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Shen, H.-B.

Publications and source records attributed to Shen, H.-B..

18 recordsLinked to original sources

Auditing Protein-Protein Interaction Signals with Sparse Autoencoder Fingerprints

Protein language models have become a dominant foundation for sequence-based protein-protein interaction (PPI) prediction, but their generalization remains limited under stringent evaluation, and benchmark accuracy alone cannot reveal whether a PPI predictor learns partner-specific biological signals or exploits contextual shortcuts. Here we introduce AuditPPI, an interpretable framework that transforms sparse-autoencoder (SAE) features from a frozen protein language model into order-invariant pair fingerprints, recasting PPI prediction as an auditable tabular-learning problem. AuditPPI achieves competitive predictive performance across diverse PPI benchmarks while enabling multiscale auditing of the information supporting its predictions. Protein-level audits show that partner-independent participation signals remain substantial in conventional benchmarks, including protein-disjoint benchmark, but are largely insufficient for pair-level discrimination in topology- and degree-controlled benchmark. Pair-level analyses further show that removing protein identity overlap does not eliminate contextual structure: protein-disjoint benchmarking retains strong subcellular co-localization signals and benchmark-dependent feature matching captured by SAE co-activation and absolute-difference. Structural analyses, however, provide little evidence that these predictive signals are specifically grounded in PPI interfaces: feature enrichment at interfaces for protein-disjoint benchmark was no longer detectable after controlling for surface exposure, and contact-specific enrichment was observed for only a small fraction of testable SAE feature pairs. AuditPPI therefore separates predictive success from mechanistic fidelity and provides a practical framework for evaluating whether sequence-based PPI predictors capture partner-specific biological signals instead of contextual shortcuts.

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Information Geometry Reconciles Discrete and Continuous Variation in Single-Cell and Spatial Transcriptomic Analysis

Single-cell and spatial transcriptomics provide high-resolution cellular characterization, yet standard analytical approaches remain theoretically misaligned with the probabilistic nature of the data. After UMI normalization, current pipelines rely on Euclidean or log-transformed Euclidean distance for similarity measurement. Both are fundamentally ill-suited to model the multinomial count data. Euclidean distance in normalized space overemphasizes high-variance genes, while log-transformation inverts this bias but at the cost of distorting subtle, continuous expression modulations. Neither approach naturally captures the dual nature of gene expression: both discrete presence/absence transitions and continuous quantitative variation. To overcome these limitations, we introduce GAIA (Geometric Analysis from an Information Aspect), an information-geometric framework for cell representation learning and inter-cell similarity measurement. By anchoring analysis in the true probabilistic model, treating cells as multinomial distributions over genes and projecting cells to a statistical manifold, GAIA organically reconciles both the presence/absence effect and the more continuous expression modulations. Mathematically, GAIA exploits the equivalence between Fisher-Rao distance in multinomial space and geodesic distance on the unit hypersphere, a property that enables both theoretical guarantees and computational efficiency. Experiments in synthetic and real scRNA-seq and spatial transcriptomic datasets demonstrate that GAIA preserves robust and consistent cell-to-cell relationships, delineates biologically nuanced sub-types, mitigates batch effects arising from sequencing depth variation, and eliminates the dependence on knowledge-restricted gene selection for learning meaningful cell representations. Overall, GAIA offers a knowledge-lean, variance-stabilizing framework for analyzing single-cell and spatial transcriptomic data, enhancing discrimination between nuanced cell sub-type and -states.

