Hope Center Member Publications

Targeted delivery of therapeutic agents to the mouse brain using a stereotactic-guided focused ultrasound device
(2023) STAR Protocols, 4 (1), art. no. 102132, . 

Hu, Z.^a , Yang, Y.^a , Ye, D.^a , Chen, S.^a , Gong, Y.^a , Chukwu, C.^a , Chen, H.^a b

^a Department of Biomedical Engineering, Washington University in St. Louis, Saint Louis, MO 63130, United States
^b Department of Radiation Oncology, Washington University School of Medicine, Saint Louis, MO 63108, United States

Abstract
Existing protocols of focused ultrasound (FUS) combined with microbubble-mediated blood-brain barrier (BBB) opening (FUS-BBBO) in preclinical research require expensive ultrasound equipment and complex operating procedures. We developed a low-cost, easy-to-use, and precise FUS device for small animal models in preclinical research. Here, we provide a detailed protocol for building the FUS transducer, attaching the transducer to a stereotactic frame for precise brain targeting, applying the integrated FUS device to perform FUS-BBBO in mice, and evaluating the FUS-BBBO outcome. For complete details on the use and execution of this protocol, please refer to Hu et al. (2022).1 © 2023 The Authors

Author Keywords
Biotechnology and bioengineering;  Cancer;  Health Sciences;  Neuroscience;  Physics

Funding details
National Institutes of HealthNIHR01EB027223, R01EB030102, R01MH116981, UG3MH126861
Office of Naval ResearchONRN00014-19-1-2335

Document Type: Article
Publication Stage: Final
Source: Scopus

The Mechanism of Enantioselective Neurosteroid Actions on GABAA Receptors
(2023) Biomolecules, 13 (2), art. no. 341, . 

Tateiwa, H.^a b , Chintala, S.M.^a , Chen, Z.^a c , Wang, L.^a d , Amtashar, F.^a , Bracamontes, J.^a , Germann, A.L.^a , Pierce, S.R.^a , Covey, D.F.^a c e f , Akk, G.^a c , Evers, A.S.^a c f

^a Department of Anesthesiology, Washington University School of Medicine, St. Louis, MO 63110, United States
^b Department of Anesthesiology and Intensive Care Medicine, Kochi Medical School, Kochi, 7838505, Japan
^c Taylor Institute for Innovative Psychiatric Research, St. Louis, MO 63110, United States
^d Department of Anesthesiology, Union Hospital, Tongji Medical College, Huazhong University of Sciences and Technology, Wuhan, 430074, China
^e Department of Psychiatry, Washington University School of Medicine, St. Louis, MO 63110, United States
^f Department of Developmental Biology (Pharmacology), Washington University School of Medicine, St. Louis, MO 63110, United States

Abstract
The neurosteroid allopregnanolone (ALLO) and pregnanolone (PREG), are equally effective positive allosteric modulators (PAMs) of GABAA receptors. Interestingly, the PAM effects of ALLO are strongly enantioselective, whereas those of PREG are not. This study was aimed at determining the basis for this difference in enantioselectivity. The oocyte electrophysiology studies showed that ent-ALLO potentiates GABA-elicited currents in α1β3 GABAA receptors with lower potency and efficacy than ALLO, PREG or ent-PREG. The small PAM effect of ent-ALLO was prevented by the α1(Q242L) mutation in the intersubunit neurosteroid binding site between the β3 and α1 subunits. Consistent with this result, neurosteroid analogue photolabeling with mass spectrometric readout, showed that ent-ALLO binds weakly to the β3-α1 intersubunit binding site in comparison to ALLO, PREG and ent-PREG. Rigid body docking predicted that ent-ALLO binds in the intersubunit site with a preferred orientation 180° different than ALLO, PREG or ent-PREG, potentially explaining its weak binding and effect. Photolabeling studies did not identify differences between ALLO and ent-ALLO binding to the α1 or β3 intrasubunit binding sites that also mediate neurosteroid modulation of GABAA receptors. The results demonstrate that differential binding of ent-ALLO and ent-PREG to the β3-α1 intersubunit site accounts for the difference in enantioselectivity between ALLO and PREG. © 2023 by the authors.

