bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 1914: Acute myeloid leukemia cell and stem-progenitor cell behavior studied in mimetic bone marrow microenvironment
Authors: Simonetti Giorgia , Malferrari Marco , Becconi Maila , Bruno Samantha , Martinelli Giovanni , Rapino Stefania 
Published: August 2019

Abstract
The bone marrow (BM) microenvironment plays a pivotal role in the survival and chemoresistance of leukemia stem cells (LSC) in acute myeloid leukemia (AML). However, conventional culture approaches do not reflect physiological conditions of crucial anatomical districts and signals of incubation-induced dysregulation may confound leukemic transcriptome results. In vivo mimetic cultures of the LSC districts will significantly aid in the disentanglement of LSC mechanisms and in the assessment of the therapy. We engineered Petri dishes able to mimic the “hypoxic” conditions found in the stem cell niches and in BM district. Our approach is based on a patented method which controls the physiological concentrations of oxygen, glucose and pH in the culture environment. Cell proliferation and hypoxia-induced apoptosis in the engineered device were investigated for OCI-AML3 and KG1 AML cell lines. We observed a slight but significant reduction in cell proliferation compared with control cells for OCI AML-3, which is in line with a more quiescent cell condition. No significant differences were obtained by culturing KG-1 cells, which are more resistant to low oxygen concentrations. We have analyzed the effects of cell culturing in the hypoxic dishes on the expression of MYC and p21. Immunofluorescence analysis of HIF-1α levels and HIF-1α nuclear localization as a function of oxygen concentration were determined. Time-lapse experiments of AML cell lines and primary BM cells were performed in the in vivo mimetic device to elucidate the response of CD34+ AML cells to different oxygen concentrations. We tested an innovative cell culturing device able to mimic the dynamical conditions of oxygen content, which are found in vivo. Our results shed light on AML cells and progenitor-stem cells behavior in response to oxygen conditions in the BM niche and allow to disentangle the LSCs complex biology. This ”in vivo mimetic” approach will generate crucial information on the molecular mechanisms preserving LSC in AML, thus providing novel potential therapeutic targets to eradicate leukemia initiating cells in their own microenvironment, contrasting therapy resistance.
Supported by: Associazione Italiana per la Ricerca sul Cancro, MFAG id. 19044.
Citation Format: Giorgia Simonetti, Marco Malferrari, Maila Becconi, Samantha Bruno, Giovanni Martinelli, Stefania Rapino. Acute myeloid leukemia cell and stem-progenitor cell behavior studied in mimetic bone marrow microenvironment [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1914.
Abstract 1927: High-throughput cell line panel drug screening in organoids and 3D systems
Authors: Scales Tim M. , Rada-Kovacs Anett , Pemberton Helen N. , Wiggins Ceri M. , McCarthy Nicola J. , Little Annette S. , Sorrell David A. 
Published: August 2019

Abstract
Cell panel screening of cancer lines grown in standard 2D conditions is a well-established component of the drug discovery toolbox. A sufficiently large cell line panel, reflecting the molecular heterogeneity within a cancer type/sub-type, is required to detect correlations with robust statistical power that aid biomarker identification and patient stratification. However, the heterogenous exposure of tumor cells to nutrients, oxygen and other physical or chemical stresses found in three-dimensional solid tumors also has a role to play in patient response and this is not well mimicked in 2D monolayer cell cultures.
To meet the need for high-throughput cell-panel screening in more complex culture systems, we have developed 3D cell-based screens to more closely model the complex physiological environment found in tumors. We have characterised our OncoSignature panel of 300 cell lines for growth as spheroids to produce a 200 strong panel of spheroid forming lines. Using our high-throughput platform to compare drug activity in 2D and 3D cultures, we have identified clear differentials for some compounds in single agent and drug combination screens. For example, MEK and EGFR inhibitors have significantly higher activity in 3D cultures than 2D cultures, with a greater than 10-fold shift in EC50 values. These results were validated in 3D soft agar assays, with similar compound potencies being observed in the HTS spheroid assay and the lower throughput soft-agar assay.
We have further extended our 3D screening into organoid cultures. Organoid cultures, with increased complexity in terms of structure and cell heterogeneity compared to spheroids, have historically been challenging to use in high-throughput screening. However, our initial results show encouragingly robust data, with compound activity that is congruent with the genetic characteristics of the organoids.
Finally, we used our integrated screening platform to evaluate the power of CRISPR approaches to examine genetic dependencies in KRAS mutant cells in 2D, 3D spheroid and organoid cell cultures. Pooled, next-generation sequencing-linked CRISPRko allows an alternative and complementary genomics approach to arrayed cell panel screening, providing a powerful and robust target identification and validation strategy.
Overall, Horizon’s 3D screening platforms will enable large scale interrogation of both drug and gene interactions in more complex cell culture systems.
Citation Format: Tim M. Scales, Anett Rada-Kovacs, Helen N. Pemberton, Ceri M. Wiggins, Nicola J. McCarthy, Annette S. Little, David A. Sorrell. High-throughput cell line panel drug screening in organoids and 3D systems [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1927.
Abstract 1933: Tissue print Vmem imaging: Visualizing bioelectric signatures in cancer
Authors: Adams Dany S. , Tracey Brian H. , Takiff Larry , Kearns James , Naber Stephen P. , Gaston Sandra M. 
Published: August 2019

