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Vafaee, F.

Publications and source records attributed to Vafaee, F..

At least 19 recordsLinked to original sources

Optimal transport reveals unresolved cells in single cell multi-omics

Single-cell multi-omics methods align cells across modalities, integrate them into one embedding and transfer labels between them. All three steps depend on a match between cells in two modalities, but this match usually stays hidden inside each tool. Its errors are hidden too, and every cell looks equally certain. Here we make the match explicit. For each pair of datasets, we build a soft coupling matrix with optimal transport and use it in three ways beyond estimating the match itself: to score how uncertain each cell's match is, to upgrade existing integration and label-transfer tools without retraining them, and to decide which cells to set aside. Across 78 dataset pairs spanning six modality combinations, we evaluated ten integration and ten label-transfer tools using matched random removal as the control. We developed an ensemble coupling achieving the most accurate recovery of true cell pairs. Feeding couplings back into the tools improved label transfer clearly but improved integration only in how well the modalities mix. The ambiguity score, which we computed as a group-vote margin, gave the larger and more consistent gain in both tasks. Ambiguous-flagged cells matched worse in almost every dataset and removing them beat random removal in 98% of integration tests and 91% of label-transfer comparisons. These hard-to-integrate cells are mostly among rare, transitional and poorly measured populations. Making correspondence explicit therefore provides a practical way to distinguish reliable cross-modality results from uncertain ones while flagging cells that might have important roles in development and disease.

bioinformatics↗

Molecular Determinants of Functional Bacterial sRNA-mRNA Interactions Revealed by Integrating RNA Interactomes and Interpretable Machine Learning

Bacterial small RNAs (sRNAs) regulate gene expression by base pairing with target mRNAs, yet transcriptome-wide interactome mapping has shown that many sRNA-mRNA interactions detected in vivo have modest or no regulatory effect using orthogonal reporter assays. The features that determine functional outcome remain poorly defined. Here, we integrated Hfq-CLASH interactome mapping with matched transcriptomic and proteomic profiling in Escherichia coli and developed an interpretable machine-learning framework to identify the determinants that distinguish functional from non-functional interactions. Using sequence, structural, thermodynamic, duplex and protein-occupancy features, transcriptomic and proteomic responses were predicted with above-chance performance, achieving AUCs of 0.78 and 0.74, respectively. Feature attribution revealed that physical pairing alone is insufficient for regulation; instead, regulatory outcome is shaped by a coordinated interplay between RNA secondary structure, thermodynamic accessibility and local protein-binding context. Target-side Hfq occupancy emerged as a positive predictor of functional regulation, whereas AR2-domain occupancy on the sRNA was associated with non-responsive interactions, suggesting that distinct ribonucleoprotein states may separate productive regulation from non-productive binding. These findings indicate that the regulatory fate of an sRNA-mRNA interaction is an emergent property of its biophysical context and protein-binding environment, rather than a direct consequence of physical pairing alone.

molecular biology↗

SCIMETAR-seq tracks immunophenotype, demethylation, mutations, and transcriptomes in single cells undergoing HMA therapy

5-azacytidine improves haematopoiesis and delays leukaemic progression in myelodysplastic neoplasms, but responses vary and are complicated by clonal mosaicism and heterogenous demethylation. Using a novel single-cell pipeline ("SCIMETAR-seq"), we found 5-azacytidine induced clonally distinct differentiation responses, with nuclear DNA demethylation occurring primarily in cycling progenitors. Despite the absence of significant nuclear DNA demethylation, quiescent stem cells underwent transcriptional remodelling in vivo, accompanied by 5-azacytidine-induced C*G-to-G*C mutations in mitochondria.

cancer biology↗

Single cell multiomics reveal clonal and functional dynamics of MDS stem/progenitor cells during hypomethylating therapy

