Transcranial focused ultrasound-induced blood‒brain barrier opening in mice without shaving hairs
(2023) Scientific Reports, 13 (1), art. no. 13500, .
Xu, L.a , Gong, Y.a , Chien, C.-Y.a , Leuthardt, E.a b c , Chen, H.a b
a Department of Biomedical Engineering, Washington University in St. Louis, Saint Louis, MO 63130, United States
b Department of Neurosurgery, Washington University School of Medicine, Saint Louis, MO 63110, United States
c Center for Innovation in Neuroscience and Technology, Washington University School of Medicine, Saint Louis, MO 63110, United States
Abstract
Acoustic coupling through hairs remains a challenge to performing transcranial-focused ultrasound procedures. Here, we demonstrated that this challenge could be addressed by using oil as the coupling medium, leveraging oil’s high affinity to hairs due to their inherent hydrophobicity. We compared focused ultrasound-induced blood–brain barrier opening (FUS-BBBO) outcomes in mice under three coupling conditions: oil with hairs (“oil + hairs”), ultrasound gel with hair shaving (“ultrasound gel + no hair”), and ultrasound gel with hairs (“ultrasound gel + hairs”). The quality of the coupling was evaluated by T 2 -weighted magnetic resonance imaging (MRI) and passive cavitation detection (PCD). The outcome of FUS-BBBO was assessed by MRI contrast agent extravasation using in vivo T 1 -weighted contrast-enhanced MRI. It was also evaluated by ex vivo fluorescence imaging of the mouse brain after intravenous injection of a model drug, Evans blue. The results showed that “oil + hairs” consistently achieved high-quality acoustic coupling without trapping air bubbles. The FUS-BBBO outcome was not significantly different between the “oil + hairs” and the “ultrasound gel + no hair” groups. These two groups had significantly higher levels of BBB opening than the “ultrasound gel + hairs” group. This study demonstrated that oil could be a coupling medium for transcranial FUS procedures without shaving hairs. © 2023, Springer Nature Limited.
Funding details
National Institutes of HealthNIHR01CA276174, R01EB027223, R01EB030102, R01MH116981, R01NS128461
Office of Extramural Research, National Institutes of HealthOER
Office of Research Infrastructure Programs, National Institutes of HealthORIP, NIH, NIH-ORIP, ORIP
Document Type: Article
Publication Stage: Final
Source: Scopus
Reliability of diurnal salivary cortisol metrics: A meta-analysis and investigation in two independent samples
(2023) Comprehensive Psychoneuroendocrinology, 16, art. no. 100191, .
Norton, S.A.a , Baranger, D.A.a , Young, E.S.b , Voss, M.a , Hansen, I.a , Bondy, E.a , Rodrigues, M.a , Paul, S.E.a , Edershile, E.c , Hill, P.L.a , Oltmanns, T.F.a , Simpson, J.c , Bogdan, R.a
a Washington University in St. Louis, Department of Psychological & Brain Sciences, United States
b Utrecht University, Department of Psychology, Netherlands
c University of Minnesota, Department of Psychology, United States
Abstract
Stress-induced dysregulation of diurnal cortisol is a cornerstone of stress-disease theories; however, observed associations between cortisol, stress, and health have been inconsistent. The reliability of diurnal cortisol features may contribute to these equivocal findings. Our meta-analysis (5 diurnal features from 11 studies; total participant n = 3307) and investigation (15 diurnal cortisol features) in 2 independent studies (St. Louis Personality and Aging Network [SPAN] Study, n = 147, ages 61–73; Minnesota Longitudinal Study of Risk and Adaptation [MLSRA] Study, n = 90, age 37) revealed large variability in the day-to-day test-retest reliability of diurnal features derived from salivary cortisol data (i.e., ICC = 0.00–0.75). Collectively, these data indicate that some commonly used diurnal cortisol features have poor reliability that is insufficient for individual differences research (e.g., cortisol awakening response) while others (e.g., area under the curve with respect to ground) have fair-to-good reliability that could support reliable identification of associations in well-powered studies. © 2023 The Authors
Author Keywords
Cortisol; Reliability; Stress; Variability
Document Type: Article
Publication Stage: Final
Source: Scopus
Finding new and better treatments for psychiatric disorders
(2023) Neuropsychopharmacology, .