Abstract
Work in models of embryonic development shows that membrane voltage, or resting potential (Vmem), regulates cell behaviors that are disrupted in cancer, including proliferation, migration, apoptosis, homeostasis, and cell-cell signaling. Normal cells and tissues maintain stereotypical patterns of Vmem and these patterns are recognizable in Vmem Imaging as bioelectric signatures. In embryonic tissues, cells that are relatively undifferentiated are usually less negative (depolarized) than fully differentiated cells, and different cell lineages have characteristic bioelectric signatures. Recently, the development of innovative imaging technologies using Voltage Sensitive Dyes (VSDs) allows the visualization of Vmem in living cells and tissues. In culture, Vmem Imaging using VSDs can readily differentiate between sister breast cancer cell lines of low and high metastatic potential (EpH4 and EpH4.2, respectively). Although the cultures are identical in bright field images, Vmem Imaging shows a dramatically more heterogeneous bioelectric signature for EpH4.2 than for EpH4 cells, consistent with reduced gap junctional communication. One of the challenges of Vmem Imaging has been the limited thickness of tissue that can be analyzed using the VSDs (about 50 microns). To overcome this limitation, we have combined innovative Tissue Print technologies with Vmem Imaging in order to visualize patterns of electric potential in living cancer cells obtained from human surgical specimens. Tissue Prints are touch preps that transfer a thin layer of viable cells to a nitrocellulose membrane as an oriented imprint; biomarkers identified on these tissues prints can be mapped back to the source tissue to establish pathology and gene expression correlations. Tissue Print Vmem Imaging of radical prostatectomy specimens shows differentiation of bioelectric signatures between cancer and adjacent benign tissue and between cancer of different grades (Gleason pattern 3 and pattern 4). Such differences in bioelectric signatures represent unexplored cancer phenotypes that are made accessible by Tissue Print Vmem Imaging technologies.
Citation Format: Dany S. Adams, Brian H. Tracey, Larry Takiff, James Kearns, Stephen P. Naber, Sandra M. Gaston. Tissue print Vmem imaging: Visualizing bioelectric signatures in cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1933.
Abstract 1936: Label-free optical imaging and characterization of cancer-associated urinary extracellular vesicles: Implications for biomarker discovery
Authors: Barkalifa Ronit , You Sixian , Tu Haohua , Masyr Alison , Kamerer Rebecca , Marjanovic Marina , Park Jaena , Selting Kimberly , Boppart Stephen 
Published: August 2019

