Biology subjects

Vestal, B.

Publications and source records attributed to Vestal, B..

4 recordsLinked to original sources

bioRxiv2025

Monocyte Subset Predict Clinical Outcomes in Fibrotic Hypersensitivity Pneumonitis

RationaleHypersensitivity pneumonitis (HP) is an immunologically mediated form of lung disease resulting from inhalational exposure to various antigens. While there is growing understanding of the immune cells involved in HP, the relationship between clinical outcomes and circulating cell types in HP is not well established. MethodsSingle-cell RNA sequencing was performed on peripheral blood mononuclear cells (PBMCs) from 40 patients with fibrotic hypersensitivity pneumonitis (fHP) in a clinical trial of pirfenidone (NCT02958917). After quality control, we compared data from 33 fHP patients with 36 sex-matched healthy controls (GSE196735). Using Seurat v5, we harmonized the data, formed clusters, and reduced dimensionality through UMAP. Cell type identities were assigned based on a published reference (GSE271789), with clusters annotated for predominant cell type(s) present in at least 30% of the cells. We compared cell type proportions between fHP and HC using the chi-squared test and performed differential expression analysis with DESeq2 on pseudobulk counts, adjusting for sex. Genes significantly upregulated in fHP versus HC at an FDR-adjusted p-value of 0.01 were used to create a multivariate predictive signature for clinical outcomes in fHP participants. Performance was assessed through leave-one-out cross-validation for prediction accuracy. ResultsCompared to HC, fHP has an increased proportion of non-classical CD16+ monocytes (4.2% fHP vs. 1.5% HC), C14+ monocytes with a myeloid-derived dendritic phenotype (2.1% fHP vs. 0.1% HC), platelets (1.0% fHP vs. 0.3% HC), plasmacytoid dendritic cells (0.5% fHP vs. 0.1% HC) and predominantly classical CD14+ monocytes (29.5% fHP vs. 3.4% HC, adjusted-p<0.001). In patients with fHP, cell clusters were associated with the presence of baseline chest CT honeycombing, increased extent of CT lung fibrosis, and worse progression-free survival. CD14+ monocytes consistently demonstrated a high prediction accuracy (>0.90) for these clinical metrics. ConclusionsPeripheral monocyte clusters may be valuable prognostic markers, potentially helping identify at-risk fHP patients.

genetics↗

bioRxiv2025

Antennal RNAseq reveals odorant receptors with sex-biased expression in the common eastern firefly, Photinus pyralis

BackgroundWith their charismatic nighttime flashes, fireflies are a classic organismal system for studying visual mating signal evolution. However, across their diversity, fireflies employ a variety of mating strategies that include both chemical and visual signals. While phylogenetic evidence points to a common ancestor that relied on long-range pheromones, behavioral evidence suggests that light-dependent flashing fireflies do not use smell for mating. We tested this hypothesis by investigating the olfactory genetics of the nocturnal, light-using common eastern firefly, Photinus pyralis. In insects, odors are primarily detected by odorant receptor (OR) proteins embedded in the dendritic membranes of olfactory receptor neurons. If pheromones are part of mate signaling in light-using fireflies, then one or more OR genes should be upregulated in the antennae of the searching sex (males). We therefore annotated the complete suite of ORs from the genome of P. pyralis and measured expression of OR genes between the sexes. ResultsWe identified 102 ORs in the Photinus pyralis genome, including the conserved single-copy Orco. Our phylogenetic analysis showed lineage-specific OR diversification in P. pyralis relative to other beetle species. Differential expression analysis of male and female antennae and hind legs revealed that a subset of ORs are upregulated in antennae as compared to legs, suggesting a role in adult olfaction. Notably, PpyrOR6 was one of two genes, and the only OR, that was significantly upregulated between male and female antennae, suggesting a role in mating. ConclusionsThese findings increase known diversity of insect ORs in an understudied beetle family and suggest that bioluminescent fireflies use multimodal signals during mating.

evolutionary biology↗

bioRxiv2024

Siglec-F Deficiency Prevents Fibrosis After Bleomycin-Induced Acute Lung Injury

Injury to the lungs causes acute inflammation that can lead to pathological lung fibrosis. Airspace macrophages (AMs) are critical for repair of injured tissue, but they can contribute fibrosis through mechanisms that are incompletely understood. Siglecs are expressed by immune cells. In mice, Siglec-F is chiefly expressed AMs where it is considered inflammosuppressive. We hypothesized that its deletion would worsen lung injury and fibrosis in response to intratracheal bleomycin challenge. We evaluated Siglec-F expression and function in mice challenged with bleomycin on days 7, 14, and 21 post-challenge (2.5 U/kg). AMs were the predominant inflammatory cells at all timepoints, and they included resident (RAM) and recruited (RecAM) subsets. Siglec-F deficiency prevented fibrosis than in Siglecf-/- mouse lungs, as evident from biochemical and histologic readouts. We performed RNAseq on pooled RAMs and RecAMs from wild type and Siglec-F deficient mice. Lung fibrosis 21 d after bleomycin challenge was associated with differentially expressed genes (DEGs) related to cholesterol synthesis and metabolism. In AMs from healthy lung lavage fluid and idiopathic pulmonary fibrosis patient tissues, the human paralogs Siglec-7 and Siglec-9 were expressed. Findings here identify novel mechanisms that control protective and detrimental functions of AMs after lung injury.

physiology↗

bioRxiv2020

Human and mouse transcriptome profiling identifies cross-species homology in pulmonary and lymph node mononuclear phagocytes

The mononuclear phagocyte (MP) system consists of macrophages, monocytes, and dendritic cells (DCs). MP subtypes play distinct functional roles in steady state and inflammatory conditions. Though murine MPs are well characterized, their pulmonary and lymph node (LN) human homologs remain poorly understood. To address this gap, we created a gene expression compendium across 15 distinct human and 9 distinct murine MPs from lung, LN, blood, and spleen. Human blood MPs and murine spleen MPs served as validation datasets, as the human-mouse MP homologs are relatively well-defined in these tissues. In-depth RNA sequencing identified corresponding human-mouse MP subtypes and determined marker genes shared and divergent across between species counterparts. Unexpectedly, at the gene expression level, only 13-23% of the top 1000 marker genes (i.e., genes not shared across species-specific MP subtypes) overlapped in corresponding human-mouse MP counterparts, indicating a need for caution when translating mouse studies to human gene targets and functions. Lastly, CD88 was useful in both species to distinguish macrophage and tissue monocytes from DCs. Our cross-species gene expression compendium serves as a resource for future translational studies to investigate beforehand whether pursuing specific MP subtypes, or genes will prove fruitful.

immunology↗