tbuckley/cabot-search huggingface dataset:
GENE-37. ATRX INACTIVATION DISRUPTS GLOBAL HETEROCHROMATIN LANDSCAPES AND ALTERS DISEASE-RELEVANT TRANSCRIPTIONAL ACTIVITY
Authors: Carla Danussi, Anand K Singh, Grant Fischer, David J Picketts, Kasthuri Kannan, Kunal Rai, Jason T Huse
Journal: Neuro-Oncology (2018)

Diffusely infiltrating gliomas feature loss-of function mutations in the SWI/SNF chromatin remodeler gene ATRX as defining molecular alterations delineating major adult and pediatric disease subtypes. So far, ATRX inactivation in cancer has been mainly correlated with alternative lengthening of telomeres, however, we recently reported that Atrx deficiency drives glioma-relevant phenotypes, such as increased motility and astrocytic differentiation profiles, by directly modulating epigenomic landscapes and the corresponding transcriptional profiles in glioma cells of origin. In particular, Atrx deficiency was associated with disruptions in H3.3 histone content at key genetic loci. To further understand the downstream epigenomic dysfunction induced by ATRX deficiency, we compared genome-wide chromatin-state maps of Atrx+ and Atrx- primary murine neuroepithelial progenitors (mNPCs). This ChIP–seq analysis revealed major differences in the localization of heterochromatin repressive marks H3K9me3 and H3K27me3. Specifically, we identified peculiar locations in the genome displaying H3K9me3 depletion and gain of H3K27me3 upon Atrx inactivation. Interestingly, these regions were flanked by Atrx binding sites and perfectly co-localized with Lamina-Associated Domains (LADs). LADs are widely involved in the control of gene expression programs during lineage commitment and terminal differentiation; typically they have a cell-specific distribution and are very dynamic during differentiation stages. Gene Set Enrichment Analysis confirmed that the genes corresponding to newly formed LADs in mNPC-to-astrocyte differentiation are significantly enriched for genes down-regulated in Atrx deficient mNPCs and belonging to Gene Ontology categories such as cell cycle, chromosome organization and DNA damage. On the other hand, genes corresponding to decreased LAD association are enriched for up-regulated genes in Atrx- mNPCs and for regulation of differentiation, adhesion and cell death. These data are in perfect agreement with our previously described glioma-relevant phenotypes associated with Atrx deficiency and indicate a novel role of Atrx in regulating the spatial organization of heterochromatin and its underlying transcriptional activity.
TMIC-32. INHIBITION OF MerTK MODULATES GLIOMA-ASSOCIATED MACROPHAGES AND MICROGLIA IN TUMOR MICROENVIRONMENT
Authors: Yu-Ting Su, Funita Phan, Lee Hwang, H Shelton Earp, Mark Gilbert, Jing Wu
Journal: Neuro-Oncology (2018)

Glioma-associated macrophages and microglia (GAMs) are predominant immune cells in a glioma microenvironment. Receptor tyrosine kinase MerTK is highly expressed in GAMs. Activation of MerTK triggers efferocytosis and polarizes GAMs to an immunosuppressive phenotype, promoting tumor growth. We hypothesize that inhibiting MerTK by UNC2371, a small molecule MerTK inhibitor, may lead to a less immunosuppressive glioma microenvironment. Monocytes were isolated from the blood of healthy human donors followed by differentiation and stimulation into proinflammatory (CD80+/CD86+) and anti-inflammatory (CD163+/CD206+) macrophages by granulocyte-macrophage colony-stimulating factor (GM-CSF)/Interferon-gamma and macrophage colony-stimulating factor (M-CSF)/Interleukin-4 respectively. Phenotyping of the macrophages was done by flow cytometry analysis of the cell surface markers. Cytotoxicity of UNC2371 in GBM cells and macrophages was determined by cell viability. Cytokine profiling from the macrophages and a human microglial cell line was performed using cytokine array analysis. The protein expressions of arginase I, interleukin-8, and MerTK were quantified by Western blotting. We demonstrate that UNC2371 inhibits the growth of GBM cells with EC50 of 95.5 nM in U251 glioma cells, but not macrophages. While cultured in U251 conditioned medium (CM), a decrease in CD163+/CD206+(anti-inflammatory) macrophages was observed in 43% of healthy donors (n=7) after UNC2371 treatment, suggesting a proinflammatory polarization. UNC2371-treated macrophages or microglia reduces secretion of Interleukin-8, a chemokine known to promote gliomagenesis, was detected by cytokine array analysis. UNC2371 decreases protein expression of MerTK, Interleukin-8, and arginase I in GAMs. These data suggest that UNC2371 inhibits MerTK signaling in GAMs and alleviates immunosuppression in the glioma microenvironment. Our findings suggest that UNC2371 is a promising drug to polarize GAMs towards proinflammatory, suppress pro-glioma cytokines and modulate the glioma microenvironment. Preclinical evaluations of UNC2371 as a tumor microenvironment modulator using animal models are currently underway.
PATH-48. FUSION TESTING IN ADULT VERSUS PEDIATRIC LOW AND HIGH GRADE BRAIN TUMORS FOR ELIGIBILITY FOR TRIALS
Authors: David Ormond, Denise Damek, Nicholas Willard, Mark Ewalt, Bette Kleinschmidt-DeMasters
Journal: Neuro-Oncology (2018)

