tbuckley/cabot-search huggingface dataset:
CSIG-09. ASSESSMENT OF TUMOR TREATING FIELDS (TTFields) COMBINED WITH TRICHOSTATIN A (TSA) IN PATIENT-DERIVED GLIOBLASTOMA (GBM) CELLS
Authors: Manxiu Ma, Sharon Michelhaugh, Sandeep Mittal
Journal: Neuro-Oncology (2021)

Abstract BACKGROUND TTFields therapy is a non-invasive biophysical approach for GBM patient treatment. TTFields produce an anti-mitotic effect by destabilizing microtubule polymerization and interrupting cell division. However, epigenetic modifications that may be induced by TTFields remain unknown. To address this, TTFields were applied to patient-derived GBM cells ± the histone deacetylase (HDAC) inhibitor trichostatin A (TSA) and cellular proliferation was assessed. HDAC activity was also measured after TTFields application. METHODS GBM cells were isolated from a newly-diagnosed patient tumor (IDH-WT, unmethylated MGMT promoter). Cells were grown in DMEM/F12 media with 10% FBS and gentamicin. Cells were plated on plastic coverslips (1×104cells/coverslip) and incubated overnight. TTFields were applied at 200 kHz (field intensity of ~1.6 V/cm) for 6 days. For the last 2 days of TTFields, cells were incubated with DMSO or 250 nM TSA. Cells were harvested and counted to assess proliferation (n=4-5/group). To determine if TTFields have a direct effect on HDAC activity, TTFields were applied as described and nuclei were extracted. Nuclear HDAC activity was measured using a commercial kit (n=2-3/group). Cell counts were compared with one-way ANOVA and Tukey multiple comparisons with p&amp;lt; 0.05 considered significant. RESULTS Cell counts for the DMSO control group, TSA only, TTFields only, and the combination were: 643,400 ± 111,384, 233,800 ± 144,200, 90,775 ± 45,209, and 41,520 ± 36,168, respectively (mean ± SD; ANOVA p&amp;lt; 0.0001). There was no statistical difference between the TTFields only and TSA only or TTFields + TSA groups, with all other comparisons statistically significant. For the HDAC activity measurements, control nuclei were 1068 ± 135 pmol/min/mg and TTFields-treated nuclei were 1301 ± 313.6 pmol/min/mg and were not statistically different. CONCLUSIONS The addition of TSA to TTFields did not alter cellular proliferation more than TTFields alone and TTFields did not alter HDAC activity in these GBM cells.
TAMI-35. DETECTING SINGLE-CELL INTERACTIONS IN ORGANOTYPIC CULTURES OF GLIOBLASTOMA USING BARCODED RABIES VIRUS
Authors: Brian Andersen, Michael Wheeler, Zhaorong Li, E Antonio Chiocca, David Reardon, Francisco Quintana
Journal: Neuro-Oncology (2021)

Abstract Cell-cell interactions are thought to drive tumor-promoting signals in the microenvironment of glioblastoma, but standard approaches for single cell analysis do not directly identify cell interactions and the mechanisms that mediate them. We recently developed a novel method to analyze cell-cell interactions—rabies barcode interaction detection followed by sequencing (RABID-seq), which combines barcoded viral tracing and single-cell RNA sequencing (scRNAseq). RABID-seq was first implemented in transgenic mice to investigate the interactions of astrocytes with other cells in the CNS enabling the study of astrocyte connectome perturbations and candidate therapeutic targets in multiple sclerosis and its pre-clinical model, experimental autoimmune encephalomyelitis (EAE). Here, we report the first use of RABID-seq in human tissues in organotypic cultures established from three IDH-wildtype glioblastoma (GBM) patients. In organotypic GBM cultures, initial infection by pseudotyped barcoded rabies virus deficient for viral glycoprotein was achieved after previous culture transduction with a lentivirus containing the avian TVA receptor and rabies glycoprotein under the human EF1a promoter. We employed this system to initially infect approximately 1,000 malignant or non malignant cells in the tumor microenvironment. After five days, infected cells were isolated from cultures and processed for single cell analysis using SMART seq. We were able to capture at least 6,000 interacting cells per tumor specimen, from which barcodes were recovered and cDNA was sent for sequencing. Here we present connectomic data from our initial cohort of three glioblastoma patients as an introduction to RABID-seq, with a focus on astrocyte-tumor interactions. Candidate mechanisms of cellular interactions will undergo functional validation in murine models of glioblastoma.
