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Alinejad-Rokny, H.

Publications and source records attributed to Alinejad-Rokny, H..

15 recordsLinked to original sources

Decoding Prokaryotic Whole Genomes with a Product-Contextualized Large Language Model

Genomes encode the instructions for life, yet their interpretation requires models capable of capturing long-range genome-wide functional context at scale. Existing genomic foundation models have primarily focused on sequence-level representations, providing powerful tools for biological prediction while leaving complementary opportunities for function-level genome modeling. Here, we present GenSyntax, a function-level genome representation learning framework that models prokaryotic genomes as ordered sequences of gene-product descriptors. By treating gene products as semantic units, GenSyntax transforms annotated replicons into interpretable "genetic paragraphs" that capture genome-wide functional context. We trained GenSyntax on 49,250 annotated prokaryotic genomes and evaluated its ability across genome-scale tasks, including plasmid host prediction, gene-product disambiguation, genome contig ordering and gene essentiality prediction. Compared with general-purpose large language models and representative genomic foundation models, GenSyntax achieved competitive performance across multiple benchmarks, with independent external evaluations supporting its generalization beyond the original training datasets. GenSyntax embeddings also captured signals associated with microbial phenotypes, and gene essentiality predictions enabled exploratory design of minimal genomes. Together with the lightweight GenSyntax-Tiny model, GenSyntax provides a complementary, post-annotation framework for function-level analysis of prokaryotic genomes.

genomics↗

Comparative Analysis of Single-Nucleus and Single-Cell RNA Sequencing in Human Bone Marrow Mononuclear Cells: Methodological Insights and Trade-offs

Bone marrow mononuclear cells (BMMCs) are a heterogeneous pool of hematopoietic progenitors and mature immune cells that collectively sustain hematopoiesis and coordinate immune responses. The bone marrow serves not only as the primary site for blood cell production but also as a niche for various disorders, including blood cancers. Advances in single-cell RNA sequencing (scRNA-seq) and single-nucleus RNA sequencing (snRNA-seq) have significantly enhanced our understanding of the cellular biology and molecular dynamics within this complex microenvironment. The choice between these two approaches, however, is often shaped or constrained by the study design, such as research objectives, sample type, and preservation conditions. Consequently, methodological differences in library preparation and transcript capture efficiency can introduce systematic biases that complicate downstream analyses and interpretation, underscoring the need to identify and account for method-specific features. In this study, we conducted a comparative analysis of matched snRNA-seq and scRNA-seq datasets from 11 pairs of healthy donor bone marrow mononuclear cell samples, generated using the popular 10x Genomics platform. We evaluated method-specific biases using multiple quality metrics and compared cell type proportions and transcriptomic signatures captured by each approach. Integrative analysis of these datasets is feasible but not advisable due to systematic gene length biases that were observed between these approaches. Our results showed that despite inherent differences in library complexity, both protocols reliably captured all major cell types. This comparative analysis highlights intrinsic differences between snRNA-seq and scRNA-seq data, providing valuable insights into their respective advantages, limitations, and trade-offs. These findings can assist researchers in selecting the optimal method tailored to specific biological questions and sample characteristics, and also enable more method-aware data analysis and interpretation.

bioinformatics↗

Unveiling multi-scale architectural features in single-cell Hi-C data using scCAFE

Single-cell Hi-C (scHi-C) has provided unprecedented insights into the heterogeneity of 3D genome organization. However, its sparse and noisy nature poses challenges for computational analyses, such as chromatin architectural feature identification. Here, we introduce scCAFE, a deep learning model for the multi-scale detection of architectural features at the single-cell level. scCAFE provides a unified framework for annotating chromatin loops, TAD-like domains (TLDs), and compartments across individual cells. Our model outperforms previous scHi-C loop calling methods and delivers accurate predictions of TLDs and compartments that are biologically consistent with previous studies. The resulting single-cell annotations also offer a measure to characterize the heterogeneity of different levels of architectural features across cell types. We leverage this heterogeneity and identify a series of marker loop anchors, which demonstrate the potential of the 3D genome data to annotate cell identities without the aid of simultaneously sequenced omics data. Overall, scCAFE not only serves as a useful tool for analyzing single-cell genomic architecture, but also paves the way for precise cell-type annotations solely based on 3D genome features.

bioinformatics↗

Genome-Wide CRISPR-Cas9 Screening Identifies a Synergy between Hypomethylating Agents and SUMOylation Blockade in MDS/AML

