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Mohammadi, P.

Publications and source records attributed to Mohammadi, P..

At least 19 recordsLinked to original sources

Silicon exaptation discovers antimicrobial peptides

Advances in machine learning (ML) have enabled the de novo design of synthetic proteins and peptides tailored for specific structural or functional properties. As ML becomes a central tool in forward design, we suggest that its value extends beyond optimizing known objectives. It can also lead to what we term here in this work as silicon exaptation, revealing unexpected functional properties that were never part of the original design goals. This reflects a principle common in biology, that repurposes traits where features evolved for one role can later enable unforeseen functions. In this context, these accidental findings are not constrained by preconceptions but arise from latent function emergence and the algorithm's gradual drift, a new process that can co-evolves with human curiosity, intuition, and interpretive reasoning, which can be used to recognize unexpected patterns and transform them into new discoveries. In this study, we discovered a set of peptides through silicon exaptation, in which the ML pipeline was originally optimized for structural characteristics unrelated to any antimicrobial functionalities. The peptides were originally designed to adopt defined secondary structures in response to environmental stimuli, with no antimicrobial properties intended or included in the training objectives. We systematically evaluated the antimicrobial potential of these peptides using broth microdilution assays and membrane integrity tests, identifying several candidates with potent and selective antibacterial activity without detectable cytotoxicity toward mammalian cells.

molecular biology↗

Data-driven RNA phenotyping captures genetically regulated dimensions of the transcriptome

Transcriptomic diversity across individuals arises from multiple modes of RNA regulation--including pre-mRNA expression, splicing, degradation, and other processes--and has been widely leveraged to map quantitative trait loci (xQTLs) and interpret GWAS signals. We recently developed a multimodal framework called Pantry that can extend discovery beyond total expression by integrating multiple transcriptomic modalities. However, Pantry and similar tools remain limited by their reliance on complete gene annotations and the statistical complexity of jointly analyzing correlated modalities. Here, we present LaDDR (Latent Data-Driven RNA phenotyping), a mechanism-agnostic framework that generates orthogonal, latent coverage features per gene, enabling xQTL discovery and GWAS integration without requiring complete gene annotations. Applied to GTEx, LaDDR identified an average of 95% more independent xQTLs per tissue than the six transcriptional regulation modes implemented in Pantry ("knowledge-driven"). Residualizing known modalities prior to LaDDR and combining with knowledge-driven phenotypes increased discovery by an additional 41% per tissue on average, while retaining the interpretability of knowledge-driven signals. In a transcriptome-wide association study (TWAS) of 114 complex traits, using LaDDR-derived phenotypes uncovered an average of 11,790 unique gene-trait pairs per tissue, versus 8,579 from knowledge-driven phenotypes. The newly captured genetic signals exhibit functional and colocalization qualities consistent with known mechanisms, suggesting that LaDDR broadens the detectable landscape of trait-relevant transcriptomic regulation by efficiently recovering regulatory variation missed by current pipelines.

genomics↗

Allele Specific Expression Quality Control Fills Critical Gap in Transcriptome Assisted Rare Variant Interpretation

Allele-specific expression (ASE) captures the functional impact of genetic variation on transcription, offering a high-resolution view of cis-regulatory effects, but its quality can be diminished by technical, biological, and analysis artifacts. We introduce aseQC, a statistical framework that quantifies sample-level ASE quality in terms of the overall expected extra-binomial variation to exclude uncharacteristically noisy samples in a cohort to improve robustness of downstream analyses. Applying aseQC to a dataset of rare mendelian muscular disorders, successfully identified previously annotated low-quality cases demonstrating clinical genomic utility. When applied to 15,253 samples in extensively quality controlled GTEx project data, aseQC uncovered 563 low-quality samples that exhibit excessive allelic imbalance. We identify these to be associated with specific processing dates but not otherwise described adequately by any other quality control measures and metadata available in GTEx data. We show that these low-quality samples lead to 23.6 and 31.6 -fold increased ASE, and splicing outliers, degrading the performance of transcriptome analysis for rare variant interpretation. In contrast, we did not observe any adverse effect associated with inclusion of these samples in common-variant analysis using quantitative traits loci mapping. By enabling quick and reliable assessment of sample quality, aseQC presents a critical step for identifying subtle quality issues that remain critical for a successful analysis of rare variant effects using transcriptome data.

