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Biology subjects

Halder, A.

Publications and source records attributed to Halder, A..

At least 19 recordsLinked to original sources

Evaluation of Cross-slice Imputation for Spatial Proteomics

Spatial single-cell protein profiling is at the forefront of translational medicine research efforts that try to decipher tissue heterogeneity. However, existing experimental approaches are limited in the number of biomarkers that they measure. This has motivated extensive research into the problem of protein imputation, where proteins or biomarkers not directly measured in a given slice of biopsy tissue are learned from those measured. In this work, we focus on the specific problem of cross-slice protein imputation (CSPI). In the CSPI problem, two sets of biomarker expressions at the single-cell level are known for physically adjacent slices of tissue. Each slice has its own known set of biomarkers, which overlaps with but does not subsume that of the adjacent. For each slice, we wish to learn the expressions of the proteins unique to the adjacent. Existing literature for protein imputation methodologies can be grouped in two categories: learning-based and alignment-based approaches. Learning-based approaches do not account for spatial and structural similarities between the training and the target sets. Alignment based approaches aim to determine an association between cells in adjacent slices, but without the explicit goal of using this alignment for imputation. In this work, we evaluate several methods from both categories and their adaptability to the cross-slice protein imputation problem. We provide a statistical analysis of all methods' performance under varying conditions of implementation. We find that learning-based methods significantly outperform alignment-based methods at the task of cross-slice protein imputation, with the latter more suited to the task of inter-slice interpolation or 3D tissue reconstruction. Moreover, inter-patient imputation is far less accurate than intra-patient imputation for all methods evaluated.

bioinformatics↗

Hypersensitive Detection of Neurotransmitters in Biological Media by Optically Enhanced Benchtop NMR Spectroscopy

Liquid-state Nuclear Magnetic Resonance (NMR) is an invaluable tool to gain atomic-resolution insights onto molecular structure and dynamics. The impact of NMR spectroscopy, however, is curtailed by high costs and hard-to-maintain high-field magnets. While low-field benchtop NMR spectrometers may address the challenge, severe trade-offs in sensitivity and resolution limit the appeal of this avenue. Here, we introduce a set of novel optically enhanced NMR pulse sequences and procedures (hyperpolarization toolkit) to readily generate 13C and 1H nuclear-spin hyperpolarization in situ on benchtop spectrometers. This approach leads to unprecedented sensitivity gains, enabling atomic-resolution conformation-dependent detection of a broad range of aromatic compounds, including clinically relevant metabolites and biomarkers. Neurotransmitters including epinephrine, serotonin and melatonin are detected at nanomolar/micromolar levels by 1D/2D photochemically-enhanced benchtop NMR in buffer and physiological media including human serum. This advance propels benchtop NMR from a predominantly pedagogical tool to a powerful bioanalytical resource for efficient neurotransmitter identification.

biophysics↗

Efficient Through-Bond Propagation of Nuclear-Spin Hyperpolarization via TOCSY-Enhanced LC-Photo-CIDNP

Nuclear magnetic resonance (NMR) spectroscopy provides atomic-resolution insights into biomolecular structure, dynamics and interactions under non-perturbative conditions. However, the widespread applicability of this technique continues to be limited by its intrinsically low sensitivity. This drawback is particularly severe in the case of concentration-limited samples. Low-concentration photochemically induced dynamic nuclear polarization (LC-photo-CIDNP) is an optically enhanced NMR hyperpolarization strategy capable of boosting NMR sensitivity in situ by several orders of magnitude under physiologically relevant environments. Yet, the inherent nature of the LC-photo-CIDNP phenomenon confines the achievable enhancements to only a subset of nuclei, thus limiting the extent of attainable residue-specific information. Here, we overcome this shortcoming by integrating LC-photo-CIDNP with Total Correlation Spectroscopy (TOCSY) to propagate initial hyperpolarization to other nuclei via scalar-coupled spin networks. First, we demonstrate efficient through-bond transfer of LC-photo-CIDNP enhanced signals throughout entire intramolecular atomic frameworks via 1D 1H- and 13C-detected experiments. Second, we extend the approach to a model protein and demonstrate efficient polarization transfer via 1D and 2D 1H-detected 13C-TOCSY LC-photo-CIDNP. Intrinsically photo-CIDNP-inactive nuclei can thus be visualized via 13C-1H correlations within minutes down to 10 M concentration. In all, this work establishes a general strategy to achieve through-bond propagation of LC-photo-CIDNP hyperpolarization. This technology enables gaining residue-specific structural insights on amino acids and proteins at previously unattainable low concentrations.

