Hope Center Member Publications: December 31, 2023

Advanced structural brain aging in preclinical autosomal dominant Alzheimer disease
(2023) Molecular Neurodegeneration, 18 (1), art. no. 98, . 

Millar, P.R.^a , Gordon, B.A.^b , Wisch, J.K.^a , Schultz, S.A.^c d , Benzinger, T.L.^b , Cruchaga, C.^e , Hassenstab, J.J.^a , Ibanez, L.^a e f , Karch, C.^e , Llibre-Guerra, J.J.^a , Morris, J.C.^a , Perrin, R.J.^a g , Supnet-Bell, C.^a , Xiong, C.^h , Allegri, R.F.ⁱ , Berman, S.B.^j , Chhatwal, J.P.^c d , Chrem Mendez, P.A.ⁱ , Day, G.S.^k , Hofmann, A.^l m , Ikeuchi, T.ⁿ , Jucker, M.^l m , Lee, J.-H.^o , Levin, J.^p q r , Lopera, F.^s , Niimi, Y.^t , Sánchez-González, V.J.^u , Schofield, P.R.^v w , Sosa-Ortiz, A.L.^x , Vöglein, J.^p q , Bateman, R.J.^a , Ances, B.M.^a b , McDade, E.M.^a

^a Department of Neurology, Washington University in St. Louis, St. Louis, MO, United States
^b Mallinckrodt Institute of Radiology, Washington University in St. Louis, St. Louis, MO, United States
^c Department of Neurology, Harvard Medical School, Boston, MA, United States
^d Department of Neurology, Massachusetts General Hospital, Boston, MA, United States
^e Department of Psychiatry, Washington University in St. Louis, St. Louis, MO, United States
^f NeuroGenomics & Informatics Center, Washington University in St. Louis, St. Louis, MO, United States
^g Department of Pathology & Immunology, Washington University in St. Louis, St. Louis, MO, United States
^h Department of Biostatistics, Washington University in St. Louis, St. Louis, MO, United States
ⁱ Instituto Neurológico Fleni, Buenos Aires, Argentina
^j Department of Neurology, University of Pittsburgh, Pittsburgh, PA, United States
^k Department of Neurology, Mayo Clinic, Jacksonville, FL, United States
^l German Center for Neurodegenerative Diseases (DZNE), Tübingen, 72076, Germany
^m Department of Cellular Neurology, Hertie Institute for Clinical Brain Research, University of Tübingen, Tübingen, 72076, Germany
ⁿ Department of Molecular Genetics, Brain Research Institute, Niigata University, Niigata, Japan
^o Department of Neurology, University of Ulsan College of Medicine, Asan Medical Center, Seoul, South Korea
^p Department of Neurology, Ludwig-Maximilians-Universität München, Munich, Germany
^q German Center for Neurodegenerative Diseases, Munich, Germany
^r Munich Cluster for Systems Neurology (SyNergy), Munich, Germany
^s Universidad de Antioquia, Medellín, Colombia
^t Unit for Early and Exploratory Clinical Development, The University of Tokyo Hospital, Bunkyo-Ku, Tokyo, Japan
^u Departamento de Clínicas, CUALTOS, Universidad de Guadalajara, Tepatitlán de Morelos, Jalisco, Mexico
^v Neuroscience Research Australia, Sydney, NSW, Australia
^w School of Biomedical Sciences, University of New South Wales, Sydney, NSW, Australia
^x Instituto Nacional de Neurologia y Neurocirugía MVS, CDMX, Ciudad de México, Mexico

Abstract
Background: “Brain-predicted age” estimates biological age from complex, nonlinear features in neuroimaging scans. The brain age gap (BAG) between predicted and chronological age is elevated in sporadic Alzheimer disease (AD), but is underexplored in autosomal dominant AD (ADAD), in which AD progression is highly predictable with minimal confounding age-related co-pathology. Methods: We modeled BAG in 257 deeply-phenotyped ADAD mutation-carriers and 179 non-carriers from the Dominantly Inherited Alzheimer Network using minimally-processed structural MRI scans. We then tested whether BAG differed as a function of mutation and cognitive status, or estimated years until symptom onset, and whether it was associated with established markers of amyloid (PiB PET, CSF amyloid-β-42/40), phosphorylated tau (CSF and plasma pTau-181), neurodegeneration (CSF and plasma neurofilament-light-chain [NfL]), and cognition (global neuropsychological composite and CDR-sum of boxes). We compared BAG to other MRI measures, and examined heterogeneity in BAG as a function of ADAD mutation variants, APOE ε4 carrier status, sex, and education. Results: Advanced brain aging was observed in mutation-carriers approximately 7 years before expected symptom onset, in line with other established structural indicators of atrophy. BAG was moderately associated with amyloid PET and strongly associated with pTau-181, NfL, and cognition in mutation-carriers. Mutation variants, sex, and years of education contributed to variability in BAG. Conclusions: We extend prior work using BAG from sporadic AD to ADAD, noting consistent results. BAG associates well with markers of pTau, neurodegeneration, and cognition, but to a lesser extent, amyloid, in ADAD. BAG may capture similar signal to established MRI measures. However, BAG offers unique benefits in simplicity of data processing and interpretation. Thus, results in this unique ADAD cohort with few age-related confounds suggest that brain aging attributable to AD neuropathology can be accurately quantified from minimally-processed MRI. © 2023, The Author(s).