Progressive somatic mutations in hematopoietic stem cells (HSCs) drive the development of myelodysplastic neoplasms (MDS). Hypomethylating agents such as azacitidine (AZA) can improve blood counts and reduce blasts, although responses are rarely durable. Determinants of AZA response are complex and incompletely understood, although accumulating evidence suggests that epigenetic rewiring of mutated HSCs underlies improved hematopoietic output. Using single cell multiomics on longitudinal bone marrow samples, we show that AZA responsiveness involves expansion of cells with transcriptomic profiles shared with hematopoietic stem and progenitor cells (HSPCs) from healthy donors. These regenerating cells are depleted of copy number variations and of TP53 mutations. We also identify patient-restricted cell populations, some of which recede through transcriptional restoration or AZA cytotoxicity, and others which expand, regardless of initial clinical response, and dominate at progression. Individual patients carried multiple patient-restricted populations which had unique surface immunophenotypes and were genetically distinct. Strikingly, sorted cells from in vivo progression clones that were AZA-refractive in patients regained AZA-sensitivity when cultured in vitro, suggesting that lack of AZA response at the cellular level can be modulated by cell-extrinsic factors in vivo. Overall, we find that AZA response involves partial hematopoietic regeneration via functional differentiation of mutated, but not cytogenetically abnormal HSPCs, and that persistence of AZA-refractive sub-populations contributes to eventual disease progression.

cancer biology↗

Deep multiplexed 50-marker imaging of circulating tumor cells expands actionable biomarker profiling for precision oncology

Liquid biopsy-derived circulating tumor cells (CTCs) offer a minimally invasive avenue for precision oncology by enabling longitudinal monitoring of actionable molecular and cell-phenotypic biomarkers. However, conventional immunofluorescence imaging-based CTC profiling captures just 4-5 markers per cell, restricting crucial insights into oncogenic and resistance drivers, as well as tumor cell heterogeneity. Here we present an integrated pipeline employing deep multiplexed imaging to profile up to 50 molecular markers per CTC, capturing expression, phosphorylation, and subcellular localization of diverse biomarkers. Validated in a multi-stage prostate cancer resistance model and applied to prostate cancer patient-derived CTCs, this approach enhances CTC classification and reveals inter- and intra-patient heterogeneity correlating with therapy response. Machine learning identified therapeutically actionable signatures, suggesting patient-specific treatment strategies. This deep multiplexed imaging pipeline advances the utility of CTCs in guiding personalized cancer therapy by providing comprehensive molecular and phenotypic insights through minimally invasive liquid biopsies. Highlights- Multiplexed immunofluorescence imaging of circulating tumor cells captures up to 50 molecular markers per cell with subcellular localisation. - Single-cell and subcellular analyses identify quantitative signatures correlating with patient outcome, as well as therapeutically actionable biomarkers. - This method transforms the utility of CTCs for future use in patient stratification and for guiding personalised therapies. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=181 SRC="FIGDIR/small/684075v1_ufig1.gif" ALT="Figure 1"> View larger version (47K): org.highwire.dtl.DTLVardef@a36ff4org.highwire.dtl.DTLVardef@b06c1org.highwire.dtl.DTLVardef@1e8a572org.highwire.dtl.DTLVardef@c60753_HPS_FORMAT_FIGEXP M_FIG Graphical Abstract C_FIG

cancer biology↗

Epithelial-mesenchymal cell state heterogeneity predetermines differential phospho-signaling responses to epidermal growth factor stimulation

Understanding why isogenic cancer cells respond differently to equivalent oncogenic stimuli is vital for optimizing anticancer therapies. Emerging evidence suggests that pre-existing differences in cell state may modulate signaling responses to new stimuli, but the interplay of specific cell states and signals remains unclear. We investigated whether epithelial-mesenchymal (E/M) state, a major axis of cancer cell heterogeneity, influences signaling responses to epidermal growth factor (EGF), a critical oncogenic stimulus in non-small cell lung cancer (NSCLC). We imaged >64,000 A549 NSCLC cells labeled for DNA, F-actin and alternate signaling markers (p-AKT-S473, p-AKT-T308, p-ERK or p-S6) after acute stimulation. Quantitative single-cell morphological and spatial profiling defined a stimulus-invariant E/M state landscape over which EGF signaling responses were compared. This revealed state-dependent differences in signal-activation magnitudes, dynamics and subcellular routing. AKT responses exhibited phosphosite- and compartment-specific dynamics across states, with epithelial cells showing strong, transient membrane-localized S473 and higher internalized T308, whereas mesenchymal cells displayed weaker but sustained nuclear and ruffle-localized S473. Regression-based computational multiplexing concurrently inferred all signaling responses per cell, mapping state-dependent divergence in multi-molecular signaling trajectories. E/M state thus pre-determines distinctive spatiotemporal profiles of EGF-induced signaling, with implications for signaling functions and anti-signaling therapy responses across E/M state-diverse tumors.

