tbuckley/cabot-search huggingface dataset:
Novel Majeed Syndrome–Causing<i>LPIN2</i>Mutations Link Bone Inflammation to Inflammatory M2 Macrophages and Accelerated Osteoclastogenesis
Authors: Farzana Bhuyan, Adriana A. de Jesus, Jacob Mitchell, Evgenia Leikina, Rachel VanTries, Ronit Herzog, Karen B. Onel, Andrew Oler, Gina A. Montealegre Sanchez, Kim A. Johnson, Lena Bichell, Bernadette Marrero, Luis Fernandez De Castro, Yan Huang, Katherine R. Calvo, Michael T. Collins, Sundar Ganesan, Leonid V. Chernomordik, Polly J. Ferguson, Raphaela Goldbach‐Mansky
Journal: Arthritis & Rheumatology (2020)

To identify novel heterozygous LPIN2 mutations in a patient with Majeed syndrome and characterize the pathomechanisms that lead to the development of sterile osteomyelitis.Targeted genetic analysis and functional studies assessing monocyte responses, macrophage differentiation, and osteoclastogenesis were conducted to compare the pathogenesis of Majeed syndrome to interleukin-1 (IL-1)-mediated diseases including neonatal-onset multisystem inflammatory disease (NOMID) and deficiency of the IL-1 receptor antagonist (DIRA).A 4-year-old girl of mixed ethnic background presented with sterile osteomyelitis and elevated acute-phase reactants. She had a 17.8-kb deletion on the maternal LPIN2 allele and a splice site mutation, p.R517H, that variably spliced out exons 10 and 11 on the paternal LPIN2 allele. The patient achieved long-lasting remission receiving IL-1 blockade with canakinumab. Compared to controls, monocytes and monocyte-derived M1-like macrophages from the patient with Majeed syndrome and those with NOMID or DIRA had elevated caspase 1 activity and IL-1β secretion. In contrast, lipopolysaccharide-stimulated, monocyte-derived, M2-like macrophages from the patient with Majeed syndrome released higher levels of osteoclastogenic mediators (IL-8, IL-6, tumor necrosis factor, CCL2, macrophage inflammatory protein 1α/β, CXCL8, and CXCL1) compared to NOMID patients and healthy controls. Accelerated osteoclastogenesis in the patient with Majeed syndrome was associated with higher NFATc1 levels, enhanced JNK/MAPK, and reduced Src kinase activation, and partially responded to JNK inhibition and IL-1 (but not IL-6) blockade.We report 2 novel compound heterozygous disease-causing mutations in LPIN2 in an American patient with Majeed syndrome. LPIN2 deficiency drives differentiation of proinflammatory M2-like macrophages and enhances intrinsic osteoclastogenesis. This provides a model for the pathogenesis of sterile osteomyelitis which differentiates Majeed syndrome from other IL-1-mediated autoinflammatory diseases.
EXTH-76. THE INOVIVO SYSTEM: A NOVEL PRECLINICAL TOOL FOR IN VIVO DELIVERY OF TUMOR TREATING FIELDS (TTFIELDS)
Authors: Shiri Davidi, Roni Blat, Anna Shteingauz, Yaara Porat, Moshe Giladi, Uri Weinberg, Yoram Palti
Journal: Neuro-Oncology (2020)