bioinformatics↗

Multimodal analysis of RNA sequencing data powers discovery of complex trait genetics

Transcriptome data is commonly used to understand genome function via quantitative trait loci (QTL) mapping and to identify the molecular mechanisms driving genome wide association study (GWAS) signals through colocalization analysis and transcriptome-wide association studies (TWAS). While RNA sequencing (RNA-seq) has the potential to reveal many modalities of transcriptional regulation, such as various splicing phenotypes, such studies are often limited to gene expression due to the complexity of extracting and analyzing multiple RNA phenotypes. Here, we present Pantry (Pan-transcriptomic phenotyping), a framework to efficiently generate diverse RNA phenotypes from RNA-seq data and perform downstream integrative analyses with genetic data. Pantry currently generates phenotypes from six modalities of transcriptional regulation (gene expression, isoform ratios, splice junction usage, alternative TSS/polyA usage, and RNA stability) and integrates them with genetic data via QTL mapping, TWAS, and colocalization testing. We applied Pantry to Geuvadis and GTEx data, and found that 4,768 of the genes with no identified expression QTL in Geuvadis had QTLs in at least one other transcriptional modality, resulting in a 66% increase in genes over expression QTL mapping. We further found that QTLs exhibit modality-specific functional properties that are further reinforced by joint analysis of different RNA modalities. We also show that generalizing TWAS to multiple RNA modalities (xTWAS) approximately doubles the discovery of unique gene-trait associations, and enhances identification of regulatory mechanisms underlying GWAS signal in 42% of previously associated gene-trait pairs. We provide the Pantry code, RNA phenotypes from all Geuvadis and GTEx samples, and xQTL and xTWAS results on the web.

genetics↗

Metastasis founder cells activate immunosuppression early in human melanoma metastatic colonization

The earliest steps of lethal metastasis in patients are incompletely understood. To dissect them, we prospectively searched for the earliest detectable disseminated cancer cells (DCC) in sentinel lymph node biopsies of 492 stage I-III patients. By visually-controlled, micromanipulator-assisted isolation and single cell transcriptome analysis of these extremely rare DCC, we identified MCSP+ melanoma cells as strong candidates for metastasis founder cells (MFC) in lymph nodes. Based on a median follow-up time of 6 years, their detection was the strongest predictor of systemic metastasis and death upon multivariable analysis. During transition from single cells to metastasis-initiating clusters, melanoma DCC were exposed to CD8 T cell attack, activated the extracellular vesicular exosomal pathway, and expressed the immunomodulatory proteins CD155 and CD276, but rarely PD-L1. CD155 and CD276-positive extracellular vesicles from patient-derived DCC models exhibited an immunosuppressive activity on CD8 T cells. Our data indicate that either direct targeting of MFC employing MCSP or their immune escape mechanisms might be key for cure of early-stage melanoma.

cancer biology↗

Recalibrating differential gene expression by genetic dosage variance prioritizes functionally relevant genes

Differential expression (DE) analysis is a widely used method for identifying genes that are functionally relevant for an observed phenotype or biological response. However, typical DE analysis includes selection of genes based on a threshold of fold change in expression under the implicit assumption that all genes are equally sensitive to dosage changes of their transcripts. This tends to favor highly variable genes over more constrained genes where even small changes in expression may be biologically relevant. To address this limitation, we have developed a method to recalibrate each genes differential expression fold change based on genetic expression variance observed in the human population. The newly established metric ranks statistically differentially expressed genes not by nominal change of expression, but by relative change in comparison to natural dosage variation for each gene. We apply our method to RNA sequencing datasets from rare disease and in-vitro stimulus response experiments. Compared to the standard approach, our method adjusts the bias in discovery towards highly variable genes, and enriches for pathways and biological processes related to metabolic and regulatory activity, indicating a prioritization of functionally relevant driver genes. With that, our method provides a novel view on DE and contributes towards bridging the existing gap between statistical and biological significance. We believe that this approach will simplify the identification of disease causing genes and enhance the discovery of therapeutic targets.

