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Yao, J.

Publications and source records attributed to Yao, J..

At least 73 records · Page 4Linked to original sources

Genomic Touchstone: Benchmarking Genomic Language Models in the Context of the Central Dogma

The emergence of genomic language models (gLMs) has revolutionized the analysis of genomic sequences, enabling robust capture of biologically meaningful patterns from DNA sequences for an improved understanding of human genome-wide regulatory programs, variant pathogenicity and therapeutic discovery. Given that DNA serves as the foundational blueprint within the central dogma, the ultimate evaluation of a gLM is its ability to generalize across this entire biological cascade. However, existing evaluations lack this holistic and consistent framework, leaving researchers uncertain about which model best translates sequence understanding into downstream biological prediction. Here we present Genomic Touchstone, a comprehensive benchmark designed to evaluate gLMs across 36 diverse tasks and 88 datasets structured along the central dogmas modalities of DNA, RNA, and protein, encompassing 5.34 billion base pairs of genomic sequences. We evaluate 34 representative models encompassing transformers and convolutional neural networks, as well as emerging efficient architectures such as Hyena and Mamba. Our analysis yield four key insights. First, gLMs achieve comparable or superior performance on RNA and protein tasks compared to models pretrained on these molecules. Second, transformer-based models continue to lead in overall performance, yet efficient sequence models show promising task-specific capabilities and deserve further exploration. Third, the scaling behavior of gLMs remains incompletely understood. While longer input sequences and more diverse pretraining data consistently improve performance, increases in model size do not always translate into better results. Fourth, pretraining strategies, including the choice of training objectives and the composition of pretraining corpora, exert substantial influence on downstream generalization across different genomic contexts. Genomic Touchstone establishes a unified evaluation framework tailored to human genomics. By spanning multiple molecular modalities and biological tasks, it offers a valuable foundation for guiding future gLM design and understanding model generalizability in complex biological contexts.

bioinformatics↗

Deepening imaging-based spatial proteomics at high spatial resolution through controlled tissue resizing and in-situ bottom-up mass spectrometry

Understanding the spatial organization of proteins in tissues is essential for elucidating biological function, but current proteomic imaging methods face limitations in resolution, sensitivity, and multiplexing capability. Here, we present IMPACT (Imaging Mass Spectrometry for Proteome Analysis on Compressed Tissue), a hydrogel-based workflow that integrates controlled vertical tissue compression following expansion to enhance protein detection while improving spatial resolution in mass spectrometry imaging (MSI). Through a 6-fold lateral expansion and 100-fold thickness reduction, analytes were concentrated at the surface to improve detection and enable analysis of thick specimen. Coupled with optimized in-gel digestion using hybrid enzymes, IMPACT increased protein identifications 3-fold compared to conventional MSI while maintaining spatial fidelity. We demonstrate IMPACTs utility in mapping low-abundance proteins and resolving fine anatomical structures. Orthogonal validation with immunofluorescence and integration with spatial transcriptomics further reveal protein-RNA distribution discrepancies, highlighting the need for direct proteomic imaging. IMPACT bridges critical gaps in spatial proteomics by enabling high-resolution, high-sensitivity, and multiplexed untargeted protein imaging, opening new avenues for tissue biology and biomarker discovery.

