Venezuelan equine encephalitis virus infection causes chronic neurobehavioral outcomes, cellular remodeling, and hippocampal single-cell transcriptomic changes.
The 3 matches
- [1] § Results › VEEV infection significantly impacts single cell gene expression within the hippocampus at 7 DPI, including multiple immune and antiviral genes ↔ scRNA-seq_code.R, lines 167–233 · score 0.83 · endothelial cells, progenitor cells, immune cell, inhibitory neurons, excitatory neurons, UMAP
- [2] § Materials and methods › scRNA-seq sample purification, processing, and analysis ↔ scRNA-seq_code.R, lines 126–165 · score 0.77 · low quality cells, quality control, nFeature_RNA, PCA, harmony, variable
- [3] § Results › VEEV infection significantly impacts single cell gene expression within the hippocampus at 7 DPI, including multiple immune and antiviral genes ↔ scRNA-seq_code.R, lines 167–233 · score 0.74 · endothelial cells, progenitor cells, inhibitory neurons, excitatory neurons, UMAP, oligodendrocytes
Paper
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The authors' code
R · 238 lines · 11 KB · no license · 3 matches
- options(stringAsFactors=F)
- library(dplyr)
- library(Seurat)
- library(patchwork)
- library(stringr)
- library(harmony)
- #### read 10X genomics scRNAseq data
- MOCK_106_DPI_Spike <- Read10X("MOCK-90-DPI_results_FRP/sample_filtered_feature_bc_matrix/sample_filtered_feature_bc_matrix/")
- VEEV_106_DPI_Spike <- Read10X("VEEV-90-DPI_results_FRP/sample_filtered_feature_bc_matrix/sample_filtered_feature_bc_matrix/")
- VEEV_neuro_106_DPI_Spike <- Read10X("VEEV-neuro-90-DPI_results_FRP/sample_filtered_feature_bc_matrix/sample_filtered_feature_bc_matrix/")
- MOCK_7_DPI_Spike <- Read10X("MOCK-7-DPI_results_FRP/sample_filtered_feature_bc_matrix/sample_filtered_feature_bc_matrix/")
- VEEV_7_DPI_Spike <- Read10X("VEEV-7-DPI_results_FRP/sample_filtered_feature_bc_matrix/sample_filtered_feature_bc_matrix/")
- VEEV_neuro_106_DPI_v2_Spike <- Read10X("VEEV-neuro-90-DPI_results_FRP_additional/sample_filtered_feature_bc_matrix/")
- MOCK_106_DPI_Spike.obj <- CreateSeuratObject(counts = MOCK_106_DPI_Spike, project = "MOCK_106_DPI_Spike", min.cells = 0, min.features = 0)
- VEEV_106_DPI_Spike.obj <- CreateSeuratObject(counts = VEEV_106_DPI_Spike, project = "VEEV_106_DPI_Spike", min.cells = 0, min.features = 0)
- VEEV_neuro_106_DPI_Spike.obj <- CreateSeuratObject(counts = VEEV_neuro_106_DPI_Spike, project = "VEEV_neuro_106_DPI_Spike", min.cells = 0, min.features = 0)
- MOCK_7_DPI_Spike.obj <- CreateSeuratObject(counts = MOCK_7_DPI_Spike, project = "MOCK_7_DPI_Spike", min.cells = 0, min.features = 0)
- VEEV_7_DPI_Spike.obj <- CreateSeuratObject(counts = VEEV_7_DPI_Spike, project = "VEEV_7_DPI_Spike", min.cells = 0, min.features = 0)
- VEEV_neuro_106_DPI_v2_Spike.obj <- CreateSeuratObject(counts = VEEV_neuro_106_DPI_v2_Spike, project = "VEEV_neuro_106_DPI_v2_Spike", min.cells = 0, min.features = 0)
- merge_obj <- merge(x=MOCK_106_DPI_Spike.obj,
- y = c(VEEV_106_DPI_Spike.obj,
- VEEV_neuro_106_DPI_Spike.obj,
- MOCK_7_DPI_Spike.obj,
- VEEV_7_DPI_Spike.obj,
- VEEV_neuro_106_DPI_v2_Spike.obj
- ),
- add.cell.ids = c("Study-2-MOCK-90-DPI",
- "Study-2-VEEV-90-DPI",
- "Study-2-VEEV-neuro-90-DPI",