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Fast Multi-objective RNA Optimization with Autoregressive Reinforcement Learning

Codon optimization is essential in mRNA vaccine development, while existing tools face limitations in the computational efficiency, sequence diversity and universality. To address these challenges, we develop RNAJog (RNA Joint Optimization with autoregressive Generative model), a framework integrating autoregressive generation with reinforcement learning to optimize codon sequences for minimum free energy (MFE), codon adaptation index (CAI) and GC content, even enabling sequence design without requiring annotated training data. Evaluations in both in silico and wet-lab experiments have confirmed RNAJogs effectiveness and efficiency, with two orders of magnitude faster than traditional algorithm (LinearDesign) for long RNA sequence and about a 10-fold increase in antibody titer compared to the wild-type mRNA for Influenza virus hemagglutinin (HA) mRNA vaccine design in mouse. RNAJog also supports biological constraints for sequence optimization. Using this feature, we minimized m6A modification motifs in Bmp2 coding sequence for enhancing the translational efficiency and RNA stability, which are validated in cell-based experiments.

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Alignment-driven Cross-Graph Modeling for 3D RNA Inverse Folding

The 3D RNA inverse folding task aims to design RNA sequences that can fold into a target tertiary structure. This task is complicated by the inherent structural complexity and flexibility of RNA molecules. Considering that conserved RNA structures often exhibit sequence similarity, we present AlignIF, a pipeline designed to generate RNA sequences conditioned on the target RNA 3D structure and its multiple structure alignment (MStA), explicitly incorporating structural evolutionary conservation. AlignIF introduces a novel MStA backbone encoder designed to model multiple geometric graphs derived from MStA, which can better capture meaningful information from MStA structures. Additionally, AlignIF uses a random-ordered autoregressive sequence decoder to generate candidate RNA sequences. In our constructed benchmark dataset, AlignIF achieves a perplexity of 2.32 and a recovery rate of 57. 08%, respectively, a relative improvement of 17.8% and 11.8% over the state- of-the-art methods. In addition, AlignIF excels in diversity and ranking capability. We further demonstrate the added value of the incorporated MStA structures along with the proposed encoder architecture.

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RBPSignal: A deep learning approach for predicting RNA-Protein binding signals

RNA-binding proteins play critical roles in post-transcriptional regulation by interacting with RNA to regulate various cellular processes. Inferring the binding signals and sequence patterns of these interactions is essential for elucidating the mechanisms underlying gene expression regulation. In this study, we present RBPSignal, a deep learning-based computational tool designed to predict RBP binding signals on RNAs and identify potential sequence motifs associated with these interactions. RBPSignal leverages the deep learning framework and trains on comprehensive eCLIP datasets, demonstrating an enhanced predictive accuracy. Furthermore, the integration of model interpretability through Integrated Gradients enables the detailed analysis of binding motif syntax. We validate the efficacy of RBPSignal on chromosome data and compare the discovered motifs with existing motif database, showcasing its ability not only to predict binding signals but also to uncover sequence patterns correlated with binding signals. Our findings provide insights into the sequence specificity of RBPs and explore the protein-protein interaction networks. RBPSignal serves as a valuable tool for exploring RBP-RNA interaction landscapes, facilitating further investigations into the regulatory networks underlying gene expression. The web server is freely available at http://www.csbio.sjtu.edu.cn/bioinf/RBPSignal/.

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TSvelo: Comprehensive RNA velocity inference by jointly modeling Transcription and Splicing

RNA velocity approaches fit gene dynamics and infer cell fate by modeling the splicing process using single-cell RNA sequencing (scRNA-seq) data. However, due to short time scale of splicing, high noise and large complexity of data, existing RNA velocity methods often fail to precisely capture the complex velocity dynamics for individual gene and single cell, which makes its downstream analysis less reliable and less robust. We propose TSvelo, a comprehensive RNA velocity mathematics framework that can model the cascade of gene regulation, Transcription and Splicing using highly interpretable neural Ordinary Differential Equations (ODEs). TSvelo can precisely capture the transcription-unspliced-spliced 3D dynamics of all genes simultaneously, infer unified latent time shared by genes within single cell, and be applied to multi-lineage datasets. Experiments on six scRNA-seq datasets, including two multi-lineage datasets, demonstrate TSvelos superiority.