biophysics↗

FRA1 drives melanoma metastasis through an actionable transcriptional network

Transcriptional dysregulation has emerged as a critical driver of melanoma progression, yet the molecular mechanisms governing this process and their potential as therapeutic targets remain inadequately characterized. Here, we identify FRA1 as a potent and actionable driver of melanoma metastasis. FRA1 enhanced both the initial seeding and subsequent outgrowth of metastatic lesions. Comprehensive multi-omics integration revealed transcriptional target genes of FRA1, with AXL, CDK6, and FSCN1 exhibiting increased expression in melanoma metastasis and a significant correlation with poor patient outcomes. Silencing AXL, CDK6, or FSCN1 abrogated FRA1-mediated invasion in vitro and reduced metastatic colonization. Furthermore, pharmacological inhibition of CDK6 and FSCN1, and to a lesser extent AXL, suppressed melanoma metastasis and prolonged overall survival. The expression of FRA1 and its target genes correlates with shortened survival across multiple cancer types, highlighting the broader clinical relevance of this pathway. This study unveils an actionable FRA1-mediated transcriptional network that drives cancer progression and metastasis, offering potential avenues for therapeutic interventions. SIGNIFICANCEFRA1 promotes melanoma metastasis by enhancing the expression of AXL, CDK6, and FSCN1 and this transcriptional network is associated with poor survival across several cancer types. Targeting these FRA1 effectors suppresses metastasis and extends survival, offering a therapeutic strategy for metastatic melanoma and potentially other aggressive cancers.

cancer biology↗

A synergistic generative-ranking framework for tailored design of therapeutic single-domain antibodies

Single-domain antibodies (sdAbs) have emerged as powerful therapeutic agents due to their small size, high stability, and superior tissue penetration. However, unlike conventional monoclonal antibodies (mAbs), sdAbs lack an Fc domain, limiting their functional versatility and manufacturability. To address this challenge, we developed TFDesign-sdAb, a deep learning-based generative-ranking framework that enables rational engineering of sdAbs with tailored functionalities. Our framework integrates a structure-aware diffusion model (IgGM) for large-scale candidate generation and a fine-tuned sorter (A2binder) that evaluates and prioritizes them based on predicted functionality. Unlike traditional CDR-focused approaches, TFDesign-sdAb optimizes both complementarity-determining regions (CDRs) and framework regions (FRs), allowing sdAbs to acquire new functional properties while maintaining antigen specificity. We validated our approach by conferring Protein A binding to human VHs and nanobodies that originally lacked this feature, achieving high expression rates, strong binding affinities, and successful purification via industry-standard Protein A affinity chromatography. High-resolution structural characterization (1.49 [A] and 2.0 [A]) of the redesigned sdAb-Protein A complexes revealed conserved FR-mediated binding motifs that recapitulate natural Fc-Protein A interactions, validating the accuracy of our model. Furthermore, our pipeline streamlined the antibody engineering process, achieving a 100% success rate in generating Protein A-binding sdAbs while maintaining their original antigen-binding affinity. This work demonstrates the power of AI-driven design in overcoming long-standing limitations in antibody engineering and presents a scalable, generalizable solution for enhancing sdAb functionality.

bioinformatics↗

Unraveling Tyrosine-Kinase Inhibitor Resistance in NSCLC Cells via Same-cell Measurement of RNA, Protein, and Morphological Responses

Non-small cell lung cancer (NSCLC) frequently develops resistance to tyrosine kinase inhibitors (TKIs), limiting the long-term success of targeted therapies. A deeper understanding of resistance mechanisms at the molecular and cellular levels may enable the development of more effective treatment strategies. Here, we applied the Teton detection assay on the AVITI24 platform to measure concurrently RNA, protein, and cellular morphology in NSCLC cell lines treated with the TKIs gefitinib and osimertinib. This single-cell, multiomic analysis revealed distinct expression and morphological profiles between drug-sensitive and resistant cells, including differences in MAPK-related pathway activity. Stratifying responses at the single-cell level uncovered subtle responses not detectable in bulk measurements. We identified CDK4/6 activity as a route of cell survival under TKI treatment and demonstrated that co-treatment with the CDK4/6 inhibitor palbociclib enhanced TKI efficacy. The ability to measure multiomics and cellular morphology in the same cells opens new avenues for future studies aimed at improving personalized treatment strategies in NSCLC and overcoming the obstacles posed by drug resistance.

genomics↗

scTransMIL Bridges Patient-Level Phenotypes and Single-Cell Transcriptomics for Cancer Screening and Heterogeneity Inference