Author Keywords
Alzheimer disease;  Brain aging;  Machine learning;  Structural MRI

Funding details
AARFD-21-851415, K01AG073526, SG-20-690363
National Institutes of HealthNIHP01 AG00399139, P01AG026276, P30 AG06644403, R01 AG05255005, R01 AG05326704, R01 AG05456705, R01 AG05777705, R01 AG05867604, R01 AG07490901, R01 NS07532111, R01 NS09779906, U19 AG024904-16, U19 AG03243811, U19 AG06970102
National Institute on AgingNIA
Alzheimer’s AssociationAASG-20-690363-DIAN
RocheK23AG064029, U01AG057195, U01NS120901, U19AG032438
BrightFocus FoundationBFFA2022014F
Foundation for Barnes-Jewish HospitalFBJH
Fondation Brain Canada
Japan Agency for Medical Research and DevelopmentAMEDAMED JP23dk0207066
Canadian Institutes of Health ResearchIRSC
Fonds de Recherche du Québec – SantéFRQS
Consejo Nacional de Investigaciones Científicas y TécnicasCONICET
Korea Health Industry Development InstituteKHIDI
Instituto de Salud Carlos IIIISCIII
Deutsches Zentrum für Neurodegenerative ErkrankungenDZNE
Fleni

Document Type: Article
Publication Stage: Final
Source: Scopus

Plasma and cerebrospinal fluid proteomic signatures of acutely sleep-deprived humans: an exploratory study
(2023) SLEEP Advances, 4 (1), art. no. zpad047, . 

Vaquer-Alicea, A.^a , Yu, J.^b , Liu, H.^a , Lucey, B.P.^a

^a Department of Neurology, Washington University School of Medicine, St Louis, MO, United States
^b Department of Genetics, Washington University School of Medicine, St Louis, MO, United States

Abstract
Study Objectives: Acute sleep deprivation affects both central and peripheral biological processes. Prior research has mainly focused on specific proteins or biological pathways that are dysregulated in the setting of sustained wakefulness. This exploratory study aimed to provide a comprehensive view of the biological processes and proteins impacted by acute sleep deprivation in both plasma and cerebrospinal fluid (CSF). Methods: We collected plasma and CSF from human participants during one night of sleep deprivation and controlled normal sleep conditions. One thousand and three hundred proteins were measured at hour 0 and hour 24 using a high-scale aptamer-based proteomics platform (SOMAscan) and a systematic biological database tool (Metascape) was used to reveal altered biological pathways. Results: Acute sleep deprivation decreased the number of upregulated and downregulated biological pathways and proteins in plasma but increased upregulated and downregulated biological pathways and proteins in CSF. Predominantly affected proteins and pathways were associated with immune response, inflammation, phosphorylation, membrane signaling, cell-cell adhesion, and extracellular matrix organization. Conclusions: The identified modifications across biofluids add to evidence that acute sleep deprivation has important impacts on biological pathways and proteins that can negatively affect human health. As a hypothesis-driving study, these findings may help with the exploration of novel mechanisms that mediate sleep loss and associated conditions, drive the discovery of new sleep loss biomarkers, and ultimately aid in the identification of new targets for intervention to human diseases. © The Author(s) 2023.

Author Keywords
mRNA and protein expression;  sleep;  sleep deprivation;  the brain

Funding details
National Institutes of HealthNIHK76 AG054863

Document Type: Article
Publication Stage: Final
Source: Scopus

Relationships of Cognitive Measures with Cerebrospinal Fluid but Not Imaging Biomarkers of Alzheimer Disease Vary between Black and White Individuals
(2023) Annals of Neurology, . 