Abstract Tumor Treating Fields (TTFields) are an antineoplastic treatment modality targeting dividing cancer cells, approved by the FDA for treatment of glioblastoma and malignant pleural mesothelioma. TTFields are delivered to the patient continuously, using a portable signal generator and 2 pairs of transducer arrays attached to the skin. While in vitro research tools for TTFields have been available, animal experiments were so far limited due to the lack of a dedicated TTFields in vivo delivery system. The main challenges associated with TTFields application to mice are the absolute requirement for adequate and continuous contact between array electrodes and animal skin throughout treatments; and the need to deliver the electric fields through multiple wired conductors connected to electrodes, which limit animal movement thus imposing stress. This work reports on development of an in vivo system to facilitate continuous delivery of TTFields to mice bearing cancer tumors in the torso or flank that addresses the aforementioned challenges. Three major developments were introduced to the system: 1) transducer array electrodes composed of delicate, flexible inner layer to improve skin adherence and a breathable, durable, outer layer to secure electrodes to the skin; 2) a novel electric swivel to prevent cable coiling; and 3) a cage constructed to house 2 mice separately while still allowing socializing. During a 1-week study, the new inovivo system demonstrated improvement compared to previous systems in the following areas: increased animal treatment compliance, less weight loss, and fewer number of array replacements. In conclusion, the new inovivo system is a comprehensive tool for continuous, 2 directional TTFields delivery to tumors in the torso or flank of mice for conducting TTFields experiments with minimal animal stress, thus allowing further elucidation of the effects of TTFields on the whole animal.
Intracellular delivery of immunoglobulin G at nanomolar concentrations with domain Z-fused multimeric α-helical cell penetrating peptides
Authors: Seung-Eun Chong, Jae Hoon Oh, Kyungjin Min, Sohyun Park, Sejong Choi, Joon Hyung Ahn, Dahyun Chun, Hyung Ho Lee, Jaehoon Yu, Yan Lee
Journal: Journal of Controlled Release (2020)
Role of the cation-chloride-cotransporters in the circadian system
Authors: Shihan Salihu, Nur Farah Meor Azlan, Sunday Solomon Josiah, Zhijuan Wu, Yun Wang, Jinwei Zhang
Journal: Asian Journal of Pharmaceutical Sciences (2020)

The circadian system plays an immense role in controlling physiological processes in our body. The suprachiasmatic nucleus (SCN) supervises this system, regulating and harmonising the circadian rhythms in our body. Most neurons present in the SCN are GABAergic neurons. Although GABA is considered the main inhibitory neurotransmitter of the CNS, recent studies have shown that excitatory responses were recorded in this area. These responses are enabled by an increase in intracellular chloride ions [Cl−]i levels. The chloride (Cl−) levels in GABAergic neurons are controlled by two solute carrier 12 (SLC12) cation-chloride-cotransporters (CCCs): Na+/K+/Cl− co-transporter (NKCC1) and K+/Cl− co-transporter (KCC2), that respectively cause an influx and efflux of Cl−. Recent works have found altered expression and/or activity of either of these co-transporters in SCN neurons and have been associated with circadian rhythms. In this review, we summarize and discuss the role of CCCs in circadian rhythms, and highlight these recent advances which attest to CCC's growing potential as strong research and therapeutic targets.
MBCL-18. ANALYSIS OF DNA METHYLATION PROFILES OF PEDIATRIC MEDULLOBLASTOMAS: EXPERIENCE AT THE BAMBINO GESÙ CHILDREN’S HOSPITAL
Authors: Evelina Miele, Giuseppe Petruzzellis, Lucia Pedace, Antonella Cacchione, Andrea Carai, Giovanna Stefania Colafati, Francesca Diomedi Camassei, Sabrina Rossi, Franco Locatelli, Angela Mastronuzzi
Journal: Neuro-Oncology (2020)