biophysics↗

A multitask single-cell analysis framework with knowledge graph as a prior

Single-cell RNA sequencing has reshaped cancer biology by revealing the cellular and molecular diversity of the tumor microenvironment, yet most analysis pipelines still treat clustering, annotation, pathway scoring, metabolic inference, and cell-cell communication as separate tasks. Here, we present scKNIFE (Knowledge graph-based NMF for Inference of Functional Entities), a unified graph-regularized non-negative matrix factorization framework that jointly reconstructs single-cell expression profiles and infers activities of diverse biological entities, including pathways, cell-type markers, metabolic reactions and tasks, ligand-receptor interactions, and cell-state programs. By embedding a heterogeneous knowledge graph directly into the factorization objective, scKNIFE propagates information between biologically related entities through Laplacian regularization while sparsity constraints preserve interpretability. The integrated prior graph spans 47,274 nodes and 506,620 edges, assembled from complementary resources including Gene Ontology, Reactome, Hallmark gene sets, Human-GEM, CellChat, PanglaoDB, Cytopus, and Tabula Muris-derived annotations. Across multiple cancer-focused single-cell datasets, scKNIFE yields competitive to leading performance for clustering and cell-type annotation, while also recovering biologically coherent metabolic programs that agree with dedicated metabolism-focused methods and capture therapy-response-associated states. In addition, the framework supports downstream inference of cell-type-specific biological activities from a single latent representation. Together, these results establish scKNIFE as a modular and extensible framework for end-to-end biological interpretation of single-cell cancer transcriptomics

cancer biology↗

Deterministic retrieval recovers biomedical associations lost by language models

Biomedical research increasingly relies on expert-curated databases to connect diseases, genes, variants, phenotypes, pathways and therapeutics. Incomplete or irreproducible retrieval can distort interpretation, mechanistic inference, variant assessment or therapeutic prioritization despite an unchanged evidence base. Agentic natural-language-to-SQL (NL2SQL) systems and Model Context Protocol-enabled agents can autonomously retrieve evidence from biomedical databases, including Open Targets and the Highly Confident Drug-Target Database, in response to natural-language questions. However, they may truncate large result sets, miss records when terminology differs from database vocabulary and return different outputs across runs. We systematically quantify these failures and introduce BioChirp, a goal-directed autonomous system for reliable retrieval from structured biomedical databases, built on interpretation-execution separation. A coordinated language-model layer interprets intent and resolves database-field assignments, after which entity resolution maps user terms to database terminology. A deterministic Steiner-tree planner and executor then retrieve matching records without further language-model involvement. Across curated biomedical databases spanning more than one million associations, BioChirp retrieved thousands of records for exhaustive queries, whereas baseline agents returned at most a few hundred or failed. Across 70 queries, five runs and three BioChirp database backends, BioChirp achieved a median cross-run Jaccard similarity of 1.0. Across 910 database-question evaluations derived from expert-written BioASQ questions and spanning ten databases, BioChirp retrieved at least one database record in 85.4% of cases and produced a fully correct answer in 51.1% of cases, compared with 44.6% and 17.4% for agentic NL2SQL. BioChirp is publicly available at https://biochirp.iiitd.edu.in.

bioinformatics↗

RSTG: Robust Generation of High Quality Spatial Transcriptomics Data using Beta Divergence Based AutoEncoder

One of the key challenges in spatial transcriptomics data analysis is the lack of sufficient data to train models. To address this shortcoming, multiple generative models have been developed to generate synthetic spatial transcriptomics samples in a controlled environment. However, these models often fail in out-of-the-box generation in the presence of noise (such as outliers). To tackle this challenge, we propose RSTG (Robust Spatial Transcriptomic Generator), an autoencoder that incorporates the {beta}-ELBO loss, to generate realistic and high-quality spatial transcriptomic sequences. Our model uncovers data intrinsic structure by approximating its underlying distribution through variational inference, resulting in more interpretable and robust density estimation. We validate the effectiveness of RSTG across multiple tasks, including the recovery of cellular positions in both the 2D spatial and location domains. Our method shows improved performance, both qualitatively and quantitatively, across multiple datasets, including brain and liver samples generated using MERFISH, MERSCOPE, and Visium technologies. We further illustrate the robustness of RSTG to outliers by contaminating a portion of the data with anomalies (such as white noise, batch effects, and dropouts) as well as on a reallife degraded sample. The results show that our proposal maintains high quality and stability even when the training data are contaminated, across a variety of experimental settings and in comparison with existing approaches.