Paul, S.M.a , Potter, W.Z.b
a Karuna Therapeutics, Washington University School of Medicine, St. Louis, MO, United States
b Independent Consultant, Philadelphia, PA, United States
Abstract
In contrast to most fields of medicine, progress to discover and develop new and improved psychiatric drugs has been slow and disappointing. The vast majority of currently prescribed drugs to treat schizophrenia, mood and anxiety disorders are arguably no more effective than the first generation of psychiatric drugs introduced well over 50 years ago. With only a few exceptions current psychiatric drugs work via the same fundamental mechanisms of action as first-generation agents. Here we describe the reasons for this slow progress and outline a number of areas of research that involve a greater reliance on experimental therapeutics utilizing recent advances in neuroscience to better understand disease biology. We exemplify the potential impact of these areas of research focus with several recent examples of novel agents that have emerged and which support our optimism that newer, more effective and better tolerated agents, are on the horizon. Together with existing drugs these newer agents and novel mechanisms could offer markedly improved functional outcomes for the millions of people still disabled by psychiatric disorders. © 2023, The Author(s).
Document Type: Article
Publication Stage: Article in Press
Source: Scopus
Mutations in atypical hemolytic uremic syndrome provide evidence for the role of calcium in complement factor I
(2023) Blood, 142 (6), pp. 607-610.
Java, A.a , Atkinson, J.b , Hu, Z.b , Pozzi, N.c
a Division of Nephrology, Department of Medicine, Washington University School of Medicine, St. Louis, MO
b Division of Rheumatology, Department of Medicine, Washington University School of Medicine, St. Louis, MO
c Department of Biochemistry and Molecular Biology, Edward A. Doisy Research Center, Saint Louis University School of Medicine, St. Louis, MO
Abstract
Atypical hemolytic uremic syndrome (aHUS) is a rare thrombotic microangiopathy. Genetic variants in complement proteins are found in ≈60% of patients. Of these patients, ≈15% carry mutations in complement factor I (CFI). Factor I (FI) is a multidomain serine protease that cleaves and thereby inactivates C3b and C4b in the presence of cofactor proteins. Crystal structures have shown that FI possesses 2 calcium-binding domains, low-density lipoprotein receptor class A (LDLRA) 1 and LDLRA2. Yet, the role of calcium in FI is unknown. We determined that 9 genetic variants identified in aHUS (N151S, G162D, G188A, V230E, A240G, G243R, C247G, A258T, and Q260D) cluster around the calcium-binding site of LDLRA1. Using site-directed mutagenesis, we established that the synthesis of all, except A258T, was impaired, implying defective protein folding, perhaps due to loss of calcium binding. To further explore this possibility, we generated 12 alanine mutants that coordinate with the calcium in LDLRA1 and LDLRA2 (K239A, D242A, I244A, D246A, D252A, E253A, Y276A, N279A, E281A, D283A, D289A, and D290A) and are expected to perturb calcium binding. Except for K239A and Y276A, none of the mutants was secreted. These observations suggest that calcium ions play key structural and functional roles in FI. © 2023 The American Society of Hematology
Funding details
National Institutes of HealthNIH
National Heart, Lung, and Blood InstituteNHLBIR01 HL150146
Foundation for Barnes-Jewish HospitalFBJH
Washington University School of Medicine in St. LouisWUSM
Document Type: Article
Publication Stage: Final
Source: Scopus
Identification and targeting of a unique NaV1.7 domain driving chronic pain
(2023) Proceedings of the National Academy of Sciences of the United States of America, 120 (32), art. no. e2217800120, .
Gomez, K.a b , Stratton, H.J.c , Duran, P.a b , Loya, S.a b , Tang, C.a b , Calderon-Rivera, A.a b , François-Moutal, L.c , Khanna, M.a b , Madura, C.L.c , Luo, S.c , McKiver, B.d , Choi, E.d , Ran, D.c , Boinon, L.c , Perez-Miller, S.a b , Imad Damaj, M.d , Moutal, A.e , Khanna, R.a b f
a Department of Molecular Pathobiology, College of Dentistry, New York University, New York, NY 10010, United States
b NYU Pain Research Center, New York, NY 10010, United States
c Department of Pharmacology, College of Medicine, The University of Arizona, Tucson, AZ 85724, United States
d Department of Pharmacology and Toxicology and Translational Research Initiative for Pain and Neuropathy, Virginia Commonwealth University, Richmond, VA 23298-0613, United States
e Department of Pharmacology and Physiology, School of Medicine, St. Louis University, St. Louis, MO 63104, United States
f Department of Neuroscience and Physiology and Neuroscience Institute, School of Medicine, New York University, New York, NY 10010, United States
Abstract
Small molecules directly targeting the voltage-gated sodium channel (VGSC) NaV1.7 have not been clinically successful. We reported that preventing the addition of a small ubiquitin-like modifier onto the NaV1.7-interacting cytosolic collapsin response mediator protein 2 (CRMP2) blocked NaV1.7 function and was antinociceptive in rodent models of neuropathic pain. Here, we discovered a CRMP2 regulatory sequence (CRS) unique to NaV1.7 that is essential for this regulatory coupling. CRMP2 preferentially bound to the NaV1.7 CRS over other NaV isoforms. Substitution of the NaV1.7 CRS with the homologous domains from the other eight VGSC isoforms decreased NaV1.7 currents. A cell-penetrant decoy peptide corresponding to the NaV1.7-CRS reduced NaV1.7 currents and trafficking, decreased presynaptic NaV1.7 expression, reduced spinal CGRP release, and reversed nerve injury-induced mechanical allodynia. Importantly, the NaV1.7-CRS peptide did not produce motor impairment, nor did it alter physiological pain sensation, which is essential for survival. As a proof-of-concept for a NaV1.7 -targeted gene therapy, we packaged a plasmid encoding the NaV1.7-CRS in an AAV virus. Treatment with this virus reduced NaV1.7 function in both rodent and rhesus macaque sensory neurons. This gene therapy reversed and prevented mechanical allodynia in a model of nerve injury and reversed mechanical and cold allodynia in a model of chemotherapy-induced peripheral neuropathy. These findings support the conclusion that the CRS domain is a targetable region for the treatment of chronic neuropathic pain. Copyright © 2023 the Author(s).
