Type-I IFNs in EAE drive synaptic transcriptional responses in neurons and astrocytes in the brain
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE346572 | GEO | 2026/09/14
REPOSITORIES: GEO
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Project description:Kv1.3, a voltage-gated potassium channel, is a therapeutic target in various neuropathological conditions. In rodent models, Kv1.3 inhibitors have been shown to inhibit disease-induced neuroinflammation. However, the cellular mechanisms of disease-modification by Kv1.3 blockers and the signaling pathways regulated by Kv1.3 and are unclear. In this -study, we aim to delineate the major protective mechanisms of Kv1.3 blockade in Alzheimer’s disease (AD) pathology by investigating the brain-derived bulk and single nuclear transcriptomes (snRNAseq) and bulk proteomes, along with neurobehavioral and pathological changes in 5xFAD mice. Following 3 months of Kv1.3 blockade via either small molecule PAP-1 or ShK-223 peptide, we observed Ab plaque reduction with both drugs and improved behavior with PAP-1. snRNAseq of brain nuclei identified neuronal and glial populations with expected AD-related changes in microglia. Pseudo-bulk analyses showed that PAP-1 disproportionately impacted oligodendrocytes and microglia while ShK-223 had more pronounced effects on glutamatergic neurons. PAP-1 also increased expression of myelination genes in oligodendrocytes and synaptic genes in neurons, with distinct effects on excitatory and inhibitory neurons. Cell-cell communication analyses suggested that PAP-1 increased crosstalk between neurons and astrocytes with endothelial cells. Bulk brain transcriptomics and proteomics with PAP-1 showed increased levels of synaptic and cognitive resilience-associated proteins while decreasing glial activation and interferon (IFN) responsive genes/proteins. PAP-1 also decreased STAT1 phosphorylation in the 5xFAD brain. To gain further mechanistic insight into Kv1.3's role in interferon signaling in microglia, we confirmed physical interactions between Kv1.3 and STAT1 and STAT3 in BV2 microglia overexpressing Kv1.3. We also found that Kv1.3 blockade (PAP-1) specifically reduced type 2 IFN but not type 1 IFN-induced phosphorylation of STAT1/3, a finding with further validated in another microglial cell line as well as in primary microglia derived from wild-type or Kv1.3-knockout mice. Lastly, we identified novel proteomic changes occurring in cerebrospinal fluid that reflect disease-modification by Kv1.3 blockers. This study reveals cell type-specific effects of Kv1.3 blockers in AD pathology that correspond with reduced neuropathology and neuroinflammation, augmentation of resilience pathways and neuro-vascular coupling and translationally relevant biofluid biomarkers of therapeutic effect. We also provide novel mechanistic insights into functional coupling between Kv1.3 channels and type-2 IFN signaling pathways in microglia.
2026-07-13 | PXD072055 | Pride
Project description:Kv1.3, a voltage-gated potassium channel, is a therapeutic target in various neuropathological conditions. In rodent models, Kv1.3 inhibitors have been shown to inhibit disease-induced neuroinflammation. However, the cellular mechanisms of disease-modification by Kv1.3 blockers and the signaling pathways regulated by Kv1.3 and are unclear. In this -study, we aim to delineate the major protective mechanisms of Kv1.3 blockade in Alzheimer’s disease (AD) pathology by investigating the brain-derived bulk and single nuclear transcriptomes (snRNAseq) and bulk proteomes, along with neurobehavioral and pathological changes in 5xFAD mice. Following 3 months of Kv1.3 blockade via either small molecule PAP-1 or ShK-223 peptide, we observed Ab plaque reduction with both drugs and improved behavior with PAP-1. snRNAseq of brain nuclei identified neuronal and glial populations with expected AD-related changes in microglia. Pseudo-bulk analyses showed that PAP-1 disproportionately impacted oligodendrocytes and microglia while ShK-223 had more pronounced effects on glutamatergic neurons. PAP-1 also increased expression of myelination genes in oligodendrocytes and synaptic genes in neurons, with distinct effects on excitatory and inhibitory neurons. Cell-cell communication analyses suggested that PAP-1 increased crosstalk between neurons and astrocytes with endothelial cells. Bulk brain transcriptomics and proteomics with PAP-1 showed increased levels of synaptic and cognitive resilience-associated proteins while decreasing glial activation and interferon (IFN) responsive genes/proteins. PAP-1 also decreased STAT1 phosphorylation in the 5xFAD brain. To gain further mechanistic insight into Kv1.3's role in interferon signaling in microglia, we confirmed physical interactions between Kv1.3 and STAT1 and STAT3 in BV2 microglia overexpressing Kv1.3. We also found that Kv1.3 blockade (PAP-1) specifically reduced type 2 IFN but not type 1 IFN-induced phosphorylation of STAT1/3, a finding with further validated in another microglial cell line as well as in primary microglia derived from wild-type or Kv1.3-knockout mice. Lastly, we identified novel proteomic changes occurring in cerebrospinal fluid that reflect disease-modification by Kv1.3 blockers. This study reveals cell type-specific effects of Kv1.3 blockers in AD pathology that correspond with reduced neuropathology and neuroinflammation, augmentation of resilience pathways and neuro-vascular coupling and translationally relevant biofluid biomarkers of therapeutic effect. We also provide novel mechanistic insights into functional coupling between Kv1.3 channels and type-2 IFN signaling pathways in microglia.