bioinformatics↗

Mechanism-aware inference of response to targeted cancer therapies

Targeted therapies like small-molecule inhibitors often work by blocking proteins that cancer cells rely on for survival. Omics based modeling of drug sensitivity alone lack mechanistic grounding. We propose FORGE (Factorization Of Response and Gene Essentiality) a simple yet powerful joint matrix factorization framework that co-models drug response and target gene essentiality, enabling the stratification of promising treatment groups for targeted therapy consideration. FORGE also provides Benefit Score -- a predictive score that estimates treatment efficacy from basal gene expression profiles. We validated the predictive performance of FORGE across multiple targeted therapies, including Erlotinib (EGFR inhibitor) and Daporinad (NAMPT inhibitor). Our meta-analysis of large scale in-vitro studies underscores FORGEs ability to identify common determinants of drug vulnerabilities and target gene essentiality. Such convergences were not observed when treatment vulnerabilities and gene essentialities were modeled independently. We also demonstrated the universality of Erlotinib Benefit Scores by transferring transformations learned from high-throughput drug response studies across other published datasets, including the TAHOE-100M single-cell perturbation atlas and patient-derived xenograft studies. FORGE successfully identified key regulators within the molecular pathways targeted by these therapies, reinforcing its potential for mechanistically grounded treatment stratification.

cancer biology↗

DrugProtAI: A guide to the future research of investigational target proteins

Drug design and development are central to clinical research, yet ninety percent of drugs fail to reach the clinic, often due to inappropriate selection of drug targets. Conventional methods for target identification lack precision and sensitivity. While various computational tools have been developed to predict the druggability of proteins, they often focus on limited subsets of the human proteome or rely solely on amino acid properties. To address the challenge of class imbalance between proteins with and without approved drugs, we propose a novel Partitioning Method. We evaluated the druggability potential of 20,273 reviewed human proteins, of which 2,636 have approved drugs. Our comprehensive analysis of 183 features, encompassing biophysical and sequence-derived properties, achieved a median AUC of 0.86 in target predictions. We utilize SHAP (Shapley Additive Explanations) scores to identify key predictors and interpret their contribution to druggability. We have reviewed and evaluated 688 investigational proteins from DrugBank (https://go.drugbank.com/) using our tool, DrugProtAI (https://drugprotai.pythonanywhere.com/). Our tool offers druggability predictions and access to 2M+ publications on drug targets and their effects, aiding in the selection of target proteins for drug development. We believe that insights into key predictors will significantly advance drug development and propel the field forward.

bioinformatics↗

Selective Activation of GPCRs: Molecular Dynamics Investigation of Siponimod's Interaction with S1PR1 and S1PR2

G Protein-Coupled Receptors (GPCRs) are central to drug discovery, accounting for nearly 40% of approved pharmaceuticals due to their regulatory role in diverse physiological processes. Given the high structural similarity among homologues, achieving receptor selectivity while minimizing off-target effects remains a major challenge in designing drugs targeting GPCRs. Sphingosine-1-phosphate receptors (S1PRs), comprising five subtypes, are therapeutically important GPCRs critical for immune and cardiovascular functions. Siponimod, an FDA-approved drug for multiple sclerosis, selectively modulates S1PR1 over S1PR2, unlike earlier S1PR modulators. However, the molecular basis for this selectivity is unclear, as cellular and biochemical assays provide limited insights. In this study, we used long-timescale molecular dynamics simulations to investigate how S1P and Siponimod binding affect S1PR1 and S1PR2 structural dynamics. Both ligands exhibited strong active site binding in both receptors. Crucially, while S1P and Siponimod induced similar activation-linked conformational changes in S1PR1, Siponimod failed to trigger these rearrangements in S1PR2. Specifically, Siponimod binding to S1PR2 led to altered side-chain dynamics of key TM7 residues (viz. Y7.37, F7.38, F7.39) and a drift of transmembrane helix 6 (TM6) towards orientations observed in inactive state. These unique structural features differentiate Siponimods behavior from S1P and explain its lack of inability to modulate S1PR2. Our findings elucidate molecular determinants of Siponimods selectivity towards S1PR1 and highlight these residues as potential differentiators for selective modulator design. This study demonstrates how structural and dynamic insights from atomistic simulations aid rational drug design for targets with high homology.