genomics↗

CRISPR/Cas9-mediated Generation of COL7A1-deficient Keratinocyte Model of Recessive Dystrophic Epidermolysis Bullosa

ObjectiveRecessive dystrophic epidermolysis bullosa (RDEB) is a genetic skin fragility and ultimately lethal blistering disease caused by mutations in the COL7A1 gene which is responsible for coding type VII collagen. Investigating the pathological mechanisms and novel candidate therapies for RDEB could be fostered by new cellular models. Here, we developed multiple immortalized COL7A1-deficient keratinocyte cell lines using CRISPR/Cas9 technology as RDEB cellular model. Materials and MethodsIn this experimental study, we used transient transfection to express COL7A1-targeting gRNA and Cas9 in HEK001 immortalized keratinocyte cell line followed by enrichment with fluorescent-activated cell sorting (FACS) via GFP expressing cells (GFP+ HEK001). Homogenous single-cell clones were then isolated, genotyped, and evaluated for type VII collagen expression. We performed a scratch assay to confirm the functional effect of COL7A1 knockout. ResultsWe achieved 46.1% (p < 0.001) efficiency of indel induction in the enriched transfected cell population. Except for 4% of single nucleotide insertions, the remaining indels were deletions of different sizes. Out of nine single clones expanded, two homozygous and two heterozygous COL7A1-deficient cell lines were obtained with defined mutation sequences. No off-target effect was detected in the knockout cell lines. Immunostaining and western blot analysis showed the lack of type VII collagen (COL7A1) protein expression in these cell lines. We also showed that COL7A1-deficient cells had higher motility compared with their wild-type counterparts. ConclusionWe reported the first isogenic immortalized COL7A1-deficient keratinocyte lines that provide a useful cell culture model to investigate aspects of RDEB biology and potential therapeutic options.

cell biology↗

A revamped rat reference genome improves the discovery of genetic diversity in laboratory rats

The seventh iteration of the reference genome assembly for Rattus norvegicus-- mRatBN7.2--corrects numerous misplaced segments and reduces base-level errors by approximately 9-fold and increases contiguity by 290-fold compared to its predecessor. Gene annotations are now more complete, significantly improving the mapping precision of genomic, transcriptomic, and proteomics data sets. We jointly analyzed 163 short-read whole genome sequencing datasets representing 120 laboratory rat strains and substrains using mRatBN7.2. We defined [~]20.0 million sequence variations, of which 18.7 thousand are predicted to potentially impact the function of 6,677 genes. We also generated a new rat genetic map from 1,893 heterogeneous stock rats and annotated transcription start sites and alternative polyadenylation sites. The mRatBN7.2 assembly, along with the extensive analysis of genomic variations among rat strains, enhances our understanding of the rat genome, providing researchers with an expanded resource for studies involving rats.

genomics↗

Genetic control of mRNA splicing as a potential mechanism for incomplete penetrance of rare coding variants

Exonic variants present some of the strongest links between genotype and phenotype. However, these variants can have significant inter-individual pathogenicity differences, known as variable penetrance. In this study, we propose a model where genetically controlled mRNA splicing modulates the pathogenicity of exonic variants. By first cataloging exonic inclusion from RNA-seq data in GTEx v8, we find that pathogenic alleles are depleted on highly included exons. Using a large-scale phased WGS data from the TOPMed consortium, we observe that this effect may be driven by common splice-regulatory genetic variants, and that natural selection acts on haplotype configurations that reduce the transcript inclusion of putatively pathogenic variants, especially when limiting to haploinsufficient genes. Finally, we test if this effect may be relevant for autism risk using families from the Simons Simplex Collection, but find that splicing of pathogenic alleles has a penetrance reducing effect here as well. Overall, our results indicate that common splice-regulatory variants may play a role in reducing the damaging effects of rare exonic variants.

genetics↗

Exponential increase in QTL detection with increased sample size.