cancer biology↗

Medlib: A Feature-Rich C/C++ Library for Exact Alignment of Nanopore Sequences Using Multiple Edit Distance

Pairwise sequence alignment is a cornerstone of bioinformatics, driving applications from variant calling to barcode demultiplexing. However, the advent of long-read, high-error sequencing platforms--such as Oxford Nanopore Technologies--demands alignment tools that not only deliver exact results but also accommodate the elevated noise inherent in these data. Here, we introduce Medlib (Multiple Edit Distance Library) that offers both high performance and novel, biologically motivated features. Medlibs threshold mode allows users to retrieve every alignment with edit distance k [≤] kth and optionally alignment score s [≥] sth, ensuring comprehensive discovery of all plausible matches. Counterintuitively, this exhaustive retrieval outpaces traditional minimum-error searches. Medlib further provides the first exact implementations of overlap-aware reporting and motif boundary treatment. Medlib supports multiple input formats, predefined and custom alphabets, affine-gap scoring applied retrospectively, built-in dedicated multithreading schemes, and user-defined function callbacks for immediate post-processing. Benchmarking against Edlib demonstrates a 1.6x speedup in minimum mode and even greater gains (2.5x) in threshold mode--while delivering up to twice as many alignments. Medlib thus offers a robust, feature-rich engine for exact pairwise alignment in modern, high-throughput sequencing workflows. Availability and ImplementationSource code, installation instructions and test data are freely available for download at https://github.com/Mehdi-Rafeie/Medlib, under the MIT license.

bioinformatics↗

Blastocoel fluid RNA predicts pregnancy outcome in assisted reproduction

Nearly one in eight couples are affected by infertility, with many relying on assisted reproductive technologies (ART) to conceive. However, selecting the highest-quality embryo in ART remains a major challenge, as current assessment methods are often subjective, or invasive, and lack precision. Here, we introduce a novel strategy that analyses embryo-derived polyadenylated RNA in blastocoel fluid to more accurately predict pregnancy outcomes. Elevated RNA levels were strongly associated with implantation failure, particularly in embryos from women over the age of 34. Our predictive model developed using our sample cohort, incorporating both RNA and maternal age, demonstrated exceptional performance, achieving 76% accuracy in the training set and 73% in independent validation in predicting implantation outcome-- highlighting a promising advancement in embryo selection and ART success.

molecular biology↗

Single-Cell Data Integration and Cell Type Annotation through Contrastive Adversarial Open-set Domain Adaptation

Single-cell sequencing technologies have enabled in-depth analysis of cellular heterogeneity across tissues and disease contexts. However, as datasets increase in size and complexity, characterizing diverse cellular populations, integrating data across multiple modalities, and correcting batch effects remain challenges. We present SAFAARI (Single-cell Annotation and Fusion with Adversarial Open-Set Domain Adaptation Reliable for Data Integration), a unified deep learning framework designed for cell annotation, batch correction, and multi-omics integration. SAFAARI leverages supervised contrastive learning and adversarial domain adaptation to achieve domain-invariant embeddings and enables label transfer across datasets, addressing challenges posed by batch effects, biological domain shifts, and multi-omics modalities. SAFAARI identifies novel cell types and mitigates class imbalance to enhance the detection of rare cell types. Through comprehensive benchmarking, we evaluated SAFAARI against existing annotation and integration methods across real-world datasets exhibiting batch effects and domain shifts, as well as simulated and multi-omics data. SAFAARI demonstrated scalability and robust performance in cell annotation via label transfer across heterogeneous datasets, detection of unknown cell types, correction of batch effects, and cross-omics data integration while leveraging available annotations for improved integration. SAFAARIs innovative approach outperformed competing methods in both qualitative and quantitative metrics, offering a flexible, accurate, and scalable solution for single-cell analysis with broad applicability to diverse biological and clinical research questions.