Abstract 114: Safety And Efficacy Of CDR132L, A Novel Antisense Therapeutic Which Targets MicroRNA-132 In Heart Failure Patients
Authors: Jorg Taubel, Wilfried Hauke, Steffen Rump, Janika Viereck, Sandor Batkai, Poetzsch Jenny, Rode Laura, Henning Weigt, Celina Genschel, Ulrike Lorch, Carmen Theek, Arthur A Levin, Johann B Bauersachs, Scott D Solomon, Thomas Thum
Journal: Circulation Research (2021)

Cardiac microRNA-132-3p (miR-132) levels are elevated in people suffering with heart failure (HF) and mechanistically drive remodelling processes in the heart. Here we present CDR132L, a specific antisense oligonucleotide. It is a first-in-class miR-132 inhibitor that attenuates and even reverses HF in preclinical models. The aim of this clinical Phase 1b study was to assess safety, pharmacokinetics, target engagement, and pharmacodynamics of CDR132L in patients on therapy for chronic ischaemic HF in a randomized, placebo-controlled, double-blind, dose-escalation study. Inclusion criteria for participant patients was left ventricular ejection fraction between 30 and 50% or levels of amino terminal fragment of pro-brain natriuretic peptide (NT-proBNP) higher than 125 ng/L at screening. Twenty-eight patients were randomized to receive CDR132L (0.32, 1, 3, and 10 mg/kg body weight) or placebo (0.9% saline) in two intravenous infusions, 4 weeks apart. Randomization separated participants into four cohorts of seven (five verum and two placebo). Results revealed that CDR132L was safe and well tolerated, and there was no detectable dose-limiting toxicity. A pharmacokinetic/pharmacodynamic dose modelling approach suggested an effective dose level of ≥1 mg/kg CDR132L. CDR132L treatment resulted in a dose-dependent, sustained miR-132 reduction in plasma. Patients who were administered ≥1 mg/kg of CDR132L displayed 23.3% NT-proBNP reduction, whereas patients receiving placebo experienced a 0.9% increase. In addition, CDR132L treatment was observed to significantly reduce duration of the QRS interval. This is the first clinical trial in which an antisense oligonucleotide was administered as a therapeutic agent to HF patients. Linear plasma pharmacokinetics were confirmed, and there were no signs of accumulation. The application of this drug suggests cardiac functional improvements and cardiac remodelling. The efficacy of this drug is encouraging, and further clinical work with larger patient populations should follow. Although the study used small patient numbers, results suggest some clinical benefit in chronic HF patients on the top of standard of care. This included an improvement in HF severity and narrowing of the QRS complex. The pharmacodynamic findings presented here are encouraging as they confirm efficacy results seen in animal studies.
