22 research outputs found
Regulatory sites for splicing in human basal ganglia are enriched for disease-relevant information
Genome-wide association studies have generated an increasing number of common genetic
variants associated with neurological and psychiatric disease risk. An improved under-
standing of the genetic control of gene expression in human brain is vital considering this is
the likely modus operandum for many causal variants. However, human brain sampling
complexities limit the explanatory power of brain-related expression quantitative trait loci
(eQTL) and allele-specific expression (ASE) signals. We address this, using paired genomic
and transcriptomic data from putamen and substantia nigra from 117 human brains, inter-
rogating regulation at different RNA processing stages and uncovering novel transcripts. We
identify disease-relevant regulatory loci, find that splicing eQTLs are enriched for regulatory
information of neuron-specific genes, that ASEs provide cell-specific regulatory information
with evidence for cellular specificity, and that incomplete annotation of the brain tran-
scriptome limits interpretation of risk loci for neuropsychiatric disease. This resource of
regulatory data is accessible through our web server, http://braineacv2.inf.um.es/
Prediction of mechanistic subtypes of Parkinsonâs using patient-derived stem cell models
Parkinsonâs disease is a common, incurable neurodegenerative disorder that is clinically heterogeneous: it is likely that different cellular mechanisms drive the pathology in different individuals. So far it has not been possible to define the cellular mechanism underlying the neurodegenerative disease in life. We generated a machine learning-based model that can simultaneously predict the presence of disease and its primary mechanistic subtype in human neurons. We used stem cell technology to derive control or patient-derived neurons, and generated different disease subtypes through chemical induction or the presence of mutation. Multidimensional fluorescent labelling of organelles was performed in healthy control neurons and in four different disease subtypes, and both the quantitative single-cell fluorescence features and the images were used to independently train a series of classifiers to build deep neural networks. Quantitative cellular profile-based classifiers achieve an accuracy of 82%, whereas image-based deep neural networks predict control and four distinct disease subtypes with an accuracy of 95%. The machine learning-trained classifiers achieve their accuracy across all subtypes, using the organellar features of the mitochondria with the additional contribution of the lysosomes, confirming the biological importance of these pathways in Parkinsonâs. Altogether, we show that machine learning approaches applied to patient-derived cells are highly accurate at predicting disease subtypes, providing proof of concept that this approach may enable mechanistic stratification and precision medicine approaches in the future
Regulatory sites for splicing in human basal ganglia are enriched for disease-relevant information
International Parkinsonâs Disease Genomics Consortium (IPDGC), UK Brain Expression Consortium (UKBEC).Genome-wide association studies have generated an increasing number of common genetic variants associated with neurological and psychiatric disease risk. An improved understanding of the genetic control of gene expression in human brain is vital considering this is the likely modus operandum for many causal variants. However, human brain sampling complexities limit the explanatory power of brain-related expression quantitative trait loci (eQTL) and allele-specific expression (ASE) signals. We address this, using paired genomic and transcriptomic data from putamen and substantia nigra from 117 human brains, interrogating regulation at different RNA processing stages and uncovering novel transcripts. We identify disease-relevant regulatory loci, find that splicing eQTLs are enriched for regulatory information of neuron-specific genes, that ASEs provide cell-specific regulatory information with evidence for cellular specificity, and that incomplete annotation of the brain transcriptome limits interpretation of risk loci for neuropsychiatric disease. This resource of regulatory data is accessible through our web server, http://braineacv2.inf.um.es/.Mina Ryten, David Zhang, and Karishma DâSa were supported by the UK Medical Research Council (MRC) through the award of Tenure-track Clinician Scientist Fellowship to Mina Ryten (MR/N008324/1). Sebastian Guelfi was supported by Alzheimerâs Research UK through the award of a PhD Fellowship (ARUK-PhD2014-16). Regina Reynolds was supported through the award of a Leonard Wolfson Doctoral Training Fellowship in Neurodegeneration
Analysis of subcellular RNA fractions demonstrates significant genetic regulation of gene expression in human brain post-transcriptionally
Abstract Gaining insight into the genetic regulation of gene expression in human brain is key to the interpretation of genome-wide association studies for major neurological and neuropsychiatric diseases. Expression quantitative trait loci (eQTL) analyses have largely been used to achieve this, providing valuable insights into the genetic regulation of steady-state RNA in human brain, but not distinguishing between molecular processes regulating transcription and stability. RNA quantification within cellular fractions can disentangle these processes in cell types and tissues which are challenging to model in vitro. We investigated the underlying molecular processes driving the genetic regulation of gene expression specific to a cellular fraction using allele-specific expression (ASE). Applying ASE analysis to genomic and transcriptomic data from paired nuclear and cytoplasmic fractions of anterior prefrontal cortex, cerebellar cortex and putamen tissues from 4 post-mortem neuropathologically-confirmed control human brains, we demonstrate that a significant proportion of genetic regulation of gene expression occurs post-transcriptionally in the cytoplasm, with genes undergoing this form of regulation more likely to be synaptic. These findings have implications for understanding the structure of gene expression regulation in human brain, and importantly the interpretation of rapidly growing single-nucleus brain RNA-sequencing and eQTL datasets, where cytoplasm-specific regulatory events could be missed