Author Keywords
enantiomers;  GABA-A receptors;  neurosteroids;  photolabeling

Funding details
NIGMS-R35 GM140947, NIMH-P50MH122379

Document Type: Article
Publication Stage: Final
Source: Scopus

Conserved gene signatures shared among MAPT mutations reveal defects in calcium signaling
(2023) Frontiers in Molecular Biosciences, 10, art. no. 1051494, . 

Minaya, M.A.^a , Mahali, S.^a , Iyer, A.K.^a , Eteleeb, A.M.^a , Martinez, R.^a , Huang, G.^a , Budde, J.^a , Temple, S.^b , Nana, A.L.^c , Seeley, W.W.^c , Spina, S.^c , Grinberg, L.T.^c d , Harari, O.^a e f , Karch, C.M.^a e f

^a Department of Psychiatry, Washington University in St Louis, St Louis, MO, United States
^b Neural Stem Cell Institute, Rensselaer, NY, United States
^c Department of Neurology, UCSF Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, CA, United States
^d Department of Pathology, University of Sao Paulo, Sao Paulo, Brazil
^e Hope Center for Neurological Disorders, Washington University in St Louis, St Louis, MO, United States
^f NeuroGenomics and Informatics Center, Washington University in St Louis, St Louis, MO, United States

Abstract
Introduction: More than 50 mutations in the MAPT gene result in heterogeneous forms of frontotemporal lobar dementia with tau inclusions (FTLD-Tau). However, early pathogenic events that lead to disease and the degree to which they are common across MAPT mutations remain poorly understood. The goal of this study is to determine whether there is a common molecular signature of FTLD-Tau. Methods: We analyzed genes differentially expressed in induced pluripotent stem cell–derived neurons (iPSC-neurons) that represent the three major categories of MAPT mutations: splicing (IVS10 + 16), exon 10 (p.P301L), and C-terminal (p.R406W) compared with isogenic controls. The genes that were commonly differentially expressed in MAPT IVS10 + 16, p.P301L, and p.R406W neurons were enriched in trans-synaptic signaling, neuronal processes, and lysosomal function. Many of these pathways are sensitive to disruptions in calcium homeostasis. One gene, CALB1, was significantly reduced across the three MAPT mutant iPSC-neurons and in a mouse model of tau accumulation. We observed a significant reduction in calcium levels in MAPT mutant neurons compared with isogenic controls, pointing to a functional consequence of this disrupted gene expression. Finally, a subset of genes commonly differentially expressed across MAPT mutations were also dysregulated in brains from MAPT mutation carriers and to a lesser extent in brains from sporadic Alzheimer disease and progressive supranuclear palsy, suggesting that molecular signatures relevant to genetic and sporadic forms of tauopathy are captured in a dish. The results from this study demonstrate that iPSC-neurons capture molecular processes that occur in human brains and can be used to pinpoint common molecular pathways involving synaptic and lysosomal function and neuronal development, which may be regulated by disruptions in calcium homeostasis. Copyright © 2023 Minaya, Mahali, Iyer, Eteleeb, Martinez, Huang, Budde, Temple, Nana, Seeley, Spina, Grinberg, Harari and Karch.

Author Keywords
calcium signaling;  frontotemporal dementia (FTD);  IPSC-derived neurons;  MAPT mutations (tau);  transcriptomics

Funding details
P01AG019724, P01AG026276, P01AG03991, P30AG062422, P30AG066444, U01AG057195, U19AG063911, UL1TR002345
National Institutes of HealthNIHK24 AG053435, OD021629, P30 AG066444, R01 AG056293, R01 AG057777, R01 AG062359, RF1 NS110890, U54 NS123985
Hope Center for Neurological Disorders
Rainwater Charitable FoundationRCF
Office of Research Infrastructure Programs, National Institutes of HealthORIP, NIH

Document Type: Article
Publication Stage: Final
Source: Scopus

Gene-environment interactions increase the risk of paediatric-onset multiple sclerosis associated with household chemical exposures
(2023) Journal of Neurology, Neurosurgery and Psychiatry, art. no. jnnp-2022-330713, . 