Abstract BACKGROUND Medulloblastoma is the most frequent malignant brain tumor in children, still resulting fatal in about one third of affected patients. An accurate diagnosis is essential for correct therapeutic stratification. The DNA methylation profile (DMP) is a combination of changes in DNA methylation and genetic features that reflect the cell of tumor origin. DMP contributed to classify Medulloblastoma into four subgroups: WNT, SHH, Group 3/4 (the latter recently further subdivided into 8 subclasses). Each Methylation is associated with different genetic, demographic and clinical characteristics. We report our experience on Medulloblastoma molecular classification based on DMP. MATERIALS AND METHODS 54 Medulloblastoma patients (28 males, 26 females) were selected. The DMP analysis was carried out via IlluminaEPICarrays. The results were obtained using the brain tumor classifier (Capper, 2018). RESULTS In all cases the DMP allowed to classify the neoplasm, with an optimal score, in a defined methylation class. 10 WNTs, 15 SHHs, 10 Group 3, 19 Group 4 were found. Groups 3/4 were further refined based on the new consensus (Sharma 2019) in group I (n = 1); group II (n = 5); group III (n = 2); group IV (n = 4); group V (n = 3); group VI (n = 2); group VII (n = 6); group VIII (n = 7). CONCLUSIONS This study carried out the first classification of Medulloblastomas diagnosed in Italy through DMP, demonstrating its high reproducibility, precision and accuracy. The molecular classification improves diagnostic accuracy and provides further information that can guide personalized treatment.
EPCT-13. CMV PP65 RNA-PULSED DENDRITIC CELL VACCINES FOR PEDIATRIC GLIOBLASTOMA AND MEDULLOBLASTOMA: PHASE I TRIAL RESULTS
Authors: Daniel Landi, Gary Archer, Timothy Driscoll, Eric Lipp, Bridget Archambault, Eric Thompson, Charlene Flahiff, Denise Jaggers, Kathleen Hahn, Patrick Healy, Luis Ramirez, James Herndon, Kristin Schroeder, John Sampson, David Ashley
Journal: Neuro-Oncology (2020)

Abstract BACKGROUND Recurrent medulloblastoma and malignant glioma are lethal tumors that are virtually incurable. The cytomegalovirus (CMV) antigen pp65 is ubiquitously expressed on medulloblastoma and malignant glioma but not on healthy brain. We evaluated autologous CMV pp65 RNA-pulsed dendritic cell (DC) vaccines in children and young adults in a phase I trial. METHODS Circulating monocytes were harvested using leukapheresis, differentiated into DCs, matured, and pulsed with pp65 RNA using electroporation. DCs were packaged into vaccines (2x107DC/vaccine) and administered intradermally following tetanus-diphtheria toxoid site preconditioning every 2 weeks x3, then monthly. The primary objectives of the study were to establish the feasibility of generating at least 3 vaccines and safety. An exploratory objective was to evaluate the ability of vaccination to create and enhance patient pp65-specific T cell responses. RESULTS Eleven patients were enrolled with medulloblastoma (n=3) or glioblastoma (n=8). Ages ranged from 9–30 years old (mean 15.5y). Ten of 11 patients (91%) generated at least 3 vaccines (mean 6.2). Eight patients received at least 3 vaccines. To date, 4 patients have received all generated vaccines without progression, 4 patients have progressed, and 2 patients are still receiving vaccines. There have not been any severe adverse events probably or definitely related to vaccines. More mature data will be presented at ISPNO. CONCLUSIONS Leukapheresis and monocyte differentiation is a feasible strategy for generating adequate DCs for active immunization in children with malignant brain tumors. CMV pp65 RNA-pulsed DCs are well-tolerated and immunogenic. Efficacy endpoints will be evaluated in a subsequent phase II trial.
Streptococcuspyogenes infects human endometrium by limiting the innate immune response
Authors: Antonin Weckel, Thomas Guilbert, Clara Lambert, Céline Plainvert, François Goffinet, Claire Poyart, Céline Méhats, Agnès Fouet
Journal: Journal of Clinical Investigation (2020)