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Discovering nuclear localization signal universe through a novel deep learning model with interpretable attention units

Nuclear localization signals (NLSs) are essential peptide fragments within proteins that play a decisive role in guiding proteins into the cell nucleus. Determining the existence and precise locations of NLSs experimentally is time-consuming and complicated, resulting in a scarcity of experimentally validated NLS fragments. Consequently, annotated NLS datasets are relatively limited, presenting challenges for data-driven approaches. In this study, we propose an innovative interpretable approach, NLSExplorer, which leverages large-scale protein language models to capture crucial biological information with a novel attention-based deep network for NLS identification. By enhancing the knowledge retrieved from protein language models with a novel attention to key area module, NLSExplorer achieves superior predictive performance compared to existing methods on two NLS benchmark datasets. Additionally, NLSExplorer is able to detect various kinds of segments highly correlated with nuclear transport, such as nuclear export signals. We employ NLSExplorer to investigate potential NLSs and other domains that are important for nuclear transport in nucleus-localized proteins in the Swiss-Prot database. Furthermore, the comprehensive pattern analysis for all these segments uncovers a potential NLS space and internal relationship of important nuclear transport segments for 416 species. This study not only introduces a powerful tool for predicting and exploring NLS space, but also offers a versatile network that is powerful for detecting characteristic domains and motifs of NLSs.

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AttSiOff: A self-attention-based approach on siRNA de-sign with inhibition and off-target effect prediction

MotivationSmall interfering RNA (siRNA) is often used for function study and expression regulation of specific genes, as well as the development of small molecule drugs. Selecting siRNAs with high inhibition and low off-target effects from massive candidates is always a great challenge. Increasing experimentally validated samples prompt the development of machine-learning-based algorithms, including Support Vector Machine (SVM), Convolutional Neural Network (CNN), and Graph Neural Network (GNN). However, these methods still suffer from limited accuracy and poor generalization to design both potent and specific siRNAs. ResultsIn this study, we propose a novel approach for siRNA inhibition and off-target effect prediction, named AttSiOff. It combines self-attention-based siRNA inhibition predictor with an mRNA searching package and an off-target filter. The predictor gives the inhibition score via analyzing the embedding of siRNA and local mRNA sequences, generated from pre-trained RNA-FM model, as well as other meaningful prior-knowledge-based features. Self-attention mechanism can detect potentially decisive features, which may determine the inhibition of siRNA. It captures global and local dependencies more efficiently than normal convolutions. The 10-fold cross-validation results indicate that our model achieves a significant improvement of correlation between prediction and label, compared with all existing methods. And it reaches better performance of generalization and robustness on cross-dataset validation. In addition, the mRNA searching package could find all mature mRNAs for given gene name from GENOMES database, and the off-target filter can calculate the amount of unwanted off-target binding sites, which affects the specificity of siRNA. Experiments on mature siRNA drugs show that our entire framework, AttSioff, have excellent convenience and operability in practical applications. Contactyuanye_auto@sjtu.edu.cn or chengjin520@sjtu.edu.cn.

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TFvelo: gene regulation inspired RNA velocity estimation

RNA velocity is closely related with cell fate and is an important indicator for the prediction of cell states with elegant physical explanation derived from single-cell RNA-seq data. Most existing RNA velocity models aim to extract dynamics from the phase delay between unspliced and spliced mRNA for each individual gene. However, unspliced/spliced mRNA abundance may not provide sufficient signal for dynamic modeling, leading to poor fit in phase portraits. Motivated by the idea that RNA velocity could be driven by the transcriptional regulation, we propose TFvelo, which expands RNA velocity concept to various single-cell datasets without relying on splicing information, by introducing gene regulatory information. Our experiments on synthetic data and multiple scRNA-Seq datasets show that TFvelo can accurately fit genes dynamics on phase portraits, and effectively infer cell pseudo-time and trajectory from RNA abundance data. TFvelo opens a novel, robust and accurate avenue for modeling RNA velocity for single cell data.

