bluuebunny/crossref_metadata_2025_split huggingface dataset:
Abstract 3530: Mechanisms underlying Ewing sarcoma-induced osteolysis
Authors: Williams James J. , Kitlinska Joanna B. 
Published: April 2023

Abstract
Ewing Sarcoma (ES) is an aggressive pediatric tumor, which often arises in bones and forms osseous metastases, both of which present as osteolytic lesions. Our previous studies indicated that neuropeptide Y (NPY), which is secreted from ES tumors, stimulates tumor-induced bone degradation via its effect on macrophage recruitment and their subsequent osteoclastic differentiation. This effect was dependent on the autocrine activation of the NPY Y5 receptor (Y5R) expressed by ES cells. However, the exact mechanisms of the osteolytic activity induced by the NPY/Y5R axis remained unknown. To identify the osteolytic factors released from ES cells due to the NPY/Y5R autocrine loop activity, we used shRNA to silence NPY or Y5R in SK-ES-1 ES cells and compared concentrations of known factors involved in the regulation of bone homeostasis in their conditioned media by an antibody array. Four secreted factors were found to be significantly decreased after silencing of NPY signaling: interleukin 8 (IL-8; p&lt;0.005), matrix metalloproteinase 9 (MMP-9; p&lt;0.05), intercellular adhesion molecule 1 (ICAM-1; p&lt;0.005), and monocyte chemoattractant protein 1 (MCP-1; p&lt;0.05). Among these proteins, IL-8 and MMP-9 were selected as the main candidates for mediators of NPY-induced osteolysis. To test their role in ES-mediated stimulation of osteolysis, we co-cultured mouse RAW 264.7 cells, that represent monocyte/macrophage lineage, with SK-ES-1 cells in the presence or absence of the MMP-9 inhibitor (JNJ-0966; 10-7M-10-5M) or anti-IL-8 neutralizing antibody (0.1-0.4µg/ml) and tested their effect on osteoclast differentiation detected by TRAP staining. Inhibiting MMP-9 activity significantly decreased osteoclastogenesis of RAW264.7 cells in dose-dependent manner, while blocking IL-8 had no significant effect on the number of formed osteoclasts. These findings implicate MMP-9 as the mediator of NPY osteolytic activity. Further studies are required to understand the exact mechanisms underlying these effects, as well as potential role of NPY-induced release of MCP-1 in macrophage recruitment to ES tumors.
Citation Format: James J. Williams, Joanna B. Kitlinska. Mechanisms underlying Ewing sarcoma-induced osteolysis. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3530.
Abstract 362: Development of PEP3-IR750 probe to identify adipose tissue and cachexia in murine models
Authors: Chakrabarty Sophia , Han Zheng , McNally Lacey R. 
Published: April 2023

Abstract
Background Cachexia is a multifactorial adipose and muscle wasting disease that is frequently identified in patients with cancer. Cachexia results from increased lipolysis, proteolysis, inflammation, and tumor/host secreted cytokines. Unfortunatly, cachexia is often identified in the clinic at late stages after substantial adipose and muscle loss has occured. Currently, the identification of cachexia in its adipose-wasting stages is difficult due to the lack of a diagnostic imaging tool that easily, objectively, and reproducibly monitors the loss of adipose tissue. Objective identification of fat stores could be used for the early identification and monitoring of cachexia.
Methods We used Fmoc chemistry to synthesize a peptide that binds to fat, i.e. PEP3, CPATAEPC, and scrambled control peptide, AQYLNPS. Following microwave peptide synthesis, we lyophilized, purified, and conjugated the peptides to the Hilyte Fluor750 succinimidyl ester using NHS reaction. Excess dye was removed using dialysis in water. IR750-PEP3 probe was assessed using HPLC and spectroscopy to determine the probe concentrations. Sixty nanomoles of probe reconstituted in 200 µL PBS (pH 7.4). Ex vivo interscapular, perirenal, and omental fat and control muscle tissues were incubated with 20 µM IR750-PEP3 and scrambled probes for 1 h in a 6-well plate, tissues washed with PBS to remove unbound probe, and imaged using near infrared fluorescence. In vivo testing assessed, 60 nmol intravenous injection into healthy female mice. Mice were imaged using MSOT 4 h post injection, images were reconstructed and spectrally unmixed based upon the spectrums of brown fat and IR750 dye. Parametric maps were generated via MATLAB to identify distributions of fat and injected probes. Secondary confirmation of probe biodistribution was conducted using NIR fluorescence of excised tissues.
Results Following injecting 60 nmol of IR750-PEP3, MSOT images confirmed that the PEP3-IR750 dye localized to areas of fat in the interscapular, and perirenal areas in vivo compared to scrambled control (p=0.05, Mann-Whitney U Test). The higher capability of PEP3-IR750 to localize to fat tissue was further confirmed by ex vivo evaluation of PEP3-IR750 and scramble probes-IR750 (p=0.05, Mann-Whitney U Test). Notably, there was a complete lack of signal in the control muscle tissue.
Conclusion PEP3-IR750 can specifically identify fat in mice, and our data supports its potential use in the early identification of cachexia.
Citation Format: Sophia Chakrabarty, Zheng Han, Lacey R. McNally. Development of PEP3-IR750 probe to identify adipose tissue and cachexia in murine models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 362.
Abstract 3621: Identification of podocalyxin-positive tumor buds in pancreatic ductal adenocarcinoma: Implications for solid tumor collective invasion
Authors: Bell Erin M. , Kalloger Steve E. , Dean Pamela M. , Kopp Janel L. , McNagny Kelly M. , Roskelley Calvin D. 
Published: April 2023