Group A Streptococcus (GAS), a Gram-positive human-specific pathogen, yields 517,000 deaths annually worldwide, including 163,000 due to invasive infections and among them puerperal fever. Before efficient prophylactic measures were introduced, the mortality rate for mothers during childbirth was approximately 10%; puerperal fever still accounts for over 75,000 maternal deaths annually. Yet, little is known regarding the factors and mechanisms of GAS invasion and establishment in postpartum infection. We characterized the early steps of infection in an ex vivo infection model of the human decidua, the puerperal fever portal of entry. Coordinate analysis of GAS behavior and the immune response led us to demonstrate that (a) GAS growth was stimulated by tissue products; (b) GAS invaded tissue and killed approximately 50% of host cells within 2 hours, and these processes required SpeB protease and streptolysin O (SLO) activities, respectively; and (c) GAS impaired the tissue immune response. Immune impairment occurred both at the RNA level, with only partial induction of the innate immune response, and protein level, in an SLO- and SpeB-dependent manner. Our study indicates that efficient GAS invasion of the decidua and the restricted host immune response favored its propensity to develop rapid invasive infections in a gynecological-obstetrical context.
IMMU-33. MULTI-ANTIGEN RECOGNITION CIRCUITS OVERCOME CHALLENGES OF SPECIFICITY, HETEROGENEITY, AND DURABILITY IN T-CELL THERAPY FOR GLIOBLASTOMA
Authors: Payal Watchmaker, Joseph Choe, Milos Simic, Ryan Gilbert, Aileen li, Nira Krasnow, Diego Carrera, Wei Yu, Kira Downey, Anna Celli, Juhyun Cho, Jessica Briones, Ruth Dannenfelser, Lia Cardarelli, Sachdev Sidhu, Kole Roybal, Wendell Lim, Hideho Okada
Journal: Neuro-Oncology (2020)

Abstract Treatment of solid cancers with chimeric antigen receptor (CAR) T-cells is challenging because of a lack of target antigens that are both tumor-specific and homogenously expressed. While epidermal growth factor receptor (EGFR)vIII represents a glioblastoma (GBM)-specific antigen, its expression is heterogeneous within the tumor resulting in tumor escape. In contrast, more homogenously expressed GBM-associated antigens (GAA), such as EphA2, are non-ideal because of expression in other normal organs, yielding potential cross-reactive toxicity. As a way to safely target GAAs in the tumor without attacking normal cells expressing the same GAAs outside of the brain, we adapted a novel synthetic Notch (synNotch) receptor system and established a “prime and kill” sequential two-receptor CAR circuit. A synNotch receptor recognizes a specific priming antigen; the heterogeneous GBM neoantigen EGFRvIII or a brain tissue-specific antigen to prime the local expression of a CAR that mediates cytotoxicity against a GAA (e.g. EphA2). In orthotopic GBM6 glioma model, a patient-derived xenograft (PDX) with heterogeneous expression of EGFRvIII, intravenous infusion of T-cells transduced with EGFRvIII synNotch→anti-IL-13Rα2/EphA2 tandem CAR circuit resulted in long-term (over 100 days) survival and eradication of the heterogeneous tumor in all of 12 mice in two independent experiments. In contrast, constitutive CARs targeting EGFRvIII or IL-13Rα2/EphA2 (as a tandem CAR) failed to exhibit long-term anti-tumor response. Moreover, T-cells transduced with synNotch-regulated CAR maintain a less differentiated state which is associated with higher durability compared with ones with constitutive CAR in vivo. T-cells transduced with a synNotch→CAR circuit primed by a brain-specific antigen, myelin oligodendrocyte glycoprotein (MOG), exhibited a precise and potent local control of intracranial PDX without evidence of priming in extracranial organs. These data support the utility of synNotch→CAR circuits in EGFRvIII-negative GBM cases. By integrating multiple imperfect but complementary antigens, we improve both the specificity and persistence of T-cells directed against GBM.
SURG-31. SURGICAL OUTCOME ANALYSES ON 244 CENTRAL NERVOUS SYSTEM HEMANGIOBLASTOMA
Authors: Hirokazu Takami, Terry C Burns, Ian Parney
Journal: Neuro-Oncology (2020)

