Search bioRxiv⌕ Search

Biology subjects

Pham, T.

Publications and source records attributed to Pham, T..

At least 19 recordsLinked to original sources

Variable sparsity non-negative matrix and tensor factorization algorithms and their applications in cancer genomics

The field of oncology has experienced significant advancements in novel therapies and molecular diagnostics to personalize initial cancer treatments. However, cancer's propensity for evolution and resistance to therapies remains a formidable challenge due to its dynamic nature. Cancer's complex and adaptive behavior surpasses the capabilities of simple models, complicating our comprehension at every level, from basic sciences to clinical practices, highlighting the urgent need for innovative approaches to improve the modeling and classification of cancer. In this context, matrix factorization algorithms have been used to generate transcriptional signatures that summarize the most salient patterns of behaviors in cancer datasets. For example, the standard non-negative matrix factorization algorithm (NMF) generates a dimensionality-reduced representation that encodes a significant amount of data relationships as a mixture of positive components and provides a working summary of the input data. The resulting NMF factor matrices, W and H, display some degree of sparseness; however, this sparseness is often a byproduct rather than an explicit objective of the NMF algorithm, and the user has limited control over the degree of sparseness of the representation and the corresponding balance between local versus global structures in the NMF results. Some sparse versions of NMF have been introduced but still do not allow a fine-grained control on the sparsity constraints. We present here a novel variable-sparsity Non-negative Matrix Factorization algorithm (vsNMF), where instead of imposing sparsity as a regularization constraint on the NMF objective function, sparsity is explicitly enforced by a projective step performed for each column of W and row of H after a standard NMF update. In this way, the algorithm takes as input the input matrix, plus two one-dimensional vectors, that specify the sparsity functions that will be imposed on the NMF solution. The more explicit control of sparsity, and the corresponding balance between local vs. global structure, at the level of individual components, allows the generation of NMF solutions that can be tuned to operate anywhere between a generative, parts-based encoding of overlapping features, and a partitional, soft-clustering model. This allows greater flexibility to represent biological subtypes, patterns of pathway activation or deregulation, and higher biological interpretability. These learned low-dimensionality representations can also be used to integrate and merge datasets, or to define groups of similar cancer samples representing putative cellular or tumor states. Adjustable/tunable NMF representations of biological data can greatly help advance our fundamental understanding of cancer biology, provide more interpretable computational models of cancer, and better functional cancer taxonomies and precision oncology paradigms. We apply the same approach to tensors and introduce a variable-sparsity Non-Negative Tensor Factorization algorithm (vsNTF) that can take an input tensor and sparsity functions for each of the tensor dimensions.

bioinformatics↗

The Multi-scale Copula Index (MCI): a new information-theoretic measure of association and its applications in functional cancer genomics

Cancer genomic analyses increasingly involve large, complex datasets representing diverse omics variables, such as gene expression, mutations, copy number alterations, genetic perturbations (e.g. CRISPR or iRNA), drug sensitivity, and clinical phenotypes, making it crucial to accurately measure associations among these variables. Traditional correlation-based methods, like the Pearson correlation coefficient, often assume linearity, Gaussian distributions, and independence among variables, leading to oversimplified assumptions, less sensitive readouts and less accurate significance estimation. Information-theoretic measures overcome these limitations by being one step removed from the data and assessing dependencies at the level of probability distributions, thus providing a more data-agnostic, robust measure for identifying associations in common analysis tasks such as feature selection, pattern recognition, and predictive modeling. Building on these foundations, we introduce the Multi-scale Copula Index (MCI), a novel information-theoretic measure of association. The MCI offers significant advantages over existing methods by eliminating the need for density estimation and parameter tuning, thereby ensuring consistent estimation with reduced computational complexity. Furthermore, it enhances statistical power at lower sample sizes, exhibits strong robustness to outliers, and successfully captures both directionality and important symmetries under input-data transformations, such as reflection or complementation. Due to these strengths, the MCI is particularly useful for functional cancer genomic applications involving, e.g., transcriptional and perturbational data, with different statistical properties across a wide range of dataset sizes and complexity. Contextual associations often arise in functional genomics datasets: two genomic variables show little or no correlation in most samples, yet display a strong linkage within the subset that reflects the active biological context. These situations are ideal for quantifying associations with the MCI. We illustrate the performance of the MCI in a variety of benchmarks and cancer genomic examples and applications. We also introduce the Cumulative Entropy Diagram (CED), a graphical display that allows the visualization of the MCI between a reference vector, such as a phenotype, pathway activation or drug response, and a complete set of genomic features.