Power analyses are often used to determine the number of animals required for a genome wide association analysis (GWAS). These analyses are typically intended to estimate the sample size needed for at least one locus to exceed a genome-wide significance threshold. A related question that is less commonly considered is the number of significant loci that will be discovered with a given sample size. We used simulations based on a real dataset that consisted of 3,173 male and female adult N/NIH heterogeneous stock (HS) rats to explore the relationship between sample size and the number of significant loci discovered. Our simulations examined the number of loci identified in sub-samples of the full dataset. The sub-sampling analysis was conducted for four traits with low (0.15 {+/-} 0.03), medium (0.31 {+/-} 0.03 and 0.36 {+/-} 0.03) and high (0.46 {+/-} 0.03) SNP-based heritabilities. For each trait, we sub-sampled the data 100 times at different sample sizes (500, 1,000, 1,500, 2,000, and 2,500). We observed an exponential increase in the number of significant loci with larger sample sizes. Our results are consistent with similar observations in human GWAS and imply that future rodent GWAS should use sample sizes that are significantly larger than those needed to obtain a single significant result.

genetics↗

Genome-Wide Association Study Finds Multiple Loci Associated with Intraocular Pressure in HS Rats

Elevated intraocular pressure (IOP) is influenced by environmental and genetic factors. Increased IOP is a major risk factor for most types of glaucoma, including primary open angle glaucoma (POAG). Investigating the genetic basis of IOP may lead to a better understanding of the molecular mechanisms of POAG. The goal of this study was to identify genetic loci involved in regulating IOP using outbred heterogeneous stock (HS) rats. HS rats are a multigenerational outbred population derived from eight inbred strains that have been fully sequenced. This population is ideal for genome-wide association studies (GWASs) owing to the accumulated recombinations among well-defined haplotypes, the relatively high allele frequencies, the accessibility to a large collection of tissue samples, and the large allelic effect size compared to human studies. Both male and female HS rats (N=1,812) were used in the study. Genotyping-by-sequencing was used to obtain ~3.5 million single nucleotide polymorphisms (SNP) from each individual. SNP heritability for IOP in HS rats was 0.32, which agrees with other studies. We performed a GWAS for the IOP phenotype using a linear mixed model and used permutation to determine a genome-wide significance threshold. We identified three genome-wide significant loci for IOP on chromosomes 1, 5, and 16. Next, we sequenced the mRNA of 51 whole eye samples to find cis-eQTLs to aid in identification of candidate genes. We report 5 candidate genes within those loci: Tyr, Ctsc, Plekhf2, Ndufaf6 and Angpt2. Tyr, Ndufaf6 and Angpt2 genes have been previously implicated by human GWAS of IOP-related conditions. Ctsc and Plekhf2 genes represent novel findings that may provide new insight into the molecular basis of IOP. This study highlights the efficacy of HS rats for investigating the genetics of elevated IOP and identifying potential candidate genes for future functional testing. Contribution to the field statementGlaucoma is the leading cause of irreversible blindness worldwide. Intraocular pressure (IOP) is the only known modifiable risk factor. This study describes results of the genome-wide association study (GWAS) performed in outbred rats that identifies known and novel genes involved in IOP regulation. To our knowledge, this is the first GWAS performed for IOP in a rat model. Identifying novel candidate genes in the rat model provides insight into the risk factors for glaucoma in humans and potential pharmacological targets for regulating IOP. The rat model is advantageous for studying natural variations in IOP, controlling environmental exposures, and providing easier access to tissue that can be used in phenotyping and gene expression in future studies.

genomics↗

A metagenomic approach to the molecular analysis of bacterial communities in tailings of a gold mine

Gold mine operations release arsenic pollutants in environment. The present study investigated the diversity of bacterial communities in the arsenic-contaminated tailings dam effluent (TDE) of Zarshuran gold mine, Takab, northwest of Iran. The bacterial communities were examined using the next-generation sequencing method (Illumina) targeting the V3-V4 region of 16S rRNA genes. Bacteroidetes (50.3%), Proteobacteria (45.49%), Actinobacteria (1.14%) and Firmicutes (1.08%) constituted dominant phyla in the TDE. Its diversity was analyzed, and compared with that of three previously-studied arsenic-contaminated groundwater (GW) microbiomes. The raw sequencing data were analyzed in QIIME2. The prevalent taxonomic groups observed in all of the samples belonged to Proteobacteria (8.06-45.49%), Bacteroidetes (1.85-50.32%), Firmicutes (1.00-6.2%), Actinobacteria (0.86-5.09%), Planctomycetes (0.05-9.37%) and Cyanobacteria (0.6-2.71%). According to Principal Coordinates Analysis (PCoA), the TDE and GW samples were divided into high and low groups, based on their arsenic content, respectively. The average numbers of observed alpha diversity indices, i.e. Pielous evenness and Faiths phylogenetic diversity, were not significantly different (P=0.18, Kruskal-Wallis test). According to the beta diversity analysis (unweighted), the observed operational taxonomic units (OTUs) and the diversity of the bacterial populations were not significantly different in the TDE, compared to the GW samples (PERMANOVA, P=0.276, 999 Permutations).