Abstract
High expression of the single-pass transmembrane sialomucin, podocalyxin, has been shown to predict poor disease outcome in a number of solid tumor types, including pancreatic ductal adenocarcinoma (PDAC) (Taniuchi et al., 2016). In addition to the classical TMN staging measures of disease progression (tumor size, lymph node involvement, and distant metastasis) local invasion at the primary tumor site is a valuable indicator of solid tumor progression. Local invasion has been observed histologically as tumor buds, defined by the presence of small cohesive clusters of tumor cells in the invasive tumor-stromal interface. To date, tumor budding has been characterized as an indicator of poor prognostic outcome in colorectal carcinoma (CRC), pancreatic ductal adenocarcinoma (PDAC), invasive ductal breast carcinoma (IDC), lung adenocarcinoma (LUA), stomach and esophageal carcinomas, oral squamous cell carcinoma, periampullary adenocarcinoma, and head and neck squamous cell carcinoma (HNSCC). Our group has previously modelled tumor budding experimentally using the MCF7 human breast cancer cell line (Graves et al., 2016). We showed that forced over-expression of podocalyxin was a driver of local collective invasion and tumor budding in vitro and in vivo. Because podocalyxin is highly expressed in PDAC we sought to determine if its expression was involved in the process of tumor budding in this tumor context as well. We assessed large format histological sections from a cohort of patient PDAC tumors and identified high podocalyxin expression in tumor buds. Further, we generated podocalyxin-null MiaPaCa2 PDAC cell lines and demonstrated that podocalyxin-expression is required for collective invasion in 3D culture conditions. We are currently evaluating in vivo xenograft tumor models with these podocalyxin-expressing and podocalyxin-null MiaPaCa2 cells to determine whether changes in 3D culture invasion are representative of tumor budding in this model. Expression of podocalyxin by tumor buds provides rationale to support investigation into the impact of therapeutically targeting these invasive cells.
Citation Format: Erin M. Bell, Steve E. Kalloger, Pamela M. Dean, Janel L. Kopp, Kelly M. McNagny, Calvin D. Roskelley. Identification of podocalyxin-positive tumor buds in pancreatic ductal adenocarcinoma: Implications for solid tumor collective invasion. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3621.
Abstract 3659: RAS drivesmalignancy through stem cell crosstalk with the microenvironment
Authors: Yuan Shaopeng 
Published: April 2023