Since January 2017, our molecular laboratory has assessed 47 pediatric and 23 adult primary brain tumors for fusions that might result in novel treatment; adults with glioblastoma (GBM) had been screened specifically for the STARTRK-2 study. This trial has open arms for patients whose tumors demonstrate ROS1, ALK, or NTRK 1/2/3 fusions. We now review our results in pediatric versus adults. MATERIALS AND Fusion analysis was performed with the Archer FUSIONPlex Solid Tumor Kit, 1/2017-5/2018. 18 adults with GBMs (17/18 IDH-wildtype) screened for the STARTRK-2 study were negative for fusions or potentially oncogenic transcripts in ALK, ROS1 or MET. However, one adult IDH-wildtype GBM showed a FGFR3-TACC3 actionable fusion that might allow erdafitinib therapy. Three more were found to have fusions of uncertain significance: an IDH-wildtype GBM with EGFR-SEPT14 fusion, an IDH-wildtype GBM with EGFR-PSPH fusion, and the sole IDH-mutant GBM with ZMIZ1-FGFR2 fusion transcript. 5 additional adults were screened for BRAF V600E mutation / BRAF-KIAA1549 fusion, of which 1 fusion and 1 mutation was identified. 42 pediatric patients with low-grade glial/glioneuronal tumors were assessed: 10 had mutation and 5 fusion, potentially allowing for BRAF or MEK inhibitors. Actionable fusions for potential trial entry were found in a 6-year-old male with pilocytic astrocytoma (PA) (GOPC-ROS1 fusion), a 4 month-old male with congenital GBM (MZT2B-ALK fusion), and a 3 year-old female with epithelioid GBM (ETV6-NTRK3 fusion). FGFR1-TACC1 fusions were found in three pediatric patients (16-year-old female with PA, 9-year-old male with extraventricular neurocytoma, 1-year-old male with spinal cord PA). CONCLUSIONS: Actionable fusions can be found in both adult and pediatric patients, although STARTRK-2 study entry criteria were not met in any of the 18 adults. The most unusual fusions thus far have been the congenital GBM (MZT2B-ALKfusion) and the epithelioid GBM (ETV6-NTRK3 fusion).
Abstract 18416: Serum Adropin Level is a Predictor of Exercise Training-Induced Improvement of Arterial Stiffness in the Middle and Older Adults
Authors: Shumpei Fujie, Natsuki Hasegawa, Kiyoshi Sanada, Takafumi Hamaoka, Motoyuki Iemitsu
Journal: Circulation (2016)

Introduction: In our recent study, aging-induced arterial stiffening was gradually decreased by aerobic exercise. Furthermore, circulating level of adropin, a regulator of endothelial nitric oxide,...
TMIC-40. INTERLEUKIN-6 IN CEREBROSPINAL FLUID AS A PROGNOSTIC MARKER FOR GLIOBLASTOMA PATIENTS
Authors: Takashi Sasayama, Kazuhiro Tanaka, Tatsuo Hori, Masamitsu Nishihara, Masahiro Maeyama, Satoshi Nakamizo, Hirotomo Tanaka, Eiji Kohmura
Journal: Neuro-Oncology (2018)

