay can be utilized for studying cancer therapeutics based on immune-oncology (e.g. pembrolizumab and atezolizumab) and to evaluate the potential of immune cells in killing cancer cells in the presence of drugs affecting the target-effector interaction.
Citation Format: Jenni H. Mäki-Jouppila, Jussi M. Halleen, Katja M. Fagerlund. Immune cell killing assay for analyzing the effects of immunomodulating agents in vitro [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 2759.
Abstract 2760: A mouse orthotopic model of pancreatic cancer and response to treatment
Authors: Rainbolt Elizabeth , Cecil-Taylor Madelyn , Wong Andrew , Ball Robin , Wood Patrick , Wood Edgar R. , de Marval Paula Miliani , Yeyeodu Susan 
Published: September 2020

Abstract
The mouse pancreatic ductal adenocarcinoma PAN02 was produced in C57BL/6 mice exposed to 3-methyl-cholanthrene. PAN02 is generally resistant to standard cancer therapeutics, thus providing an appropriately rigorous model for the development of clinically effective agents to treat aggressive and intractable pancreatic ductal adenocarcinoma. Subcutaneous implants of syngeneic PAN02 provide the dual advantages of being able to closely monitor changes in tumor growth without compromising the complex pro- and anti-tumor immune responses of the host that collectively determine the outcome of tumor progression. Unfortunately, subcutaneous PAN02 tends to grow slowly and tumor ulcerations are common which may result in tumor regression or loss of animals in a study. Here we report the growth properties and response to therapy of orthotopically implanted PAN02. To establish the orthotopic model, we implanted cells by direct injection into the pancreas through an incision in the abdominal wall at two implant densities, 3 × 105 and 3 × 106 PAN02 cells in 30 μl. Both cell inocula demonstrated progressive growth with median survival of thirty-seven and thirty days post implant respectively. We chose 3 × 106 PAN02 cells to evaluate efficacy of cytotoxic agents gemcitabine and oxaliplatin. Dosing was initiated 7 days after implant; gemcitabine was dosed at 120 mg/kg, by intraperitoneal injection (IP) every third day for four doses and oxaliplatin was dosed at 10 mg/kg IP once a week for 3 weeks. Gemcitabine and oxaliplatin increased life span by 9.0 days and 20.5 days respectively over vehicle control. The morphology of the tumors revealed by histological staining is also presented. The development of this orthotopic pancreatic model provides a more clinically relevant model for evaluation of novel therapies compared to subcutaneous implantation.
Citation Format: Elizabeth Rainbolt, Madelyn Cecil-Taylor, Andrew Wong, Robin Ball, Patrick Wood, Edgar R. Wood, Paula Miliani de Marval, Susan Yeyeodu. A mouse orthotopic model of pancreatic cancer and response to treatment [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 2760.
Abstract 2763: Identification of mouse embryo genes in human cancer using cBioportal
Authors: Xiong Miranda , Wang Christina , Chen Eric , Hu Arthur , Luo Raymond 
Published: September 2020

Abstract
Cancer is the second leading cause of death, often without a cure. There are many genetic factors that can either cause cancer, worsen existing conditions, or shorten a person's life span; however, it is not fully understood. During embryogenesis, many genes are expressed during a short window of the developmental stages and then switched off in adults. It is known that some of the embryogenesis specific genes are switched on in tumors. The purpose of this study was to identify genes that are exclusively expressed in the central nervous system (CNS) of the E14.5 mouse embryo (Dlez-Roux, G. et al) associated with cancer to provide a foundation for future research.Using cBioPortal, we queried the TCGA PanCancer Atlas Studies (32 studies and 10,967 samples) by each of the 80 genes expressed in the CNS of E14.5 mouse embryo. The data types analyzed included cancer type percentages and survival estimate (overall and disease/progression-free Kaplan-Meier estimate). To identify genes that are relevant in human cancer, the 80 genes were filtered based on three criteria: whether the gene mutations were found in at least 10% of various cancer types, whether the p-value for either the overall survival Kaplan-Meier Estimate, and of the disease/progression-free Kaplan-Meier Estimate was equal or less than 0.05. Fourteen genes carried mutations in more than 10% of various cancer types. Mutations in nine genes significantly decreased cancer patients' overall survival, but mutations in one gene significantly improved overall survival. Mutations in ten genes also significantly decreased the patients' disease/progression-free survival. Three genes demonstrated a high mutation rate (&gt;15%) and significantly reduced patients' survival in both overall survival and disease/progression-free estimates. None of these genes have been identified as cancer driver genes. We report here that many of the CNS E14.5 mouse embryonic genes are mutated in human cancers and some of the mutations may predict cancer patient outcome. Using the same approach, we will evaluate whether these genes are correlated with the manifestation of all cancers or a subset of cancer types. In the future, we would like to obtain the opportunity to study the role of these genes in cancer development or cancer treatment in a laboratory. Dlez-Roux, G. et al: A high-resolution anatomical atlas of the transcriptome in the mouse embryo. PLoS Biol. 2011This study was supervised by Dr. Yangjun Qin, and all authors contributed equally.
Citation Format: Miranda Xiong, Christina Wang, Eric Chen, Arthur Hu, Raymond Luo. Identification of mouse embryo genes in human cancer using cBioportal [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 2763.
Abstract 2776: Real-time measurement of temperature and tissue damage during laser photothermal ablation
Authors: Fathi Faraneh , Maddirela Dilip R. , Gogineni Venkateswara Rao , White Sarah B. , Yu Bing 
Published: September 2020

