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Biology subjects

Marr, C.

Publications and source records attributed to Marr, C..

At least 19 recordsLinked to original sources

TopoMIL: Topology Improves Multiple Instance Learning in Diagnostic Microscopic Images

Microscopic images of cells and tissues are central to disease diagnosis. In computational pathology, multiple instance learning (MIL) has emerged as a key paradigm for analyzing numerous images within a single patient sample. While the representative distribution of cells in a sample is important for diagnosis, existing MIL frameworks largely overlook it. We introduce TopoMIL, a framework that extracts the representative topological structure of the sample and integrates it into the MIL classifier. Three topological representations are assessed, each with distinct advantages and computational costs. We evaluate TopoMIL on four histopathology and cytomorphology datasets, each presenting unique challenges. Integrating the samples topological information into MIL enhances classification across average, max, attention-based, and transformer pooling, yielding AUCROC gains of 3.3%, 4.2%, 5.9%, and 0.5%, respectively, with moderate computational cost. Our work underscores the potential of TopoMIL as a scalable extension to existing morphology-based models in computational pathology.

bioinformatics↗

BaSiCPy: Scalable and Robust Shading Correction for Optical Microscopy Images

Quantitative fluorescence microscopy is frequently confounded by spatially varying illumination and temporal intensity drift. Although BaSiC is a widely adopted retrospective correction method, it can fail when foreground content is strongly correlated across images--a common regime in time-lapse, tiled and volumetric acquisitions--and its application often requires manual parameter tuning that limits reproducibility and scalability. We introduce BaSiCPy, a foreground-aware implementation of BaSiC that improves illumination profile estimation under correlated foreground structures, provides automatic hyperparameter selection and accelerates large-scale processing through GPU support. BaSiCPy is distributed as an open-source Python package with graphical and programmatic interfaces, facilitating integration into contemporary bioimage analysis workflows.

bioengineering↗

Leveraging Twin Information for Inference of Gene Regulatory Networks

Gene regulatory networks (GRNs) underlie maintenance of cellular phenotypes and responses to stimuli. Modern single-cell profiling methods offer high-throughput datasets to infer GRNs en masse but do not capture dynamic information. Cell-state heterogeneity further confounds correlation-based inference approaches. We addressed these challenges with TwINFER, a conceptual framework leveraging information from recently divided sister cells, or "twins," identifiable via recently developed barcoding techniques. We show that twin information discriminates regulatory from non-regulatory correlations and resolves interaction direction and type (activation/repression). We performed a diverse set of simulations, covering common network motifs and large-scale networks, where TwINFER outperforms state-of-the-art inference capabilities. Crucially, TwINFER resolved the commonly observed false positives in fan-out and feed-forward loop motifs where most methods perform poorly. Lastly, we applied TwINFER to a lineage-barcoded hematopoiesis dataset which refined the network inference, flagged multi-state genes, and determined causal relations. Our work exploits cellular twins as untapped information, readily complementing existing inference approaches.

systems biology↗

EchoVisuALL: From Echocardiography to Gene Discovery

Cardiovascular diseases are a major global health burden, demanding phenotyping frame-works that can match the scale and complexity of contemporary mouse genetics. Here, we introduce EchoVisuALL, an AI-enabled pipeline for automated high-throughput transthoracic echocardiography (TTE) coupling deep-learning-based left-ventricular segmentation with data reporting. Across 65,000 recordings from over 18,000 mice, including single-gene knockouts from the International Mouse Phenotyping Consortium, the framework quantified cardiac morphology and function with minimal operator dependency and high reliability, validated against an expert-curated gold standard dataset. By extracting quantitative parameters across the cardiac cycle, EchoVisuALL in combination with multi-dimensional clustering uncovered nonlinear phenotypic relationships and revealed 37 of 715 genes associated with significant cardiac abnormalities, encompassing well-known human disease genes as well as 12 previously unrecognized candidates, including Cep70, Acot12, Atp8b3, Eea1, Kctd2, and Tspan15. These genotype-phenotype associations are involved in myocardial energetics, membrane biology, and cardiac remodeling. We demonstrate the potential of EchoVisuALL to move beyond image segmentation by delivering a standardized, quantitative foundation for scalable downstream analyses, enabling the discovery of novel cardiac disease genes.

bioinformatics↗

Live high-content imaging with automated analysis reveals mitochondrial changes during vascular calcification