Hypomethylating agents (HMAs) are frontline therapies effective at altering the natural course of Myelodysplastic Neoplasms (MDS) and Acute Myeloid Leukemia (AML). However, acquired resistance and treatment failure are hallmarks of HMA therapy. To address this clinical need, we performed a genome-wide CRISPR-Cas9 screen in a human MDS-derived cell line, MDS-L, and identified TOPORS as a highly ranked loss-of-function target that synergizes with HMAs, reducing leukemic burden and improving survival in xenograft models. We demonstrate that the depletion of TOPORS mediates sensitivity to HMAs by predisposing leukemic blasts to an impaired DNA damage response (DDR) accompanied by an accumulation of SUMOylated DNMT1 in HMA-treated TOPORS-depleted cells. Importantly, the combination of HMAs with targeting of TOPORS did not functionally impair healthy hematopoiesis. While inhibitors of TOPORS are currently unavailable, we show that inhibition of protein SUMOylation (upstream of TOPORS functions) with TAK-981 partially phenocopies HMA-sensitivity and DDR impairment. Overall, our data suggest that the combination of HMAs with the inhibition of SUMOylation or TOPORS demonstrates a favourable therapeutic index and is a rational treatment framework for High-Risk MDS (HR-MDS) or AML.

molecular biology↗

DeepGenePrior: A deep learning model to prioritize genes affected by copy number variants

The genetic etiology of neurodevelopmental disorders is highly heterogeneous. They are characterized by abnormalities in the development of the central nervous system, which lead to diminished physical or intellectual capabilities. Determining which gene is the driver of disease (not just a passenger), termed gene prioritization, is not entirely known. In terms of disease-gene associations, genome-wide explorations are still underdeveloped due to the reliance on previous discoveries when spotting new genes and other evidence sources with false positive or false negative relations. This paper introduces DeepGenePrior, a model based on deep neural networks that prioritizes candidate genes in Copy Number Variant (CNV) mediated diseases. Based on the well-studied Variational AutoEncoder (VAE), we developed a score to measure the impact of the genes on the target diseases. Unlike other methods that use prior data on gene-disease associations to prioritize candidate genes (using the guilt by association principle), the current study exclusively relies on copy number variants. Therefore, the procedure can identify disease-associated genes regardless of prior knowledge or auxiliary data sources. We identified genes that distinguish cases from disorders (autism, schizophrenia, and developmental delay). A 12% increase in fold enrichment was observed in brain-expressed genes compared to previous studies, while 15% more fold enrichment was found in genes associated with mouse nervous system phenotypes. We also explored sex dimorphism for the disorders and discovered genes that overexpress more in one gender than the other. Additionally, we investigated the gene ontology of the putative genes with WebGestalt and the associations between the causative genes and the other phenotypes in the DECIPHER dataset. Furthermore, some genes were jointly present in the top genes associated with the three disorders in this study (i.e., autism spectrum disorder, schizophrenia, and developmental delay); namely, deletions in ZDHHC8, DGCR5, and CATG00000022283 were common between them. These findings suggest the common etiology of these clinically distinct conditions. With DeepGenePrior, we address the obstacles in existing gene prioritization studies. This study identified promising candidate genes without prior knowledge of diseases or phenotypes using deep learning.

bioinformatics↗

A benchmarking of deep neural network models for cancer subtyping using single point mutations

It is now well-known that genetic mutations contribute to development of tumors, in which at least 15% of cancer patients experience a causative genetic abnormality including De Novo somatic point mutations. This highlights the importance of identifying responsible mutations and the associated biomarkers (e.g., genes) for early detection in high-risk cancer patients. The next-generation sequencing technologies have provided an excellent opportunity for researchers to study associations between De Novo somatic mutations and cancer progression by identifying cancer subtypes and subtype-specific biomarkers. Simple linear classification models have been used for somatic point mutation-based cancer classification (SMCC); however, because of cancer genetic heterogeneity (ranging from 50% to 80%), high data sparsity, and the small number of cancer samples, the simple linear classifiers resulted in poor cancer subtypes classification. In this study, we have evaluated three advanced deep neural network-based classifiers to find and optimized the best model for cancer subtyping. To address the above-mentioned complexity, we have used pre-processing clustered gene filtering (CGF) and indexed sparsity reduction (ISR), regularization methods, a Global-Max-Pooling layer, and an embedding layer. We have evaluated and optimized the three deep learning models CNN, LSTM, and a hybrid model of CNN+LSTM on publicly available TCGA-DeepGene dataset, a re-formulated subset of The Cancer Genome Atlas (TCGA) dataset and tested the performance measurement of these models is 10-fold-cross-validation accuracy. Evaluating all the three models using a same criterion on the test dataset revealed that the CNN, LSTM, and CNN+LSTM have 66.45% accuracy, 40.89% accuracy, and 41.20% accuracy in somatic point mutation-based cancer classification. Based on our results, we propose the CNN model for further experiments on cancer subtyping based on DNA mutations.