Abstract
The discovery of extracellular vesicles (EVs) in all bodily fluids is rapidly increasing the fundamental knowledge of disease mechanisms and the ways in which cells communicate systemically to facilitate tumor progression and metastasis. The type, content, and magnitude of tumor-associated EVs have been linked to tumor invasiveness both in-vitro and in-vivo. In this study we utilized a novel label-free multimodal multiphoton optical imaging method to detect and characterize isolated urinary EVs and classify their optical signatures. Blinded analysis conducted on urinary EVs isolated from 4 healthy dogs and 19 dogs diagnosed with multiple types of cancer showed different optical signatures; cancer-associated urinary EVs were found to have significantly higher NADPH concentrations in comparison to those isolated from healthy dogs. These results suggest a potential label-free optical methodology to detect and characterize EVs by their optical signatures, which can be utilized as possible diagnostic and prognostic biomarkers for cancer.
Citation Format: Ronit Barkalifa, Sixian You, Haohua Tu, Alison Masyr, Rebecca Kamerer, Marina Marjanovic, Jaena Park, Kimberly Selting, Stephen Boppart. Label-free optical imaging and characterization of cancer-associated urinary extracellular vesicles: Implications for biomarker discovery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1936.
Abstract 1937: pH-responsive tumor-targeted mesoporous silica nanoparticle for the identification of pancreatic cancer using optoacoustic tomography
Authors: MacCuaig William , Samykutty Abhilash , McNally Molly , Nairon Kylie , Banks Surya , Grizzle William , McNally Lacey R. 
Published: August 2019

Abstract
Purpose: Due to inadequate early detection and inability to operate at advanced stages, pancreatic ductal adenocarcinoma (PDAC) has remained one of the most difficult types of cancer to treat. A small range of non-specific symptoms coupled with quick metastasis rate result in a poor 5-year survival rate; 14% for those diagnosed within stage IA, and as low as 1% for those diagnosed during stage IV. Nanoparticles have recently emerged as a potential delivery agent for diagnostic and therapeutic agents, and although clinical success has not been ample due to targeting accuracy issues. This work shows a nanoparticle that has been functionalized with a pancreatic cancer-specific targeting ligand and exhibits specific particle release in pancreatic malignant environment (pH 6.6) as compared to non-malignant environments (pH 7.4). Methods: Wormhole-pored mesoporous silica nanoparticles were formed at 80°C using Tetrapropyl orthosilicate (TPOS) and a scaffold of hexadecyltrimethyl-ammonium bromide (CTAB). Acetic acid and ethanol were used in dialysis procedures to remove the CTAB scaffold and create worm-like pores. Chitosan was added to coat the silica particles and serve as a gatekeeper. IR-780 was added before the solution was acidified to load the dye into the worm-hole particles created. Shortly after, the solution pH was raised back to physiological levels (pH=7.4) to trap the dye within the particle. The particle surfaces were functionalized to attach a targeting ligand pH-low insertion peptide (V7) to conjugate the dye-loaded nanoparticles. Pancreatic adenocarcinoma cells (S2VP10 line) were plated in pH-7.4,6.8, and 6.6 PBS solutions with the loaded particles to assess uptake via near-infrared fluorescence and multispectral optoacoustic imaging. Results: Zeta potential and dynamic light scattering were used to ensure the 63nm size nanoparticle and proper coating. Near-infrared fluorescence imaging showed ~10X increased signal at pH 6.6 as compared to pH=7.4. MSOT imaging ~5X increased signal in the malignant microenvironment environment that is acidic as compared to the non-malignant environment at pH 7.4. Conclusion: The functionalized wormhole mesoporous silica nanoparticles coated with chitosan demonstrated pH-sensitivity in terms of cellular uptake via NIR fluorescence and MSOT imaging.
Citation Format: William MacCuaig, Abhilash Samykutty, Molly McNally, Kylie Nairon, Surya Banks, William Grizzle, Lacey R. McNally. pH-responsive tumor-targeted mesoporous silica nanoparticle for the identification of pancreatic cancer using optoacoustic tomography [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1937.
Abstract 1956: ICG pHLIP: A novel agent for fluorescence-guided surgery
Authors: Crawford Troy , Moshnikova Anna , Roles Sean , Carter Lukas M. , Lewis Jason S. , Engelman Donald M. , Andreev Oleg A. , Reshetnyak Yana K. 
Published: August 2019