Author Keywords
chronic pain; CRMP2; gene therapy; NaV1.7; SUMO
Funding details
National Institutes of HealthNIH
National Institute on Drug AbuseNIDADA042852, NS119263
National Institute of Neurological Disorders and StrokeNINDSNS098772, NS120663
Document Type: Article
Publication Stage: Final
Source: Scopus
Sulfenylation links oxidative stress to protein disulfide isomerase oxidase activity and thrombus formation
(2023) Journal of Thrombosis and Haemostasis, 21 (8), pp. 2137-2150.
Yang, M.a , Chiu, J.b , Scartelli, C.a , Ponzar, N.c , Patel, S.a , Patel, A.a , Ferreira, R.B.d , Keyes, R.F.e , Carroll, K.S.d , Pozzi, N.c , Hogg, P.J.b , Smith, B.C.e f , Flaumenhaft, R.a
a Division of Hemostasis and Thrombosis, Beth Israel Deaconess Medical Center and Harvard Medical School, Boston, MA, United States
b The Centenary Institute and University of Sydney, Sydney, NSW, Australia
c Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St. Louis, MO, United States
d Department of Chemistry, UF Scripps Biomedical Research, Jupiter, FL, United States
e Program in Chemical Biology, Medical College of Wisconsin, Milwaukee, WI, United States
f Department of Biochemistry, Medical College of Wisconsin, Milwaukee, WI, United States
Abstract
Background: Oxidative stress contributes to thrombosis in atherosclerosis, inflammation, infection, aging, and malignancy. Oxidant-induced cysteine modifications, including sulfenylation, can act as a redox-sensitive switch that controls protein function. Protein disulfide isomerase (PDI) is a prothrombotic enzyme with exquisitely redox-sensitive active-site cysteines. Objectives: We hypothesized that PDI is sulfenylated during oxidative stress, contributing to the prothrombotic potential of PDI. Methods: Biochemical and enzymatic assays using purified proteins, platelet and endothelial cell assays, and in vivo murine thrombosis studies were used to evaluate the role of oxidative stress in PDI sulfenylation and prothrombotic activity. Results: PDI exposure to oxidants resulted in the loss of PDI reductase activity and simultaneously promoted sulfenylated PDI generation. Following exposure to oxidants, sulfenylated PDI spontaneously converted to disulfided PDI. PDI oxidized in this manner was able to transfer disulfides to protein substrates. Inhibition of sulfenylation impaired disulfide formation by oxidants, indicating that sulfenylation is an intermediate during PDI oxidation. Agonist-induced activation of platelets and endothelium resulted in the release of sulfenylated PDI. PDI was also sulfenylated by oxidized low-density lipoprotein (oxLDL). In an in vivo model of thrombus formation, oxLDL markedly promoted platelet accumulation following an arteriolar injury. PDI oxidoreductase inhibition blocked oxLDL-mediated augmentation of thrombosis. Conclusion: PDI sulfenylation is a critical posttranslational modification that is an intermediate during disulfide PDI formation in the setting of oxidative stress. Oxidants generated by vascular cells during activation promote PDI sulfenylation, and interference with PDI during oxidative stress impairs thrombus formation. © 2023 International Society on Thrombosis and Haemostasis
Author Keywords
cysteine; disulfide; oxidation-reduction; protein disulfide isomerase; sulfenylation; thrombosis
Funding details
National Institutes of HealthNIHK99HL164888, R01GM102187, R01HL150146, R35GM128840, R35HL135775, T32HL007917, T32HL134643, U01HL143365
National Institutes of HealthNIH
American Society of HematologyASH
Sarnoff Cardiovascular Research Foundation
Cardiovascular Center, Medical College of WisconsinCVC, MCW
Document Type: Article
Publication Stage: Final
Source: Scopus