Single-cell sequencing technology has revolutionized cancer research by revealing unprecedented insights into tumor heterogeneity. However, reliably connecting patient-level cancer phenotypes with single-cell transcriptomic profiles remains challenging, due to technical constraints and labeling ambiguity. This further hinders the precise cancer screening and intensive study of tumor mechanism based on single-cell sequencing. To bridge this gap, we introduce scTransMIL, a scRNA-seq Transformer-based Multi-Instance Learning framework that learns whole-genome context to deliver comprehensive cancer insights at the sample, cell, and gene levels across three biological scales: (1) accurate patient-level cancer phenotype prediction, (2) precise single-cell disease scoring (validated on 4 million single cells), and (3) genome-wide biomarker discovery. Benchmark experiments demonstrate scTransMILs exceptional performance, including robust out-of-distribution generalization and clinically relevant prediction of metastatic tissue-of-origin, a crucial capability for identifying cancers of unknow primary. At single-cell resolution, scTransMIL identified both known and novel biomarkers for tumor B cells that conventional differential expression analysis failed to detect while maintaining consistent concordance with malignant cell annotations. scTransMILs adaptability is exemplified in acute myelocytic leukemia, where minimal fine-tuning with only a few patient sample labels enabled: (i) discovery of novel disease subtypes with distinct clinical outcomes, (ii) reconstruction of differentiation trajectories at the single-cell resolution, and (iii) identification of subtype-specific gene signatures. In summary, by systematically linking cellular and molecular profiles with clinical disease phenotypes, scTransMIL emerges as a transformative tool poised to advance both basic cancer research and precision oncology applications.

bioinformatics↗

Palmdelphin facilitates R-spondin2 secretion to activate Wnt signaling and promote colorectal cancer stemness and tumorigenesis

The Wnt signaling pathway is a key driver of stemness and progression which contributes to mortality in colorectal cancer (CRC). R-spondins bind to LGR receptors to inhibit ubiquitin E3 ligases, thus protecting Frizzled from degradation and activating downstream Wnt signaling. Herein, we identify Palmdelphin (PALMD) as a functional marker of CRC stem cells, interspersed between intestinal and colonic crypt base epithelial cells, and predictive of aggressive CMS4 CRC and poor survival. Gene knockdown and overexpression studies revealed that PALMD initiates paracrine activation of Wnt/{beta}-catenin signaling via interacting with and facilitating the secretion of R-spondin2 (RSPO2), resulting in enhanced stemness and tumor growth in vitro and in vivo. Physiologic or pharmacologic inhibition of the PALMD-RSPO2 axis using R-spondin2-specific antibody or 6-methyl-1,3,8-trihydroxyanthraquinone (emodin), respectively, attenuates PALMD- mediated Wnt reporter activation, self-renewal, and tumorigenesis in cell and patient-derived organoid models. Together, these findings identify PALMD as a previously unknown player in Wnt signaling in CRC, and underscore the pro-tumorigenic role of R-spondin2 in this context.

cancer biology↗

Evaluating genetic-ancestry inference from single-cell RNA-seq data

Characterizing the ancestry of donors in single-cell transcriptomic studies is crucial to ensure genetic homogeneity, reduce biases in analyses, identify ancestry-specific regulatory mechanisms and their downstream roles in disease, and ensure that existing datasets are representative of human genetic diversity. While these datasets are now widely available, information on the ancestry of donors is often missing, hindering further analysis. Here, we propose a framework to evaluate methods for inferring genetic-ancestry from genetic polymorphisms detected in single-cell sequencing reads. We demonstrate that widely used tools (e.g., ADMIXTURE) provide accurate inference of genetic-ancestry and admixture proportions, despite the limited number of genetic polymorphisms identified and imperfect variant calling from sequencing reads. We infer genetic-ancestry for 401 donors from ten Human Cell Atlas datasets and report a high proportion of donors of European ancestry in this resource. For researchers generating single-cell transcriptomic datasets, we recommend reporting genetic-ancestry inference for all donors and generating datasets that represent diverse ancestries.

bioinformatics↗

De Novo Negatively Charged Binders Targeting MMLVRT Nucleic Acid Binding Sites Overcome Stability-Activity Trade-offs