Abstract
Hepatocellular carcinoma (HCC) is the sixth most common cancer and tumor resection is feasible in only 25±30% of these patients. Thermal ablation therapies using lasers, radiofrequency waves and microwaves are considered as an alternative treatment for unresectable tumors. Our goal was to develop optical fiber-based laser ablation while monitoring the changes in temperature and tissue properties in real-time. A portable diffuse reflectance spectroscopy (DRS) system consisting of a DRS probe with self-calibration and fiber optic temperature sensor was made. Tissue mimicking phantoms using human hemoglobin powders and polystyrene particles with fixed total number of scatterers were used to calibrate the system. In vivo studies were performed in 8 rats orthotopically implanted with HCC. Temperature sensor was inserted in the center of the tumor via hypodermic needle and DRS fiber was overlaid on the temperature sensor. Ablation was performed by a fiber optic diffuser connected to an 808nm laser using different power settings in pulse mode. The tissue temperature and optical properties were continuously monitored until both plateaued using a custom LabVIEW program. Rats were immediately euthanized, and the ablated liver tissues were collected for histological analysis. The DRS showed average errors of 6.31% and 4.74% in measuring the absorption (μa) and reduced scattering coefficients (μs) of the phantoms respectively. Tumor bearing liver tissues showed a decreasing trend in both μa and μs irrespective of the laser power (1W-2.5W) and the temperatures recorded (50°C to 88°C) in different animals. Histological analysis showed discrete zones of tissue damage from treated regions with hemorrhage in the outer most zone. Changes in optical properties of the ablated tumors with temperatures and histological changes needs further correlation by performing more in vivo studies.
Citation Format: Faraneh Fathi, Dilip R. Maddirela, Venkateswara Rao Gogineni, Sarah B. White, Bing Yu. Real-time measurement of temperature and tissue damage during laser photothermal 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 2776.
Abstract 2790: Evaluation of S100A9 targeted nanoparticles containing rapamycin to treat pancreatic adenocarcinoma
Authors: Thomas Karl , McNally Molly , Samykutty Abhilash , McNally Lacey R. 
Published: September 2020

Abstract
Despite Pancreatic cancer being expected to contribute to only 3% of new cancer cases in both males and females in 2019, it is expected to cause an estimated 45,750 deaths making it the third highest contributor to cancer deaths. This is due to the low survivability associated with pancreatic cancer which has a 5-year survival rate of only 9%. In order to address the poor prognosis common with pancreatic cancer, biomarkers are being used to create therapies targeted to specific cancer cells. The S100A8/A9 complex is a heterodimer, known for modulating the inflammatory response, and it is upregulated in a variety of cancers including pancreatic cancers. For this reason, we selected it as a targeting agent to deliver Rapamycin to pancreatic cancer cells via a mesoporous silica-encased gold nanorod (MS-GNR). The use of this nanoparticle allows an anticancer molecule to be encapsulated within the mesoporous silica matrix by further encapsulation of the nanoparticle with the pH sensitive polymer chitosan. A low pH tumor microenvironment induces relaxation of the chitosan surrounding this chitosan-capped mesoporous silica gold nanorod (CMG) allowing for the anticancer molecule to be released only in cancer cells. The gold nanorod at the core of this enables these nanoparticles to be imaged in real time via Multispectral Optoacoustic Tomography (MSOT). Using these tools, we linked S100A9 to a mesoporous silica-encased gold nanorod which contained Rapamycin held in via the chitosan polymer (S100A9-Rap-CMG). MSOT imaging revealed a strong uptake in S2013Q and MiaPaca pancreatic cancer cells. Further tests showed that at a pH 6.8 S100A9-Rap-CMG treatment resulted in an increase in cell death by 69% compared to treatment at physiologic pH of 7.4 by the S100A9-Rap-CMG's which did not induce cell death. This data indicates the feasibility of S100A9 as a targeting agent for pancreatic cancer cells and the potential benefits of nano-drug delivery of Rapamycin to treat pancreatic adenocarcinoma. The ultimate benefit of the nano-drug delivery includes both increased tumor targeting and mitigating premature drug release and offsite delivery.
Citation Format: Karl Thomas, Molly McNally, Abhilash Samykutty, Lacey R. McNally. Evaluation of S100A9 targeted nanoparticles containing rapamycin to treat pancreatic adenocarcinoma [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 2790.
Abstract 2824: In vitro tumor killing assays for predicting cellular therapeutic efficacy
Authors: Nunan Robert , Yap Michelle , Garcia-Alvarez Enrique , Carkill Marie , Pascoe Anna , Brackenbury Louise , Self-Fordham Jezrom , Jenkinson S. Rhiannon 
Published: September 2020