Abstract
Squamous cell carcinomas (SCCs) are triggered by marked elevation of RAS/MAPK signaling and progression from benign papilloma to invasive malignancy. At tumor-stromal interfaces, a subset of tumor-initiating progenitors, the cancer stem cells (CSCs), obtain increased resistance to chemo and immunotherapy along this path. The distribution and changes in CSCs during progression from a benign state to invasive SCC remain elusive. Here, we show that following HRASG12V activation, CSCs rewire their gene expression program and trigger a self-propelling, aberrant signaling crosstalk with their tissue microenvironment that drives their malignant progression. Surprisingly, the non-genetic, dynamic cascade of intercellular exchanges involves downstream pathways often mutated in advanced metastatic SCCs with high mutational burden. Coupling our clonal skin HRASG12V model with single-cell transcriptomics, chromatin-landscaping, lentiviral reporters and lineage-tracing, we show that aberrant crosstalk between CSCs and their microenvironment triggers angiogenesis and TGFβ-signaling, creating conditions ripe for hijacking leptin and leptin receptor (LEPR)-signaling, which in turn launches downstream PI3K-AKT-mTOR signaling at the benign-malignant transition. By functionally interrogating each step in this pathway, we unravel how dynamic temporal crosstalk with the microenvironment and orchestrated by the stem cells (SCs) profoundly fuels this path to malignancy. These new insights suggest broad implications for cancer therapeutics.
Citation Format: Shaopeng Yuan. RAS drivesmalignancy through stem cell crosstalk with the microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3659.
Abstract 3665: Umbilical cord blood endothelial progenitor cell-derived extracellular vesicles control important endothelial cell functions
Authors: Fraj Sawssen Ben , Naserian Sina , Lorenzini Bileyle , Goulinet Sylvie , Mauduit Philippe , Uzan Georges , Haouas Houda 
Published: April 2023

Abstract
Background: Circulating endothelial progenitor cells (EPCs) play a pivotal role in vascular regeneration and the repair of diseases in which angiogenesis is required, including ischemia and tissue transplantation. Although they are a potentially valuable cell therapy tool, their clinical use remains limited due to suboptimal storage conditions and especially, immune rejection. Even if these cells have been shown to be tolerated by the immune system, their in vivo long-term effects still remain to be elucidated. EPC-derived extracellular vesicles (EPC-EVs) may be an attractive alternative to EPCs given their key role in cell-cell communication and expression of the same parental markers. In this study, we investigated the regenerative effects of umbilical cord blood EPC-EVs on EPCs in vitro.
Methods: EPCs were obtained from human umbilical cord blood (CB) and characterized by flow cytometry. After amplification, EPCs were cultured in a medium containing an EVs-depleted serum (EV-free medium). Then the conditioned medium was used to isolate the EVs with Tangential Flow Filtration (TFF). The EVs were characterized by a series of tests defining their size, number, and protein content. The regenerative effects of EPC-EVs on endothelial cells were investigated by analyzing cell migration, wound healing, and tube formation. We also analyzed the effect of EPC-EVs on endothelial cell inflammation and NO production.
Results: We showed that adding different doses of EPC-EVs on EPCs does not alter the basal expression of the endothelial cell markers CD31, CD144, and KDR, nor change their proliferative potential and NO production level. Furthermore, we demonstrated that EPC-EVs, when used at a higher than physiologic dose, create a mild inflammatory condition that activates EPCs and boosts their regenerative features like migration and tube formation functions.
Conclusions: Our results reveal for the first time that EPC-EVs, when used at a high dose, could enhance EPC regenerative functions without altering their endothelial identity.
Citation Format: Sawssen Ben Fraj, Sina Naserian, Bileyle Lorenzini, Sylvie Goulinet, Philippe Mauduit, Georges Uzan, Houda Haouas. Umbilical cord blood endothelial progenitor cell-derived extracellular vesicles control important endothelial cell functions. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3665.
Abstract 3678: Rewiring of amino acid metabolism in neurofibromatosis type 1-related tumors
Authors: La Spina Martina 
Published: April 2023