The limited thermostability and storage-induced inactivation of Moloney murine leukemia virus reverse transcriptase (MMLV RT) have constrained its applications. In this study, a high-stability mutant, MMLV RT-SV, was generated through a multi-site mutagenesis strategy targeting the nucleic acid-binding pocket. Through de novo protein design, three highly negatively charged binders with affinities of 45 nM, 362 nM, and 829 nM were developed to specifically target the positively charged nucleic acid-binding region on the surface of MMLV RT-SV. Experimental results demonstrated that these binders formed complexes with the enzyme via electrostatic interaction, significantly enhancing the thermostability and long-term storage stability of MMLV RT-SV while not affecting its RNA- dependent DNA polymerase function. This work pioneers the rational design of negatively charged binders targeting strongly positively charged nucleic acid-binding sites, overcoming the traditional stability-activity trade-off inherent in site-directed mutagenesis and directed evolution. The proposed strategy not only provides an innovative solution to address the thermal sensitivity and storage instability of reverse transcriptase but also establishes a novel paradigm for De Novo-based precision engineering of enzyme functions, demonstrating significant potential in molecular diagnostics, gene editing, and related fields.

bioengineering↗

National Scale Genomic Engine for Precision Medicine: Singapore PRECISE-SG100K Experience

The whole-of-nation National Precision Medicine (NPM) programme in Singapore aims to integrate a genomics backbone into standard healthcare. Phase II recently completed the sequencing of 102,202 genomes from the diverse Asian population of Singapore, generating the PRECISE-SG100K dataset. The scaling from the pilot SG10K_Health phase to national production scale demanded a central shift: from earlier generation workflows to an integrated, industrial grade "Genomic Engine". The initiative was co-developed under a public-private partnership model. Our Genomic Engine was designed with an automation-first approach, as a modular platform linking a purpose-built, centralized biospecimen processing and DNA extraction facility (EXTRACT Lab), a high-throughput sequencing operation (Illumina Data Generation Infrastructure, IDGI), with the cloud-native analytics (GRIDS) running on a fully integrated PRECISE Sample Tracking System (STS). Here we describe the key design features of our Genomic Engine. Highlighting the Stress Test phase where the automated DNA extraction yields failed to meet expectations. We share the operational adjustments which were implemented to stabilize production and resulted in a three-fold reduction in sample triage due to improved DNA recovery. Without establishing the operational guardrails during the Stress Test, the Engine would not have reached a steady-state production evidenced by >3,000 genomes per month, median turnaround time 11 days, 99.6% operational success, within in six months. This framework offers a practical blueprint for other population-scale initiatives seeking to bridge the gap between scientific piloting and industrial delivery.

genomics↗

Asymmetric Histone Inheritance Regulates Differential Transcription Re-initiation and Cell Fate Decisions in Mouse Olfactory Horizontal Basal Cells

The olfactory epithelium (OE) possesses an adult stem cell population, the horizontal basal cells (HBCs), to permit lifelong tissue regeneration. Here we show that HBCs exhibit asymmetric inheritance of histone H4 but not H2A-H2B during OE regeneration in mice. Primary HBC cultures further revealed asymmetric histone inheritance for H3 and H3.3. Upon mitotic exit, asymmetric histone inheritance correlates with asynchronous transcription re-initiation and differential enrichment of p63, a key transcription factor for HBC cell fate. Disruption of asymmetric histone inheritance abolished these asymmetric cellular features and attenuated OE regeneration and smell behavior recovery. Single-cell RNA sequencing of paired HBC daughters in culture further supported asymmetric multilineage cell fate priming. Together, these findings reveal asymmetric histone inheritance in a mammalian adult stem cell lineage and highlight its biological significance in neural tissue regeneration and animal behavior.

developmental biology↗

Identification of pyroptosis-related diagnostic model relate to immune infiltration in osteoarthritis