- "Study-3-MOCK-7-DPI",
- "Study-3-VEEV-7-DPI",
- "VEEV_neuro_106_DPI_add"
- ) )
- merge_obj <- JoinLayers(merge_obj)
- #################################################
- ######################## merge two batches of VEEV_neuro_106_DPI
- length(unique(c(colnames(VEEV_neuro_106_DPI_Spike) , colnames(VEEV_neuro_106_DPI_v2_Spike))))
- all(rownames(VEEV_neuro_106_DPI_Spike) == rownames(VEEV_neuro_106_DPI_v2_Spike))
- VEEV_neuro_106_DPI_merge_Spike <- VEEV_neuro_106_DPI_v2_Spike
- cells <- intersect(colnames(VEEV_neuro_106_DPI_Spike) , colnames(VEEV_neuro_106_DPI_v2_Spike))
- VEEV_neuro_106_DPI_merge_Spike <- VEEV_neuro_106_DPI_v2_Spike
- VEEV_neuro_106_DPI_merge_Spike[,cells] <- VEEV_neuro_106_DPI_merge_Spike[,cells] + VEEV_neuro_106_DPI_Spike[,cells]
- VEEV_neuro_106_DPI_merge_Spike <- cbind(VEEV_neuro_106_DPI_merge_Spike,
- VEEV_neuro_106_DPI_Spike[,-which(colnames(VEEV_neuro_106_DPI_Spike) %in% cells)]
- )
- VEEV_neuro_106_DPI_merge_Spike.obj <- CreateSeuratObject(counts = VEEV_neuro_106_DPI_merge_Spike, project = "VEEV_neuro_106_DPI_merge_Spike", min.cells = 0, min.features = 0)
- merge_obj <- merge(x=MOCK_106_DPI_Spike.obj,
- y = c(VEEV_106_DPI_Spike.obj,
- VEEV_neuro_106_DPI_merge_Spike.obj,
- MOCK_7_DPI_Spike.obj,
- VEEV_7_DPI_Spike.obj
- ),
- add.cell.ids = c("Study-2-MOCK-90-DPI",
- "Study-2-VEEV-90-DPI",
- "Study-2-VEEV-neuro-90-DPI",
- "Study-3-MOCK-7-DPI",
- "Study-3-VEEV-7-DPI"
- ) )
- merge_obj <- JoinLayers(merge_obj)
- #######################################################################
- ##### summary parameters after merge two batches
- table(merge_obj$orig.ident)
- tapply(merge_obj$nCount_RNA,merge_obj$orig.ident,median)
- tapply(merge_obj$nFeature_RNA,merge_obj$orig.ident,median)
- length(which(rowSums(MOCK_106_DPI_Spike)>0))
- length(which(rowSums(VEEV_106_DPI_Spike)>0))
- length(which(rowSums(VEEV_neuro_106_DPI_merge_Spike)>0))
- length(which(rowSums(MOCK_7_DPI_Spike)>0))
- length(which(rowSums(VEEV_7_DPI_Spike)>0))
- ###########################################
- ## summary infected cells
- merge_obj$probe1=merge_obj[["RNA"]]$counts["Probe1",]
- merge_obj$probe2=merge_obj[["RNA"]]$counts["Probe2",]
- merge_obj$probe3=merge_obj[["RNA"]]$counts["Probe3",]
- merge_obj$probe4=merge_obj[["RNA"]]$counts["Probe4",]
- merge_obj$probe5=merge_obj[["RNA"]]$counts["Probe5",]
- merge_obj$probe6=merge_obj[["RNA"]]$counts["Probe6",]
- merge_obj$probe7=merge_obj[["RNA"]]$counts["Probe7",]
- merge_obj$probe9=merge_obj[["RNA"]]$counts["Probe9",]
- merge_obj$probe10=merge_obj[["RNA"]]$counts["Probe10",]
- merge_obj$probe11=merge_obj[["RNA"]]$counts["Probe10",]
- merge_obj$probe12=merge_obj[["RNA"]]$counts["Probe12",]
- probe_mat <- data.frame(probe1=merge_obj[["RNA"]]$counts["Probe1",],
- probe2=merge_obj[["RNA"]]$counts["Probe2",],
- probe3=merge_obj[["RNA"]]$counts["Probe3",],
- probe4=merge_obj[["RNA"]]$counts["Probe4",],
- probe5=merge_obj[["RNA"]]$counts["Probe5",],
- probe6=merge_obj[["RNA"]]$counts["Probe6",],
- probe7=merge_obj[["RNA"]]$counts["Probe7",],
- probe9=merge_obj[["RNA"]]$counts["Probe9",],
- probe10=merge_obj[["RNA"]]$counts["Probe10",],
- probe11=merge_obj[["RNA"]]$counts["Probe11",],
- probe12=merge_obj[["RNA"]]$counts["Probe12",]
- )