2026-07-13 | PXD071599 | Pride
Project description:Abstract Inhibition of voltage-gated potassium channel Kv1.3 is a therapeutic strategy to curb microglia-mediated neuroinflammation in neurodegeneration, although the cellular and signaling mechanisms of disease-modification by Kv1.3 blockers are unclear. In this study, we delineate protective mechanisms of Kv1.3 blockade in a mouse model of Alzheimer's disease (AD) pathology using comprehensive transcriptomics and proteomics profiling of brain, corresponding with neuropathological effects of two translationally relevant Kv1.3 blockers, namely small molecule PAP-1 and peptide ShK-223. Following 3 months of treatment, both molecules reduced Ab plaque burden. Single nuclear RNA seq (snRNA seq) of brain nuclei showed that PAP-1 disproportionately impacted oligodendrocytes and microglia and increased crosstalk between neurons and astrocytes with endothelial cells. In contrast, ShK-223 had pronounced effects on glutamatergic neurons and astrocytes. Both blockers increased expression of myelination genes in oligodendrocytes and synaptic genes in neurons. Neuroprotective effects of PAP-1 were further confirmed by bulk brain transcriptomics and proteomics whereby PAP-1 increased levels of synaptic, cognitive resilience and mitochondrial proteins, while decreasing glial and immune pathways including STAT1/3 phosphorylation. Using proximity labeling and co-immunoprecipitation, we found that Kv1.3 interacts with STAT1/3 in microglia. Using microglial cell lines and primary microglia, we discovered a preferential functional coupling between Kv1.3 and type 2 but not type 1 IFN signaling. Brain-level disease modification by Kv1.3 blockade was reflected in the cerebrospinal fluid (CSF) via reduced levels of neurofilament-light (NEFL) and resilience protein RPH3A, both of which are increased in human AD CSF. Together, this study demonstrates functional links between Kv1.3 channels and type 2 IFN signaling and reveals distinct cellular effects of Kv1.3 blockers in AD pathology that correspond with reduced neuropathology and neuroinflammation, augmentation of resilience and neuro-vascular pathways, along with biomarkers of therapeutic effect.
2026-07-15 | GSE338689 | GEO
Project description:Type I interferons (IFN-I) are crucial for effective antimicrobial defence in the central nervous system (CNS) but also can cause severe neurological disease (termed cerebral interferonopathy) as exemplified by Aicardi-Goutières Syndrome and chronic viral infection. In the CNS, microglia and astrocytes have essential roles in host responses to infection and injury, with both cell types responding to IFN-I. However, the extent to which the IFN-I responses of these cells differ, if at all, is still unknown. Here we determined the global transcriptional responses of astrocytes and microglia to the IFN-I, IFN-alpha. MGCs were prepared from 2–4 day-old C57BL/6 mice. Purified primary astrocytes were obtained from the MGCs by magnetic activated cell sorting using anti-CD11b beads. Microglia were obtained from mixed glial cell cultures by mechanical shaking for 4 h. After treating astrocytes and microglia with IFN-alpha for 12 h, microarray using Affymetrix mouse genome array 430 2.0 array was performed on total RNA extracted from these cells. We found that under basal conditions, each cell type has a unique gene expression pattern reflective of its developmental origin and biological function. Following stimulation with IFN-alpha for 12 h, astrocytes and microglia also displayed a common core response that was characterized by the increased expression of genes required for pathogen detection and elimination. Microglia had a more extensive and diverse response to IFN-alpha with twice the number of genes upregulated (282 vs. 141 genes) when compared with astrocytes. Validation of the findings in vivo further suggested that astrocytes and microglia play important but distinct roles in the development of IFN-alpha-driven cerebral interferonopathies.
2018-03-18 | E-MTAB-6301 | biostudies-arrayexpress
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2024-10-04 | GSE252454 | GEO
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2025-07-01 | GSE282917 | GEO
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2024-05-25 | GSE233717 | GEO
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