BackgroundOsteoarthritis (OA) is a common chronic diseases which were associated with aging and progressive joint dysfunction. Pyroptosis, a novel mode of cell death. The effect of pyroptosis on osteoarthritis (OA) progression is still in the exploratory stage, and the specific mechanism is unclear. MethodsOA data sets were obtained from the GEO databases website. Bioinformatics analysis was conducted to identify pyroptosis-related genes (PRGs), construct a pyroptosis-related diagnostic model, and identify subtypes. We established the random forest (RF) model as well as a nomogram of differentially expressed PRGs. The association between immunity and pyroptosis was comprehensively explored. Finally, the expression levels of 3 OA characteristic genes were verified by qRT-PCR and immunohistochemistry. ResultsSeven differentially expressed PRGs (CASP3, GPX4, NOD1, TIRAP, SCAF11, CASP4, and CASP9) were obtained by differential analysis to establish RF models, and three important OA genes (CASP9, TIRAP, and SCAF11) were obtained. ROC curve proved that the established model had high diagnostic accuracy. Based on the above three important PRGs, a high-precision nomogram model is constructed. Two PRG models are determined by consensus clustering method. The enrichment degree of immune cells in cluster A was higher than that in cluster B, suggesting that cluster A might be related to the occurrence and development of OA. GPX4 was found to be positively correlated with higher immune infiltration. Finally, the expression levels of 3 OA characteristic genes were verified by qRT-PCR and immunohistochemistry. ConclusionWe constructed an effective prognostic model according to PRGs in OA and integrated explored the association between pyroptosis and immunity-related elements. This article provides a strong point for investigating directed immunotherapy methods with novelty for OA.

immunology↗

Application and mechanism of anticancer peptides in organoid models of intrahepatic cholangiocarcinoma

BackgroundThe clinical manifestations of intrahepatic cholangiocarcinoma (ICC) are non-specific, and only a small number of patients are eligible for surgical resection at the time of diagnosis, limiting treatment options. Anticancer peptides (ACPs) exhibit strong inhibitory activity against tumour cell lines, exhibit minimal side effects, are easily modified and optimised, and have low production costs. These attributes make ACPs a prospect for clinical applications. Simultaneously, the development of patient-derived three-dimensional organoids as a novel disease model has enabled the replication of the structure and heterogeneity of solid tumours. These organoids provide valuable tools for understanding disease mechanisms, conducting drug sensitivity tests, and developing targeted therapies. However, the effect of ACPs at the organoid level in ICC remains unknown. Therefore, this study aims to explore the potential of ACPs in treating ICC using patient-derived organoids as a model system to evaluate their efficacy and optimize therapeutic strategies. MethodsWe designed and synthesised a novel ACP sequence and applied it to a patient-derived organoids (PDOs) model. Organoids were evaluated using histochemistry analysis and whole-exome sequencing. ACPs were compared with representative chemotherapeutic drugs, including sorafenib and gemcitabine, to assess treatment differences. Additionally, scanning electron microscopy and flow cytometry were performed for further mechanistic analyses. ResultsOrganoid models exhibited histological characteristics similar to those of maternal tumours, especially showing strong positivity for EpCAM and KRT19 expression. Furthermore, whole-exome sequencing of the primary tumour and derived organoids revealed that PDOs closely recapitulated the genomic similarities of the primary tumour across multiple levels. Drug response analyses revealed that ACPs exhibited highly efficient anti-tumour effects, with specific differences observed between patients. ACPs affected the growth of tumour cells and exerted anticancer effects through direct membrane disruption and indirect induction of apoptosis. ConclusionIn this study, organoids derived from patients with ICC were established. This in vitro model provides valuable tools for evaluating the therapeutic response of ACPs and offers novel insight for the study of ICC.

cancer biology↗

Unifying Genetic and Chemical Perturbagen Representation through a Hybrid Deep Learning Framework