- index <- apply(probe_mat,1,function(x){return(all(x>0))} )
- merge_obj$infected2 <- as.numeric(0)
- merge_obj$infected2[which(index == TRUE )] <- 1
- ##############################################
- ######### quality control
- merge_obj[["percent.mt"]] <- PercentageFeatureSet(merge_obj, pattern = "mt-")
- # Visualize QC metrics as a violin plot
- VlnPlot(merge_obj, features = c("nFeature_RNA", "nCount_RNA", "percent.mt"), ncol = 3)
- plot1 <- FeatureScatter(merge_obj, feature1 = "nCount_RNA", feature2 = "percent.mt")
- plot2 <- FeatureScatter(merge_obj, feature1 = "nCount_RNA", feature2 = "nFeature_RNA")
- plot1 + plot2
- ####################################
- ######### filter low quality cells
- mt.cutoff <- 10
- merge_obj <- merge_obj[,which(merge_obj$percent.mt <= mt.cutoff)]
- merge_obj <- merge_obj[,which(merge_obj$nFeature_RNA >= 200)]
- #######################################################################
- ##### summary parameters after quality control
- table(merge_obj$orig.ident)
- tapply(merge_obj$nCount_RNA,merge_obj$orig.ident,median)
- tapply(merge_obj$nFeature_RNA,merge_obj$orig.ident,median)
- #######################################################################
- ## clustering
- merge_obj <- NormalizeData(merge_obj)
- merge_obj <- FindVariableFeatures(merge_obj, selection.method = "vst", nfeatures = 2000)
- top10 <- head(VariableFeatures(merge_obj), 10)
- # plot variable features with and without labels
- plot1 <- VariableFeaturePlot(merge_obj)
- plot2 <- LabelPoints(plot = plot1, points = top10, repel = TRUE)
- plot1 + plot2
- all.genes <- rownames(merge_obj)
- merge_obj <- ScaleData(merge_obj, features = all.genes)
- merge_obj <- RunPCA(merge_obj, features = VariableFeatures(object = merge_obj))
- merge_obj <- RunHarmony(merge_obj, group.by.vars = "orig.ident")
- DimPlot(merge_obj, reduction = "pca") + NoLegend()
- DimHeatmap(merge_obj, dims = 1:3, cells = 500, balanced = TRUE)
- ElbowPlot(merge_obj,ndims =30)
- merge_obj <- FindNeighbors(merge_obj, dims = 1:20,reduction='harmony')
- merge_obj <- FindClusters(merge_obj, resolution = 0.2)
- merge_obj <- RunUMAP(merge_obj, dims = 1:20,reduction = "harmony")
- DimPlot(merge_obj, reduction = "umap",label = T,pt.size=0.5)
- save(merge_obj,file=merge_obj,cluster.Rdata)
- #################################################################
- ############################## cell type annotation
- merge_obj$celltype <- ""
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(0,11,14))] = "Inhibitory_neurons"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(4:6,13))] = "Excitatory_neurons"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(1))] = "Oligodendrocytes"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(2))] = "Microglia"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(3))] = "Astrocytes"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(7))] = "Pericytes "
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(8,16))] = "Immune_cells"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(9))] = "Endothelial_cells"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(10))] = "OPCs"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(12))] = "Neural_progenitor_cell"
