Thrombin generation as a predictor of outcomes in patients with non-traumatic intracerebral hemorrhage

Lóczi, Linda; Orbán-Kálmándi, Rita; Árokszállási, Tamás; Fekete, István; Fekete, Klára; Héja, Máté; Tóth, Judit; Csiba, László; Bagoly, Zsuzsa

doi:10.3389/fneur.2022.912664

Cited by 6 publications

(1 citation statement)

References 28 publications

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“…Thrombin is a serine protease that is formed during the clotting cascade in response to trauma . Its transience and role in wound healing make it an attractive biomarker for thrombotic diseases like cerebral aneurism and pulmonary embolism, − and as such, it was one of the first reported protein targets of aptamers. , HD22 forms a distorted G-quadruplex structure that is stabilized by binding to exosite II of thrombin, with outward-facing bases in the protein-binding loops (T9, A10, T18, and T19) and duplex (G21, G22, T24, and C28) providing the main contacts (Figure , Table S1). , Of these, the loop residues are promising targets for modification because mutating them does not stop the G-quadruplex from folding. , …”

Section: Resultsmentioning

confidence: 99%

Terminal Alkyne-Modified DNA Aptamers with Enhanced Protein Binding Affinities

Kohn,

Konovalov,

Gomez

et al. 2023

ACS Chem. Biol.

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Nucleic acid-based receptors, known as aptamers, are relatively fast to discover and manufacture but lack the diverse functional groups of protein receptors (e.g., antibodies). The binding properties of DNA aptamers can be enhanced by attaching abiotic functional groups; for example, aromatic groups such as naphthalene slow dissociation from proteins. Although the terminal alkyne is a πelectron-rich functional group that has been used in small molecule drugs to enhance binding to proteins through noncovalent interactions, it remains unexplored for enhancing DNA aptamer binding affinity. Here, we demonstrate the utility of the terminal alkyne for improving the binding of DNA to proteins. We prepared a library of 256 terminalalkyne-bearing variants of HD22, a DNA aptamer that binds the protein thrombin with nanomolar affinity. After a one-step thrombin-binding selection, a high-affinity aptamer containing two alkynes was discovered, exhibiting 3.2-fold tighter thrombin binding than the corresponding unmodified sequence. The tighter binding was attributable to a slower rate of dissociation from thrombin (5.2-fold slower than HD22). Molecular dynamics simulations with enhanced sampling by Replica Exchange with Solute Tempering (REST2) suggest that the π-electron-rich alkyne interacts with an asparagine side chain N−H group on thrombin, forming a noncovalent interaction that stabilizes the aptamer− protein interface. Overall, this work represents the first case of terminal alkynes enhancing the binding properties of an aptamer and underscores the utility of the terminal alkyne as an atom economical π-electron-rich functional group to enhance binding affinity with minimal steric perturbation.

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Section: Resultsmentioning

confidence: 99%

Terminal Alkyne-Modified DNA Aptamers with Enhanced Protein Binding Affinities

Kohn,

Konovalov,

Gomez

et al. 2023

ACS Chem. Biol.

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Blood prognostic biomarker signatures for hemorrhagic cerebral cavernous malformations (CCMs)

Croft

Grajeda

Abou‐Fadel

et al. 2023

Preprint

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Background: Cerebral cavernous malformations (CCMs) is a neurological disorder that causes enlarged intracranial capillaries in the brain, leading to an increased risk of hemorrhagic strokes, which is a leading cause of death and disability worldwide. Objectives: The current treatment options for CCMs are limited, highlighting the need for prognostic biomarkers to predict the risk of hemorrhagic events to better inform treatment decisions and identify future pharmacological targets. Eligibility Criteria: The study is centered on a comparative proteomic analysis between hemorrhagic CCMs (HCs) and healthy controls, while excluding patients with non-hemorrhagic CCMs (NHCs) from the analysis due to the experimental design. Sources of Evidence: Recent research has identified several serum biomarkers and blood circulating biomarkers in a selected cohort of homogeneous CCM patients and animal models. Method: Proteomic profiles from both human and mouse CCM models were examined, and pathway enrichment analyses were performed using three approaches (GO, KEGG, and DOSE). To account for multiple comparisons, t-tests were employed to evaluate differences. A p-value below 0.05 was deemed statistically significant. Results: The authors have developed the first panel of candidate biomarker signatures, featuring both etiological and prognostic markers in two distinct pathways. This panel of biomarker signatures demonstrates a robust correlation with the likelihood of hemorrhagic CCMs. Conclusions: This groundbreaking biomarker panel paves the way for further investigation of potential blood biomarkers to determine the risk of hemorrhagic CCMs.

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Effect of anti-SARS-CoV-2 BNT162b2 mRNA vaccination on thrombin generation in children with inflammatory bowel disease

Stercel,

Lóczi,

Kadenczki

et al. 2023

Front. Immunol.

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BackgroundInflammatory bowel disease (IBD) including Crohn’s disease (CD) and ulcerative colitis (UC), are associated with higher thrombotic risk and enhanced thrombin generation (TG) in adults. Despite encouraging data reporting vaccine safety and low IBD flare rates in adults with IBD, vaccine hesitancy was demonstrated to be high in families of children with IBD. We aimed to find out whether TG is increased in children with IBD as compared to healthy controls and whether TG parameters show significant changes following SARS-CoV-2 mRNA vaccination.Patients and methodsIn this observational case-control study, 38 children with IBD (CD:18, UC: 20) aged 12-18 years and 62 healthy age-and sex-matched children were enrolled. Blood was collected before the first dose and 2-6 weeks after the second dose of BNT162b2 (Pfizer-BioNTech) mRNA vaccine dose. Blood cell counts, fibrinogen, inflammatory markers (hsCRP, ferritin), anti-SARS-CoV-2 antibody levels were investigated, TG assay was carried-out using platelet-poor plasma. Detailed clinical parameters including disease activity scores (PUCAI, PCDAI) were registered pre-and post- vaccination. A guided questionnaire was used to collect data on adverse reactions (AEs) post- vaccination.ResultsBaseline TG parameters did not differ between patients and controls. Endogenous thrombin potential showed a significant positive correlation with markers of inflammation and with PCDAI. Inflammatory parameters and TG did not increase in patients and controls post-vaccination. Vaccination significantly increased antibody levels in all three investigated groups, but post-vaccination anti-SARS-CoV-2 S IgG/IgM levels were below the 5th percentile value of healthy children in more than one third of patients. Those receiving TNFα inhibitor therapy presented significantly lower SARS-CoV-2 S IgG/IgM levels as compared to patients on other immunosuppressive regimens. Systemic AEs did not differ between patients and controls while lower rate of local symptoms was found post-vaccination in children with IBD. Only 2 IBD flares were detected 2-6 weeks after the second dose of vaccination.ConclusionOur study is the first to support the safety and efficacy of anti-SARS-CoV-2 BNT162b2 vaccination in children with IBD with detailed pre-and post-vaccination laboratory data including TG. Results of this study may further increase confidence and reduce vaccine hesitancy in caretakers of pediatric IBD patients.

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Thrombin generation as a predictor of outcomes in patients with non-traumatic intracerebral hemorrhage

Cited by 6 publications

References 28 publications

Terminal Alkyne-Modified DNA Aptamers with Enhanced Protein Binding Affinities

Terminal Alkyne-Modified DNA Aptamers with Enhanced Protein Binding Affinities

Blood prognostic biomarker signatures for hemorrhagic cerebral cavernous malformations (CCMs)

Effect of anti-SARS-CoV-2 BNT162b2 mRNA vaccination on thrombin generation in children with inflammatory bowel disease

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