Artificial Intelligence virtual cell (AIVC) holds transformative potential for biomedical research. Central to this vision is the systematic modeling of genetic and chemical perturbation phenotypes to accurately predict cellular dynamic states from diverse interventions. However, disparities in screening agents, library scales, experimental technologies, and data production efficiency hinder the integration, modeling, and analysis of the cross-data. Here we present UniPert-G2CP, a two-phase deep learning approach comprising i) UniPert, a multimodal molecular representation model that bridges genetic and chemical domains, and ii) G2CP (Genetic-to-Chemical Perturbation transfer learning), which systematically transforms CRISPR screen-based genetic insights into chemical perturbation modeling for cost-effective in silico drug screening. UniPert not only encodes multimodal perturbagens into a unified functionally interpretable sematic embedding space, but also improves phenotypic effect prediction for previously unseen gene perturbations and drug treatments. Building upon UniPert, G2CP successfully modeled large-scale cellular post-perturbation states spanning 4,994 gene and 7,821 compound perturbagens, while reducing modeling data costs by over 60%. We demonstrate that UniPert-G2CP enables efficient, generalizable simulations of multicellular, multi-domain perturbation cause-effect spaces, revealing differential cellular biological causality and informing mechanism-driven therapy. UniPert-G2CP opens new avenues for biological causal foundation model building, AIVC creation, and AI-powered precision medicine.

bioinformatics↗

Extracting knowledge from scientific texts on patient-derived cancer models using Large Language Models: algorithm development and validation

Patient-derived cancer models (PDCMs) have emerged as indispensable tools in both cancer research and preclinical studies. The number of publications on PDCMs increased significantly in the last decade. Developments in Artificial Intelligence (AI), particularly Large Language Models (LLMs), hold promise for extracting knowledge from scientific texts at scale. This study investigates the use of LLM-based systems for automatically extracting PDCM-related entities from scientific texts. We evaluated two approaches: direct prompting and soft prompting using LLMs. For direct prompting, we manually create prompts to guide the LLMs to output PDCM-related entities from texts. The prompt consists of an instruction, definitions of entity types, gold examples and a query. We automatically train soft prompts - a novel line of research in this domain - as continuous vectors using machine learning approaches. Our experiments utilized state-of-the-art LLMs - proprietary GPT4-o and a series of open LLaMA3 family models. In our experiments, GPT4-o with direct prompts maintained competitive results. Our results demonstrate that soft prompting can effectively enhance the capabilities of smaller open LLMs, achieving results comparable to proprietary models. These findings highlight the potential of LLMs in domain-specific text extraction tasks and emphasize the importance of tailoring approaches to the task and model characteristics.

cancer biology↗

Fast and accurate modeling of TCR-peptide-MHC complexes using tFold-TCR

Alpha-beta T cell receptor ({beta}TCR) recognition of peptide-major histocompatibility complexes (pMHCs) is a corner- stone of the adaptive immune system. Fast and accurate modeling of TCR-pMHC structures is crucial for understanding TCR recognition of pMHCs at the molecular level, which is essential for the development of TCR-based therapeutics and vaccines. Despite significant interest, this challenge remains unresolved due to the diversity of TCR-pMHC interactions and limited structural data. Here, we present tFold-TCR, a high-throughput, end-to-end universal model for predicting three-dimensional (3D) atomic-level structures of TCR-pMHC complexes, capable of predicting TCRs of different classes and MHC structures from diverse systems. tFold-TCR leverages a specially trained, protein-protein interaction-sensitive large protein language model to extract intra- and inter-chain residue contact information and evolutionary relationships, bypassing the need for multiple sequence alignment (MSA) searches. It also features innovative structure prediction and flexible docking modules to enhance accuracy, particularly for interacting contacts. Compared to existing methods, including AlphaFold-3, tFold-TCR demonstrates a 30.7% increase in prediction success rate evaluated by DockQ and is over 25 times faster. These advancements enable large-scale structural characterization of TCRs and their interactions with pMHCs. Utilizing this capability, we constructed TCRStructDB, the largest database of TCR-pMHC structures to date, encompassing 2.2 million TCRs, 0.8 million pMHCs, and 45,000 TCR-pMHC complexes. TCRStructDB provides unprecedented insights into one of the most diverse receptor-ligand interactions in biology.