NCOG-05. RISK FACTORS FOR PROLONGED LENGTH OF STAY AND POSTOPERATIVE COMPLICATIONS IN PATIENTS WITH INTRAMEDULLARY SPINAL CORD TUMORS
Authors: Romaric Waguia Kouam, Timothy Wang
Journal: Neuro-Oncology (2021)

Abstract INTRODUCTION Primary spine tumors are rare neoplasms that affect about 0.62 per 100,000 individuals in the US. Intramedullary spinal cord tumors (IMSCTs) are the rarest of all primary tumors involving the spine and can cause pain, imbalance, urinary dysfunction and neurological deficits. These types of tumors oftentimes necessitate surgical treatment, yet there is a lack of data on length of stay and complication rates following treatment. Given that treatment candidacy, quality of life, and outcomes are tied so closely to these metrics, it is important to illustrate the risk factors for prolonged length of stay and their association with post-op complications with the ultimate goal of improving patient selection and preoperative optimization. METHODS The National Surgical Quality Improvement Program (NSQIP) database was queried for all patients undergoing surgery for treatment of intramedullary spinal cord tumors between 2005 and 2017. Univariate and multivariate analysis were performed to assess patient risk factor influencing prolonged length of stay and post-op complications. RESULTS A total of 638 patients were included in the analysis. Pre-operative American Society of Anesthesiology (ASA) physical status classification of 3 and above ( OR 1.89; p=0.0005), dependent functional status ( OR 2.76; p=0.0035) and transfer from facilities other than home ( OR 8.12; p&amp;lt; 0.0001) were independent predictors of prolonged length of stay ( &amp;gt;5 days). The most commonly reported complications were pneumonia (5.7%), urinary tract infection (9.4%), septic shock (3.8%), superficial incisional infection (5.7%), organ or space infection (5.7%), pulmonary embolism (11.3%), DVT requiring therapy (15.1%) and wound dehiscence (5.7%). CONCLUSION Our study demonstrated the significant influence of clinical variables on prolonged hospitalization of IMSCT patients. This should be factored into clinical and surgical decision making and when counseling patients of their expected outcomes.
Abstract P508: Hyperglycemia Exacerbated Sars-cov-2 Spike Protein-induced Electrophysiological Dysfunctions In Hipsc-derived Cardiomyocytes
Authors: Chloe Wong, Thong B Nguyen, Michael Tong, Takashi Matsui, Yiqiang Zhang
Journal: Circulation Research (2021)

Background: Cardiac arrhythmias, heart failure, and myocarditis are common symptoms associated with coronavirus 2 (SARS-CoV-2) infection. The cytopathic effect of SARS-CoV-2 is initiated by spike protein (S-protein) binding to the ACE2 receptor. However, cellular mechanisms of SARS-CoV-2 causing cardiac complications, particularly in a subset of the population such as diabetic patients, remain unclear. Aims: To determine electrophysiological cytopathic effects of SARS-CoV-2 S-protein in human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs). Methods and Results: CMs were differentiated from WTC-11, then re-plated on 48-well microelectrode array (MEA) plates and cultured under normal or hyperglycemic conditions (Glucose 5 mM (NG), and Glucose 20 mM (HG)) for three days. These HiPSC-CMs were treated with recombinant S-protein with or without insulin supplementation. Cardiac modules in AxIS MEA program were used to acquire and analyze electric and contractile functions. Beat period and field potential duration were significantly prolonged, while conductance velocity and spike slope were accelerated in HG without insulin compared to that in HG with insulin (p&lt;0.05). S-protein-induced perturbations in local extracellular action potential (LEAP) matrices (e.g., LPD30, 50, 90) and Contractility matrices in HG were mitigated by insulin treatment. Higher concentration S-protein induced perturbations in normal hiPSC-CMs (NG+insulin). One day after S-protein treatment, CM functions appeared comparable to the pretreatment state, although changes in some electro-contractile matrices persisted. Conclusion: These results demonstrate SARS-CoV-2 viral structural protein can induce acute electrophysiological dysfunctions in hiPSC-CMs, which is influenced by glucose level. This establishes a new model for dissecting the mechanism of cardiac complications in viral infection.