Inflammatory cells are key components of the tumor microenvironment, and cytokines could create a favorable microenvironment to support tumorigenesis. Several studies have reported that IL-6 levels in glioma tissues are associated with the prognosis of patients with glioma; however, IL-6 levels in cerebrospinal fluid (CSF) of glioma patients has not been sufficiently studied yet. The patients with gliomas were treated at the Department of Neurosurgery, University of Kobe, between January 2007 and December 2016. CSF was withdrawn from the patients via lumbar puncture, and IL-6 concentration was measured by ELISA. after obtaining informed consent when not contraindicated. Infiltration of tumor-associated macrophages (TAMs) were analyzed by CD163 staining. Seventy-six patients with gliomas were analyzed in this study. The median follow-up period was 25 months. At the end of the follow-up period, 22 patients were alive, and 53 patients were dead. In the pre-treatment CSF examination, mean concentration of IL-6 was 26.3 ± 88.5 pg/ml. In glioblastoma alone, the mean value of CSF IL-6 was 35.6 ± 102.5 pg/ml, which was significantly higher than that of other grades of gliomas. The concentration of CSF IL-6 was correlated with the concentration of CSF LDH. IL-6 protein was expressed in the area where TAMs were predominantly existed. In double immunostaining examination, IL-6 protein was expressed in TAMs, and the concentration of CSF IL-6 was statistically correlated with the percentage of TAMs infiltration in the tumor (p=0.014). In multivariate analysis, high levels of CSF IL-6 were significantly associated with shorter overall survival (HR=1.007, 95% CI: 1.002~-.012, p=0.0049) These results indicated that the concentration of CSF IL-6 may reflect the microenvironment of the tumor and may be a useful prognostic biomarker for glioma patients.
Insulin receptor signaling regulates renal collecting duct and intercalated cell antibacterial defenses
Authors: Matthew J. Murtha, Tad Eichler, Kristin Bender, Jackie Metheny, Birong Li, Andrew L. Schwaderer, Claudia Mosquera, Cindy James, Laura Schwartz, Brian Becknell, John David Spencer
Journal: Journal of Clinical Investigation (2018)

People with diabetes mellitus have increased infection risk. With diabetes, urinary tract infection (UTI) is more common and has worse outcomes. Here, we investigate how diabetes and insulin resistance impact the kidney's innate defenses and urine sterility. We report that type 2 diabetic mice have increased UTI risk. Moreover, insulin-resistant prediabetic mice have increased UTI susceptibility, independent of hyperglycemia or glucosuria. To identify how insulin resistance affects renal antimicrobial defenses, we genetically deleted the insulin receptor in the kidney's collecting tubules and intercalated cells. Intercalated cells, located within collecting tubules, contribute to epithelial defenses by acidifying the urine and secreting antimicrobial peptides (AMPs) into the urinary stream. Collecting duct and intercalated cell-specific insulin receptor deletion did not impact urine acidification, suppressed downstream insulin-mediated targets and AMP expression, and increased UTI susceptibility. Specifically, insulin receptor-mediated signaling regulates AMPs, including lipocalin 2 and ribonuclease 4, via phosphatidylinositol-3-kinase signaling. These data suggest that insulin signaling plays a critical role in renal antibacterial defenses.
Abstract 17695: SCN5A H558R Polymorphism Counteracts Effects of SCN5A Mutations by Decrease Risk Allele Expression Level
Authors: Hiroya Matsumura, Yukiko Nakano, Kazuyoshi Suenari, Takehito Tokuyama, Akinori Sairaku, Hiroshi Kawazoe, Shunsuke Tomomori, Yasuki Kihara
Journal: Circulation (2015)

The common SCN5A polymorphism, H558R (c.1673A&gt;G,rs1805124), has been reported to be a genetic modulator of Brugada syndrome (BrS) carrying a SCN5A mutation. H558R has been known to improve the sodium channel activity in mutated channels by restoring the abnormal channel gating kinetics or membrane trafficking. The mechanisms by which H558R rescues the SCN5A mutations remain incompletely defined. We investigated the possible mechanism of why H558R modifies the effects brought by an SCN5A mutation. We investigated rs1805124 in 95 BrS patients (91 males, mean age 42 ± 14 years) and 1,875 normal controls (1546 male, mean age 54±18 years). We obtained the right atrial sections during aortic aneurysm operations from 21 patients and rs1805124 was also genotyped in them. Using cDNA of these patients, we examined the expression level of SCN5A by real time RT-PCR and whether splicing of exon 9 to exon 10 in SCN5A was correct by RT-PCR.and evaluate the difference between the genotypes of rs1805124. Additionally, we analyzed the expression ratio of the A allele and G allele separately by specific probes in heterozygous patients. H558R was less frequent in patients with BrS than in normal controls (4.7% vs. 10.3%, P = 0.01). We found strong Linkage disequibrium (LD) in c1141-3C&gt;A (Intron 9) and H558R (r2=0.946). The genotype distribution of the rs1805124 of AA, AG, and GG genotypes in the 21 patients were 13 (61.9 %), 8 (38.1 %), and 0 (0%), respectively. The expression level of SCN5A was similar in the patients with and without H558R. There were not any splicing abnormalities of SCN5A despite of H558R existence. The A allele expression level was mean 1.38 fold higher than that for the G allele (P=0.004). The result suggested that the mutation effect was relieved if the heterozygous mutation rode on the risk allele G. We may have elucidated one reason why H558R modifies the effects brought on by SCN5A mutations.
Loss of the BCR-FGFR1 GEF Domain Suppresses RHOA Activation and Enhances B-Lymphomagenesis in Mice
Authors: Tianxiang Hu, Yating Chong, Sumin Lu, Haiyan Qin, Mingqiang Ren, Natasha M. Savage, Chang-Sheng Chang, John K Cowell
Journal: Cancer Research (2018)