bioengineering↗

Somatic point mutations are enriched in long non-coding RNAs with possible regulatory function in breast cancer

De novo somatic point mutations identified in breast cancer are predominantly non-coding and typically attributed to altered regulatory elements such as enhancers and promoters. However, while the non-coding RNAs (ncRNAs) form a large portion of the mammalian genome, their biological functions are mostly poorly characterized in cancers. In this study, using a newly developed tool, SomaGene, we reanalyze de novo somatic point mutations from the International Cancer Genome Consortium (ICGC) whole-genome sequencing data of 1,855 breast cancers. We identify 929 candidates of ncRNAs that are significantly and explicitly mutated in breast cancer samples. By integrating data from the ENCODE regulatory features and FANTOM5 expression atlas, we show that the candidate ncRNAs in breast cancer samples significantly enrich for active chromatin histone marks (1.9 times), CTCF binding sites (2.45 times), DNase accessibility (1.76 times), HMM predicted enhancers (2.26 times) and eQTL polymorphisms (1.77 times). Importantly, we show that the 929 ncRNAs contain a much higher level (3.64 times) of breast cancer-associated genome-wide association (GWAS) single nucleotide polymorphisms (SNPs) than genome-wide expectation. Such enrichment has not been seen with GWAS SNPs from other diseases. Using breast tissue related Hi-C data we then show that 82% of our candidate ncRNAs (1.9 times) significantly interact with the promoter of protein-coding genes, including previously known cancer-associated genes, suggesting the critical role for candidate ncRNA genes in activation of essential regulators of development and differentiation in breast cancer. We provide an extensive web-based resource (http://ncrna.ictic.sharif.edu), to communicate our results with the research community. Our list of breast cancer-specific ncRNA genes has the potential to provide a better understanding of the underlying genetic causes of breast cancer. Lastly, the tool developed in this study can be used in the analysis of somatic mutations in all cancers.

cancer biology↗

Signatures of Mutational Processes in Human DNA Evolution

The human genome contains over 100 million SNPs, most of which are C/T (G/A) variations. The type and sequence context of these SNPs are not random, suggesting that they are caused by distinct mutational processes. Deciphering the mutational signatures is a crucial step to discovering the molecular processes responsible for DNA variations across human populations, and potentially for causing genetic diseases. Our analyses of the 1000 Genomes Project SNPs and germline de novo mutations suggest that at least four mutational processes are responsible for human genetic variations. One process is European-specific and no longer active. The remaining three processes are currently active in all human populations. Two of the active processes co-occur and leave a single joint mutational signature in human nuclear DNA. The third active process is specific to mitochondrial DNA, and inflicts C-to-T mutations at mostly non-CG sites. We found neither evidence of APOBEC-induced cytosine deamination in the human germline, nor de novo mutation enrichment within certain regions of the human genome.

evolutionary biology↗

Seeing the forest through the trees: Identifying functional interactions from Hi-C

Eukaryotic genomes are highly organised within the nucleus of a cell, allowing widely dispersed regulatory elements such as enhancers to interact with gene promoters through physical contacts in three-dimensional space. Recent chromosome conformation capture methodologies such as Hi-C have enabled the analysis of interacting regions of the genome providing a valuable insight into the three-dimensional organisation of the chromatin in the nucleus, including chromosome compartmentalisation and gene expression. Complicating the analysis of Hi-C data however is the massive amount of identified interactions, many of which do not directly drive gene function, thus hindering the identification of potentially biologically functional 3D interactions. In this review, we collate and examine the downstream analysis of Hi-C data with particular focus on methods that identify significant functional interactions. We classify three groups of approaches; structurally-associated domain discovery methods e.g. topologically-associated domains and compartments, detection of statistically significant interactions via background models, and the use of epigenomic data integration to identify functional interactions. Careful use of these three approaches is crucial to successfully identifying functional interactions within the genome.