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Decoding protein binding landscape on circular RNAs with base-resolution Transformer models

Circular RNAs (circRNAs) interact with RNA-binding proteins (RBPs) to modulate gene expression. To date, most computational methods for predicting RBP binding sites on circRNAs focus on circRNA fragments instead of circRNAs. These methods detect whether a circRNA fragment contains binding sites, but cannot determine where are the binding sites and how many binding sites are on the circRNA transcript. We report a hybrid deep learning-based tool, CircSite, to predict RBP binding sites at single-nucleotide resolution and detect key contributed nucleotides on circRNA transcripts. CircSite takes advantage of convolutional neural networks (CNNs) and Transformer for learning local and global representations of circRNAs binding to RBPs, respectively. We construct 37 datasets of RBP-binding circRNAs for benchmarking and the experimental results show that CircSite offers accurate predictions of RBP binding nucleotides and detects key subsequences aligning well with known binding motifs.

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High-accuracy protein model quality assessment using attention graph neural networks

Great improvement has been brought to protein tertiary structure prediction through deep learning. It is important but very challenging to accurately rank and score decoy structures predicted by different models. CASP14 results show that existing quality assessment (QA) approaches lag behind the development of protein structure prediction methods, where almost all existing QA models degrade in accuracy when the target is a decoy of high quality. How to give an accurate assessment to high-accuracy decoys is particularly useful with the available of accurate structure prediction methods. Here we propose a fast and effective single-model QA method, QATEN, which can evaluate decoys only by their topological characteristics and atomic types. Our model uses graph neural networks and attention mechanisms to evaluate global and amino acid level scores, and uses specific loss functions to constrain the network to focus more on high-precision decoys and high-precision protein domains. On the CASP14 evaluation decoys, QATEN performs better than other QA models under all correlation coefficients when targeting average LDDT. QATEN shows promising performance when considering only high-accuracy decoys. Compared to the embedded evaluation modules of predicted C-RMSD (pRMSD) in RosettaFold and predicted LDDT (pLDDT) in AlphaFold2, QATEN is complementary and capable of achieving better evaluation on some decoy structures generated by AlphaFold2 and RosettaFold themselves. These results suggest that the new QATEN approach can be used as a reliable independent assessment algorithm for high-accuracy protein structure decoys.

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Leveraging Scaffold Information to Predict Protein-ligand Binding Affinity with an Empirical Graph Neural Network

MotivationProtein-ligand binding affinity prediction is an important task in structural bioinformatics for drug discovery and design. Although various scoring functions have been proposed, it remains challenging to accurately evaluate the binding affinity of a protein-ligand complex with known bound structure due to the potential preference of scoring system. In recent years, deep learning techniques have been applied to scoring functions without sophisticated feature engineering. Nevertheless, existing methods cannot model the differential contribution of atoms in various regions of proteins, and the relationship between atom properties and intermolecular distance is also not fully explored. ResultsWe propose a novel empirical graph neural network for accurate protein-ligand binding affinity prediction (EGNA). Graphs of protein, ligand and their interactions are constructed based on different regions of each bound complex. Proteins and ligands are effectively represented by graph convolutional layers, enabling the EGNA to capture interaction patterns precisely by simulating empirical scoring functions. The contributions of different factors on binding affinity can thus be transparently investigated. EGNA is compared with the state-of-the-art machine learning-based scoring functions on two widely used benchmark datasets. The results demonstrate the superiority of EGNA and its good generalization capability. Availability and implementationThe web server and source code of EGNA is available at www.csbio.sjtu.edu.cn/bioinf/EGNA and https://github.com/chunqiux/EGNA. Contacthbshen@sjtu.edu.cn or 2008xypan@sjtu.edu.cn Supplementary informationSupplementary data are available at Bioinformatics online.