Abstract
Cellular immunotherapy is emerging as a fifth pillar of cancer treatment. Clinical efficacy has been proven for a variety of cancers; however, its true potential has yet to be unlocked. Development of reliable and mechanistically appropriate in vitro screening assays that recapitulate in vivo immune-tumor interactions have been outpaced by in vivo living cell therapy efficacy models. Here we describe a range of in vitro primary human assays to model tumor killing by immune cells. This platform addresses multiple anti-tumor immune effector pathways and has been validated with standard of care therapeutics. Fluorescently labelled target cells (PDX-cell lines, tumor cell lines or human primary/iPSC derived cells) screened for relevant antigen expression were cultured in monolayer or spheroid forms and co-cultured with the cellular therapeutic. On target killing by effectors is driven by binding of the TCR or CAR to its cognate antigen (MHC/peptide or protein, respectively) with readouts of cell killing utilising live cell imaging (IncuCyte) and flow cytometry based approaches to measure target cell growth and apoptosis. Additional analysis of soluble mediators can determine the extent of effector cell activation. To discriminate between ‘on' and ‘off' target killing by cellular therapeutics, primary/iPSC-derived normal cell types representative of different tissue types were screened for expression of an HLA subtype capable of binding exogenously applied tumor-associated antigen (TAA) prior to co-culture. Under these conditions in the case of a TCR based cellular therapeutic, efficacy can be screened for, or in the absence of the putative TAA expression, the extent of off-target killing determined. Enhancement of tumor targeting and killing by immune cells is an attractive therapeutic modality partly due to the clinical success of checkpoint inhibitors which can enhance the activation and effector function of endogenous T/NK cells or cellular therapeutics. The presented data highlight the importance of assays which can recapitulate tumor-immune interactions and include immune effector pathways known to potentiate anti-tumor immune responses. These assays represent valuable screening tools capable of informing cellular therapeutic target selection and refinement before moving to more complex in vivo models.
Citation Format: Robert Nunan, Michelle Yap, Enrique Garcia-Alvarez, Marie Carkill, Anna Pascoe, Louise Brackenbury, Jezrom Self-Fordham, S. Rhiannon Jenkinson. In vitro tumor killing assays for predicting cellular therapeutic efficacy [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 2824.
Abstract 2898: A biparatopic HER2xHER2 antibody drug conjugate with potent anti-tumor activity
Authors: Bay Andres Perez , Faulkner Devon , Tiwari Shiwani , Castanaro Carla , Zheng Xiang , Kunz Arthur , Nittoli Thomas , Han Amy , Olson William , Thurston Gavin , Daly Christopher , Andreev Julian 
Published: September 2020

Abstract
A biparatopic HER2xHER2 antibody drug conjugate with potent anti-tumor activity.
The antibody drug conjugate (ADC) Trastuzumab-emtansine (T-DM1) targets the well-characterized breast cancer oncogene HER2 and is active in tumors that express very high levels of HER2 protein. However, tumors expressing intermediate levels of HER2 remain resistant to T-DM1 therapy, apparently due to insufficient lysosomal trafficking of T-DM1. We generated a series of biparatopic HER2xHER2 antibodies in which each arm recognizes a distinct epitope of HER2. One of our HER2xHER2 antibodies bound to cells with greater avidity than Trastuzumab and formed visible cell surface clusters that were efficiently internalized. While some of the internalized HER2xHER2 antibody trafficked to lysosomes and promoted HER2 degradation, a significant fraction recycled back to the plasma membrane through recycling endosomes. To exploit the increased internalization and recycling of the HER2xHER2 antibody, ADCs were generated by site-specific conjugation of Tubulysin A payload through a protease-cleavable linker. Recent data suggest that cleavage of this linker does not require lysosomal trafficking, but can occur in recycling endosomes as well. HER2xHER2-Tubulysin killed cell lines expressing high (IHC3+) and intermediate (IHC2+) HER2 levels with subnanomolar IC50, while exhibiting very weak killing of cells expressing low (IHC1+) HER2 levels (IC50 greater than 10nM). HER2xHER2-Tubulysin induced complete and durable tumor regression and outperformed T-DM1 in a collection of IHC3+ and 2+ tumor xenograft models (including PDX). Our work shows that a detailed understanding of antibody trafficking can enable the design of more effective ADCs and suggests that HER2xHER2-Tubulysin is a potential clinical candidate for HER2 IHC2+ breast cancer.
Citation Format: Andres Perez Bay, Devon Faulkner, Shiwani Tiwari, Carla Castanaro, Xiang Zheng, Arthur Kunz, Thomas Nittoli, Amy Han, William Olson, Gavin Thurston, Christopher Daly, Julian Andreev. A biparatopic HER2xHER2 antibody drug conjugate with potent anti-tumor activity [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 2898.