Additional file 4 of An additional k-means clustering step improves the biological features of WGCNA gene co-expression networks
Heat-maps showing âl o g 10(p-values) from Ficherâs Exact test on significant concentration of specific cell marker gene sets (rows) on each tissue module (columns). The one within the Additional file 1 corresponds to the standard WGCNA. (JPG 1198 kb
An integrated genomic approach to dissect the genetic landscape regulating the cell-to-cell transfer of α-synuclein
Neuropathological and experimental evidence suggests that the cell-to-cell transfer of α-synuclein has an important role in the pathogenesis of Parkinsonâs disease (PD). However, the mechanism underlying this phenomenon is not fully understood. We undertook a small interfering RNA (siRNA), genome-wide screen to identify genes regulating the cell-to-cell transfer of α-synuclein. A genetically encoded reporter, GFP-2A-αSynuclein-RFP, suitable for separating donor and recipient cells, was transiently transfected into HEK cells stably overexpressing α-synuclein. We find that 38 genes regulate the transfer of α-synuclein-RFP, one of which is ITGA8, a candidate gene identified through a recent PD genome-wide association study (GWAS). Weighted gene co-expression network analysis (WGCNA) and weighted protein-protein network interaction analysis (WPPNIA) show that those hits cluster in networks that include known PD genes more frequently than expected by random chance. The findings expand our understanding of the mechanism of α-synuclein spread
Characterizing the Relation Between Expression QTLs and Complex Traits:Exploring the Role of Tissue Specificity
Measurement of gene expression levels and detection of eQTLs (expression quantitative trait loci) are difficult in tissues with limited sample availability, such as the brain. However, eQTL overlap between tissues might be high, which would allow for inference of eQTL functioning in the brain via eQTLs detected in readily accessible tissues, e.g. whole blood. Applying Stratified Linkage Disequilibrium Score Regression (SLDSR), we quantified the enrichment in polygenic signal of blood and brain eQTLs in genome-wide association studies (GWAS) of 11 complex traits. We looked at eQTLs discovered in 44 tissues by the Genotype-Tissue Expression (GTEx) consortium and two other large representative studies, and found no tissue-specific eQTL effects. Next, we integrated the GTEx eQTLs with regions associated with tissue-specific histone modifiers, and interrogated their effect on rheumatoid arthritis and schizophrenia. We observed substantially enriched effects of eQTLs located inside regions bearing modification H3K4me1 on schizophrenia, but not rheumatoid arthritis, and not tissue-specific. Finally, we extracted eQTLs associated with tissue-specific differentially expressed genes and determined their effects on rheumatoid arthritis and schizophrenia, these analysis revealed limited enrichment of eQTLs associated with gene specifically expressed in specific tissues. Our results pointed to strong enrichment of eQTLs in their effect on complex traits, without evidence for tissue-specific effects. Lack of tissue-specificity can be either due to a lack of statistical power or due to the true absence of tissue-specific effects. We conclude that eQTLs are strongly enriched in GWAS signal and that the enrichment is not specific to the eQTL discovery tissue. Until sample sizes for eQTL discovery grow sufficiently large, working with relatively accessible tissues as proxy for eQTL discovery is sensible and restricting lookups for GWAS hits to a specific tissue for which limited samples are available might not be advisable
Additional file 5 of Mitochondrial dysfunction is a key pathological driver of early stage Parkinsonâs
Additional file 5:Table S2. Biological processes represented across brain regions for early PD cases compared to controls. Shaded box reflects GO term represented in that brain region
Additional file 1 of Mitochondrial dysfunction is a key pathological driver of early stage Parkinsonâs
Additional file 1: Figure S1. GO terms altered in early PD compared to controls and mitochondrial gene expression patterns (a) Bar charts indicating number of Gene Ontology (GO) terms that are represented in the dataset for each region as determined by Webgestalt and DAVID databases. Top chart shows Biological processes, middle the molecular functions and lower the cellular components. The pie chart adjacent to each chart shows the number of GO terms that overlap or are uniquely represented in the dataset for a region of severe pathology (substantia nigra), mild pathology (parahippocampus) and a region unaffected at Braak stage 3/4 (frontal cortex) as determined by GOview. Colours indicate the level each region is affected at Braak stage 3/4 as determined in Figure 1b. Protein heatmaps from (b) first mass spectrometry run and (c) second mass spectrometry run from IPA showing the level of expression change per protein in the Mitochondrial Dysfunction pathway across each brain region in Braak stage 3/4 compared to controls. Red indicated upregulation and green indicated downregulation compared to controls. Intensity of colour shows level of expression change with deeper colour indicating higher up- or down- regulation