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OUGene 2.0: An updated disease-associated over- and under-expressed gene database by mining full-text articles

Many diseases are closely associated with over- or under-expressed genes. In order to cover more up to date associations between over- or under-expressed genes and various diseases, we develop an updated database OUGENE 2.0 for disease-associated over- and under-expressed genes by automatic full-text mining. In total, the new OUGene 2.0 includes 197,236 associations between 12,672 diseases and 11,542 over- or under-expressed genes, which increases by about 5 folds compared to the previous version of OUGene. A novel method for rescaling the raw score based on support evidences is designed to prioritize the mined associations. OUGene 2.0 provides a holistic view of disease-gene associations and it supports user-friendly data exploration at www.csbio.sjtu.edu.cn/bioinf/OUGene for academic use.

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SPRI: Spatial Pattern Recognition using Information based method for spatial gene expression data

The rapid development of spatially resolved transcriptomics has made it possible to analyze spatial gene expression patterns in complex biological tissues. To identify spatially differential expressed genes, we propose a novel and robust nonparametric information-based approach, SPRI. SPRI converts the problem of identifying spatial gene expression patterns into the detection of dependencies between spatial coordinates with observed frequencies measured by read counts. It directly models spatial transcriptome raw count without assuming a parametric model. SPRI was applied to spatial datasets with different resolutions, suggesting that SPRI outperforms previous methods, by robustly detecting more genes with significant spatial expression patterns, and revealing biological insights that cannot be identified by other methods.

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Inferring disease-associated microRNAs using semi-supervised multi-label graph convolutional networks

MicroRNAs (miRNAs) play crucial roles in many biological processes involved in diseases. The associations between diseases and protein coding genes (PCGs) have been well investigated, and further the miRNAs interact with PCGs to trigger them to be functional. Thus, it is imperative to computationally infer disease-miRNA associations under the context of interaction networks.\n\nIn this study, we present a computational method, DimiG, to infer miRNA-associated diseases using semi-supervised Graph Convolutional Network model (GCN). DimiG is a multi-label framework to integrate PCG-PCG interactions, PCG-miRNA interactions, PCG-disease associations and tissue expression profiles. DimiG is trained on disease-PCG associations and a graph constructed from interaction networks of PCG-PCG and miRNA-PCG using semi-supervised GCN, which is further used to score associations between diseases and miRNAs. We evaluate DimiG on a benchmark set collected from verified disease-miRNA associations. Our results demonstrate that the new DimiG yields promising performance and outperforms the best published baseline method not trained on disease-miRNA associations by 11% and is also superior to two state-of-the-art supervised methods trained on disease-miRNA associations. Three case studies of prostate cancer, lung cancer and Inflammatory bowel disease further demonstrate the efficacy of DimiG, where the top miRNAs predicted by DimiG for them are supported by literature or databases.

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Predicting circRNA-RBP interaction sites using a codon-based encoding and hybrid deep neural networks

Circular RNAs (circRNAs), with their crucial roles in gene regulation and disease development, have become a rising star in the RNA world. A lot of previous wet-lab studies focused on the interaction mechanisms between circRNAs and RNA-binding proteins (RBPs), as the knowledge of circRNA-RBP association is very important for understanding functions of circRNAs. Recently, the abundant CLIP-Seq experimental data has made the large-scale identification and analysis of circRNA-RBP interactions possible, while no computational tool based on machine learning has been developed yet.\n\nWe present a new deep learning-based method, CRIP (CircRNAs Interact with Proteins), for the prediction of RBP binding sites on circRNAs, using only the RNA sequences. In order to fully exploit the sequence information, we propose a stacked codon-based encoding scheme and a hybrid deep learning architecture, in which a convolutional neural network (CNN) learns high-level abstract features and a recurrent neural network (RNN) learns long dependency in the sequences. We construct 37 datasets including sequence fragments of binding sites on circRNAs, and each set corresponds to one RBP. The experimental results show that the new encoding scheme is superior to the existing feature representation methods for RNA sequences, and the hybrid network outperforms conventional classifiers by a large margin, where both the CNN and RNN components contribute to the performance improvement. To the best of our knowledge, CRIP is the first machine learning-based tool specialized in the prediction of circRNA-RBP interactions, which is expected to play an important role for large-scale function analysis of circRNAs.