Bonomi, S.^a , Lu, R.^b , Schindler, S.E.^a c , Bui, Q.^b , Lah, J.J.^d e , Wolk, D.^f , Gleason, C.E.^g h i , Sperling, R.^j , Roberson, E.D.^k , Levey, A.I.^d e , Shaw, L.^f l , Van Hulle, C.^g h , Benzinger, T.^c m , Adams, M.^f , Manzanares, C.^d e , Qiu, D.^e , Hassenstab, J.^a c , Moulder, K.L.^a c , Balls-Berry, J.E.^a c , Johnson, K.ⁿ , Johnson, S.C.^g h i , Murchison, C.F.^k , Luo, J.^b , Gremminger, E.^a , Agboola, F.^b c , Grant, E.A.^b c , Hornbeck, R.^m , Massoumzadeh, P.^m , Keefe, S.^m , Dierker, D.^m , Gray, J.^c , Henson, R.L.^a c , Streitz, M.^a c , Mechanic-Hamilton, D.^f , Morris, J.C.^a c , Xiong, C.^b c

^a Department of Neurology, Washington University School of Medicine in St. Louis, St. Louis, MO, United States
^b Division of Biostatistics, Washington University School of Medicine in St. Louis, St. Louis, MO, United States
^c Knight Alzheimer Disease Research Center, Washington University School of Medicine in St. Louis, St. Louis, MO, United States
^d Department of Neurology, Emory University School of Medicine, Atlanta, GA, United States
^e Goizueta Alzheimer’s Disease Research Center, Emory University, Atlanta, GA, United States
^f Department of Neurology, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, United States
^g Division of Geriatrics and Gerontology, Department of Medicine, University of Wisconsin School of Medicine and Public Health, Madison, WI, United States
^h Wisconsin Alzheimer’s Disease Research Center, Madison, WI, United States
ⁱ Geriatric Research, Education and Clinical Center, William S. Middleton Memorial Veterans Hospital, Madison, WI, United States
^j Department of Neurology, Center for Alzheimer Research and Treatment, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA, United States
^k Center for Neurodegeneration and Experimental Therapeutics, Alzheimer’s Disease Center, Department of Neurology, University of Alabama at Birmingham, Birmingham, AL, United States
^l Department of Pathology and Laboratory Medicine, Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, United States
^m Mallinckrodt Institute of Radiology, Washington University School of Medicine in St. Louis, St. Louis, MO, United States
ⁿ Department of Neurology, Massachusetts General Hospital, Harvard Medical School, Boston, MA, United States

Abstract
Objective: Biomarkers of Alzheimer disease vary between groups of self-identified Black and White individuals in some studies. This study examined whether the relationships between biomarkers or between biomarkers and cognitive measures varied by racialized groups. Methods: Cerebrospinal fluid (CSF), amyloid positron emission tomography (PET), and magnetic resonance imaging measures were harmonized across four studies of memory and aging. Spearman correlations between biomarkers and between biomarkers and cognitive measures were calculated within each racialized group, then compared between groups by standard normal tests after Fisher’s Z-transformations. Results: The harmonized dataset included at least one biomarker measurement from 495 Black and 2,600 White participants. The mean age was similar between racialized groups. However, Black participants were less likely to have cognitive impairment (28% vs 36%) and had less abnormality of some CSF biomarkers including CSF Aβ42/40, total tau, p-tau181, and neurofilament light. CSF Aβ42/40 was negatively correlated with total tau and p-tau181 in both groups, but at a smaller magnitude in Black individuals. CSF Aβ42/40, total tau, and p-tau181 had weaker correlations with cognitive measures, especially episodic memory, in Black than White participants. Correlations of amyloid measures between CSF (Aβ42/40, Aβ42) and PET imaging were also weaker in Black than White participants. Importantly, no differences based on race were found in correlations between different imaging biomarkers, or in correlations between imaging biomarkers and cognitive measures. Interpretation: Relationships between CSF biomarkers but not imaging biomarkers varied by racialized groups. Imaging biomarkers performed more consistently across racialized groups in associations with cognitive measures. ANN NEUROL 2023. © 2023 American Neurological Association.

Funding details
National Institutes of HealthNIH067505
National Institute on AgingNIAP01 AG026276, P01 AG036694, P01 AG03991, P20 AG068024, P30 AG066444, P30 AG066511, P30 AG072979, R01 AG053550, R01 AG054059, R01 AG067505, R24 AG074915‐02

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

Longitudinal clinical, cognitive and biomarker profiles in dominantly inherited versus sporadic early-onset Alzheimer’s disease
(2023) Brain Communications, 5 (6), art. no. fcad280, . 

