JAK2-mutant hematopoietic cells display metabolic alterations that can be targeted to treat myeloproliferative neoplasms

Increased energy requirement and metabolic reprogramming are hallmarks of cancer cells. We show that metabolic alterations in hematopoietic cells are fundamental to the pathogenesis of mutant JAK2-driven myeloproliferative neoplasms (MPNs). We found that expression of mutant JAK2 augmented and subverted metabolic activity of MPN cells, resulting in systemic metabolic changes in vivo, including hypoglycemia, adipose tissue atrophy, and early mortality. Hypoglycemia in MPN mouse models correlated with hyperactive erythropoiesis and was due to a combination of elevated glycolysis and increased oxidative phosphorylation. Modulating nutrient supply through high-fat diet improved survival, whereas high-glucose diet augmented the MPN phenotype. Transcriptomic and metabolomic analyses identified numerous metabolic nodes in JAK2-mutant hematopoietic stem and progenitor cells that were altered in comparison with wild-type controls. We studied the consequences of elevated levels of Pfkfb3, a key regulatory enzyme of glycolysis, and found that pharmacological inhibition of Pfkfb3 with the small molecule 3PO reversed hypoglycemia and reduced hematopoietic manifestations of MPNs. These effects were additive with the JAK1/2 inhibitor ruxolitinib in vivo and in vitro. Inhibition of glycolysis by 3PO altered the redox homeostasis, leading to accumulation of reactive oxygen species and augmented apoptosis rate. Our findings reveal the contribution of metabolic alterations to the pathogenesis of MPNs and suggest that metabolic dependencies of mutant cells represent vulnerabilities that can be targeted for treating MPNs

RoBuster: A Corpus Annotated with Risk of Bias Text Spans in Randomized Controlled Trials (Preprint)

BACKGROUND Background: Risk of bias (RoB) assessment of randomized clinical trials (RCTs) is vital to answering systematic review questions accurately. Manual RoB assessment for hundreds of RCTs is a cognitively demanding and lengthy process. Automation has the potential to assist reviewers in rapidly identifying text descriptions in RCTs that indicate potential risks of bias. However, no RoB text span annotated corpus could be used to fine-tune or evaluate large language models (LLMs), and there are no established guidelines for annotating the RoB spans in RCTs. OBJECTIVE Objective: The revised Cochrane RoB Assessment 2 (RoB 2) tool provides comprehensive guidelines for RoB assessment; however, due to the inherent subjectivity of this tool, it cannot be directly used as RoB annotation guidelines. Our objective was to develop precise RoB text span annotation instructions that could address this subjectivity and thus aid the corpus annotation. METHODS Methods: We leveraged RoB 2 guidelines to develop visual instructional placards that serve as text annotation guidelines for RoB spans and risk judgments. Expert annotators employed these visual placards to annotate a dataset named RoBuster, consisting of 41 full-text RCTs from the domains of physiotherapy and rehabilitation. We report inter-annotator agreement (IAA) between two expert annotators for text span annotations before and after applying visual instructions on a subset (9 out of 41) of RoBuster. We also provide IAA on bias risk judgments using Cohen's Kappa. Moreover, we utilized a portion of RoBuster (10 out of 41) to evaluate an LLM using a straightforward evaluation framework. This evaluation aimed to gauge the performance of LLM (here GPT 3.5) in the challenging task of RoB span extraction and demonstrate the utility of this corpus using a straightforward evaluation framework. RESULTS Results: We present a corpus of 41 RCTs with fine-grained text span annotations comprising more than 28,427 tokens belonging to 22 RoB classes. The IAA at the text span level calculated using the F1 measure varies from 0% to 90%, while Cohen's kappa for risk judgments ranges between -0.235 and 1.0. Employing visual instructions for annotation increases the IAA by more than 17 percent points. LLM (GPT-3.5) shows promising but varied observed agreements with the expert annotation across the different bias questions. CONCLUSIONS Conclusions: Despite having comprehensive bias assessment guidelines and visual instructional placards, RoB annotation remains a complex task. Utilizing visual placards for bias assessment and annotation enhances IAA compared to cases where visual placards are absent; however, text annotation remains challenging for the subjective questions and the questions for which annotation data is unavailable in RCTs. Similarly, while GPT-3.5 demonstrates effectiveness, its accuracy diminishes with more subjective RoB questions and low information availability

Proteomic Profiling in Patients With Peripartum Cardiomyopathy:A Biomarker Study of the ESC EORP PPCM Registry