bioinformatics↗

The regulatory landscape of multiple brain regions in outbred heterogeneous stock rats

Heterogeneous Stock (HS) rats are a genetically diverse outbred rat population that is widely used for studying genetics of behavioral and physiological traits. Mapping Quantitative Trait Loci (QTL) associated with transcriptional changes would help to identify mechanisms underlying these traits. We generated genotype and transcriptome data for five brain regions from 88 HS rats. We identified 21,392 cis-QTLs associated with expression and splicing changes across all five brain regions and validated their effects using allele specific expression data. We identified 80 cases where eQTLs were colocalized with genome-wide association study (GWAS) results from nine physiological traits. Comparing our dataset to human data from the Genotype-Tissue Expression (GTEx) project, we found that the HS rat data yields twice as many significant eQTLs as a similarly sized human dataset. We also identified a modest but highly significant correlation between genetic regulatory variation among orthologous genes. Surprisingly, we found less genetic variation in gene regulation in HS rats relative to humans, though we still found eQTLs for the orthologs of many human genes for which eQTLs had not been found. These data are available from the RatGTEx data portal (RatGTEx.org) and will enable new discoveries of the genetic influences of complex traits.

genetics↗

Haplotype-aware modeling of cis-regulatory effects highlights the gaps remaining in eQTL data

Expression Quantitative Trait Loci (eQTLs) are critical to understanding the mechanisms underlying disease-associated genomic loci. Nearly all protein-coding genes in the human genome have been associated with one or more eQTLs. Here we introduce a multi-variant generalization of allelic Fold Change (aFC), aFC-n, to enable accurate quantification of the cis-regulatory effects in genes with multiple conditionally independent eQTLs. Applying aFC-n to 458,465 eQTLs in the Genotype-Tissue Expression (GTEx) project data, we demonstrate significant improvement in accuracy over the current tools for estimating the eQTL effect size and predicting genetically regulated gene expression. We characterize some of the empirical properties of the eQTL data and use this framework to assess the current state of eQTL data in terms of characterizing cis-regulatory landscape in individual genomes. Notably, we show that 77.4% of the genes with an allelic imbalance in a sample show 0.5 log2 fold or more of residual imbalance after accounting for the eQTL data underlining the remaining gap in characterizing regulatory landscape in individual genomes. We further contrast this gap across tissue types, and ancestry backgrounds to identify its correlates and guide future studies.