Nasr, Z.^a , Schoeps, V.A.^a , Ziaei, A.^a , Virupakshaiah, A.^a , Adams, C.^b , Casper, T.C.^c , Waltz, M.^c , Rose, J.^c , Rodriguez, M.^d , Tillema, J.-M.^d , Chitnis, T.^e , Graves, J.S.^f , Benson, L.^g , Rensel, M.^h , Krupp, L.ⁱ , Waldman, A.T.^j , Weinstock-Guttman, B.^k , Lotze, T.^l , Greenberg, B.^m , Aaen, G.ⁿ , Mar, S.^o , Schreiner, T.^p , Hart, J.^a , Simpson-Yap, S.^q r s , Mesaros, C.^t , Barcellos, L.F.^b u , Waubant, E.^a

^a UCSF Weill Institute for Neurosciences, University of California San Francisco, San Francisco, CA 94117, United States
^b Genetic Epidemiology and Genomics Laboratory, Divisions of Epidemiology and Biostatistics, School of Public Health, University of California Berkeley, Berkeley, CA, United States
^c University of Utah Health, Salt Lake City, UT, United States
^d Mayo Clinic, Rochester, MN, United States
^e Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, United States
^f University of California San Diego, San Diego, CA, United States
^g Childrens Hospital Boston, Boston, MA, United States
^h Cleveland Clinic, Cleveland, OH, United States
ⁱ New York University Medical Center, New York City, NY, United States
^j Division of Child Neurology, Children’s Hospital of Philadelphia, Philadelphia, PA, United States
^k Jacobs School of Medicine and Biomedical Sciences, University of Buffalo, Buffalo, NY, United States
^l Texas Children’s Hospital, Houston, TX, United States
^m University of Texas Southwestern Medical Center, Dallas, TX, United States
ⁿ Loma Linda University Children’s Hospital, Loma Linda, CA, United States
^o Washington University in St. Louis, St Louis, MO, United States
^p Denver Children’s Hospital, Denver, CO, United States
^q Neuroepidemiology Unit, University of Melbourne School of Population and Global Health, Carlton, Melbourne, Australia
^r Clinical Outcomes Research Unit (CORe), Royal Melbourne Hospital, University of Melbourne, Parkville, Melbourne, Australia
^s Multiple Sclerosis Flagship, Menzies Institute for Medical Research, University of Tasmania, Hobart, TAS, Australia
^t Department of Systems Pharmacology and Translational Therapeutics (SPATT), University of Pennsylvania, Philadelphia, PA, United States
^u Department of Integrative Biology, University of California Berkeley, Berkeley, CA, United States

Abstract
Background: We previously reported an association between household chemical exposures and an increased risk of paediatric-onset multiple sclerosis. Methods: Using a case-control paediatric multiple sclerosis study, gene-environment interaction between exposure to household chemicals and genotypes for risk of paediatric-onset multiple sclerosis was estimated. Genetic risk factors of interest included the two major HLA multiple sclerosis risk factors, the presence of DRB1∗15 and the absence of A∗02, and multiple sclerosis risk variants within the metabolic pathways of common household toxic chemicals, including IL-6 (rs2069852), BCL-2 (rs2187163) and NFKB1 (rs7665090). Results: 490 paediatric-onset multiple sclerosis cases and 716 controls were included in the analyses. Exposures to insect repellent for ticks or mosquitos (OR 1.47, 95% CI 1.06 to 2.04, p=0.019), weed control products (OR 2.15, 95% CI 1.51 to 3.07, p<0.001) and plant/tree insect or disease control products (OR 3.25, 95% CI 1.92 to 5.49, p<0.001) were associated with increased odds of paediatric-onset multiple sclerosis. There was significant additive interaction between exposure to weed control products and NFKB1 SNP GG (attributable proportions (AP) 0.48, 95% CI 0.10 to 0.87), and exposure to plant or disease control products and absence of HLA-A∗02 (AP 0.56; 95% CI 0.03 to 1.08). There was a multiplicative interaction between exposure to weed control products and NFKB1 SNP GG genotype (OR 2.30, 95% CI 1.00 to 5.30) but not for other exposures and risk variants. No interactions were found with IL-6 and BCL-2 SNP GG genotypes. Conclusions: The presence of gene-environment interactions with household toxins supports their possible causal role in paediatric-onset multiple sclerosis. © 2023 Author(s). Published by BMJ.