Transformation of hematopoietic stem cells by the BCR-FGFR1 fusion kinase found in a variant of stem cell leukemia/lymphoma (SCLL) syndrome leads to development of B-lymphomas in syngeneic mice and humans. In this study, we show that the relatively rapid onset of this leukemia is potentially related to oncogenic domains within the BCR component. BCR recruited a guanidine nucleotide exchange factor (GEF) domain to the fusion kinase to facilitate activation of small GTPases such as the Ras homology gene family, member A (RHOA). Deletion of this GEF domain increased leukemogenesis, enhanced cell survival and proliferation, and promoted stem cell expansion and lymph node metastasis. This suggests that, in an SCLL context, the presence of the endogenous GEF motif leads to reduced leukemogenesis. Indeed, loss of the GEF domain suppressed activation of RHOA and PTEN, leading to increased activation of AKT. Loss of the GEF domain enhanced cell proliferation and invasion potential, which was also observed in cells in which RHOA is knocked down, supported by the observation that overexpression of RHOA leads to reduced viability and invasion. In vivo depletion of RHOA in SCLL cells significantly increased disease progression and shortened latency. Collectively, these data show that the BCR GEF domain affects phenotypes associated with progression of SCLL through suppression of RHOA signaling. SIGNIFICANCE: RHOA activation is a critical event in the progression of BCR-FGFR1-driven leukemogenesis in stem cell leukemia and lymphoma syndrome and is regulated by the BCR GEF domain.
CMET-28. IMPACT OF DISEASE SITE, SIZE AND SURGICAL RESECTION ON SURVIVAL FROM METASTATIC CNS NEUROBLASTOMA
Authors: Kim Kramer, Sofia Haque, Suzzane Wolden, Jeffrey Greenfield, Brian Kushner, Shakeel Modak, Nai-Kong Cheung, Maria Donzelli, Mahmoud Eljalby, Mark Souweidane
Journal: Neuro-Oncology (2018)

To evaluate the impact of site of disease, size of the metastases, and impact of surgical resection of CNS metastases in patients with neuroblastoma. METHODS AND MATERIALS: Patients with neuroblastoma metastatic to the CNS were considered for surgical resection of the lesion(s) prior to undergoing conventional external beam radiation therapy (EBRT) and intraventricular radioimmunotherapy (cRIT). Lesions were classified as unifocal, multifocal, or leptomeningeal. Post-surgical EBRT consisted of 18 or 21 Gy craniospinal irradiation with a boost of 30–36 Gy to the metastatic site(s). A Mantel-Cox analysis was performed to assess the prognostic significance of disease site, size, and extent of resection on overall survival (OS). 93 patients with CNS neuroblastoma were assessed (2003–2017) with a follow-up of 24–177 (median 48) months since the detection of CNS disease. Lesions were unifocal parenchymal (N=54), leptomeningeal (N=6) or multifocal +/-leptomeningeal (N=33). As regards unifocal lesions, 40 (74%) were > 2 cm in largest diameter, and 44 (81%) underwent gross total resection (GTR). Following GTR, EBRT and cRIT, local control with no progression at the site was achieved in 39/44 (89%) patients. Systemic neuroblastoma in the absence of CNS progression occurred in 19/93 (20%). Improved OS was noted for those with unifocal vs multifocal disease (p=.01). Although a trend towards improved OS was associated with GTR, it was not statistically significant (p=0.12). Neither the size nor the site of the unifocal lesion impacted OS. Local control of the metastatic lesion is successfully achieved with the combined approach of surgical resection, EBRT and cRIT in 89% of patients with unifocal CNS neuroblastoma, with a trend towards improved OS in those undergoing GTR of metastases. There is no impact on the size or location of the metastatic lesion(s).
Regulation of Fatty Acid Synthase (FAS) gene expression by Peroxisome Proliferator-activated receptor-gamma (PPARγ) is dependent of Her-2/neu (erbB-2) signaling in breast cancer cells
Authors: Javier A. Menendez, Gershon Locker, Ruth Lupu
Journal: Cancer Research (2005)