biophysics↗

Selective promotion of oligonucleotides in the course of evolution

Evolution shapes the structure and content of genomes, yet the contribution of local sequence composition to variant selection remains poorly understood. While traditional models emphasize protein function or cross-species conservation, we propose that intra-genomic patterns of oligonucleotide (k-mer) frequencies also reflect selective forces. To explore this, we developed kGain score, a metric that quantifies the frequency shift of a k-mer upon single-nucleotide substitution, using the surrounding genomic context as a baseline. We hypothesize that variants arising in high-kGain contexts are more likely to persist due to evolutionary favorability. We validated this hypothesis across multiple systems. In E. coli and S. cerevisiae long-term evolution experiments, we found that fixed, essential, and parallel mutations consistently show elevated kGain scores. This trend held in SARS-CoV-2 variants of concern and in an in-house antibiotic adaptation experiment, where a high-kGain fusA Y515N mutation conferred resistance and maintained fitness when overexpressed, demonstrating a causal link between kGain and adaptive potential. To enable cross-species generalization, we trained a transformer-based neural network regressor on LTEE-derived mutations to predict kGain from sequence alone. The model achieved high correlation in held-out in-domain data (Pearson r = 0.81) and accurately predicted kGain trends in out-of-domain data (Pearson r = 0.82), demonstrating that k-mer-based sequence constraints learned from one genome can be effectively transferred to others. Together, our results establish kGain as a biologically meaningful, scalable metric for probing within-genome sequence selection, offering a complementary lens to existing conservation-based frameworks for understanding evolutionary fitness and variant persistence.

evolutionary biology↗

The Hidden Potential of PDE4 Inhibitor Rolipram: A Multifaceted Examination of its Inhibition of MMP2/9 Reveals Theraputic Implications

A PDE4 inhibitor, Rolipram, was previously found to down-regulate (in a manner dependent on cAMP-PKA) MMP2 and MMP9 levels, important markers of epithelial-to-mesenchymal transition in human breast cancer cell lines. However, zymographic studies revealed that rolipram could also alter the enzymatic activities of these MMPs, even in the presence of the PKA inhibitor H89. This calls for more detailed investigations of the inhibitory mechanism of rolipram on MMP2 and MMP9. The prediction of ligand-based targets through online reverse screening indicated that proteases are likely targets of rolipram. Computational molecular docking also demonstrated significant binding affinities of rolipram for both MMP2 and MMP9 proteins. Concurrently, a well-known inhibitor of MMPs, SB3CT, was utilized as a positive control for this study. The best models of the docked complexes were used as initial conditions for molecular dynamics (MD) simulations to explore their dynamic behavior and stability. In particular, both the MMP2-rolipram and MMP9-rolipram complexes were found to be stable and compact for the duration of the simulation (300 ns). Several stable hydrogen bonds were also detected between the proteins and rolipram. In vitro experiments using primary cells from patients with breast cancer also showed that rolipram could alter the enzymatic activities of MMP2 and MMP9, independent of the cAMP-PKA signaling pathway. These observations indicate the ability of rolipram to control breast cancer by regressing the functions of MMP2 and MMP9, thus having off-targets other than PDE4 to have direct control over proteins that are involved in the advancement of metastasis.

cancer biology↗

Investigating spatial dynamics in spatial omics data with StarTrail

Spatial omics technologies revolutionize our view of biological processes within tissues. However, existing methods fail to capture localized, sharp changes characteristic of critical events (e.g. tumor development). Here, we present StarTrail, a novel gradient based method that powerfully defines rapidly changing regions and detects "cliff genes", genes exhibiting drastic expression changes at highly localized or disjoint boundaries. StarTrail, the first to leverage spatial gradients for spatial omics data, also quantifies directional dynamics. Across multiple datasets, StarTrail accurately delineates boundaries (e.g., brain layers, tumor-immune boundaries), and detects cliff genes that may regulate molecular crosstalk at these biologically relevant boundaries but are missed by existing methods. For instance, StarTrail precisely pinpointed the cancer-immune interface in a HER2+ breast cancer dataset, unveiled key cliff genes including a potential prognostic biomarker IGSF3, highlighting NK-, B-cell mediated immunity, and B cell receptor signaling pathways missed by all spatial variable gene methods attempted. StarTrail, filling important gaps in current literature, enables deeper insights into tissue spatial architecture.