bioinformatics↗

A robust and scalable image analysis framework for phenotyping human thoracic aorta

Large-scale phenotyping of the thoracic aorta requires a workflow in which every step, from segmentation to phenotype extraction, runs without manual intervention. We present a robust and scalable framework that converts contrast-enhanced CT into anatomical landmarks, a patient-specific quadrilateral mesh, a wall-stress field and regional phenotypes of the thoracic aorta. Four nnU-Net segmentation tasks delineate the aorta, its anatomical regions and two partitions of the aortic root. A surface-curve-point procedure derives the annulus, the sinotubular junction, the ostia of the supra-aortic branches, the cusp nadirs and the commissures from the interfaces between segmented regions. We fit a structured template of 24,882 quadrilateral elements to the segmented aortic surface by staged, differentiable mesh deformation and compute the transmural mean wall stress with PyTorch-FEA. The structured mesh provides node-to-node correspondence across patients, anchored to the anatomy by the landmarks, from which the framework extracts regional morphological and biomechanical phenotypes.

bioengineering↗

Loss of short-form Ron promotes B and T cell cooperation to prevent outgrowth of breast cancer bone metastasis

While immunotherapy has great potential to eradicate tumors, success in metastatic breast cancer is limited. Bone metastasis, the most common site for breast cancer metastasis, has been associated with weakened anti-tumor immunity, both in the bone and in other metastatic sites. Additional strategies promoting anti-tumor immunity are needed. Using the MMTV-PyMT intratibial model of breast cancer bone metastasis, we discovered that deletion of host short-form Ron (Ron SF-/-) protected against bone metastatic outgrowth by promoting strong infiltration of lymphocytes, including B cells, into tumors at the micrometastasis stage. Transcriptionally, Ron SF-/- B cells were more active and inflammatory compared to wildtype. Elimination of mature B cells (JH-/-) rescued tumor growth in Ron SF-/- mice, indicating that mature B cells were necessary for anti-tumor immunity and protection against bone metastasis. In Ron SF-/- hosts, T follicular helper (Tfh) cells were abundant in tumor-enriched regions of bone, suggesting interactions between B and T cells. B-T cell cooperation was further supported by attenuation of CD4 and CD8 T cell tumor infiltration in Ron SF-/-JH-/- hosts and, conversely, diminished B cell tumor infiltration and rescue of tumor growth in Ron SF-/- hosts lacking CD4 or CD8 T cells. B cells were not required to protect Ron SF-/- mice from lung metastasis, indicating a tissue-dependent anti-tumor role for B cells. These data support a positive feedback loop between B and T cells regulated by SF-Ron and reveal the anti-tumor potential of B cells in the bone.

cancer biology↗

Multimodal Imaging of the Cellular and Extracellular Microenvironment on the Same Formalin-Fixed Paraffin-Embedded Tissue Section

Same-tissue section multimodal imaging is a powerful strategy that spatially profiles tissue histology, cell populations and molecular composition while maximizing tissue economy, preserving spatial molecular relationships, and increasing co-registration capacity. However, performing multiple modalities on the same tissue section can destroy or chemically alter the tissue, compromising downstream data. Here, we systematically assess integration of picrosirius red staining, hematoxylin and eosin staining, and multiplexed immunofluorescence into N-glycan and extracellular matrix peptide matrix-assisted laser/desorption ionization imaging mass spectrometry (IMS) workflows. We evaluate alterations in tissue morphology, stain efficiency, IMS feature intensity as well as IMS feature localization after upstream modality integration. We propose an optimized multimodal sequence that maximizes data quality and follows a very specific order of: autofluorescence microscopy, multiplexed immunofluorescence, picrosirius red staining, N-glycan IMS, hematoxylin and eosin staining, and extracellular matrix peptide IMS. Overall, this work develops an optimized multimodal workflow that comprehensively images tissue morphology, collagen fibers, and cell populations at single-cell resolution as well as multiplexed N-glycan composition and multiplexed extracellular matrix peptides with post-translational modification status from a single tissue section.