Background: Peripartum cardiomyopathy (PPCM) remains an important cause of maternal morbidity and mortality globally. The pathophysiology remains incompletely understood, and the diagnosis is often missed or delayed. Objectives: This study explored the serum proteome profile of patients with newly diagnosed PPCM, as compared with matched healthy postpartum mothers, to unravel novel protein biomarkers that would further an understanding of the pathogenesis of PPCM and improve diagnostic precision. Methods: Study investigators performed untargeted serum proteome profiling using data-independent acquisition–based label-free quantitative liquid chromatography–tandem mass spectrometry on 84 patients with PPCM, as compared with 29 postpartum healthy controls (HCs). Significant changes in protein intensities were determined with nonpaired Student's t-tests and were further classified by using the Boruta algorithm. The proteins’ diagnostic performance was evaluated by area under the curve (AUC) and validated using the 10-fold cross-validation. Results: Patients with PPCM presented with a mean left ventricular ejection fraction of 33.5% ± 9.3% vs 57.0% ± 8.8% in HCs (P &lt; 0.001). Study investigators identified 15 differentially up-regulated and 14 down-regulated proteins in patients with PPCM compared with HCs. Seven of these proteins were recognized as significant by the Boruta algorithm. The combination of adiponectin, quiescin sulfhydryl oxidase 1, inter-α-trypsin inhibitor heavy chain, and N-terminal pro–B-type natriuretic peptide had the best diagnostic precision (AUC: 0.90; 95% CI: 0.84-0.96) to distinguish patients with PPCM from HCs. Conclusions: Salient biologic themes related to immune response proteins, inflammation, fibrosis, angiogenesis, apoptosis, and coagulation were predominant in patients with PPCM compared with HCs. These newly identified proteins warrant further evaluation to establish their role in the pathogenesis of PPCM and potential use as diagnostic markers.</p

Impaired immune response mediated by prostaglandin E2 promotes severe COVID-19 disease

The SARS-CoV-2 coronavirus has led to a pandemic with millions of people affected. The present study finds that risk-factors for severe COVID-19 disease courses, i.e. male sex, older age and sedentary life style are associated with higher prostaglandin E2 (PGE2) serum levels in blood samples from unaffected subjects. In COVID-19 patients, PGE2 blood levels are markedly elevated and correlate positively with disease severity. SARS-CoV-2 induces PGE2 generation and secretion in infected lung epithelial cells by upregulating cyclo-oxygenase (COX)-2 and reducing the PG-degrading enzyme 15-hydroxyprostaglan- din-dehydrogenase. Also living human precision cut lung slices (PCLS) infected with SARS- CoV-2 display upregulated COX-2. Regular exercise in aged individuals lowers PGE2 serum levels, which leads to increased Paired-Box-Protein-Pax-5 (PAX5) expression, a master regulator of B-cell survival, proliferation and differentiation also towards long lived memory B-cells, in human pre-B-cell lines. Moreover, PGE2 levels in serum of COVID-19 patients lowers the expression of PAX5 in human pre-B-cell lines. The PGE2 inhibitor Taxi- folin reduces SARS-CoV-2-induced PGE2 production. In conclusion, SARS-CoV-2, male sex, old age, and sedentary life style increase PGE2 levels, which may reduce the early anti-viral defense as well as the development of immunity promoting severe disease courses and multiple infections. Regular exercise and Taxifolin treatment may reduce these risks and prevent severe disease courses

JAK2-V617F and interferon-α induce megakaryocyte-biased stem cells characterized by decreased long-term functionality.

We studied a subset of hematopoietic stem cells (HSCs) that are defined by elevated expression of CD41 (CD41hi) and showed bias for differentiation toward megakaryocytes (Mks). Mouse models of myeloproliferative neoplasms (MPNs) expressing JAK2-V617F (VF) displayed increased frequencies and percentages of the CD41hi vs CD41lo HSCs compared with wild-type controls. An increase in CD41hi HSCs that correlated with JAK2-V617F mutant allele burden was also found in bone marrow from patients with MPN. CD41hi HSCs produced a higher number of Mk-colonies of HSCs in single-cell cultures in vitro, but showed reduced long-term reconstitution potential compared with CD41lo HSCs in competitive transplantations in vivo. RNA expression profiling showed an upregulated cell cycle, Myc, and oxidative phosphorylation gene signatures in CD41hi HSCs, whereas CD41lo HSCs showed higher gene expression of interferon and the JAK/STAT and TNFα/NFκB signaling pathways. Higher cell cycle activity and elevated levels of reactive oxygen species were confirmed in CD41hi HSCs by flow cytometry. Expression of Epcr, a marker for quiescent HSCs inversely correlated with expression of CD41 in mice, but did not show such reciprocal expression pattern in patients with MPN. Treatment with interferon-α further increased the frequency and percentage of CD41hi HSCs and reduced the number of JAK2-V617F+ HSCs in mice and patients with MPN. The shift toward the CD41hi subset of HSCs by interferon-α provides a possible mechanism of how interferon-α preferentially targets the JAK2 mutant clone

Erratum to: Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition) (Autophagy, 12, 1, 1-222, 10.1080/15548627.2015.1100356

non present