genetics↗

Improved Python Package for DNA Sequence Encoding using Frequency Chaos Game Representation

SummaryFrequency Chaos Game Representation (FCGR), an extended version of Chaos Game Representation (CGR), emerges as a robust strategy for DNA sequence encoding. The core principle of the CGR algorithm involves mapping a onedimensional sequence representation into a higher-dimensional space, typically in the two-dimensional spatial domain. This paper introduces a use case wherein FCGR serves as a kmer frequency-based encoding method for motif classification using a publicly available dataset. Availability and implementationThe FCGR python package, use case, along with additional functionalities, is available in the GitHub. Our FCGR package demonstrates superior accuracy and computational efficiency compared to a leading R-based FCGR library [1], which is designed for versatile tasks, including proteins, letters, and amino acids with user-defined resolution. Nevertheless, it is important to note that our Python package is specifically designed for DNA sequence encoding, where the resolution is predetermined based on the kmer length. It is a drawback of our current package compared to the state-of-the-art R-based kaos package [1].

bioinformatics↗

BrainProt(™) 3.0: Understanding Human Brain Diseases using comprehensively curated & Integrated OMICS datasets

BrainProt 3.0 is an integrative and simplified omics-based knowledge base of the human brain and its associated diseases. The current version of BrainProt includes six domains, which provide simplified, robust, and comprehensive data visualization to understand the human brain and its diseases/disorders based on proteomics, transcriptomics, public data curation, and integration strategies. Firstly, the HBDA (Human Brain Disease Atlas), index and navigator of BrainProt provides a resource table for 56 brain diseases. Secondly, Brain Disease Marker Curator (BDMC) and Brain Disease Drug Finder (BDDF) include a total of 20,202 diseases associated genes, more than 1,30,000 Chemical Target interactions, and around 2,145 Clinical Trial Information for more than 50 Brain Diseases. Thirdly, Brain Disease Transcriptome Map (BDTM) and Brain Disease Proteome Map (BDPM) integrate multi-omics data for 11 and 6 alarming brain diseases respectively. Currently, these two domains feature an expressional profile of 52 datasets, information of 1,868 samples, 3,657 DEPs, and 6,256 DEGs. Lastly, BrainProt also modifies and integrates the proteome and phosphoproteome data of the Inter-hemispheric Brain Proteome Map (IBPM). Overall, BrainProt is the first knowledgebase that connects different omics level information of brain diseases and provides a powerful scoring-based ranking platform to select and identify brain disease-associated markers, along with exploration of clinical trials, and drugs/chemical compounds to accelerate the identification of new disease markers and novel therapeutic strategies. The objectives of BrainProt are to support and follow the footsteps of the HBPP (Human Brain Proteome Project) by integrating different datasets to unravel the complexity of Human Brain and its associated diseases.

neuroscience↗

Full bandwidth electrophysiology of seizures and epileptiform activity enabled by flexible graphene micro-transistor depth neural probes

Mapping the entire frequency bandwidth of neuronal oscillations in the brain is of paramount importance for understanding physiological and pathological states. The ability to record simultaneously infraslow activity (<0.1 Hz) and higher frequencies (0.1-600 Hz) using the same recording electrode would particularly benefit epilepsy research. However, commonly used metal microelectrode technology is not well suited for recording infraslow activity. Here we use flexible graphene depth neural probes (gDNP), consisting of a linear array of graphene microtransistors, to concurrently record infraslow and high frequency neuronal activity in awake rodents. We show that gDNPs can reliably record and map with high spatial resolution seizures, post-ictal spreading depolarisation, and high frequency epileptic activity through cortical laminae to the CA1 layer of the hippocampus in a mouse model of chemically-induced seizures. We demonstrate functionality of chronically implanted devices over 10 weeks by recording with high fidelity spontaneous spike-wave discharges and associated infraslow activity in a rat model of absence epilepsy. Altogether, our work highlights the suitability of this technology for in vivo electrophysiology research, in particular, to examine the contributions of infraslow activity to seizure initiation and termination.