Mitochondrial dysfunction is implicated in a wide range of disorders, including cancer, neurodegeneration, and cardiovascular diseases. Conventional assays typically assess mitochondrial function by measuring bulk respiration rates across thousands of cells. While informative, these approaches cannot resolve the behavior of individual mitochondria, may overlook rare mitochondrial events, and often lack sensitivity to subtle changes. Fluorescence microscopy provides single-organelle resolution but is usually low throughput; for example, imaging 1000 cells can require from 20 minutes to an entire day depending on imaging mode. Additionally, analysis of fluorescence images frequently relies on manually selected thresholds, introducing potential bias and variability due to differences in signal-to-background ratios across cells. Here, we present a High-throughput Mitochondrial Imaging Platform (H-MIP) combined with a deep learning segmentation model, enabling rapid imaging and automated characterization of mitochondrial morphology in millions of mitochondria from tens of thousands of cells. We demonstrate the utility of H-MIP by assessing mitochondrial morphology following treatment with Mdivi-1, an inhibitor of mitochondrial division. Furthermore, we employ an in vitro disease model (vascular calcification) to show that mitochondria become elongated in vascular smooth muscle cells undergoing pathological calcification. Overall, H-MIP provides a scalable and robust tool for investigating mitochondrial structure, with the potential to accelerate therapeutic discovery targeting mitochondrial health. HighlightsO_LIH-MIP enables high-throughput imaging of mitochondrial morphology and automated single-mitochondrion analysis at scale. C_LIO_LIH-MIP links mitochondrial morphology to an established disease model, with exciting implications for drug discovery. C_LI

cell biology↗

Leonardo: a toolset to correct sample-induced artifacts in light sheet microscopy images

Selective plane illumination microscopy (SPIM, also known as light sheet fluorescence microscopy) is the method of choice for studying morphogenesis and function in biological specimens over extended periods, as it permits gentle and rapid volumetric imaging. In inhomogeneous samples, however, sample-induced artifacts, including light absorption, scattering, and refraction, can impact the image quality, particularly as the focal plane gets deeper into the sample. Here, we present Leonardo, the first toolbox designed to address the major sample-induced artifacts by using two modules: (1) DeStripe removes stripe artifacts in SPIM caused by light absorption while preserving fine sample structures; (2) Fuse reconstructs a single high-quality image from dualsided illumination and/or dual-sided detection, while eliminating blur and optical distortions caused by light scattering and refraction. The efficacy of Leonardo is validated on a wide range of biological samples, from minimally invasive experiments on sensitive specimens (translucent embryonic and optically opaque larval zebrafish) to cleared mouse samples up to two centimeters in size. We provide model code and a Napari-based graphical user interface, enabling the SPIM community to easily apply Leonardo to advance light sheet imaging of inhomogeneous and complex specimens.

bioinformatics↗

Predictive value of preclinical models for CAR-T cell therapy clinical trials: a systematic review and meta-analysis

Experimental mouse models are indispensable for the preclinical development of cancer immunotherapies, whereby complex interactions in the tumor microenvironment (TME) can be somewhat replicated. Despite the availability of diverse models, their predictive capacity for clinical outcomes remains largely unknown, posing a hurdle in the translation from preclinical to clinical success. This study systematically reviews and meta-analyzes clinical trials of chimeric antigen receptor (CAR-) T cell monotherapies with their corresponding preclinical studies. Adhering to PRISMA guidelines, a comprehensive search of PubMed and ClinicalTrials.gov was conducted, identifying 422 clinical trials and 3157 preclinical studies. From these, 105 clinical trials and 180 preclinical studies, accounting for 44 and 131 distinct CAR constructs, respectively, were included. Patient[s] responses varied based on the target antigen, expectedly with higher efficacy and toxicity rates in hematological cancers. Preclinical data analysis revealed homogenous and antigen-independent efficacy rates. Our analysis revealed that only 4 % (n = 12) of mouse studies used syngeneic models, highlighting their scarcity in research. Three logistic regression models were trained on CAR structures, tumor entities, and experimental settings to predict treatment outcomes. While the logistic regression model accurately predicted clinical outcomes based on clinical or preclinical features (Macro F1 and AUC > 0.8), it failed in predicting preclinical outcomes from preclinical features (Macro F1 < 0.5, AUC < 0.6), indicating that preclinical studies may be influenced by experimental factors not accounted for in the model. These findings underscore the need for better understanding the experimental factors enhancing the predictive accuracy of mouse models in preclinical settings.