Abstract
Neurofibromatosis type 1 (NF1) is a tumor-predisposing genetic disorder caused by heterozygous mutations in the NF1 gene encoding for the tumor suppressor Neurofibromin. Biallelic NF1 inactivation in Schwann cells leads to the onset of neurofibromas, benign neoplasms that can cause severe medical complications and give rise to malignant peripheral nerve sheath tumors (MPNSTs), for which no treatment exists. Neurofibroma cells undergo bioenergetic changes required to sustain their malignant transformation. Our group has shown that inhibition of the mitochondrial chaperone TRAP1, a master metabolic regulator, hampers MPNST cell tumorigenicity. A growing number of evidence indicates that urea cycle (UC) dysregulation can redirect metabolic substrates to specific biosynthetic routes in many tumor cell types. We have found that a complete UC is absent in MPNST cell models, as they lack ornithine transcarbamylase (OTC), a key UC enzyme that converts ornithine into citrulline. In the absence of TRAP1, MPNST cells strongly reduce arginine levels and accumulate ornithine through the activity of the mitochondrial enzymes arginase 2 (ARG2) and ornithine aminotransferase (OAT). These changes can reverberate on glutamine and proline levels ultimately affecting extracellular matrix deposition, motility and ROS homeostasis, as well as on ornithine-dependent polyamine biosynthesis that can regulate cell growth and proliferation. Surprisingly, in the absence of TRAP1 MPNST cells also accumulate citrulline, even though they do not express either OTC or any nitric oxide synthase (NOS) isoform that converts arginine into citrulline. We ascribe citrulline production to the activity of dimethylarginine dimethylaminohydrolase (DDAH) enzymes, involved in the catabolism of asymmetric dimethylarginines. Induction of DDAHs indicates an elevated protein catabolism that could contribute to tune MPNST cell tumorigenicity. A thorough comprehension of the amino acid metabolism rewiring in NF1-related tumors may unveil new targetable liabilities of cancer cells to be exploited for the design of new effective therapeutic approaches.
Citation Format: Martina La Spina. Rewiring of amino acid metabolism in neurofibromatosis type 1-related tumors. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3678.
Abstract 3679: DNAJB1-PRKACA fusion in fibrolamellar hepatocellular carcinoma induces glutamine addiction and an immunosuppressive tumor microenvironment
Authors: Kamdar Zeal , Lopez-Vidal Tamara , Howe Kathryn , Munjal Kabeer , Saeed Ali , Zabransky Daniel , Shu Daniel , Longway Gabriella , Kartalia Emma , Leatherman James , Mohan Aditya , Khare Pratik , Zhang Cissy , Le Anne , Pearce Erika , Furth Mark , Baretti Marina , Leone Robert , Jaffee Elizabeth , Yarchoan Mark 
Published: April 2023

Abstract
Fibrolamellar hepatocellular carcinoma (FLC) is a rare and often lethal form of liver cancer that primarily affects children and young adults. A fusion between DNAJB1, a heat shock chaperone protein, and PRKACA, the catalytic domain of protein kinase A (PKA) has been identified as a signature genomic event in FLC, but the effect of this fusion on the tumor immune microenvironment is not understood. We created an orthotopic, syngeneic model of FLC (TIBx-FLC) by inducing the DNAJB1-PRKACA fusion in a murine hepatoblastoma-derived cell line (TIBx). CD8 T cells isolated from TIBx-FLC tumors demonstrated markedly impaired activation as compared to CD8 T cells isolated from control TIBx tumors. We investigated metabolic programming as a potential mechanism for DNAJB1-PRKACA immunosuppression in FLC. Labeled glucose metabolomics performed on TIBx-FLC and TIBx tumor cells demonstrated a metabolic shift away from aerobic metabolism to an increased glucose contribution towards the hexosamine biosynthetic pathway and purine synthesis, which requires glutamine as a nitrogen source. As compared to the parental TIBx cell line, the TIBx-FLC cell line demonstrated high sensitivity to glutamine antagonism in vitro, consistent with glutamine addiction. Systemic treatment of BALB/c mice bearing TIBx-FLC tumors with JHU-083, a glutamine antagonist, in combination with immune checkpoint inhibitor therapy enhanced survival as compared to vehicle or monotherapy. These data identify altered glutamine metabolism as a target in FLC, and may provide an explanation for immune suppression seen in the FLC tumor microenvironment.
Citation Format: Zeal Kamdar, Tamara Lopez-Vidal, Kathryn Howe, Kabeer Munjal, Ali Saeed, Daniel Zabransky, Daniel Shu, Gabriella Longway, Emma Kartalia, James Leatherman, Aditya Mohan, Pratik Khare, Cissy Zhang, Anne Le, Erika Pearce, Mark Furth, Marina Baretti, Robert Leone, Elizabeth Jaffee, Mark Yarchoan. DNAJB1-PRKACA fusion in fibrolamellar hepatocellular carcinoma induces glutamine addiction and an immunosuppressive tumor microenvironment. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3679.
Abstract 3717: Defining the mechanism of ER receptor transcription complex activity and its effect on epigenetic modulations using super resolution microscopy
Authors: Akhshi Tara , Brown Myles 
Published: April 2023