biochemistry↗

Juvenile influenza can impair myelin development and adult behavior through chemokine signaling in mice

Brain development, especially developmental myelination, continues through young adulthood. Concordantly, children may be particularly vulnerable to neural-immune challenges. To investigate the consequences of major childhood immune challenges, juvenile mice were exposed to respiratory influenza (H1N1) infection. White matter-specific microglial reactivity accompanied by oligodendrocyte loss was evident until two months following infection. Mice exhibited hyperlocomotion and impaired attention, but not anxiety-like behavior, at one month following infection. Linking the oligodendroglial and behavioral deficits, genetic disruption of oligodendrocyte development at the same juvenile timepoint recapitulated this behavioral phenotype. Microglial reactivity and oligodendrocyte numbers normalized by young adulthood. However, myelin development was disrupted, with persistently decreased myelinated axon density and reduced myelin sheath thickness. Hyperlocomotion resolved, but anxiety-related behaviors emerged at two months after infection. At 6 months, anxiety resolved but cognitive deficits persisted. Elevated CSF chemokines and microglial chemokine expression prompted testing the role of the multi-chemokine receptor CCR3. CCR3 inhibition rescued these cellular and behavioral aberrations after juvenile H1N1 infection. Together, these findings underscore the potential for disruption of myelin development and lasting cognitive and neuropsychiatric sequelae following major immune challenges during the juvenile period and highlight chemokine signaling as an important therapeutic target.

neuroscience↗

Learning with Recurrence Geometric AI in Spatial Transcriptomics

Cross-domain analysis of spatial transcriptomics is challenging because tissues from different organs, diseases and experimental platforms exhibit distinct cellular compositions, spatial organisations and technical biases, making direct comparison of tissue states difficult. Existing methods primarily focus on domain integration or batch correction but generally do not explicitly model the intrinsic geometry underlying tissue-state organisation across biological systems. This paper presents recurrence geometric artificial intelligence (RGAI), a geometric deep-learning framework for discovering and aligning latent tissue states across heterogeneous spatial transcriptomic domains. RGAI first learns domain-specific latent representations using variational graph autoencoders while simultaneously estimating a Riemannian metric tensor that captures the local geometry of each latent manifold. Geodesic distances induced by the learned metric are used to construct multiscale recurrence graphs that characterise intrinsic tissue-state organisation independently of the original measurement space. Cross-domain manifold correspondence is then established through entropy-regularised Gromov-Wasserstein alignment, after which fuzzy clustering identifies latent tissue states and optimal transport aligns tissue-state signatures across domains. Evaluation on six human spatial transcriptomic datasets spanning wound healing, periodontitis, oral squamous cell carcinoma, head and neck squamous cell carcinoma, cardiac tissue and colorectal cancer shows that RGAI automatically determines biologically meaningful latent tissue-state complexity and identifies coherent recurrence-based tissue states within each domain. The learned geometric representations enable cross-domain alignment of latent manifolds while preserving biologically interpretable tissue-state correspondences despite substantial differences in cellular composition and tissue architecture, demonstrating that integrating learned Riemannian geometry, recurrence analysis and optimal transport provides a robust and interpretable framework for cross-domain tissue-state discovery and comparison in spatial transcriptomics.

bioinformatics↗

Rac1 deficiency reduces mitochondrial respiratory capacity, impairs fatty acid metabolism and causes muscle wasting

