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Okeke, C.

Publications and source records attributed to Okeke, C..

3 recordsLinked to original sources

Variation in enhancer activity across brain regions defines neurological disease risk and shapes cellular pathology

Genetic variants associated with neurological traits are enriched in enhancers active in neurons. However, there are many types of neurons with varying functions across the brain, making it difficult to pinpoint insights into disease pathology. We set out to investigate whether cell type and brain region specificity of transcriptional enhancers could reveal new insights into the cellular pathology of neurological traits. We performed H3K27ac ChIP-seq on neurons and glia sorted from post-mortem tissue for six brain regions in triplicate and integrated these datasets with existing single-cell ATAC-seq data. While 87% of neuronal enhancers had similar activity across brain regions, we identified 40,049 neuronal regulatory elements that vary in activity across brain regions, which we termed Brain Region Variable Elements (BRVEs). Some BRVEs reflect differences in cell composition, such as a high proportion of medium spiny neurons (MSNs) in the nucleus accumbens, whereas others capture developmental trajectories, including enhancers with robust activity across telencephalic regions that are inactive in the diencephalic hypothalamus. Genetic variants within BRVEs are disproportionately enriched for the heritability of neurological traits, capturing far more heritability than neuronal enhancers with uniform activity across the brain. Additionally, genes linked to BRVEs are also more likely to be implicated in neuropsychiatric disorders by differential expression studies. Finally, we present a new method, GWAS-LOCATE, that leverages variation in enhancer activity across the brain to assign 23,494 GWAS loci to specific cell types and brain regions, including 9.8% linked to MSNs. This includes a BMI risk locus linked to the transcription factor ISL1. ISL1 knockdown in iPSC-derived MSNs revealed differentially expressed genes enriched for BMI heritability, supporting a role for ISL1-mediated MSN pathways with BMI. Collectively, these findings demonstrate that the enhancer specificity across the brain provides a powerful framework for dissecting complex trait biology and revealing cellular pathology.

genetics↗

Rationally Minimizing Natural Product Libraries Using Mass Spectrometry

Natural product libraries are crucial to drug development, but large libraries drastically increase the time and cost during initial high throughput screens. Here, we developed a method that leverages liquid chromatography-tandem mass spectrometry spectral similarity to dramatically reduce library size, with minimal bioactive loss. This method offers a broadly applicable strategy for accelerated drug discovery with cost reductions, which enable implementation in resource-limited settings.

bioinformatics↗

High dose IFN-β activates GAF to enhance expression of ISGF3 target genes in epithelial cells

Interferon {beta} (IFN-{beta}) signaling activates the transcription factor complex ISGF3 to induce gene expression programs critical for antiviral defense and host immune responses. It has also been observed that IFN-{beta} activates a second transcription factor, {gamma}-activated factor (GAF), but the significance of this coordinated activation is unclear. We report that in respiratory epithelial cells high doses of IFN-{beta} indeed activate both ISGF3 and GAF, which bind to distinct genomic locations defined by their respective DNA sequence motifs. In contrast, low doses of IFN-{beta} preferentially activate ISGF3 but not GAF. Surprisingly, in epithelial cells GAF binding does not induce nearby gene expression even when strongly bound to the promoter. Yet expression of interferon stimulated genes is enhanced when GAF and ISGF3 are both active compared to ISGF3 alone. Our data suggest that GAF enhances ISGF3 target gene expression by co-localizing with ISGF3 at some promoters and facilitating chromosome looping between distal enhancers and other promoters. We propose that GAF may function as a dose-sensitive amplifier of ISG expression to enhance antiviral immunity and establish pro-inflammatory states in respiratory epithelial cells. One sentence summaryGAF is transcriptionally inactive in epithelial cells but enhances expression of ISGF3 target genes, thus functioning as a dose-sensitive amplifier of the IFN-{beta} response.

molecular biology↗