- merge_obj$celltype[which(merge_obj$seurat_clusters %in% c(15))] = "Ependymal_cells"
- ###### plot cell type markers and UMAP
- library(ggplot2)
- colors <- c( "#FB8072", "#45BFC0", "#96B85D", "#C790E0","#DEB068", "#4EAEE3","#FF8DC6", "#52B77C", "#7DACD8", "#FFB090", "#B89FC8"
- )
- DimPlot(merge_obj,group.by = "celltype", cols = colors,label=F )
- selected_features = c(
- "Aqp4", "Slc1a3", ## Astrocytes
- "Slc17a7","Slc17a6", ## EXC neuron
- "Gad1", "Gad2", ## INH neuron
- "Ttr","Ecrg4", ## NPC
- "Itgam","Aif1", ## Microglia
- "Mog","Mag","Ermn", ## Oligodendrocytes
- "Cspg4", "Vcan", ## OPC
- "Cd3e" , "Nkg7" , ## Immune cells
- "Pecam1", "Cldn5" , ## Endothelial
- "Col1a2","Pdgfrb", ## Pericytes
- "Foxj1", "Ccdc153","Cfap65","Pax6","Hes5" ## Ependymal cells
- )
- merge_obj$celltype <- factor(merge_obj$celltype,
- levels = c("Astrocytes",
- "Excitatory_neurons",
- "Inhibitory_neurons",
- "NPC",
- "Microglia",
- "Oligodendrocytes",
- "OPCs",
- "Immune_cells",
- "Endothelial_cells",
- "Pericytes",
- "Ependymal_cells"
- ))
- p1 <- DoHeatmap(merge_obj, features = selected_features,group.by="celltype",
- group.colors = colors
- )+
- scale_fill_gradientn(colors = c("white","grey","firebrick3"))
- VlnPlot(merge_obj,
- features = selected_features,group.by="celltype",raster=T,
- flip = F,stack = T,fill.by ="ident", cols = rev(colors))
- p2 <- DotPlot(object = merge_obj,
- features = selected_features, cols =c("white","firebrick3"),
- group.by = 'celltype',scale.min=30) + RotatedAxis()
- cowplot::plot_grid(p1,p2,ncol=1)
scRNA-seq_code.R at commit 22f1db8, no license · at the source
Overview
- Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, United States of America
- Center for Emerging, Zoonotic, and Arthropod-borne Pathogens, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, United States of America
- Department of Statistics, Virginia Polytechnic Institute and State University, Blacksburg, Virginia, United States of America
- United States Army Medical Research Institute of Chemical Defense, Aberdeen Proving Ground, Maryland, United States of America
Abstract
The abstract is not reproduced here: the paper's license (none stated) does not allow it. Read it in the paper, at the publisher or on Europe PMC.
Repository
Its files are read in the Code ↔ Paper reader above, with 3 matches between paragraphs and lines of code.
Gavin-Yinld/VEEV_scRNA-seq
22f1db873ab296d9fb35877330a427335e41cde6, 14 February 2025Availability: 1 check, the latest on 29 September 2026: the link answers
- 29 September 2026: the link answers
2 files
- scRNA-seq_code.R, R, 238 lines, 3 matches
- README.md, Text, 1 line
The paper's code and data availability statement is in the Data section.
Tracing map
Proposed by the machine: these links were found in the paper and verified at the source, without human review. The map will receive a Zenodo DOI once one of the paper's authors has validated it with their ORCID.
What the map holds:
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- 3 matches between paragraphs of the paper and lines of the code (method lexical-v1);
- neither the text of the paper nor the code itself.
Its JSON (tracing-map.json) is deposited on Zenodo with its DOI once the map is validated.