BackgroundThe age-related progressive decline in skeletal muscle function is characterised by declining mitochondrial quality control and perturbed fatty acid metabolism, contributing to frailty and increased mortality. The actin cytoskeleton, a key structural component of skeletal muscle, has recently been implicated in mitochondrial anchoring and dynamics. However, the role of actin-regulating proteins, including the Rho GTPase Rac1, in mitochondrial function and age-associated metabolic and functional muscle deterioration remains undefined. MethodsSkeletal muscle from mice with inducible muscle-specific deletion of Rac1 (Rac1 imKO) underwent unbiased mass spectrometry-based proteomic profiling. Mitochondrial morphology was assessed by transmission electron microscopy, and physiological parameters, including muscle mass and contraction-stimulated palmitate oxidation in isolated soleus muscle, were evaluated. Mitochondrial respiratory function was determined by high-resolution respirometry in permeabilised gastrocnemius skeletal muscle fibre bundles. Biochemically, muscular triacylglycerol (TG) content, mRNA (qPCR) and protein (immunoblotting) content were determined. In vastus lateralis muscle biopsies from healthy, untrained young (20-30 years) and old, sarcopenic (83-94 years) men, Rac1 and mitochondrial respiratory protein abundances were measured. A complementary human genetic association analysis was performed using the FinnGen dataset. ResultsRac1 deficiency triggered muscle wasting in middle-aged mice (Gastrocnemius: -10%; Quadriceps: -7%). Preceding muscle wasting, gene set enrichment analysis identified enrichment in fatty acid metabolism and oxidative phosphorylation pathways, consistent with increased mitochondrial volume density in Rac1 imKO muscle (subsarcolemmal: +467%; intermyofibrillar: +166%). Despite mitochondrial expansion at this stage, Rac1 deficiency attenuated the increase in palmitate oxidation in response to muscle contraction (-62%). At the muscle-wasting stage, Rac1 imKO muscle exhibited reduced mitochondrial respiratory capacity (-25-32%). Additionally, the mitochondrial dysfunction was associated with an accumulation of muscle TG (+78%, p = 0.096) and upregulation of fatty acid transporter, CD36 protein content (+25%), indicative of altered fatty acid handling. In humans, Rac1 muscle protein content was increased in old, sarcopenic subjects compared to young (+41%), and negatively correlated with quadriceps cross-sectional area (CSA) (r = -0.475) and type II fibre CSA (r = -0.466). In old, sarcopenic muscle, Rac1 protein content correlated negatively with protein content of multiple mitochondrial respiratory complexes (CI: r = -0.690, CIV: r = -0.938, CV: r = -0.704). GWAS further identified associations between Rac1 SNP variants and lipid metabolic and muscle-wasting diseases. ConclusionsMuscle Rac1 deficiency reduces mitochondrial respiratory capacity and metabolic flexibility through impaired fatty acid metabolism, leading to muscle wasting and highlighting a potential therapeutic target in age-related functional decline.

Molecular Biology↗

Satellite DNA dynamics across phylogenetic scales in ground beetles and other insects

Satellite DNAs (satDNAs) are tandemly repeated sequences that can comprise large fractions of eukaryotic genomes. For decades, we have known that satellite DNAs are among the most rapidly evolving genomic components, yet few studies have examined their dynamics across broad evolutionary frameworks--likely in part because their rapid evolution complicates or precludes homology identification. We investigated satDNA dynamics across 50 ground beetle (Carabidae) species, sampling above and below the species level in the subgenus Plataphus of Bembidion. We used RepeatExplorer2 and a novel homology detection pipeline, which we validated using species of Drosophila for which satDNAs are well-documented, to study satDNA dynamics in a comparative framework. We show that satDNAs comprise large genomic fractions (mean = 25.7%, range = 3.2%-53%), with individual satDNA motifs accounting for >30% of total DNA within some species. We quantify major satDNA turnover among closely related species which drives extensive repatterning of repeat content over short evolutionary scales. To place our findings in a broader context, we also surveyed satDNA diversity across the beetle suborder (Adephaga), the beetle order (Coleoptera), and the next six largest insect orders (Lepidoptera, Diptera, Hymenoptera, Hemiptera, Orthoptera, Trichoptera), representing 400 additional species. While satDNA abundance was significantly elevated in the focal subgenus (Plataphus), high genomic proportions were also found in species across Adephaga, Coleoptera, Hymenoptera, and Diptera. Conversely, satDNA proportions were notably reduced in Lepidoptera and Trichoptera. Our investigation of satDNA dynamics at narrow and broad taxonomic scales provides a quantitative illustration of satDNA turnover across a well-sampled clade. It also identifies new high-potential systems for gaining mechanistic insights into the drivers and impacts of satDNA evolution, and raises questions regarding potential links between satDNA turnover, speciation, and core strategies of chromosome and genome architecture in diverse clades.