Llibre-Guerra, J.J.^a , Iaccarino, L.^b , Coble, D.^c , Edwards, L.^b , Li, Y.^c , Mcdade, E.^a , Strom, A.^b , Gordon, B.^d , Mundada, N.^b , Schindler, S.E.^a , Tsoy, E.^b , Ma, Y.^c , Lu, R.^c , Fagan, A.M.^a , Benzinger, T.L.S.^d , Soleimani-Meigooni, D.^b , Aschenbrenner, A.J.^a , Miller, Z.^b , Wang, G.^c , Kramer, J.H.^b , Hassenstab, J.^a , Rosen, H.J.^b , Morris, J.C.^a , Miller, B.L.^b , Xiong, C.^c , Perrin, R.J.^a e , Allegri, R.^f , Chrem, P.^f , Surace, E.^f , Berman, S.B.^g , Chhatwal, J.^h , Masters, C.L.ⁱ , Farlow, M.R.^j , Jucker, M.^k l , Levin, J.^m n o , Fox, N.C.^p , Day, G.^q , Gorno-Tempini, M.L.^b , Boxer, A.L.^b , La Joie, R.^b , Rabinovici, G.D.^b r , Bateman, R.^a

^a Department of Neurology, Washington University in St Louis, St Louis, MO 63108, United States
^b Department of Neurology, UCSF Weill Institute for Neurosciences, University of California, San Francisco, CA 94158, United States
^c Division of Biostatistics, Washington University in St Louis, St Louis, MO 63108, United States
^d Malinckrodt Institute of Radiology, Washington University in St Louis, St Louis, MO 63108, United States
^e Department of Pathology and Immunology, Washington University in St Louis, St. Louis, MO 63108, United States
^f Department of Cognitive Neurology, Institute for Neurological Research Fleni, Buenos Aires, Argentina
^g Department of Neurology, University of Pittsburgh, Pittsburgh, PA 15213, United States
^h Massachusetts General Hospital, Harvard Medical School, Boston, MA 02114, United States
ⁱ Florey Institute, The University of Melbourne, Melbourne, 3052, Australia
^j Neuroscience Center, Indiana University School of Medicine, Indianapolis, IN 46202, United States
^k DZNE-German Center for Neurodegenerative Diseases, Tübingen, 72076, Germany
^l Hertie-Institute for Clinical Brain Research, University of Tübingen, Tübingen, 72076, Germany
^m Department of Neurology, Ludwig-Maximilians-University, Munich, 80539, Germany
ⁿ German Center for Neurodegenerative Diseases, Munich, 81377, Germany
^o Munich Cluster for Systems Neurology (SyNergy), Munich, 81377, Germany
^p Dementia Research Centre, Department of Neurodegenerative Disease, University College London, Institute of Neurology, London, WC1N 3BG, United Kingdom
^q Department of Neurology, Mayo Clinic Florida, Jacksonville, FL 33224, United States
^r Department of Radiology and Biomedical Imaging, University of California, San Francisco, CA 94158, United States