genetics↗

Transcription factor regulation of eQTL activity across individuals and tissues

Tens of thousands of genetic variants associated with gene expression (cis-eQTLs) have been discovered in the human population. These eQTLs are active in various tissues and contexts, but the molecular mechanisms of eQTL variability are poorly understood, hindering our understanding of genetic regulation across biological contexts. Since many eQTLs are believed to act by altering transcription factor (TF) binding affinity, we hypothesized that analyzing eQTL effect size as a function of TF level may allow discovery of mechanisms of eQTL variability. Using GTEx Consortium eQTL data from 49 tissues, we analyzed the interaction between eQTL effect size and TF level across tissues and across individuals within specific tissues and generated a list of 6,262 TF-eQTL interactions across 1,598 genes that are supported by at least two lines of evidence. These TF-eQTLs were enriched for various TF binding measures, supporting with orthogonal evidence that these eQTLs are regulated by the implicated TFs. We also found that our TF-eQTLs tend to overlap genes with gene-by-environment regulatory effects and to colocalize with GWAS loci, implying that our approach can help to elucidate mechanisms of context-specificity and trait associations. Finally, we highlight an interesting example of IKZF1 TF regulation of an APBB1IP gene eQTL that colocalizes with a GWAS signal for blood cell traits. Together, our findings provide candidate TF mechanisms for a large number of eQTLs and offer a generalizable approach for researchers to discover TF regulators of genetic variant effects in additional QTL datasets. Author SummaryGene expression is regulated by local genomic sequence and can be affected by genetic variants. In the human population, tens of thousands of cis-regulatory variants have been discovered that are associated with altered gene expression across tissues, cell types, or environmental conditions. Understanding the molecular mechanisms of how these small changes in the genome sequence affect genome function would offer insight to the genetic regulatory code and how gene expression is controlled across tissues and environments. Current research efforts suggest that many regulatory variants effects on gene expression are mediated by them altering the binding of transcription factors, which are proteins that bind to DNA to regulate gene expression. Here, we exploit the natural variation of TF activity among 49 tissues and between 838 individuals to elucidate which TFs regulate which regulatory variants. We find 6,262 TF-eQTL interactions across 1,598 genes that are supported by at least two lines of evidence. We validate these interactions using functional genomic and experimental approaches, and we find indication that they may pinpoint mechanisms of environment-specific genetic regulatory effects and genetic variants associated to diseases and traits.

genomics↗

Transcriptome variation in human tissues revealed by long-read sequencing

Regulation of transcript structure generates transcript diversity and plays an important role in human disease. The advent of long-read sequencing technologies offers the opportunity to study the role of genetic variation in transcript structure. In this paper, we present a large human long-read RNA-seq dataset using the Oxford Nanopore Technologies platform from 88 samples from GTEx tissues and cell lines, complementing the GTEx resource. We identified just under 100,000 new transcripts for annotated genes, and validated the protein expression of a similar proportion of novel and annotated transcripts. We developed a new computational package, LORALS, to analyze genetic effects of rare and common variants on the transcriptome via allele-specific analysis of long reads. We called allele-specific expression and transcript structure events, providing novel insights into the specific transcript alterations caused by common and rare genetic variants and highlighting the resolution gained from long-read data. We were able to perturb transcript structure upon knockdown of PTBP1, an RNA binding protein that mediates splicing, thereby finding genetic regulatory effects that are modified by the cellular environment. Finally, we use this dataset to enhance variant interpretation and study rare variants leading to aberrant splicing patterns.

genomics↗

EdiTyper: a high-throughput tool for analysis of targeted sequencing data from genome editing experiments

Genome editing experiments are generating an increasing amount of targeted sequencing data with specific mutational patterns indicating the success of the experiments and genotypes of clonal cell lines. We present EdiTyper, a high-throughput command line tool specifically designed for analysis of sequencing data from polyclonal and monoclonal cell populations from CRISPR gene editing. It requires simple inputs of sequencing data and reference sequences, and provides comprehensive outputs including summary statistics, plots, and SAM/BAM alignments. Analysis of simulated data showed that EdiTyper is highly accurate for detection of both single nucleotide mutations and indels, robust to sequencing errors, as well as fast and scalable to large experimental batches. EdiTyper is available in github (https://github.com/LappalainenLab/edityper) under the MIT license.

genomics↗

Diverse transcriptomic signatures across human tissues identify functional rare genetic variation

Rare genetic variation is abundant in the human genome, yet identifying functional rare variants and their impact on traits remains challenging. Measuring aberrant gene expression has aided in identifying functional, large-effect rare variants. Here, we expand detection of genetically driven transcriptome abnormalities by evaluating and integrating gene expression, allele-specific expression, and alternative splicing from multi-tissue RNA-sequencing data. We demonstrate that each signal informs unique classes of rare variants. We further develop Watershed, a probabilistic model that integrates multiple genomic and transcriptomic signals to predict variant function. Assessing rare variants prioritized by Watershed in the UK Biobank and Million Veterans Program, we identify large effects across 34 traits, and 33 rare variant-trait combinations with both high Watershed scores and large trait effect sizes. Together, we provide a comprehensive analysis of the transcriptomic impact of rare variation and a framework to prioritize functional rare variants and assess their trait relevance.\n\nOne-sentence summaryIntegrating expression, allelic expression and splicing across tissues identifies rare variants with relevance to traits.

genomics↗