evolutionary biology↗

Replication fork remodeling proteins, Smc5/6 and Rtt107, promote palindrome-mediated genome instability

Palindromic sequences are a potent source of genomic instability that can lead to cancer and hereditary diseases in humans. Molecular evidence shows that palindrome instability results from the formation of secondary structures, such as hairpins and cruciforms, which are cleaved by structure-specific nucleases. However, the mechanisms underlying cruciform formation and cleavage at palindromic sequences in eukaryotic cells remain incompletely understood. Here, we describe a pathway for secondary structure formation and chromosomal breakage at palindromes involving DNA helicase Mph1, Rad51 recombinase, Rad54 ATPase DNA strand remodeler, Rtt107 scaffold protein, and the multifunctional Smc5/6 complex. Deletion or mutation of any of these components results in a similar reduction in double-strand breaks at an Alu palindrome and a significant decrease in chromosomal rearrangements. We propose that Mph1, Rad51, and Rad54 work together at stalled replication forks to generate cruciform structures via fork remodeling, while Smc5/6 and Rtt107 mark the cruciforms, indicating an appropriate substrate for nuclease cleavage. As members of this pathway are conserved in humans, the uncovered mechanisms may underlie genomic instability at palindrome sites involved in disease etiology.

molecular biology↗

Predicting Antibody Self-Association with Sequence Structure Fusion Models: The Central Role of CSI-BLI in Early Developability Screening

Antibody-based biologics are expanding rapidly, yet challenges in development from self-association, high viscosity, aggregation, and unfavorable clearance underscore the need for accurate in silico screening. Clone self-interaction biolayer interferometry (CSI-BLI) is a plate-based, low-material assay of weak, reversible self-association that serves as an early proxy for high-concentration viscosity and a complementary predictor of in vivo clearance. In a 246-mAb panel, CSI-BLI moderately correlates with viscosity; further, in hFcRn Tg32 mice (41 antibodies), CSI-BLI strongly associates with clearance. Here, we present an end-to-end framework that distinguishes high versus low self-interacting clones (CSI-BLI class) by coupling a fine-tuned protein language model (ESM-2) with residue-aligned 3D context from AlphaFold-predicted structures encoded as residue graphs. Disentangled multi-stream attention fuses sequence content, chain-aware positional information, and structural signals to capture spatially proximate interactions that are distant in sequence. Edit-distance-controlled splits across 1499 IgGs and 988 VHHs assess generalization. The structure-aware model achieves the highest hold-out performance (VHH-Fc F1 = 0.76; IgG F1 = 0.57), while a sequence-only disentangled variant outperforms a standard PLM baseline without structural inputs. Complementary biophysical feature-based models, built from AlphaFold structures and sequence/structure-derived physicochemical descriptors with cluster-aware selection, deliver robust, interpretable performance (VHH F1 = 0.72; IgG F1 = 0.57), with SHAP analyses highlighting charge/dipole, hydrophobicity, and aggregation-propensity drivers across CDRs and frameworks. This interaction-aware sequence-structure framework, supported by interpretable feature models, is extensible to other developability endpoints and broader protein classification tasks where joint modeling of language-derived representations and residue-level geometry is advantageous.

bioinformatics↗

Decreased food intake contributes to elevated insulin-responsiveness in pre-clinical cancer cachexia