Abstract
Approximately 5% of Alzheimer’s disease cases have an early age at onset (<65 years), with 5-10% of these cases attributed to dominantly inherited mutations and the remainder considered as sporadic. The extent to which dominantly inherited and sporadic early-onset Alzheimer’s disease overlap is unknown. In this study, we explored the clinical, cognitive and biomarker profiles of early-onset Alzheimer’s disease, focusing on commonalities and distinctions between dominantly inherited and sporadic cases. Our analysis included 117 participants with dominantly inherited Alzheimer’s disease enrolled in the Dominantly Inherited Alzheimer Network and 118 individuals with sporadic early-onset Alzheimer’s disease enrolled at the University of California San Francisco Alzheimer’s Disease Research Center. Baseline differences in clinical and biomarker profiles between both groups were compared using t-tests. Differences in the rates of decline were compared using linear mixed-effects models. Individuals with dominantly inherited Alzheimer’s disease exhibited an earlier age-at-symptom onset compared with the sporadic group [43.4 (SD ± 8.5) years versus 54.8 (SD ± 5.0) years, respectively, P < 0.001]. Sporadic cases showed a higher frequency of atypical clinical presentations relative to dominantly inherited (56.8% versus 8.5%, respectively) and a higher frequency of APOE-ϵ4 (50.0% versus 28.2%, P = 0.001). Compared with sporadic early onset, motor manifestations were higher in the dominantly inherited cohort [32.5% versus 16.9% at baseline (P = 0.006) and 46.1% versus 25.4% at last visit (P = 0.001)]. At baseline, the sporadic early-onset group performed worse on category fluency (P < 0.001), Trail Making Test Part B (P < 0.001) and digit span (P < 0.001). Longitudinally, both groups demonstrated similar rates of cognitive and functional decline in the early stages. After 10 years from symptom onset, dominantly inherited participants experienced a greater decline as measured by Clinical Dementia Rating Sum of Boxes [3.63 versus 1.82 points (P = 0.035)]. CSF amyloid beta-42 levels were comparable [244 (SD ± 39.3) pg/ml dominantly inherited versus 296 (SD ± 24.8) pg/ml sporadic early onset, P = 0.06]. CSF phosphorylated tau at threonine 181 levels were higher in the dominantly inherited Alzheimer’s disease cohort (87.3 versus 59.7 pg/ml, P = 0.005), but no significant differences were found for t-tau levels (P = 0.35). In summary, sporadic and inherited Alzheimer’s disease differed in baseline profiles; sporadic early onset is best distinguished from dominantly inherited by later age at onset, high frequency of atypical clinical presentations and worse executive performance at baseline. Despite these differences, shared pathways in longitudinal clinical decline and CSF biomarkers suggest potential common therapeutic targets for both populations, offering valuable insights for future research and clinical trial design. © 2023 The Author(s). Published by Oxford University Press on behalf of the Guarantors of Brain.

Author Keywords
dominantly inherited;  early-onset Alzheimer’s disease;  sporadic

Funding details
K01AG073526
K99AG065501, P01-AG019724, P30 AG062422, R01-AG045611, R35-AG072362
National Institutes of HealthNIH
National Institute on AgingNIAU19AG032438
National Institute of Neurological Disorders and StrokeNINDSR01-NS050915
Michael J. Fox Foundation for Parkinson’s ResearchMJFFMJFF-020770
Alzheimer’s AssociationAAAARFD-21-851415, SG-20-690363, SG-20-690363-DIAN, ZEN-21-848216
Foundation for Barnes-Jewish HospitalFBJH
Fondation Brain Canada
Japan Agency for Medical Research and DevelopmentAMED
Avid RadiopharmaceuticalsP01AG003991, P01AG026276, P30 AG066444, U19 AG024904, U19 AG032438
Canadian Institutes of Health ResearchIRSC
Fonds de Recherche du Québec – SantéFRQS
Korea Health Industry Development InstituteKHIDI
Instituto de Salud Carlos IIIISCIII
Deutsches Zentrum für Neurodegenerative ErkrankungenDZNE
Fleni

Document Type: Article
Publication Stage: Final
Source: Scopus

Antisense oligonucleotides targeting the miR-29b binding site in the GRN mRNA increase progranulin translation
(2023) Journal of Biological Chemistry, 299 (12), art. no. 105475, . 

Aggarwal, G.^a b c , Banerjee, S.^a b c , Jones, S.A.^a b c , Benchaar, Y.^d , Bélanger, J.^d , Sévigny, M.^d , Smith, D.M.^a b c , Niehoff, M.L.^a e , Pavlack, M.^b c , de Vera, I.M.S.^b c , Petkau, T.L.^f , Leavitt, B.R.^f g h , Ling, K.ⁱ , Jafar-Nejad, P.ⁱ , Rigo, F.ⁱ , Morley, J.E.^a , Farr, S.A.^a b c e , Dutchak, P.A.^d , Sephton, C.F.^d , Nguyen, A.D.^a b c

^a Division of Geriatric Medicine, Department of Internal Medicine, Saint Louis University School of Medicine, St Louis, MO, United States
^b Department of Pharmacology and Physiology, Saint Louis University School of Medicine, St Louis, MO, United States
^c Institute for Translational Neuroscience, Saint Louis University, St Louis, MO, United States
^d Department of Psychiatry and Neuroscience, CERVO Brain Research Centre, Laval University, Quebec City, QC, Canada
^e Veterans Affairs Medical Center, St Louis, MO, United States
^f Department of Medical Genetics, Centre for Molecular Medicine & Therapeutics, B.C. Children’s Hospital, University of British Columbia, Vancouver, BC, Canada
^g Division of Neurology, Department of Medicine, University of British Columbia Hospital, Vancouver, BC, Canada
^h Center for Brain Health, University of British Columbia, Vancouver, BC, Canada
ⁱ Ionis Pharmaceuticals, Carlsbad, CA, United States