bioinformatics↗

Evidence for ZAP-independent CpG reduction in SARS-CoV-2 genome, and pangolin coronavirus origin of 5'UTR

SARS-CoV-2, the causative agent of COVID-19, has an RNA genome, which is, overall, closely related to the bat coronavirus sequence RaTG13. However, the ACE2-binding domain of this virus is more similar to a coronavirus isolated from a Guangdong pangolin. In addition to this unique feature, the genome of SARS-CoV-2 (and its closely related coronaviruses) has a low CpG content. This has been postulated to be the signature of an evolutionary pressure exerted by the host antiviral protein ZAP. Here, we analyzed the sequences of a wide range of viruses using both alignment-based and alignment free approaches to investigate the origin of SARS-CoV-2 genome. Our analyses revealed a high level of similarity between the 5UTR of SARS-CoV-2 and that of the Guangdong pangolin coronavirus. This suggests bat and pangolin coronaviruses might have recombined at least twice (in the 5UTR and ACE2 binding regions) to seed the formation of SARS-CoV-2. An alternative hypothesis is that the lineage preceding SARS-CoV-2 is a yet to be sampled bat coronavirus whose ACE2 binding domain and 5UTR are distinct from other known bat coronaviruses. Additionally, we performed a detailed analysis of viral genome compositions as well as expression and RNA binding data of ZAP to show that the low CpG abundance in SARS-CoV-2 is not related to an evolutionary pressure from ZAP.

genomics↗

Integrative analysis of mutated genes and mutational processes reveals seven colorectal cancer subtypes

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths in the world. It has been reported that [~]10%-15% of individuals with colorectal cancer experience a causative mutation in the known susceptibility genes, highlighting the importance of identifying mutations for early detection in high risk individuals. Through extensive sequencing projects such as the International Cancer Genome Consortium (ICGC), a large number of somatic point mutations have been identified that can be used to identify cancer-associated genes, as well as the signature of mutational processes defined by the tri-nucleotide sequence context (motif) of mutated sites. Mutation is the hallmark of cancer genome, and many studies have reported cancer subtyping based on the type of frequently mutated genes, or the proportion of mutational processes, however, none of these cancer subtyping methods consider these features simultaneously. This highlights the need for a better and more inclusive subtype classification approach to enable biomarker discovery and thus inform drug development for CRC. In this study, we developed a statistical pipeline based on a novel concept gene-motif, which merges mutated gene information with tri-nucleotide motif of mutated sites, to identify cancer subtypes, in this case CRCs. Our analysis identified for the first time, 3,131 gene-motif combinations that were significantly mutated in 536 ICGC colorectal cancer samples compared to other cancer types, identifying seven CRC subtypes with distinguishable phenotypes and biomarkers. Interestingly, we identified several genes that were mutated in multiple subtypes but with unique sequence contexts. Taken together, our results highlight the importance of considering both the mutation type and mutated genes in identification of cancer subtypes and cancer biomarkers.

cancer biology↗

Evidence for enhancer noncoding RNAs (enhancer-ncRNAs) with gene regulatory functions relevant to neurodevelopmental disorders

Noncoding RNAs (ncRNAs) comprise a significant proportion of the mammalian genome, but their biological significance in neurodevelopment disorders is poorly understood. In this study, we identified 908 brain-enriched noncoding RNAs comprising at least one nervous system-related eQTL polymorphism that is associated with protein coding genes and also overlap with chromatin states characterised as enhancers. We referred to such noncoding RNAs with putative enhancer activity as brain enhancer-ncRNAs. By integrating GWAS SNPs and Copy Number Variation (CNV) data from neurodevelopment disorders, we found that 265 enhancer-ncRNAs were either mutated (CNV deletion or duplication) or contain at least one GWAS SNPs in the context of such conditions. Of these, the eQTL-associated gene for 82 enhancer-ncRNAs did not overlap with either GWAS SNPs or CNVs suggesting in such contexts that mutations to neurodevelopment gene enhancers disrupt ncRNA interaction. Taken together, we identified 49 novel NDD-associated ncRNAs that influence genomic enhancers during neurodevelopment, suggesting enhancer mutations may be relevant to the functions for such ncRNAs in neurodevelopmental disorders.