bioinformatics↗

Genes in Humans and Mice: Insights from Deep learning of 777K Bulk Transcriptomes

Mice are widely used as animal models in biomedical research, favored for their small size, ease of breeding, and anatomical and physiological similarities to humans1,2. However, discrepancies between mouse gene experimental results and the actual behavior of human genes are not uncommon, despite their shared DNA sequence similarity3-8. This suggests that DNA sequence similarity does not always reliably predict functional similarity. On the other hand, RNA-level gene expression could offer additional information about gene function9,10. In this study, we undertook characterization and inter-species comparison of human and mouse genes by applying innovative deep learning methodologies to a large dataset of 410K human and 366K mouse bulk RNA-seq samples. This was achieved by using gene representations from our Transformer-based GeneRAIN model11,12. These gene representations aggregate information from large gene expression datasets, and provide insights beyond DNA sequence similarity. We identified 2,407 human-mouse homologous genes with high DNA similarity but distinct RNA characteristics, and showed that these genes are more likely to have differing disease/phenotype associations between the two species. Additionally, we found 3,070 homologous genes with low similarity at both the DNA and RNA levels, suggesting the highest risk of discrepancies in study results between the two species. We propose that this approach will support future decision making around whether the mouse will be an appropriate model for studying specific human genes, and whether the results of specific mouse gene studies are likely to be recapitulated in humans. Our methodological innovations offer valuable lessons for future deep learning applications in cross-species omics data. The interspecies gene relationship findings from our study also contribute valuable insights into the gene biology and evolution of the two species.

bioinformatics↗

Multifaceted Representation of Genes via Deep Learning of Gene Expression Networks

Accurate predictive modeling of human gene relationships would fundamentally transform our ability to uncover the molecular mechanisms that underpin key biological and disease processes. Recent studies have employed advanced AI techniques to model the complexities of gene networks using large gene expression datasets1-11. However, the extent and nature of the biological information these models can learn is not fully understood. Furthermore, the potential for improving model performance by using alternative data types, model architectures, and methodologies remains underexplored. Here, we developed GeneRAIN models by training on a large dataset of 410K human bulk RNA-seq samples, rather than single-cell RNA-seq datasets used by most previous studies. We showed that although the models were trained only on gene expression data, they learned a wide range of biological information well beyond gene expression. We introduced GeneRAIN-vec, a state-of-the-art, multifaceted vectorized representation of genes. Further, we demonstrated the capabilities and broad applicability of this approach by making 4,797 biological attribute predictions for each of 13,030 long non-coding RNAs (62.5 million predictions in total). These achievements stem from various methodological innovations, including experimenting with multiple model architectures and a new Binning-By-Gene normalization method. Comprehensive evaluation of our models clearly demonstrated that they significantly outperformed current state-of-the-art models3,12. This study improves our understanding of the capabilities of Transformer and self-supervised deep learning when applied to extensive expression data. Our methodological advancements offer crucial insights into refining these techniques. These innovations are set to significantly advance our understanding and exploration of biology.

bioinformatics↗

Cell Type-Specific Regulation by a Heptad of Transcription Factors in Human Hematopoietic Stem and Progenitor Cells