Abstract BACKGROUND Surgical resection of the central nervous system hemangioblastoma is often challenging, with vascular-rich nature and surrounding critical structures. von Hippel-Lindau (VHL) disease cases make the management more difficult with repeated surgeries required for multiple lesions. Here we investigated the surgical outcome of CNS HGBs on a large cohort to clarify the clinical and radiological parameters pertaining to the surgical success. METHODS Retrospective analysis of consecutive, neurosurgically managed CNS HGB at Mayo Clinic, 1988-2018. RESULTS Total 244 surgeries were performed for 172 patients. Gross total resection (GTR) was achieved in 90.0%, which was lower in cases brainstem lesions (78.3%) and was associated with the tumor, especially solid-portion volume (p=0.017). Intraoperative blood loss correlated with the size of solid portions, and the transfusion was performed in 7.5%. Postoperative complication was observed in 52.2%, including new/worsening neurological deficit in 45.4%, wound complication in 9.1% and systemic complication in 4.0%. Postoperative rehabilitation was introduced in 61.7% of the patients, which was statistically associated with age, non-GTR and tumor location (brainstem and spine) on multivariate analysis (p=0.0031, 0.027 and 0.0066, respectively). Treatment-free survival was longer in VHL (vs sporadic) cases and GTR (vs non-GTR) cases, and multivariate analysis showed GTR was the only factor associated with treatment-free survival (p=0.0015). CONCLUSIONS Surgery for CNS HGBs was shown to be challenging, with abundant intraoperative bleeding and high risk of postoperative complications necessitating rehabilitation. GTR of the lesion is of utmost importance, which increases the chance of long treatment-free survival as well as favorable direct postoperative clinical course.
Impact of Population Growth and Aging on Estimates of Excess U.S. Deaths During the COVID-19 Pandemic, March to August 2020
Authors: Meredith S. Shiels, Jonas S. Almeida, Montserrat García-Closas, Paul S. Albert, Neal D. Freedman, Amy Berrington de González
Journal: Annals of Internal Medicine (2020)

Excess death estimates quantify the full impact of the coronavirus disease 2019 (COVID-19) pandemic. Widely reported U.S. excess death estimates have not accounted for recent population changes, especially increases in the population older than 65 years.To estimate excess deaths in the United States in 2020, after accounting for population changes.Surveillance study.United States, March to August 2020.All decedents.Age-specific excess deaths in the United States from 1 March to 31 August 2020 compared with 2015 to 2019 were estimated, after changes in population size and age were taken into account, by using Centers for Disease Control and Prevention provisional death data and U.S. Census Bureau population estimates. Cause-specific excess deaths were estimated by month and age.From March through August 2020, 1 671 400 deaths were registered in the United States, including 173 300 COVID-19 deaths. An average of 1 370 000 deaths were reported over the same months during 2015 to 2019, for a crude excess of 301 400 deaths (128 100 non-COVID-19 deaths). However, the 2020 U.S. population includes 5.04 million more persons aged 65 years and older than the average population in 2015 to 2019 (a 10% increase). After population changes were taken into account, an estimated 217 900 excess deaths occurred from March through August 2020 (173 300 COVID-19 and 44 600 non-COVID-19 deaths). Most excess non-COVID-19 deaths occurred in April, July, and August, and 34 900 (78%) were in persons aged 25 to 64 years. Diabetes, Alzheimer disease, and heart disease caused the most non-COVID-19 excess deaths.Provisional death data are underestimated because of reporting delays.The COVID-19 pandemic resulted in an estimated 218 000 excess deaths in the United States between March and August 2020, and 80% of those deaths had COVID-19 as the underlying cause. Accounting for population changes substantially reduced the excess non-COVID-19 death estimates, providing important information for guiding future clinical and public health interventions.National Cancer Institute.
ATRT-28. SINGLE NUCLEI SEQUENCING REVEALS THE DIFFERENT PHENOTYPIC COMPOSITION OF THE ATRT SUBGROUPS
Authors: Pascal Johann, Annette Büllesbach, Kati Ernst, Konstantin Okonechnikov, David T W Jones, Stefan M Pfister, Martin Hasselblatt, Marcel Kool
Journal: Neuro-Oncology (2020)