PurposeCancer cachexia is a life-threatening complication of advanced malignancies, driven by anorexia and profound systemic metabolic reprogramming. Insulin action in skeletal muscle is markedly impaired in patients with cancer and may contribute directly to cachexia pathogenesis. However, the interplay between reduced nutrient intake and cancer-associated metabolic rewiring in cachexia remains poorly defined. Clarifying this relationship is essential for identifying the fundamental drivers of cachexia and for developing effective therapeutic strategies. MethodsWe assessed metabolic rewiring by glucose tolerance test and isotopic tracers to determine muscle insulin-stimulated glucose uptake in male cachectic and non-cachectic C26- and KPC-tumor-bearing, as well as mice towards C26 cachectic mice. ResultsCachectic C26-tumor-bearing mice displayed reduced body weight, lean, and fat mass, and food intake (-20%, -15%, -75%, -40%, respectively). Cachectic C26- and KPC-tumor mice showed improved glucose tolerance compared to non-cachectic mice, correlating inversely with tumor size. Ex vivo insulin-stimulated glucose uptake was elevated in soleus (+78%) and extensor digitorum longus (+35%) muscle from cachectic C26-cancer mice compared to non-cachectic and control mice. This increase was associated with enhanced AKT signaling. This was phenocopied in pair-fed non-tumor-bearing mice to match the food intake of cachectic mice, where glucose tolerance, insulin-stimulated glucose uptake ex vivo, and AKT signaling were all enhanced by food restriction. ConclusionsOur findings suggest that enhanced skeletal muscle insulin responsiveness in cachectic tumor-bearing mice is due to anorexia-induced adaptations, highlighting AKT signaling as a key node connecting nutrient status to muscle glucose metabolism in cancer cachexia. HighlightsO_LIC26 and KPC cancer-induced weight loss (cachexia) increases glucose tolerance in mice C_LIO_LIInsulin responsiveness is increased in cachectic, but not in non-cachectic, tumor-bearing mice. C_LIO_LILowered food intake drives elevated muscle insulin responsiveness in cachectic mice C_LI

molecular biology↗

Comparative profiling of SARS-CoV-2 variant infections reveals diverse impacts on host cell RNA and RNA binding protein distribution and regulation

RNA viruses perturb cellular RNA metabolism to promote viral transcript expression and genome replication. However, the extent of host cell RNA reprogramming and the consequences on RNA binding protein (RBP) expression and localization remain poorly understood. Here, we utilized an integrated set of transcriptomics, proteomics, and high-content imaging approaches to compare the impact of different SARS-CoV-2 variants, including an early viral isolate and later emerging variants of concern (Alpha, Delta and Omicron) on the host cellular RNA and RBP landscapes. While all variants deeply remodelled the host transcriptome in a subcellular compartment-specific manner, we found that Omicron variants express lower levels of viral RNA products compared to earlier variants, while still driving efficient production of viral proteins. The surge in viral RNA expression correlated with altered subcellular RNA biotype distribution, cytoplasmic mRNA exclusion, the selective disruption of the localization of polyA-tail interacting proteins, and the nuclear accumulation of non-coding transcripts such as RN7SL family members and MALAT-1. Unexpectedly, we found many RBPs that exhibiting altered subcellular localization following abiotic innate immune stimulation with poly(I:C) but remained unaltered in cells infected with SARS-CoV-2 variants, revealing a capacity for these viruses to limit the overall impact on the RBP subcellular distribution. Our results further indicate that Omicron infection sustains higher expression levels of proteins components of the signal recognition particle and translation factors, which may enable a more effective production of immune response and viral components. Altogether, this work highlights the deep impact of SARS-CoV-2 infection on host cell RNA regulatory landscape and provides new insights to explain the differential impact of Omicron compared to early SARS-CoV-2 variants. HighlightsO_LISARS-CoV-2 variants drive varying degrees of viral RNA expression in infected Calu-3 cells C_LIO_LIInfection reshapes the host cell RNA landscape in a cell-compartment specific manner C_LIO_LIRN7SL RNA exhibits a strong increase in relative abundance and accumulates in the nucleus of infected cells C_LIO_LISARS-CoV-2 infections impact the localization of polyA-tail binding proteins while blocking the nuclear accumulation of immune response-sensitive RBPs C_LIO_LIOmicron infections are less disruptive to cellular function and sustain the production of translation factors and components of the SRP complex C_LI

molecular biology↗

Quantitative Substrate Kinetics Screening: One-Pot LC/MS Based Approach to Map Protease Specificity