Hematopoietic stem and progenitor cells (HSPCs) rely on a complex interplay of transcription factors (TFs) to regulate differentiation into mature blood cells. A heptad of TFs - FLI1, ERG, GATA2, RUNX1, TAL1, LYL1, LMO2 - bind regulatory elements in bulk CD34+ HSPCs. However, whether specific heptad-TF combinations have distinct roles in regulating hematopoietic differentiation remained unknown. We mapped genome-wide chromatin contacts and TF binding profiles in HSPC subsets (HSC, CMP, GMP, MEP) and found that heptad occupancy and enhancer-promoter interactions varied significantly across cell types and were associated with cell-type-specific gene expression. Distinct regulatory elements were enriched with specific heptad-TF combinations, including stem-cell-specific elements with ERG, and myeloid- and erythroid-specific elements with combinations of FLI1, RUNX1, GATA2, TAL1, LYL1, and LMO2. These findings suggest that specific heptad-TF combinations play critical roles in regulating hematopoietic differentiation and provide a valuable resource for development of targeted therapies to manipulate specific HSPC subsets.

developmental biology↗

Omics Imagification: Transforming High-throughput Molecular Representation of a Cell into an Image

Different omics profiles, depending on the underlying technology, encompass measurements of several hundred to several thousand molecules in a biological sample or a cell. This study develops upon the concept of "omics imagification" as a process of transforming a vector representing these numerical measurements into an image with a one-to-one relationship with the corresponding sample. The proposed imagification process transforms a high-dimensional vector of molecular measurements into a two-dimensional RGB image to enable holistic molecular representation of a biological sample and to improve the classification of different biological phenotypes using automated image recognition methods in computer vision. A transformed image represents 2D coordinates of molecules in a neighbour-embedded space representing molecular abundance and gene intensity. The proposed method was applied to a single-cell RNA sequencing (scRNA-seq) data to "imagify" gene expression profiles of individual cells. Our results show that a simple convolutional neural network trained on single-cell transcriptomics images accurately classifies diverse cell types outperforming the best-performing scRNA-seq classifiers such as support vector machine and random forest.

bioinformatics↗

Computer-aided diagnosis of reflectance confocal images to differentiate between lentigo maligna (LM) and atypical intraepidermal melanocytic proliferation (AIMP)

Lentigo maligna (LM), a form of melanoma in situ that predominantly affects sun-exposed areas such as the face, has an ill-defined clinical border and has a high rate of recurrence. Atypical Intraepidermal Melanocytic Proliferation (AIMP) is a term used to describe the melanocytic proliferation of an uncertain malignant potential. Clinically and histologically, AIMP can be difficult to distinguish from LM, and indeed AIMP may in some cases progress to LM. Reflectance Confocal Microscopy (RCM) is often used to investigate these lesions non-invasively, however, RCM is often not readily available nor is the associated expertise for RCM image interpretation. Here, we demonstrate machine learning architectures that can correctly classify lesions between LM and AIMP on stacks of RCM images. Overall, our methods showcase the potential for computer-aided diagnosis in dermatology, which in conjunction with the remote acquisition, can expand the range of diagnostic tools in the community.

bioinformatics↗

Blood-based transcriptomic signature panel identification for cancer diagnosis: Benchmarking of feature extraction methods

Liquid biopsy has shown promise for cancer diagnosis due to its minimally invasive nature and the potential for novel biomarker discovery. However, the low concentration of relevant blood-based biosources and the heterogeneity of samples (i.e. the variability of relative abundance of molecules identified), pose major challenges to biomarker discovery. Moreover, the number of molecular measurements or features (e.g., transcript read counts) per sample could be in the order of several thousand, whereas the number of samples is often substantially lower, leading to the curse of dimensionality. These challenges, among others, elucidate the importance of a robust biomarker panel identification or feature extraction step wherein relevant molecular measurements are identified prior to classification for cancer detection. In this work, we performed a benchmarking study on eleven feature extraction methods using transcriptomic profiles derived from different blood-based biosources. The methods were assessed both in terms of their predictive performance and the robustness of the biomarker panels in diagnosing cancer or stratifying cancer subtypes. While performing the comparison, the feature extraction methods are categorised into feature subset selection methods and transformation methods. A transformation feature extraction method, namely PLS-DA, was found to perform consistently superior in terms of classification performance, while a feature subset selection method, namely Ranger, selected feature sets that were the most robust to sub-sampling in terms of consistently selecting the same features. As part of the benchmarking study, a generic pipeline has been created and made available as an R package to ensure reproducibility of the results and allow for easy extension of this study to other datasets.