Abstract
Heterozygous GRN (progranulin) mutations cause frontotemporal dementia (FTD) due to haploinsufficiency, and increasing progranulin levels is a major therapeutic goal. Several microRNAs, including miR-29b, negatively regulate progranulin protein levels. Antisense oligonucleotides (ASOs) are emerging as a promising therapeutic modality for neurological diseases, but strategies for increasing target protein levels are limited. Here, we tested the efficacy of ASOs as enhancers of progranulin expression by sterically blocking the miR-29b binding site in the 3′ UTR of the human GRN mRNA. We found 16 ASOs that increase progranulin protein in a dose-dependent manner in neuroglioma cells. A subset of these ASOs also increased progranulin protein in iPSC-derived neurons and in a humanized GRN mouse model. In FRET-based assays, the ASOs effectively competed for miR-29b from binding to the GRN 3′ UTR RNA. The ASOs increased levels of newly synthesized progranulin protein by increasing its translation, as revealed by polysome profiling. Together, our results demonstrate that ASOs can be used to effectively increase target protein levels by partially blocking miR binding sites. This ASO strategy may be therapeutically feasible for progranulin-deficient FTD as well as other conditions of haploinsufficiency. © 2023 The Authors

Author Keywords
antisense oligonucleotides;  frontotemporal dementia;  haploinsufficiency;  microRNA;  progranulin

Funding details
National Institutes of HealthNIHAG047339, AG064069
National Institutes of HealthNIH
National Center for Advancing Translational SciencesNCATSUL1TR002345
National Center for Advancing Translational SciencesNCATS
Saint Louis UniversitySLU
Natural Sciences and Engineering Research Council of CanadaNSERCDGECR-2018-00093, DGECR-2020-00060, RGPIN-2018-06227, RGPIN-2020-06376
Natural Sciences and Engineering Research Council of CanadaNSERC

Document Type: Article
Publication Stage: Final
Source: Scopus

Neuropilin-1 is essential for vascular endothelial growth factor A-mediated increase of sensory neuron activity and development of pain-like behaviors
(2023) Pain, 164 (12), pp. 2696-2710. 

Gomez, K.^a b , Duran, P.^a b , Tonello, R.^a b , Allen, H.N.^a b , Boinon, L.^c , Calderon-Rivera, A.^a b , Loya-López, S.^a b , Nelson, T.S.^a b , Ran, D.^c , Moutal, A.^d , Bunnett, N.W.^a b e , Khanna, R.^a b e

^a Department of Molecular Pathobiology, College of Dentistry, New York University, 433 1st Ave Room 720, New York City, NY 10010, United States
^b NYU Pain Research Center, New York, NY, United States
^c Department of Pharmacology, College of Medicine, University of Arizona, Tucson, AZ, United States
^d Department of Pharmacology and Physiology, School of Medicine, Saint Louis University, Saint Louis, MO, United States
^e Department of Neuroscience and Physiology, New York University Grossman School of Medicine, New York, NY, United States

Abstract
Neuropilin-1 (NRP-1) is a transmembrane glycoprotein that binds numerous ligands including vascular endothelial growth factor A (VEGFA). Binding of this ligand to NRP-1 and the co-receptor, the tyrosine kinase receptor VEGFR2, elicits nociceptor sensitization resulting in pain through the enhancement of the activity of voltage-gated sodium and calcium channels. We previously reported that blocking the interaction between VEGFA and NRP-1 with the Spike protein of SARS-CoV-2 attenuates VEGFA-induced dorsal root ganglion (DRG) neuronal excitability and alleviates neuropathic pain, pointing to the VEGFA/NRP-1 signaling as a novel therapeutic target of pain. Here, we investigated whether peripheral sensory neurons and spinal cord hyperexcitability and pain behaviors were affected by the loss of NRP-1. Nrp-1 is expressed in both peptidergic and nonpeptidergic sensory neurons. A CRIPSR/Cas9 strategy targeting the second exon of nrp-1 gene was used to knockdown NRP-1. Neuropilin-1 editing in DRG neurons reduced VEGFA-mediated increases in CaV2.2 currents and sodium currents through NaV1.7. Neuropilin-1 editing had no impact on voltage-gated potassium channels. Following in vivo editing of NRP-1, lumbar dorsal horn slices showed a decrease in the frequency of VEGFA-mediated increases in spontaneous excitatory postsynaptic currents. Finally, intrathecal injection of a lentivirus packaged with an NRP-1 guide RNA and Cas9 enzyme prevented spinal nerve injury-induced mechanical allodynia and thermal hyperalgesia in both male and female rats. Collectively, our findings highlight a key role of NRP-1 in modulating pain pathways in the sensory nervous system. © 2023 Lippincott Williams and Wilkins. All rights reserved.