Abstract
Introduction: Fluorescence imaging has applications in medicine for many image-guided procedures such as mapping and visualization of lymph nodes, targeting and marking cancerous lesions and assessment of tumor margins. Several fluorescent molecules are approved for clinical use including the ICG (indocyanine green) near infrared (NIR) fluorescent dye. We conjugated ICG to a pH Low Insertion Peptide (pHLIP®), which senses and targets the acidity at the surfaces of cancer cells, and evaluated ICG pHLIP® as a targeted imaging agent for tumor visualization. Our construct is progressing toward clinical trials.
Experimental Procedures: ICG pHLIP® was manufactured for proof-of-concept tumor-targeting imaging studies in mice, and toxicity evaluation in mice, dogs and rats. Tumor targeting was assessed in various human and murine tumor models. Biodistribution was evaluated at different time points from 5 min to 48 hours after a single intravenous administration of 0.5 mg/kg of ICG pHLIP®. The distribution of the agent within tumors was evaluated in tumor sections.
Results: The fluorescence of ICG is enhanced about 15 times when pHLIP® tethers it to a cell membrane, significantly improving the contrast index. No toxicity is seen at 25-30x of the proposed human dose, evaluated in mice and dogs. ICG pHLIP® demonstrates excellent tumor targeting, including targeting of 1-3 mm sized tumors in breast. About 10-15% ID/g was observed within tumors at 4 hours p.i., and the signal persists for up to 24 hours with a slight decline at 48 hours. The background signal in major organs decayed after agent administration. The highest uptake, of about 50% ID/g, was observed in liver at 4 hrs p.i. and decayed after that. An excellent correlation was established between ICG pHLIP® NIRF imaging and H&E histopathology at the indicated tumor location.
Conclusions: ICG pHLIP® is a promising new imaging agent for fluorescence-guided surgical applications. It is on the path to clinical translation for accurate resection of breast tumors. Since pHLIP® targeting of tumors is based on the general principle of cancer cell surface acidity, ICG pHLIP® is an agent that may be used in a range of fluorescence-guided surgical interventions.
Citation Format: Troy Crawford, Anna Moshnikova, Sean Roles, Lukas M. Carter, Jason S. Lewis, Donald M. Engelman, Oleg A. Andreev, Yana K. Reshetnyak. ICG pHLIP: A novel agent for fluorescence-guided surgery [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1956.
Abstract 1957: Novel substrates for NanoLuc luciferase with improved brightness and signal duration for bioluminescence imaging in vivo
Authors: Walker Joel R. , Park Yunhee , Lin Michael , Kirkland Thomas A. , Hall Mary P. , Encell Lance P. , Oh Younghee , Liu Lan 
Published: August 2019

Abstract
NanoLuc® luciferase, and its substrate furimazine, are becoming a preferred bioluminescent system in cellular and biochemical assays due to its brightness and glow-type kinetics. The construct Antares, made by fusing NanoLuc to the cyan-excitable orange-red fluorescent protein CyOFP, combines the high catalytic rate of NanoLuc with the high quantum yield and red-shifted emission of CyOFP (584 nm), which is well suited for in vivo imaging. Transgenic mice were created to constituently express Antares and these mice were used to screen coelenterazine analogues for a substrate that showed appropriate distribution and brightness in vivo. Our new Antares (NanoLuc) substrates are administered i.p., have improved solubility over furimazine, exhibit long (&gt;30 min) signal kinetics, and are brighter than D-luciferin/Luc2 expressed on the same promoters.
Citation Format: Joel R. Walker, Yunhee Park, Michael Lin, Thomas A. Kirkland, Mary P. Hall, Lance P. Encell, Younghee Oh, Lan Liu. Novel substrates for NanoLuc luciferase with improved brightness and signal duration for bioluminescence imaging in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1957.
Abstract 1966: Obesity and prostate cancer progression: Role of the chemokine CXCL12
Authors: Ahn Songyeon , Saha Achinto , Kolonin Mikhail G. , DiGiovanni John 
Published: August 2019