bioinformatics↗

dSeqSb: A systems biology approach to decipher dynamics of 1 host-pathogen interactions using temporal dual RNA-seq data

Infection triggers a dynamic cascade of reciprocal events between host and pathogen wherein the host activates complex mechanisms to recognise and kill pathogens while the pathogen adjusts its virulence and fitness to avoid eradication by the host. The interaction between the pathogen and the host results in large-scale changes in gene expression in both organisms. Dual RNA-seq, the simultaneous detection of host and pathogen transcripts, has become a leading approach to unravel complex molecular interactions between the host and the pathogen and is particularly informative for intracellular organisms. The amount of in vitro and in vivo dual RNA-seq data is rapidly growing which demands computational pipelines to effectively analyse such data. In particular, holistic, systems-level, and temporal analyses of dual RNA-seq data are essential to enable further insights into the host-pathogen transcriptional dynamics and potential interactions. Here, we developed an integrative network-driven bioinformatics pipeline, dRNASb, a systems biology-based computational pipeline to analyse temporal transcriptional clusters, incorporate molecular interaction networks (e.g., protein-protein interactions), identify topologically and functionally key transcripts in host and pathogen, and associate host and pathogen temporal transcriptome to decipher potential between-species interactions. The pipeline is applicable to various dual RNA-seq data from different species and experimental conditions. As a case study, we applied dRNASb to analyse temporal dual RNA-seq data of Salmonella-infected human cells, which enabled us to uncover genes contributing to the infection process and their potential functions and to identify potential host-pathogen interactions between host and pathogen genes. Overall, dRNASb has the potential to identify key genes involved in bacterial growth or host defence mechanisms for future uses as therapeutic targets.

microbiology↗

Disentangling single-cell omics representation with a power spectral density-based feature extraction

Emerging single-cell technologies provide high-resolution measurements of distinct cellular modalities opening new avenues for generating detailed cellular atlases of many and diverse tissues. The high dimensionality, sparsity, and inaccuracy of single cell sequencing measurements, however, can obscure discriminatory information, mask cellular subtype variations and complicate downstream analyses which can limit our understanding of cell function and tissue heterogeneity. Here, we present a novel pre-processing method (scPSD) inspired by power spectral density analysis that enhances the accuracy for cell subtype separation from large-scale single-cell omics data. We comprehensively benchmarked our method on a wide range of single-cell RNA-sequencing datasets and showed that scPSD pre-processing, while being fast and scalable, significantly reduces data complexity, enhances cell-type separation, and enables rare cell identification. Additionally, we applied scPSD to transcriptomics and chromatin accessibility cell atlases and demonstrated its capacity to discriminate over 100 cell types across the whole organism and across different modalities of single-cell omics data.

bioinformatics↗

Ageing impairs the airway epithelium defence response to SARS-CoV-2

Age-dependent differences in the clinical response to SARS-CoV-2 infection is well-documented1-3 however the underlying molecular mechanisms involved are poorly understood. We infected fully differentiated human nasal epithelium cultures derived from healthy children (1-12 years old), young adults (26-34 years old) and older adults (56-62 years old) with SARS-COV-2 to identify age-related cell-intrinsic differences that may influence viral entry, replication and host defence response. We integrated imaging, transcriptomics, proteomics and biochemical assays revealing age-related changes in transcriptional regulation that impact viral replication, effectiveness of host responses and therapeutic drug targets. Viral load was lowest in infected older adult cultures despite the highest expression of SARS-CoV-2 entry and detection factors. We showed this was likely due to lower expression of hijacked host machinery essential for viral replication. Unlike the nasal epithelium of young adults and children, global host response and induction of the interferon signalling was profoundly impaired in older adults, which preferentially expressed proinflammatory cytokines mirroring the "cytokine storm" seen in severe COVID-194,5. In silico screening of our virus-host-drug network identified drug classes with higher efficacy in older adults. Collectively, our data suggests that cellular alterations that occur during ageing impact the ability for the host nasal epithelium to respond to SARS-CoV-2 infection which could guide future therapeutic strategies.

immunology↗