Proteases play critical roles in many biological processes and diseases. Proteases tend to have limited and overlapping specificities, which hinders our understanding of their roles in physiology. We designed a naive protease substrate library with the aim of fully utilizing modern mass spectrometry proteomics tools to quantitatively map protease substrate specificity. A library of approximately 200,000 peptides was efficiently produced in E. coli with sufficient library diversity to identify cleavage efficiency of thousands of substrates for most proteases in a single assay. Each experiment results in ample substrate kinetics data to build a positional specificity matrix that quantitatively maps protease specificity. We apply this method to two different proteases, GluC commonly used as a proteomics tool, and Fibroblast Activation Protein Alpha that is overexpressed in activated fibroblasts of epithelial cancers and fibrotic diseases. This research quantitatively maps substrate specificity for GluC and FAP in a single assay and can be applied to map substrate specificity for most proteases. Graphical Abstract O_FIG O_LINKSMALLFIG WIDTH=122 HEIGHT=200 SRC="FIGDIR/small/680748v1_ufig1.gif" ALT="Figure 1"> View larger version (44K): org.highwire.dtl.DTLVardef@1496d11org.highwire.dtl.DTLVardef@1979fc1org.highwire.dtl.DTLVardef@17967e8org.highwire.dtl.DTLVardef@194b741_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

Integrative Spatial Omics for Systems-Level Mapping of Pathological Niches

Spatial omics technologies are a powerful tool for mapping the relationship between cellular organization and molecular distributions in healthy and diseased tissue microenvironments. Here, we describe a novel multimodal pipeline that represents experimental and computational advances for spatiomolecular analysis of tissue samples across molecular classes. This adaptable method integrates matrix-assisted laser desorption/ionization imaging mass spectrometry spatial lipidomics, spatial transcriptomics, protein imaging via multiplexed immunofluorescence microscopy, and histopathological staining to uncover spatiomolecular profiles associated with unique cellular niches and pathological features. We demonstrate the power of this approach using two different complex human disease systems: Alzheimers disease in human brain tissue and type 2 diabetes mellitus in the human pancreas. This work establishes and demonstrates a generalizable framework for multimodal spatial integration, enabling precise mapping of molecular mechanisms that underlie complex tissue pathologies.

systems biology↗

Streamlining Spatial Transcriptomics for Human Kidney Tissue

Single-cell spatial technologies have emerged in recent years, enabling tissue architecture and organization characterization at unprecedented resolution. However, computational analysis of spatial transcriptomics data is often a bottleneck for scientific discoveries in the absence of a dedicated bioinformatician. Here, we describes a workflow to annotate cell types from a dataset generated using NanoStrings CosMx single-cell resolution spatial transcriptomic technology, enabling a comparison between healthy kidney biopsies and diseased tissue. We validated our pipelines accuracy with both gene expression analysis and pathological changes associated with biopsy-proven diabetic kidney disease (DKD). Through precise cell type annotation, we observed significant changes in the proportions of podocytes and immune cells in DKD, with DKD tissue showing regional enrichment of immune cells and differential gene expression. Notably, injured proximal tubules had the expected increased expression of HAVCR1 and VCAM1 and genes associated with diabetes, including IL18, ITGA3 and ITGB1. The entire workflow, now fully integrated into the BioTuring SpatialX (Lens V2.0), is available as a platform designed for users with no formal bioinformatics training.

bioinformatics↗

Statistical Causal Discovery in Developing and Refining Adverse Outcome Pathway (AOP)