Immunomodulatory and/or immunosuppressive drugs should not be stopped prior to skin tests for the assessment of drug allergy
Authors: F. Dezoteux, S. El Mesbahi, B. Tedbirt, J. Grosjean, S. Gautier, D. Lannoy, C. Nassar, F. Tétart, D. Staumont‐Sallé
Journal: British Journal of Dermatology (2021)
Abstract P329: Exosomes Derived From Podoplanin Positive Cells Alter The Cellular Composition Of Healthy Mouse Hearts
Authors: Maria Cimini, Venkata Garikipati, Andrea Elia, Chunlin Wang, MAY TRUONGCAO, Anna Maria Lucchese, Grace Huang, Vandana Mallaredy, Carolina Gonzalez, Cindy Benedict, Raj Kishore
Journal: Circulation Research (2021)

Fibrosis and blood hypoperfusion stimulated by paracrine signals enhances the ventricular dysfunctionafter myocardial infarction (MI). We have earlier reported that within 2 days post-MI a cohort ofpodoplanin (PDPN) positive cells populate injured heart and enhance inflammatory response by physicalinteractions with monocytes. Here we explored whether exosomes from these cells could independentlyalter healthy heart physiology and structure. PDPN+ cells were isolated 2 days after MI, culture expandedand activated with TNFα and Angiotensin II. Exosomes derived from activated PDPN+cells conditionedmedia (PDPN+exo) were used in vitro for the treatment of mouse cardiac endothelial cells (mCECs) andmouse fibroblast (3T3) and in vivo for the treatment of healthy mouse hearts. In vitro, PDPN+exoinfluenced the phenotype of mCECs, stimulating their lineage into lymphatic endothelial cells andfacilitated fibroblasts transition to myofibroblast. Characterization of the protein content of PDPN+exoshowed high concentration of Notch receptors and γ-Secretase, suggesting these cellular transitions maydepend on exosome-mediated Notch translocation and cleavage. In fact, after exosomes treatmentcleaved notch (NICD) translocated in the nuclei of mCECs and 3T3 as early as 1h of treatment and eitherHes-1 or Hey-1, major transcription factors activated by NICD were enhanced within 2d of treatment.Using DAPT, a γSecretase inhibitor, notch cleavage was inhibited, and no phenotype switching in responseto exosome treatment was observed. In vivo, PDPN+exo were injected into the left ventricle of healthymouse hearts followed by boosters delivered by retro-orbital vein injection. Treated mice developed anextended epicardial fibrosis with a subsequent impairment in the contractility and increase of the enddiastolic and systolic volumes. The fibrotic area was characterized by vessels double positive toendothelial and lymphatic endothelial markers, and infiltrating CD45+ cells. Podoplanin positive cellsrepresent 80% of the scar’s cells of a chronic infarcted myocardium and the specific exosomes cargo highlyinfluence the lineage of cardiac cells altering the biology of endothelial cells and fibroblasts which mayfacilitate adverse remodeling.
Abstract 9989: Safety of Neurohormonal Antagonist Initiation Among Older Adults Following a Heart Failure Hospitalization
Authors: Chukwuma Onyebeke, Andrew Zullo, Barbara Gladders, Min Ji Kwak, Larry A Allen, Emily B Levitan, Monika M Safford, Lauren Gilstrap, Parag Goyal
Journal: Circulation (2021)

Introduction: Heart Failure (HF) hospitalizations may be an opportunity for clinicians to initiate neurohormonal antagonists (NHAs), but there is concern that initiation of multiple NHAs during the post-hospitalization period could worsen outcomes in older patients. We examined the safety of starting multiple NHAs in older adults following a hospitalization for HF with reduced ejection fraction (HFrEF). Hypothesis: Number of NHAs initiated following HFrEF hospitalization is associated with higher mortality and readmissions. Methods: The study population included beneficiaries aged &gt;= 65 years with HFrEF from a random 40% national sample of Medicare Part A, B, and D discharged following a HF admission between 2008 and 2015. The exposure variable was the count of NHAs initiated within 90 days of discharge. NHAs included ACE inhibitors, Angiotensin II Receptor Blockers, Beta blockers, and Mineralocorticoid Receptor Antagonists. The primary outcomes were mortality and readmission at 90 days. We calculated the hazard ratios of number of NHAs initiated (0-3) for each outcome with 95% confidence intervals using probability weighting to account for group differences and generated cumulative incidence plots for both outcomes in each group. Results: We examined 211,885 patients, among whom 14% had 0 NHAs initiated, 33% had 1, 42% had 2, and 11% had 3. Mortality and readmission at 90 days were lowest among those with higher NHA initiation counts (Figures 1, 2). Conclusion: Greater NHA initiation following HFrEF hospitalization in older adults was associated with lower 90-day mortality and readmission rates. On average, the benefits of starting multiple NHAs may outweigh risks.