neuroscience↗

Occurrences of protonated base triples in RNA are determined by their cooperative binding energies and specific functional requirements

With wide ranging diversity in their geometries, binding strengths and chemical properties, noncanonical base pairs are equipped to intricately regulate and control the structural dynamics of RNA molecules. Protonation of nucleobases adds to the diversity. Compared to the unprotonated scenario, on one hand they open up new alternatives for base pairing interactions (Class I) while on the other, they modulate the geometry and stability of existing base pairing interactions (Class II). In both cases, compensation of the energetic cost associated with nucleobase protonation at physiological pH, can be understood in terms of protonation induced restructuring of charge distribution. This not only leads to modifications in existing base-base interactions but often also leads to additional stabilizing interactions, resulting in the formation of protonated base triples. Here we report our detailed quantum chemical studies, in conjunction with structural bioinformatics based analysis of RNA crystal and NMR structure datasets, probing into the contribution of such protonated triples in the structural dynamics of RNA. Our studies revealed more than 55 varieties of protonated triples in RNA, some of which occur recurrently within conserved structural motifs present in rRNAs, tRNAs and in other synthetic RNAs. Our studies suggest that high occurrence frequencies are associated with protonated triples which satisfy the specific structural requirements of conserved motifs where they occur. For example, protonated triples with flexible geometries are involved in the formation of tertiary contacts between different distant motifs. Stabilization of protonated base pairs, through the induction of additional energetically cooperative interactions, appears to be another factor. These results provide significant insights into the sequence-structure-function relationships in RNA.

biophysics↗

The metal cofactor zinc and interacting membranes modulate SOD1 conformation-aggregation landscape in an in vitro ALS Model

Aggregation of Cu-Zn superoxide dismutase (SOD1) is implicated in the motor neuron disease, ALS. Although more than 140 disease mutations of SOD1 are available, their stability or aggregation behaviors in membrane environment are not correlated with disease pathophysiology. Here, we use multiple mutational variants of SOD1 to show that the absence of Zn, and not Cu, significantly impacts membrane attachment of SOD1 through two loop regions facilitating aggregation driven by lipid induced conformational changes. These loop regions influence both the primary (through Cu intake) and the gain of function (through aggregation) of SOD1 presumably through a shared conformational landscape. Combining experimental and theoretical frameworks using representative ALS disease mutants, we develop a co-factor derived membrane association model wherein mutational stress closer to the Zn (but not to the Cu) pocket is responsible for membrane association mediated toxic aggregation and survival time scale after ALS diagnosis.

biophysics↗

Mg2+ Sensing by an RNA Fragment: Role of Mg2+ Coordinated Water Molecules

RNA molecules selectively bind to specific metal ions to populate their functional active states making it important to understand their source of ion selectivity. In large RNA systems, metal ions interact with the RNA at multiple locations making it difficult to decipher the precise role of ions in folding. To overcome this complexity, we studied the role of different metal ions (Mg2+, Ca2+ and K+) in the folding of a small RNA hairpin motif (5'-ucCAAAga-3') using unbiased all-atom molecular dynamics simulations. The advantage in studying this small system is that it requires specific binding of a single metal ion to fold to its native state. We find that even for this small RNA, the folding free energy surface (FES) is multidimensional as different metal ions present in the solution can simultaneously facilitate folding. The FES shows that specific binding of a metal ion is indispensable for its folding. We further show that in addition to the negatively charged phosphate groups, spatial organization of electronegative nucleobase atoms drive the site specific binding of the metal ion. Even though the binding site cannot discriminate between different metal ions, RNA folds efficiently only in Mg2+ solution. We show that the rigid network of Mg2+ coordinated water molecules facilitate the formation of important interactions in the transition state. The other metal ions such as K+ and Ca2+ cannot facilitate the formation of such interactions. These results allow us to hypothesize possible metal sensing mechanisms in large metallo-riboswitches and they also provide useful insights for the design of appropriate collective variables for studying large RNA molecules using enhanced sampling methods.

biochemistry↗