neuroscience↗

MaxHiC: robust estimation of chromatin interaction frequency in Hi-C and capture Hi-C experiments

Hi-C is a genome-wide chromosome conformation capture technology that detects interactions between pairs of genomic regions, and exploits higher order chromatin structures. Conceptually Hi-C data counts interaction frequencies between every position in the genome and every other position. Biologically functional interactions are expected to occur more frequently than random (background) interactions. To identify biologically relevant interactions, several background models that take biases such as distance, GC content and mappability into account have been proposed. Here we introduce MaxHiC, a background correction tool that deals with these complex biases and robustly identifies statistically significant interactions in both Hi-C and capture Hi-C experiments. MaxHiC uses a negative binomial distribution model and a maximum likelihood technique to correct biases in both Hi-C and capture Hi-C libraries. We systematically benchmark MaxHiC against major Hi-C background correction tools and demonstrate using published Hi-C and capture Hi-C datasets that 1) Interacting regions identified by MaxHiC have significantly greater levels of overlap with known regulatory features (e.g. active chromatin histone marks, CTCF binding sites, DNase sensitivity) and also disease-associated genome-wide association SNPs than those identified by currently existing models, and 2) the pairs of interacting regions are more likely to be linked by eQTL pairs and more likely to link known regulatory features than any of the existing methods. We also demonstrate that interactions between different genomic region types have distinct distance distribution only revealed by MaxHiC. MaxHiC is publicly available as a python package for the analysis of Hi-C and capture Hi-C data. Author summaryMaxHiC is a robust machine learning based tool for identifying significant interacting regions from both Hi-C and capture Hi-C data. All the current existing models are designed for either Hi-C or capture Hi-C data, however we developed MaxHiC to be applicable for both Hi-C and capture Hi-C libraries (two different models have been used for Hi-C and capture Hi-C data). MaxHiC is also able to analyse very deep Hi-C libraries (e.g., MicroC) without any computational issues. MaxHiC significantly outperforms current existing tools in terms of enrichment of interactions between known regulatory regions as well as biologically relevant interactions.

bioinformatics↗

Brain-enriched coding and long non-coding RNA genes are overrepresented in recurrent autism spectrum disorder CNVs

Autism spectrum disorder (ASD) is a neurodevelopmental disorder with substantial phenotypic and etiological heterogeneity. It is estimated that 10-20% of cases are due to copy number variations (CNVs). Here we apply a newly developed CNV association tool (SNATCNV) to reanalyse CNV data from 19,663 autistic and 6,479 control subjects from the AutDB database. We demonstrate that SNATCNV outperforms existing CNV association methods by finding smaller genomic regions that better discriminate cases and controls. By integrating data from the FANTOM5 expression atlas we show that both known ASD causal genes identified by the SFARI and MSSNG consortia and genes within the CNVs identified by SNATCNV have brain enriched expression patterns; both brain-enriched coding and long-non-coding RNA genes are over-represented. We provide full lists of these brain enriched coding and lncRNA genes as a resource to the research community. We also go on to show that each CNV region is associated with a distinct set of phenotypes, that some are sex biased and highlight one deleted region where a brain-enriched lncRNA is the only gene present. Our analyses identify 47 high confidence ASD associated CNV regions and identifies brain-enriched genes which underlie this neurodevelopmental disorder.

neuroscience↗

CANCERSIGN: a user-friendly and robust tool for identification and classification of mutational signatures and patterns in cancer genomes

Analyses of large somatic mutation datasets, using advanced computational algorithms, have revealed at least 30 independent mutational signatures in tumor samples. These studies have been instrumental in identification and quantification of responsible endogenous and exogenous molecular processes against cancer. The quantitative approach used to deconvolute mutational signatures is becoming an integral part of cancer research. Therefore, development of a stand-alone tool with a user-friendly graphical interface for analysis of cancer mutational signatures is necessary. In this manuscript, we introduce CANCERSIGN as an open access1 bioinformatics tool that uses raw mutation data (BED files) as input, and generates 3-mer and 5-mer mutational signatures. Additionally, this tool enables users to perform clustering on tumor samples based on the raw mutation counts as well as using the proportion of mutational signatures in each sample. Using this tool, we analysed all the whole genome somatic mutation datasets of International Cancer Genome Consortium (ICGC) samples and identified a number of novel signatures.

bioinformatics↗