Comparable Outcomes Among Adult Patients Allotransplanted for Myelodysplastic Syndrome Using Haploidentical, Matched Unrelated or Matched Sibling Donors: A Single-Center Study
Authors: Alice Garnier, Maxime Jullien, Thierry Guillaume, Pierre Peterlin, Amandine Le Bourgeois, Béatrice Mahé, Viviane Dubruille, Nicolas Blin, Cyrille Touzeau, Thomas Gastinne, Anne Lok, Benoit Tessoulin, Sophie Vantyghem, Marie C Bene, Philippe Moreau, Steven Le Gouill, Patrice Chevallier
Journal: Blood (2021)
Association of L-α Glycerylphosphorylcholine With Subsequent Stroke Risk After 10 Years
Authors: Gyeongsil Lee, Seulggie Choi, Jooyoung Chang, Daein Choi, Joung Sik Son, Kyuwoong Kim, Sung Min Kim, Seogsong Jeong, Sang Min Park
Journal: JAMA Network Open (2021)

<h3>Importance</h3> L-α glycerylphosphorylcholine (α-GPC, choline alphoscerate) is used globally by individuals older than 50 years based on its potential function as a precursor of acetylcholine. However, choline has previously been linked to a higher risk of cardiovascular disease via trimethylamine-<i>N</i>-oxide, a metabolite of choline by microbiota. <h3>Objective</h3> To investigate the association between α-GPC use and subsequent 10-year stroke risk. <h3>Design, Setting, and Participants</h3> A population-based, retrospective cohort study was conducted using data from the National Health Insurance Service of South Korea. Participants included men and women aged 50 years or older without underlying stroke or Alzheimer disease (N = 12 008 977). <h3>Main Outcomes and Measures</h3> All participants were divided into whether they were prescribed α-GPC during 2006-2008. α-GPC users were matched with nonusers for all covariates to create a matched cohort. α-GPC use was further divided into durations less than 2, 2 to 6, 6 to 12, and more than 12 months of α-GPC prescriptions. The adjusted hazard ratios (aHRs) and 95% CIs for total stroke, ischemic stroke, and hemorrhagic stroke from January 1, 2009, to January 31, 2018, were calculated by multivariate Cox proportional hazards regression. <h3>Results</h3> A total of 12 008 977 individuals (6 401 965 [53.3%] women) aged 50 years or older were included in the study. The mean (SD) age was 61.6 (9.4) years for nonusers and 68.3 (10.0) years for users, and that of the matching cohort was 68.2 (9.9) years for both groups. Compared with α-GPC nonusers (n = 11 900 100), users (n = 108 877) had a higher risk for total stroke (aHR, 1.46; 95% CI, 1.43-1.48), ischemic stroke (aHR 1.36; 95% CI, 1.33-1.39), and hemorrhagic stroke (aHR, 1.36; 95% CI, 1.28-1.44). After matching for all covariates, α-GPC users had a higher risk for total stroke (aHR, 1.43; 95% CI, 1.41-1.46), ischemic stroke (aHR, 1.34; 95% CI, 1.31-1.37), and hemorrhagic stroke (aHR, 1.37; 95% CI, 1.29-1.46). Increasing intake of α-GPC was associated with a higher risk for total stroke in a dose-response manner. <h3>Conclusions and Relevance</h3> In this cohort study, use of α-GPC was associated with a higher 10-year incident stroke risk in a dose-response manner after adjusting for traditional cerebrovascular risk factors. Future studies are needed to determine the possible mechanisms behind the potential cerebrovascular risk–elevating effects of α-GPC.