bioinformatics↗

1,3- and 1,4-linked Polysaccharides Uptake in Intestinal Cells Relies on Clathrin/Dynamin 1/Rab5-dependent Endocytosis

Natural polysaccharides are considered to be a promising feedstock for manufacturing sustainable materials. However, polysaccharides are believed by most biologists that they could not be absorbed by intestinal mucosa to enter circulation system owing to their high hydrophilicity and large molecule size. Herein, {beta}-1,3-linked glucan (GFPBW1) and -1,4-linked glucan (WGE) is used as the model polysaccharide and the transport experiments show that different linked and charged polysaccharides can permeate Caco-2 cell monolayers. The in vivo experiments demonstrate that GFPBW1 and WGE are detectable in rodent plasma and liver after oral administration the isotope (99mTc, 3H) and fluorescein labeled polysaccharides. Further study show that polysaccharide uptake requires clathrin heavy chain (CLTC) and its associated factors Rab5 and dynamin1 in intestinal cells using gene-knockdown strategies and inhibitors. Strikingly, polysaccharide absorption is attenuated in both CLTC intestine deficient mice and Rab5, dynamin1 conventional-knockout mice. Importantly, membrane receptors bone morphogenetic protein receptor type IA (BMPRIA), Dectin-1 and epidermal growth factor receptor (EGFR) are also critical for specific structural polysaccharide internalization. These findings provide novel insight to understand polysaccharide absorption mechanism and lay foundation for oral polysaccharide- based new drugs development.

pharmacology and toxicology↗

Phages-bacteria interactions underlying the dynamics of polyhydroxyalkanoates-producing mixed microbial cultures via meta-omics study

The dynamics of the structure of polyhydroxyalkanoates-producing mixed microbial cultures (PHA-MMCs) during enrichment and maintenance is an unsolved problem. The effect of phages has been proposed as a cause of dynamic changes in community structure, but evidence is lacking. To address this question, we enriched five PHA-MMCs and sampled temporally to study the interactions between phage and prokaryotic members by combining metagenomics and metatranscriptomics. 963 metagenome-assemble genomes and 4294 phage operational taxonomic units were assembled from bulk metagenomic data. There were complex interactions between the phages in Casadabanvirus and Unclassified Hendrixvirinae and the dominant species in Azomonas, Paracoccus, Thauera and Breundimonas. The dynamic change of the structure of phage and prokaryotic communities were remarkably consistent. Further structural equation modeling analysis showed that phage communities could significantly affect the activity and community structure of prokaryotic microorganisms. In addition, seven key auxiliary metabolic genes (phaC, fadJ, acs, ackA, phbB, acdAB and fadD) in the PHA synthesis pathway were identified from phage sequences. Importantly, these auxiliary metabolic genes were expressed at the transcriptional level, indicating that they were in an active functional state. This meta-analysis provides the first catalog of phages in PHA-MMCs and the auxiliary metabolic genes they carry, as well as how they play a role in the dynamic changes of prokaryotic communities. This study provides a reference for subsequent studies on understanding and regulating the microbial community structure of artificial open microbial systems. ImportanceThe synthesis of biodegradable plastic PHA from organic waste through mixed microbial cultures (MMCs), with extremely low cost, has the potential for expanded production. However, there is a gap of understanding of the dynamics of dominant species in MMCs to better control community structure. Our results demonstrate for the first time the impact of phages on the structure of prokaryotic communities in the MMCs. There are complex interactions between the PHA producers (Azomonas, Paracoccus, Thauera) and phages (Casadabanvirus and Unclassified Hendrixvirinae). Phage communities can negatively regulate the activity and structure of prokaryotic communities. In addition, the auxiliary metabolic genes in the PHA synthesis pathways carried by phages may promote the PHA synthesis ability of prokaryotic members. This study highlights the impact of phages on prokaryotic community structure, suggesting that phages have the potential to become a tool for better controlling the microbial community structure of PHA-MMCs.

microbiology↗