Abstract
Obesity is a significant risk factor for several cancers in both men and women. A number of studies have shown that obesity is associated with increased prostate cancer (PCa) progression and higher mortality. However, the mechanism(s) for obesity-driven PCa remain unclear. In earlier studies, we showed that obesity enhances PCa progression in the HiMyc mouse model by changing the surrounding tumor microenvironment, especially the white adipose tissue (WAT). WAT consists of many cells including inflammatory cells, mature adipocytes and adipose stromal cells (ASCs). Here, we further investigated the role of ASCs and ASC derived CXCL12 in obesity-driven PCa progression. In HMVP2 cells, a PCa cell line derived from HiMyc mice expressing high levels of CXCR4, we observed activation of MAPKs (JNK and ERK), AKT, STAT3 and NFkB following treatment with CXCL12. These CXCL12-induced signaling pathways were inhibited by treating with AMD3100, a CXCR4 antagonist, and also by CXCR4 knock-down. To confirm this in vivo, we compared PCa tumor growth in lean and obese mice using the HMVP2 allograft model. Our data demonstrates that obesity promotes EMT in tumor cells and increases invasion of surrounding WAT into the tumor stoma. Treatment of mice with AMD3100 significantly inhibited the growth of HMVP2 tumors and suppressed EMT in both the obese and control diet groups. Furthermore, D-CAN, a hunter-killer peptide developed for targeting ASCs, also suppressed obesity-induced EMT and tumor growth in HMVP2 tumor allografts. Lastly, AMD3100 in combination with D-CAN, showed a significant reduction in PCa growth in obese mice compared to either agent used alone. These results provide evidence that ASCs activate the CXCL12/CXCR4 signaling as well as other pathways playing an important role in obesity-driven PCa progression. Our data suggests this pathway as a novel therapeutic target for inhibiting obesity-driven PCa growth and progression. Research supported by NCI Grant CA196259.
Citation Format: Songyeon Ahn, Achinto Saha, Mikhail G. Kolonin, John DiGiovanni. Obesity and prostate cancer progression: Role of the chemokine CXCL12 [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1966.
Abstract 198: The T3R domain of thrombospondin-1 delays tumor growth and improves tumor response to chemotherapy by remodeling the tumor vasculature
Authors: Pinessi Denise , Resovi Andrea , Morosi Lavinia , Borsotti Patrizia , Berndt Alexander , Giavazzi Raffaella , Zucchetti Massimo , Taraboletti Giulia 
Published: August 2019

Abstract
The disorganized and inefficient nature of the tumor vasculature affects the delivery and activity of antineoplastic treatments. Certain antiangiogenic agents induce a transient normalization of tumor vessels, hence representing a promising strategy to improve the delivery and efficacy of cytotoxic therapy. Our previous studies demonstrated the antiangiogenic activity of the type III repeats (T3R) domain of thrombospondin-1 (TSP-1), a major endogenous inhibitor of angiogenesis. Aim of this study was to investigate the role of the T3R domain in tumor growth and tumor response to chemotherapy. Ectopic tumor cell expression of T3R delayed in vivo growth in two tumor models, the human ovarian carcinoma A2780-1A9 and endometrial adenocarcinoma HEC-1B. This was associated with vascular remodeling, decreased cell proliferation and increased apoptosis. T3R was active in preventing tumor growth also when expressed by the host cells (rather than the tumor cells), following pretreatment of mice with adeno-associated virus (rAAV)-T3R before tumor cell implantation. Expression of T3R also resulted in improved tumor response to paclitaxel (PTX 10 mg/kg Q4x3, i.v.) and cisplatin (DDP 4 mg/kg Q4x3, i.v.). Through the pharmacokinetic study, conducted by HPLC and MALDI imaging mass spectrometry analysis, a higher concentration and a more homogeneous drug distribution were observed in tumors expressing T3R 4h after a single administration of PTX (60 mg/kg, i.v.). This was associated with a marked reorganization of the tumor vasculature that presented decreased vessel area and vessel diameter and increased number of CD31-positive vessels. These findings indicate the ability of T3R to reduce tumor growth and improve tumor response to chemotherapy through the remodeling of tumor vasculature. This sets the basis for the development of new therapeutic strategies based on T3R to be used in combination therapies.
Supported by AIRC. DP is recipient of the AIRC fellowship “Lina e Giovanni Giasini”
Citation Format: Denise Pinessi, Andrea Resovi, Lavinia Morosi, Patrizia Borsotti, Alexander Berndt, Raffaella Giavazzi, Massimo Zucchetti, Giulia Taraboletti. The T3R domain of thrombospondin-1 delays tumor growth and improves tumor response to chemotherapy by remodeling the tumor vasculature [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 198.