Abstract 12922: Electrocardiogram-Based Deep Learning and Clinical Risk Factors to Predict Incident Atrial Fibrillation
Authors: Shaan Khurshid, Samuel Friedman, Christopher Reeder, Paolo Di Achille, Nathaniel Diamant, Pulkit Singh, Lia Harrington, Xin Wang, Mostafa Al-alusi, Gopal Sarma, Patrick T Ellinor, Christopher D Anderson, Jennifer E Ho, Anthony A Philippakis, Puneet Batra, Steven A Lubitz
Journal: Circulation (2021)

Introduction: Deep learning-derived representations of 12-lead electrocardiograms (ECGs) may allow for atrial fibrillation (AF) risk prediction. However, it remains unclear whether ECG-based artificial intelligence improves prediction beyond established clinical risk factors for AF and whether predictions are generalizable. Methods: Within a dataset comprising over 500,000 individuals receiving regular primary care at a multi-institutional network, we trained a convolutional neural network to predict incident AF using 12-lead ECGs (“ECG-AI”). ECG-AI was trained in individuals with ≥1 ECG performed at Massachusetts General Hospital (MGH) within 3 years prior to start of follow-up. We then fit a Cox proportional hazards model with incident AF as the outcome and a) logit-transformed ECG-AI AF probability, and b) the Cohorts for Aging and Genomic Epidemiology AF (CHARGE-AF) score, as covariates (“CH-AI”). We compared the discrimination and calibration of CHARGE-AF versus CH-AI in three independent samples: MGH (n=4,166), Brigham and Women’s Hospital (BWH, n=37,963) and the UK Biobank (n=41,034). Based on available follow-up, AF was evaluated at 5 years in MGH and BWH, and 2 years in the UK Biobank. Results: ECG-AI was trained in 36,081 individuals with an ECG performed at MGH (mean age 55±17, 53% female). CH-AI had substantially better discrimination (area under the receiver operating characteristic curve [AUROC]: MGH 0.838, BWH 0.777, UK Biobank 0.746; average precision [AP] 0.30, 0.21, 0.06) versus CHARGE-AF (AUROC: 0.802, 0.752, 0.732; AP 0.21, 0.17, 0.02, Figure ). CH-AI was well-calibrated in MGH (calibration error 0.012) and BWH (0.019), but overestimated AF risk in the UK Biobank (0.068). Calibration in the UK Biobank was excellent after recalibration to the sample-level 2-year AF hazard (error 7.1x10 -5 ). Conclusions: A model combining clinical AF risk factors with deep learning-derived ECG-based AF risk is favorable for predicting 5-year risk of AF.
PATH-18. INTEGRATING TUMOR MICROENVIRONMENT WITH GENOMIC ABERRATIONS AND TUMOR CLASS IN ADULT GLIAL/GLIONEURONAL TUMORS BY DECONVOLUTION OF BULK METHYLATION DATA
Authors: Omkar Singh, Kenneth Aldape, Drew Pratt
Journal: Neuro-Oncology (2021)

Abstract It is increasingly recognized that the tumor microenvironment (TME) plays a critical role in the biology of cancer. To better understand the role of non-neoplastic immune cellular components in CNS tumors, we applied a deconvolution approach to bulk DNA methylation array data using methylCIBERSORT on 450k/850k methylation data from a set (n= 4057) of high- and low-grade glial and glioneuronal tumors. Using the cell type proportion data as input, we used dimension reduction (UMAP) to visualize sample-wise patterns of that emerge from the cell type proportion estimations. In glioblastomas (n= 2076) we identified distinct tumor clusters based on immune cell proportion and, interestingly, TME-based cluster groups demonstrated an association with specific genetic alterations such as EGFR amplification and/or CDKN2A/B homozygous deletion. Among 1178 IDH-mutant gliomas, clustering of tumors according to immune cell proportions led to 2 major subgroups, which largely aligned with 1p/19q co-deletion status. Among the non-codeleted tumors (IDH-mutant astrocytomas, N=734), clustering of immune cell decomposition revealed clusters which showed distinct proportions of a key genomic aberration in these tumors (CDKN2A/B loss). To investigate the possible role of monocyte proportion-relative gene expression and promoter methylation of the immune checkpoint PD-L1 and PD-L2 genes, we used a data subset (n=594) samples with matched gene expression profiles. We observed significantly high positive correlations (R=0.54 and 0.68, respectively) between monocyte proportion and expression of PD-L1 and PD-L2, in line with prior reports that monocytic cells can express these immune markers. Consistent with this, we found high negative correlations (R= -0.51 and -0.61, respectively) between monocytes and promoter methylation of PD-L1 and PD-L2, respectively. Overall, the findings highlight specific roles of the TME in biology and classification of adult CNS tumors, where specific immune cell admixtures correlate with tumor types and genomic aberrations.