3722  Peroxisome proliferator-activated receptor γ (PPARγ), a member of the nuclear hormone receptor superfamily with a prominent role in the regulation of differentiation of cell types with active lipid metabolism, has become a potential target for the prevention and treatment of cancer. However, the promise of PPARγ-based differentiation therapy has been questioned by studies demonstrating that an increased PPARγ signaling exacerbates tumor development in breast cancer animal models. Since a biologically aggressive subset of breast carcinomas exhibits high levels of Fatty Acid Synthase (FAS)-dependent endogenous fatty acid metabolism, we envisioned that a molecular explanation of this PPARγ-dependent tumor-promoting effect may be an increase in FAS signaling. Interestingly, it has recently been reported that Her-2/neu oncogene, an up-stream up-regulator of FAS, up-regulates PPARγ expression and modulates the sensitivity of breast cancer cells to PPARγ ligand therapy. In the present study we have found that two distinct ligands of PPARγ, 15dPG-J2 and triglitazone (TGZ), significantly increased the expression levels of FAS protein in MCF-7 breast cancer cells. When transient transfection experiments with a reporter construct containing the FAS promoter were carried out in MCF-7 cells, 15dPG-J2 or TGZ treatment induced two to three-fold increases in FAS promoter activity. Interestingly, the increase in luciferase activity observed in response to PPARγ ligands was completely abolished when MCF-7 cells were transfected with a deletion construct lacking the SREBP binding site of the FAS promoter. An opposite picture emerged when PPARγ and FAS signalings were evaluated in MCF-7 cells engineered to overexpress Her-2/neu oncogene (MCF-7/Her2-18 cells). As expected, Her-2/neu overexpression resulted in a significant up-regulation of PPARγ, while pharmacological inhibition of Her-2/neu-activated MAPK and PI-3K/AKT signaling cascades led to decreased levels of PPARγ in MCF-7/Her2-18 cells. Interestingly, the activation of PPARγ through 15dPG-J2 and TGZ caused a dramatic inhibition of Her-2/neu tyrosine phosphorylation and abolished FAS overexpression in MCF-7/Her2-18 cells. Moreover, pharmacological FAS blockade using the novel small compound C75 completely mimicked the effects of PPARγ ligands by suppressing phosphorylated Her-2/neu receptor as assessed by using a phosphorylation state-dependent anti-Her-2/neu monoclonal antibody. Moreover, pharmacological FAS blockade suppressed PPARγ overexpression in MCF-7/Her2-18 cells. These findings support the possibility that PPARγ-regulated FAS signaling, in a Her-2/neu-related manner, could serve as a breast cancer promotor as FAS overexpression is known to maintain and enhance a malignant phenotype in cancer cells.
Initial microbial community of the neonatal stomach immediately after birth
Authors: Sarah Bajorek, Leslie Parker, Nan Li, Kathryn Winglee, Michael Weaver, James Johnson, Michael Sioda, Josee Gauthier, Dominick J. Lemas, Christian Jobin, Graciela Lorca, Josef Neu, Anthony A Fodor
Journal: Gut Microbes (2018)

The purpose of this prospective cross-sectional cohort pilot study is to explore the initial microbial community of gastric aspirate fluid as collected immediately after birth and its relationships with mode of delivery and preterm birth. Twenty-nine gastric aspirate samples collected immediately after birth from infants born between 24–40 weeks gestation were analyzed for microbial composition. Total microbial content was low in many samples, with a substantial number sharing taxonomic composition with negative controls. qPCR targeting the 16S rRNA gene showed that infants delivered vaginally had a higher microbial load than infants delivered by C-section. Some pre-term samples showed high relative abundance of genus Ureaplasma, consistent with previous literature that has implicated infections with this taxon as a potential cause of pre-term birth. Vaginally born term infant samples, by contrast, had significantly higher levels of genus Lactobacillus with Lactobacillus crispatus the most dominant species. Microbial evaluation showed that vaginally born term infant gastric aspirate samples had higher levels of lactobacilli than pre-terms. Samples from many infants had low microbial load near the edge of the detection limit.