Data
Datasets cited
- geo:GSE289596, at NCBI GEO; found in “Data Availability”
Code and data availability statement
The paper has a code and data availability statement. Its license (none stated) does not allow reproducing it here; in short, from what the harvester recognized in it:
- it points to a dataset: NCBI GEO GSE289596
- it points to the authors' code: Gavin-Yinld/
VEEV_scRNA-seq
Read it in the paper: doi.org/10.1371/journal.ppat.1014115.
Versions
The history of this record: each version stored by the harvester or made by a correction of its authors or of the maintainers of its code, and what changed in its facts. The texts of the paper (its abstract, its availability statements) are not part of it; versions that changed only those are not listed.
Version 1, 29 September 2026: the first record
Recorded: type, language, journal, volume, issue, pages, dates, 11 authors, 10 MeSH terms, 2 funders, 117 references.
Cite
This paper
VanderGiessen, M., Harris, E., Yin, L., Heath, B., Carney, S. K., Woodson, C. M., Wu, X., Johnson, E., Xie, H., Theus, M., & Kehn-Hall, K. (2026). Venezuelan equine encephalitis virus infection causes chronic neurobehavioral outcomes, cellular remodeling, and hippocampal single-cell transcriptomic changes. PLoS pathogens, 22(4), e1014115. https://
BibTeX
@article{vandergiessen20
author = {VanderGiessen, Morgen and Harris, Elizabeth and Yin, Liduo and Heath, Brittany and Carney, Shannon K and Woodson, Caitlin M and Wu, Xiaowei and Johnson, Erik and Xie, Hehuang and Theus, Michelle and Kehn-Hall, Kylene},
title = {{Venezuelan equine encephalitis virus infection causes chronic neurobehavioral outcomes, cellular remodeling, and hippocampal single-cell transcriptomic changes}},
journal = {PLoS pathogens},
year = {2026},
month = apr,
volume = {22},
number = {4},
pages = {e1014115},
publisher = {PLOS},
issn = {1553-7366},
doi = {10.1371/
url = {https://
pmid = {41950299},
pmcid = {PMC13089902}
}
RIS
TY - JOUR
AU - VanderGiessen, Morgen
AU - Harris, Elizabeth
AU - Yin, Liduo
AU - Heath, Brittany
AU - Carney, Shannon K
AU - Woodson, Caitlin M
AU - Wu, Xiaowei
AU - Johnson, Erik
AU - Xie, Hehuang
AU - Theus, Michelle
AU - Kehn-Hall, Kylene
TI - Venezuelan equine encephalitis virus infection causes chronic neurobehavioral outcomes, cellular remodeling, and hippocampal single-cell transcriptomic changes
T2 - PLoS pathogens
J2 - PLoS Pathog
PY - 2026
DA - 2026/
VL - 22
IS - 4
SP - e1014115
SN - 1553-7366
PB - PLOS
DO - 10.1371/
UR - https://
LA - en
ER -
CSL-JSON
{
"id": "10.1371/
"type": "article-journal",
"title": "Venezuelan equine encephalitis virus infection causes chronic neurobehavioral outcomes, cellular remodeling, and hippocampal single-cell transcriptomic changes",
"container-title": "PLoS pathogens",
"author": [
{
"family": "VanderGiessen",
"given": "Morgen"
},
{
"family": "Harris",
"given": "Elizabeth"
},
{
"family": "Yin",
"given": "Liduo"
},
{
"family": "Heath",
"given": "Brittany"
},
{
"family": "Carney",
"given": "Shannon K"
},
{
"family": "Woodson",
"given": "Caitlin M"
},
{
"family": "Wu",
"given": "Xiaowei"
},
{
"family": "Johnson",
"given": "Erik"
},
{
"family": "Xie",
"given": "Hehuang"
},
{
"family": "Theus",
"given": "Michelle"
},
{
"family": "Kehn-Hall",
"given": "Kylene"
}
],
"container-title-short":
"volume": "22",
"issue": "4",
"page": "e1014115",
"DOI": "10.1371/
"PMID": "41950299",
"PMCID": "PMC13089902",
"ISSN": "1553-7366",
"publisher": "PLOS",
"URL": "https://
"language": "en",
"issued": {
"date-parts": [
[
2026,
4,
8
]
]
}
}
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