Author Keywords
genetics;  multiple sclerosis;  paediatric neurology

Funding details
National Institutes of HealthNIH
National Multiple Sclerosis SocietyNMSSHC-1509-06233
Patient-Centered Outcomes Research InstitutePCORI
Multiple Sclerosis International FederationMSIF

Document Type: Article
Publication Stage: Article in Press
Source: Scopus

Top-down and bottom-up propagation of disease in the neuronal ceroid lipofuscinoses
(2022) Frontiers in Neurology, 13, art. no. 1061363, . 

Ostergaard, J.R.^a , Nelvagal, H.R.^b c , Cooper, J.D.^b d e

^a Department of Child and Adolescencet, Centre for Rare Diseases, Aarhus, Denmark
^b Department of Pediatrics, School of Medicine, Washington University in St Louis, St Louis, MO, United States
^c UCL School of Pharmacy, University College London, London, United Kingdom
^d Department of Genetics, School of Medicine, Washington University in St Louis, St Louis, MO, United States
^e Department of Neurology, School of Medicine, Washington University in St Louis, St Louis, MO, United States

Abstract
Background: The Neuronal Ceroid Lipofuscinoses (NCLs) may be considered distinct neurodegenerative disorders with separate underlying molecular causes resulting from monogenetic mutations. An alternative hypothesis is to consider the NCLs as related diseases that share lipofuscin pathobiology as the common core feature, but otherwise distinguished by different a) initial anatomic location, and b) disease propagation. Methods: We have tested this hypothesis by comparing known differences in symptomatology and pathology of the CLN1 phenotype caused by complete loss of PPT1 function (i.e., the classical infantile form) and of the classical juvenile CLN3 phenotype. These two forms of NCL represent early onset and rapidly progressing vs. late onset and slowly progressing disease modalities respectively. Results: Despite displaying similar pathological endpoints, the clinical phenotypes and the evidence of imaging and postmortem studies reveal strikingly different time courses and distributions of disease propagation. Data from CLN1 disease are indicative of disease propagation from the body, with early effects within the spinal cord and subsequently within the brainstem, the cerebral hemispheres, cerebellum and retina. In contrast, the retina appears to be the most vulnerable organ in CLN3, and the site where pathology is first present. Pathology subsequently is present in the occipital connectome of the CLN3 brain, followed by a top-down propagation in which cerebral and cerebellar atrophy in early adolescence is followed by involvement of the peripheral nerves in later adolescence/early twenties, with the extrapyramidal system also affected during this time course. Discussion: The propagation of disease in these two NCLs therefore has much in common with the “Brain-first” vs. “Body-first” models of alpha-synuclein propagation in Parkinson’s disease. CLN1 disease represents a “Body-first” or bottom-up disease propagation and CLN3 disease having a “Brain-first” and top-down propagation. It is noteworthy that the varied phenotypes of CLN1 disease, whether it starts in infancy (infantile form) or later in childhood (juvenile form), still fit with our proposed hypothesis of a bottom-up disease propagation in CLN1. Likewise, in protracted CLN3 disease, where both cognitive and motor declines are delayed, the initial manifestations of disease are also seen in the outer retinal layers, i.e., identical to classical Juvenile NCL disease. Copyright © 2022 Ostergaard, Nelvagal and Cooper.

Author Keywords
Body-first;  Brain-first;  CLN1;  CLN3;  connectome;  disease propagation;  neurodegeneration;  neuronal ceroid lipofuscinoses

Document Type: Article
Publication Stage: Final
Source: Scopus