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Prediction of RNA-protein sequence and structure binding preferences using deep convolutional and recurrent neural networks

RNA regulation is significantly dependent on its binding protein partner, which is known as the RNA-binding proteins (RBPs). Unfortunately, the binding preferences for most RBPs are still not well characterized, especially on the structure point of view. Informative signals hiding and interdependencies between sequence and structure specificities are two challenging problems for both predicting RBP binding sites and accurate sequence and structure motifs mining.\n\nIn this study, we propose a deep learning-based method, iDeepS, to simultaneously identify the binding sequence and structure motifs from RNA sequences using convolutional neural networks (CNNs) and a bidirectional long short term memory network (BLSTM). We first perform one-hot encoding for both the sequence and predicted secondary structure, which are appropriate for subsequent convolution operations. To reveal the hidden binding knowledge from the observations, the CNNs are applied to learn the abstract motif features. Considering the close relationship between sequences and predicted structures, we use the BLSTM to capture the long range dependencies between binding sequence and structure motifs identified by the CNNs. Finally, the learned weighted representations are fed into a classification layer to predict the RBP binding sites. We evaluated iDeepS on verified RBP binding sites derived from large-scale representative CLIP-seq datasets, and the results demonstrate that iDeepS can reliably predict the RBP binding sites on RNAs, and outperforms the state-of-the-art methods. An important advantage is that iDeepS is able to automatically extract both binding sequence and structure motifs, which will improve our transparent understanding of the mechanisms of binding specificities of RBPs. iDeepS is available at https://github.com/xypan1232/iDeepS.

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RNA-protein binding motifs mining with a new hybrid deep learning based cross-domain knowledge integration approach

BackgroundRNAs play key roles in cells through the interactions with proteins known as the RNA-binding proteins (RBP) and their binding motifs enable crucial understanding of the post-transcriptional regulation of RNAs. How the RBPs correctly recognize the target RNAs and why they bind specific positions is still far from clear. Machine learning-based algorithms are widely acknowledged to be capable of speeding up this process. Although many automatic tools have been developed to predict the RNA-protein binding sites from the rapidly growing multi-resource data, e.g. sequence, structure, their domain specific features and formats have posed significant computational challenges. One of current difficulties is that the cross-source shared common knowledge is at a higher abstraction level beyond the observed data, resulting in a low efficiency of direct integration of observed data across domains. The other difficulty is how to interpret the prediction results. Existing approaches tend to terminate after outputting the potential discrete binding sites on the sequences, but how to assemble them into the meaningful binding motifs is a topic worth of further investigation.\n\nResultsIn viewing of these challenges, we propose a deep learning-based framework (iDeep) by using a novel hybrid convolutional neural network and deep belief network to predict the RBP interaction sites and motifs on RNAs. This new protocol is featured by transforming the original observed data into a high-level abstraction feature space using multiple layers of learning blocks, where the shared representations across different domains are integrated. To validate our iDeep method, we performed experiments on 31 large-scale CLIP-seq datasets, and our results show that by integrating multiple sources of data, the average AUC can be improved by 8% compared to the best single-source-based predictor; and through cross-domain knowledge integration at an abstraction level, it outperforms the state-of-the-art predictors by 6%. Besides the overall enhanced prediction performance, the convolutional neural network module embedded in iDeep is also able to automatically capture the interpretable binding motifs for RBPs. Large-scale experiments demonstrate that these mined binding motifs agree well with the experimentally verified results, suggesting iDeep is a promising approach in the real-world applications.\n\nConclusionThe iDeep framework not only can achieve promising performance than the state-of-the-art predictors, but also easily capture interpretable binding motifs. iDeep is available at http://www.csbio.sjtu.edu.cn/bioinf/iDeep

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