Statistical causal discovery (SCD) has the potential to advance the development and evaluation of Adverse Outcome Pathways (AOPs) by inferring causal relationships directly from data. However, ecotoxicology data often has challenges for SCD applications, such as missing data and violation of SCD algorithm assumptions. As a proof-of-concept, we applied a linear non-Gaussian acyclic model (LiNGAM), a representative SCD method, to three types of ecotoxicology datasets: (1) bivariate dose-response relationships, (2) bivariate response- response relationships, and (3) a multivariate dataset with a known causal structure. Missing data were addressed through multiple imputation followed by causal estimation using DirectLiNGAM, a direct method for estimating LiNGAM. DirectLiNGAM identified correct causal directions with high statistical reliabilities in three of four bivariate dose-response cases, even when assumptions such as linearity and non-Gaussianity were partially violated. In contrast, response-response cases did not yield a single dominant direction, likely due to the limited number of replicates. In the multivariate case, the inferred graphs closely resembled the expert-curated causal graph, achieving high recall (0.50-0.75), despite relatively low precision (0.31-0.40). These results demonstrate the utility of SCD, combined with multiple imputation, in identifying relevant key events, revealing missing links, and refining existing AOP and quantitative AOP (qAOP) models, under realistic ecotoxicological constraints. SynopsisStatistical causal discovery can advance the development of adverse outcome pathways in a data-driven manner, enabling efficient chemical risk assessment. TOC Art O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=165 SRC="FIGDIR/small/672289v1_ufig1.gif" ALT="Figure 1"> View larger version (54K): org.highwire.dtl.DTLVardef@117bd64org.highwire.dtl.DTLVardef@19318cdorg.highwire.dtl.DTLVardef@414aecorg.highwire.dtl.DTLVardef@9e068e_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

A translatable evaluation tool to study the biodistribution of clinically-available doxorubicin liposomes: PET imaging of Zr-Talidox

INTRODUCTIONDoxil/Caelyx is a PEGylated liposomal formulation of the chemotherapeutic doxorubicin used in the clinic for Kaposis sarcoma, advanced ovarian cancer, progressive multiple myeloma and metastatic breast cancer. Talidox(R), a smaller doxorubicin PEGylated liposome is undergoing clinical trials and has been proposed as an improvement on previous liposomal formulations for the treatment of advanced solid tumors. We aimed to validate an easily translatable radiolabeling method using zirconium-89 (89Zr) that enables quantitative whole-body PET imaging of these formulations to study their biodistribution and pharmacokinetics. METHODS[89Zr][Zr(oxinate)4] was produced using a kit-based approach followed by use as a direct radiolabeling agent of the liposomal formulations. DFT studies were performed to elucidate the mechanism behind the radiolabeling stability observed within the liposomes. Purified 89Zr-labelled Doxil/Talidox(R) liposomes (5 mg/kg doxorubicin dose) were administered in female BALB/c mice bearing 4T1 tumors. PET/CT imaging was acquired at 20 min, 24 h, 48 h, and 72 h, followed by post-mortem biodistribution at 72 h. RESULTS and DISCUSSIONBoth formulations were radiolabeled efficiently with high stability in serum in vitro for 72 h. In vivo, both formulations showed high tumor uptake at 72 h (18.5 {+/-} 2.4 % IA/g for Doxil and 20.2 {+/-} 2.3 % IA/g for Talidox). In general, ex vivo biodistribution showed similar uptake values for both formulations with high spleen/liver uptake and low bone uptake, confirming stability. Talidox(R) showed significantly lower spleen uptake and higher uptake in bone than Doxil. DFT studies confirmed that doxorubicin can form complexes with 89Zr that are more stable than [89Zr][Zr(oxinate)4], explaining the radiolabeling mechanism and stability results in vitro and in vivo. CONCLUSIONSClinically available PEGylated liposomes containing doxorubicin can be efficiently radiolabelled with 89Zr for PET imaging studies, using a clinically translatable radiolabelling method. HighlightsO_LIDoxorubicin-containing liposomes can be labeled with the positron-emitting radionuclide 89Zr with no impact on their original physicochemical properties. C_LIO_LIRadiolabeling is stable in vivo and enables imaging and biodistribution studies of the liposomes using positron emission tomography (PET). C_LIO_LIThe radiolabeling method is clinically translatable and would allow early assessment of existing and novel doxorubicin liposome biodistribution in humans or personalized medicine (nanotheranostic) approaches. C_LI

bioengineering↗