Author Keywords
CaV2.2;  Chronic pain;  NaV1.7;  Neuropilin 1;  VEGFA

Funding details
National Institutes of HealthNIHDA042852, DE026806, DK118971, NS098772, NS102722, NS120663, NS122545, R01DE029951
National Institutes of HealthNIH
U.S. Department of DefenseDODW81XWH1810431, W81XWH2210238
U.S. Department of DefenseDOD
Takeda Pharmaceuticals International

Document Type: Article
Publication Stage: Final
Source: Scopus

From haemadin to haemanorm: Synthesis and characterization of full-length haemadin from the leech Haemadipsa sylvestris and of a novel bivalent, highly potent thrombin inhibitor (haemanorm)
(2023) Protein Science, 32 (12), art. no. e4825, . 

Acquasaliente, L.^a , Pierangelini, A.^a , Pagotto, A.^a , Pozzi, N.^a b , De Filippis, V.^a

^a Laboratory of Protein Chemistry & Molecular Hematology, Department of Pharmaceutical and Pharmacological Sciences, School of Medicine, University of Padova, Padua, Italy
^b Department of Biochemistry and Molecular Biology, Edward A. Doisy Research Center, Saint Louis University, St. Louis, MO, United States

Abstract
Hirudin from Hirudo medicinalis is a bivalent α-Thrombin (αT) inhibitor, targeting the enzyme active site and exosite-I, and is currently used in anticoagulant therapy along with its simplified analogue hirulog. Haemadin, a small protein (57 amino acids) isolated from the land-living leech Haemadipsa sylvestris, selectively inhibits αT with a potency identical to that of recombinant hirudin (KI = 0.2 pM), with which it shares a common disulfide topology and overall fold. At variance with hirudin, haemadin targets exosite-II and therefore (besides the free protease) it also blocks thrombomodulin-bound αT without inhibiting the active intermediate meizothrombin, thus offering potential advantages over hirudin. Here, we produced in reasonably high yields and pharmaceutical purity (&gt;98%) wild-type haemadin and the oxidation resistant Met5 → nor-Leucine analogue, both inhibiting αT with a KI of 0.2 pM. Thereafter, we used site-directed mutagenesis, spectroscopic, ligand-displacement, and Hydrogen/Deuterium Exchange-Mass Spectrometry techniques to map the αT regions relevant for the interaction with full-length haemadin and with the synthetic N- and C-terminal peptides Haem(1–10) and Haem(45–57). Haem(1–10) competitively binds to/inhibits αT active site (KI = 1.9 μM) and its potency was enhanced by 10-fold after Phe3 → β-Naphthylalanine exchange. Conversely to full-length haemadin, haem(45–57) displays intrinsic affinity for exosite-I (KD = 1.6 μM). Hence, we synthesized a peptide in which the sequences 1–9 and 45–57 were joined together through a 3-Glycine spacer to yield haemanorm, a highly potent (KI = 0.8 nM) inhibitor targeting αT active site and exosite-I. Haemanorm can be regarded as a novel class of hirulog-like αT inhibitors with potential pharmacological applications. © 2023 The Authors. Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.

Author Keywords
coagulation;  haemadin;  HDX-MS;  hirudin;  molecular recognition;  natural anticoagulants;  noncoded amino acids;  peptide synthesis;  protease inhibitors;  thrombin

Funding details
52012

Document Type: Article
Publication Stage: Final
Source: Scopus

Low circulating adropin levels in late-middle aged African Americans with poor cognitive performance
(2023) npj Aging, 9 (1), art. no. 24, . 