Abstract Atypical teratoid/rhabdoid tumors (ATRT) represents a genomically homogeneous disease characterized by loss of SMARCB1 protein in the vast majority of cases. In recent years, it has become clear that these tumors display a high degree of intertumoral heterogeneity with three molecularly distinct subgroups. However, the degree of intratumoral heterogeneity and the information on cellular subpopulations currently remains largely an unchartered territory. To explore the transcriptomic composition of ATRTs, we performed single nuclei RNA sequencing for 16 ATRTs representing all three molecular subgroups (5 ATRT-TYR, 7 ATRT-SHH, 4 ATRT-MYC). By performing tSNE cluster analyses of all the single cell data (~50.000 cells have been sequenced), we were able to gain unprecedented insights into the phenotypic composition of ATRTs and unravelled substantial differences between the three subgroups. Integrating transcriptomic information from non-neoplastic brain cells and the data derived from single nuclei sequencing, we found an OPC like gene signature in ATRT-SHH. In contrast, ATRT-TYR subpopulations overexpressed more markers of neuronal stem cells suggesting a larger fraction of undifferentiated cells in this subgroup. We also identified a subpopulation of cells with a clear overexpression of cell cycle associated genes (CDK4, CDKN3), predominantly present in ATRT-MYC samples, a finding which may harbour important consequences for a targeted therapy with e.g. CDK inhibitors. In summary, our analyses reveal different cellular compartments in ATRT and provide important insights into the cellular differentiation of the three ATRT-subgroups. Further analyses to achieve a specific mapping of ATRT to its physiological cell of origin are currently being pursued.
Cigarette Smoke Directly Promotes Pulmonary Arterial Remodeling and Kv7.4 Channel Dysfunction
Authors: Javier Sevilla-Montero, David Labrousse-Arias, Cintia Fernández-Pérez, Laura Fernández-Blanco, Bianca Barreira, Gema Mondéjar-Parreño, Elvira Alfaro-Arnedo, Icíar P. López, Sandra Pérez-Rial, Germán Peces-Barba, José G. Pichel, Víctor Ivo Peinado, Ángel Cogolludo, María J. Calzada
Journal: American Journal of Respiratory and Critical Care Medicine (2020)

Rationale: Cigarette smoke is considered the chief leading cause of chronic obstructive pulmonary disease (COPD). Its impact on the progressive deterioration of airways has been extensively studied, but its direct effects on the pulmonary vasculature are less known. Objectives: To prove that pulmonary arterial remodeling in patients with COPD is not just a consequence of alveolar hypoxia but also due to the direct effects of cigarette smoke on the pulmonary vascular bed. Methods: We have used different molecular and cell biology approaches, as well as traction force microscopy, wire myography, and patch-clamp techniques in human cells and freshly isolated pulmonary arteries. In addition, we relied on in vivo models and human samples to analyze the effects of cigarette smoke on pulmonary vascular tone alterations. Measurements and Main Results: Cigarette smoke extract exposure directly promoted a hypertrophic, senescent phenotype that in turn contributed, through the secretion of inflammatory molecules, to an increase in the proliferative potential of nonexposed cells. Interestingly, these effects were significantly reversed by antioxidants. Furthermore, cigarette smoke extract affected cell contractility and dysregulated the expression and activity of the voltage-gated K+ channel Kv7.4. This contributed to the impairment of vasoconstriction and vasodilation responses. Most importantly, the levels of this channel were diminished in the lungs of smoke-exposed mice, smokers, and patients with COPD. Conclusions: Cigarette smoke directly contributes to pulmonary arterial remodeling through increased cell senescence, as well as vascular tone alterations because of diminished levels and function in the Kv7.4 channel. Strategies targeting these pathways may lead to novel therapies for COPD.