Abstract
Estrogen Receptor α (ERα) is expressed in ER+ breast cancers, one of the leading causes of death among women. Despite effectiveness of endocrine therapy, a significant number of patients experience resistance to these treatments. One of the major hurdles is lack of comprehensive information about ERα transcriptional complex activity. While biochemical assays have improved our knowledge significantly, it is critical to imply techniques that allow direct visualization of ERα activity and gain spatiotemporal information. We used super resolution microscopy to examine the mechanism of ERα transcriptional activity. We showed that ERα interactions with its co-activators (P300, SRC-3), is influenced upon administration of Tamoxifen and Fulvestrant, an effect that is lost in cells carrying multination in ERα ligand binding site. Additionally mutant cells are no longer able to recruit ERα co-inhibitor (NCORI). Using STORM microscopy, we showed estrogen treatment caused an “open” chromatin (H3K27ac) structure which may represent active areas of gene expression, while Tamoxifen and Fulvestrant treatments caused chromatin to form “closed” structures which likely represent close and inactive phase of chromatin.
Citation Format: Tara Akhshi, Myles Brown. Defining the mechanism of ER receptor transcription complex activity and its effect on epigenetic modulations using super resolution microscopy. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3717.
Abstract 3748: Epstein-Barr virus BART lncRNAs induce IKZF3/Aiolos to maintain EBV latency and promote tumorigenicity in nasopharyngeal carcinoma
Authors: He Songtao , Liu Jiayan , Wing-Yee Mok Bobo , Tsao Sai Wah , Chen Honglin 
Published: April 2023

Abstract
Epstein-Barr virus (EBV) maintains latency in its associated tumors, including nasopharyngeal carcinoma (NPC), expressing very few viral proteins but abundant levels of noncoding RNAs (mainly EBERs and BARTs) in NPC cells. We found that IKZF3/Aiolos is a downstream target of BART lncRNAs in NPC cells. The functions of BART lncRNAs and IKZF3 in EBV latency and pathogenesis of NPC remain elusive. In this study, meta-analysis of eight EBV-associated studies showed that IKZF3/Aiolos was consistently upregulated in EBV-infected NPC cells. Our study further showed that transcription of IKZF3/Aiolos is highly upregulated in EBV-infected NPC cells and clinical tissue samples due to the action of EBV encoded BART lncRNAs, with IKZF3/Aiolos was expressed at higher levels in late stage (stage III & IV) NPC patients than in early stage (stage I & II) disease. Secondly, we found that IKZF3/Aiolos was upregulated in BART-activated NPC cells but downregulated in BART-knockdown NPC cells. In addition, this study further demonstrated that IKZF3/Aiolos modulates transcription of EBV BZLF1 and the cellular gene, LRIG1, to maintain EBV latency and promote NPC tumorigenesis in vitro and in vivo. Our results showed that the EBV lytic reactivator BZLF1 was upregulated in IKZF3/Aiolos knockout or Aiolos inhibitor-treated NPC cells. ChIP-qPCR, immunoprecipitation and immunofluorescence analyses further revealed that IKZF3/Aiolos induces H3K27 deacetylation to silence expression of BZLF1 and maintain EBV latency in NPC cells. Moreover, functional analyses and western blotting showed that IKZF3/Aiolos inhibited the tumor suppressor, LRIG1, to upregulate expression and phosphorylation of the cellular proto-oncogene Erb-B2 for NPC pathogenesis. Sphere- and colony-formation assays demonstrated that IKZF3/Aiolos enhances growth of NPC cells in vitro, and in vivo experiments further showed that IKZF3/Aiolos promotes tumorigenicity of NPC cells in NOD mice. Mechanically, we found that the expression of EBV lytic reactivator BZLF1 and cellular LRIG1 were negatively regulated by BART lncRNA/IKZF3/Aiolos regulatory machinery in NPC cells, while Erb-B2 was positively regulated, which indicated that IKZF3/Aiolos is a new biomarker for NPC and may lead to the development of novel diagnostic tests and treatments for NPC.
Citation Format: Songtao He, Jiayan Liu, Bobo Wing-Yee Mok, Sai Wah Tsao, Honglin Chen. Epstein-Barr virus BART lncRNAs induce IKZF3/Aiolos to maintain EBV latency and promote tumorigenicity in nasopharyngeal carcinoma. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3748.