Aggarwal, G.^a b c , Malmstrom, T.K.^b d , Morley, J.E.^a , Miller, D.K.^e , Nguyen, A.D.^a b c , Butler, A.A.^b c

^a Division of Geriatric Medicine, Department of Internal Medicine, Saint Louis University School of Medicine, St. Louis, MO, United States
^b Institute for Translational Neuroscience, Saint Louis University, St. Louis, MO, United States
^c Department of Pharmacology and Physiology, Saint Louis University School of Medicine, St. Louis, MO, United States
^d Department of Psychiatry and Behavioral Neuroscience, Saint Louis University School of Medicine, St. Louis, MO, United States
^e Regenstrief Institute, Inc., Indianapolis, IN, United States

Abstract
We recently reported accelerated cognitive decline in Europeans aged > 70 years with low circulating adropin levels. Adropin is a small, secreted peptide that is highly expressed in the human nervous system. Expression profiling indicate relationships between adropin expression in the human brain and pathways that affect dementia risk. Moreover, increased adropin expression or treatment using synthetic adropin improves cognition in mouse models of aging. Here we report that low circulating adropin concentrations associate with poor cognition (worst quintile for a composite score derived from the MMSE and semantic fluency test) in late-middle aged community-dwelling African Americans (OR = 0.775, P < 0.05; age range 45–65 y, n = 352). The binomial logistic regression controlled for sex, age, education, cardiometabolic disease risk indicators, and obesity. Previous studies using cultured cells from the brains of human donors suggest high expression in astrocytes. In snRNA-seq data from the middle temporal gyrus (MTG) of human donors, adropin expression is higher in astrocytes relative to other cell types. Adropin expression in all cell-types declines with advance age, but is not affected by dementia status. In cultured human astrocytes, adropin expression also declines with donor age. Additional analysis indicated positive correlations between adropin and transcriptomic signatures of energy metabolism and protein synthesis that are adversely affected by donor age. Adropin expression is also suppressed by pro-inflammatory factors. Collectively, these data indicate low circulating adropin levels are a potential early risk indicator of cognitive impairment. Declining adropin expression in the brain is a plausible link between aging, neuroinflammation, and risk of cognitive decline. © 2023, The Author(s).

Funding details
National Institutes of HealthNIHR21NS108138
National Institutes of HealthNIH
National Institute on AgingNIAR01 AG010436
National Institute on AgingNIA
National Institute of Neurological Disorders and StrokeNINDS
Saint Louis UniversitySLU

Document Type: Article
Publication Stage: Final
Source: Scopus

Structural analyses of β2-glycoprotein I: is there a circular conformation?
(2023) Journal of Thrombosis and Haemostasis, 21 (12), pp. 3511-3521. 

Kumar, S.^a , Wulf, J., II^b , Basore, K.^b , Pozzi, N.^a

^a Edward A. Doisy Department of Biochemistry and Molecular Biology, Saint Louis University School of Medicine, St Louis, MO, United States
^b Washington University Center for Cellular Imaging, Washington University School of Medicine, St Louis, MO, United States

Abstract
Background: Antiphospholipid antibodies targeting β2-glycoprotein I (β2GPI) cause thrombosis and pregnancy morbidity in antiphospholipid syndrome (APS) patients. How these antibodies recognize β2GPI remains controversial. Objectives: This study aimed to elucidate the structure of β2GPI and evaluate how pathogenic anti–domain I (DI) antibodies recognize it in human plasma. Methods: β2GPI was made recombinant and purified from human plasma using different protocols. Structural and functional analyses were conducted using orthogonal techniques, namely, electron microscopy, size-exclusion chromatography, single-molecule Förster resonance energy transfer, and microfluidic diffusional sizing. Results: Electron microscopy and size-exclusion chromatography showed that the structure of β2GPI produced recombinantly and purified from plasma is elongated, even when subjected to conditions previously reported to favor circularization. Single-molecule Förster resonance energy transfer analyses of β2GPI labeled at positions 88 in DII and 278 in DV showed that these residues are located &gt;90 Å apart, consistent with an elongated form. They also documented that the distance between these 2 residues did not change when the protein was reconstituted in human plasma. Microfluidic diffusional sizing documented that β2GPI binds with moderate affinity to a prototypical anti-DI antibody targeting the epitope G40-R43 despite being elongated. Conclusion: Circulating β2GPI is elongated and, therefore, fully capable of binding to anti-DI antibodies. Binding of β2GPI to negatively charged phospholipids drives autoantibody recognition by increasing the local concentration of the antigen and not by dramatically changing its conformation. These findings clarify the structural properties of β2GPI, which have important implications for understanding APS pathogenesis and the development of APS diagnostics and therapeutics. © 2023 International Society on Thrombosis and Haemostasis

Author Keywords
antigen–antibody recognition;  antiphospholipid syndrome;  biophysics;  structural biology;  structure–function studies;  thrombosis

Funding details
National Heart, Lung, and Blood InstituteNHLBIR01 HL150146

Document Type: Article
Publication Stage: Final
Source: Scopus