cancer biology↗

Single-cell multiome uncovers differences in glycogen metabolism underlying species-specific speed of development

Embryos from different mammalian species develop at characteristic timescales. These timescales are recapitulated during the differentiation of pluripotent stem cells in vitro. Specific genes and molecular pathways that modulate cell differentiation speed between mammalian species remain to be determined. Here we use single-cell multi-omic analysis of neural differentiation of mouse, cynomolgus and human pluripotent cells to identify regulators for differentiation speed. We demonstrate that species-specific transcriptome dynamics are mirrored at the chromatin level, but that the speed of neural differentiation is insensitive to manipulations of cell growth and cycling. Exploiting the single-cell resolution of our data, we identify glycogen storage levels regulated by UDP-glucose pyrophosphorylase 2 (UGP2) as a species-dependent trait of pluripotent cells, and show that lowered glycogen storage in UGP2 mutant cells is associated with accelerated neural differentiation. The control of energy storage could be a general strategy for the regulation of cell differentiation speed.

developmental biology↗

A scalable computational framework for annotation-independent combinatorial target identification in scRNA-seq databases

The applicability of novel targeted cell therapies is constrained by on-target off-tumor toxicities, posing significant challenges for safe and effective treatment. To develop strategies for mitigating these toxicities, a comprehensive understanding of the distribution of single-cell profiles across human tissues is essential. Single-cell RNA sequencing (scRNA-seq) provides detailed insights into cellular diversity and function. However, as data collection expands and RNA expression atlases comprise tens of millions of cells, effectively querying these databases to identify cell populations matching specific gene expression patterns becomes challenging. Key challenges include managing data volume, handling batch effects, navigating study-dependent data curation and annotation, and addressing the combinatorial complexity of possible target patterns. Userfriendly screening tools that allow for complex query target patterns without relying on existing annotations are currently missing. Here we present otopia, a computational framework that enables fast queries of large scRNA-seq databases, enhancing the precision of cell population identification. Using precomputed neighborhood graphs and boolean logic for expression patterns, otopia enables efficient queries addressing arbitrary combinatorial target patterns. otopia does not depend on cell type annotations and by exploiting unbiased expression pattern matching scores, it uncovers critical populations that annotation based methods overlook. We exemplify otopias capabilities by identifying the largest B cell populations in the CELLxGENE census data using a CD19+CD22+CD5-query target pattern. Moreover, we show that otopia can provide a concise overview of cell populations matching hypothetical acute myeloid leukemia (AML) dual targets, realizing logic AND gates, across different tissues. otopia enables the estimation of ontarget off-tumor toxicities for drugs targeting both single markers and complex marker combinations, surpassing previous methods that focus mainly on single markers. With the expanding availability of scRNA-seq data, otopia represents a scalable, unbiased, and automated approach to refine putative therapeutic targets in order to minimize toxicities.

bioinformatics↗

Cytosolic S100A8/A9 promotes Ca2+ supply at LFA-1 adhesion clusters during neutrophil recruitment

S100A8/A9 is an endogenous alarmin secreted by myeloid cells during many acute and chronic inflammatory disorders. Despite increasing evidence of the proinflammatory effects of extracellular S100A8/A9, little is known about its intracellular function. Here, we show that cytosolic S100A8/A9 is indispensable for neutrophil post-arrest modifications during outside-in signaling under flow conditions in vitro and neutrophil recruitment in vivo, independent of its extracellular functions. Mechanistically, genetic deletion of S100A9 in mice (Mrp14-/-, functional S100a8/a9-/-) caused dysregulated Ca2+ signatures in activated neutrophils resulting in reduced Ca2+ availability at the formed LFA-1/F-actin clusters with defective {beta}2 integrin outside-in signaling during post-arrest modifications. Consequently, we observed impaired cytoskeletal rearrangement, cell polarization and spreading, as well as cell protrusion formation in Mrp14-/- compared to WT neutrophils, making Mrp14-/- cells more susceptible to detach under flow, thereby preventing efficient neutrophil recruitment and extravasation into inflamed tissue. One-sentence summaryintracellular S100A8/A9 is indispensable for firm leukocyte adhesion under flow

physiology↗

PXPermute: Unveiling staining importance in multichannel fluorescence microscopy

Imaging Flow Cytometry (IFC) enables rapid acquisition of thousands of single-cell images per second, capturing information from multiple fluorescent channels. However, the conventional process of staining cells with fluorescently labeled conjugated antibodies for IFC analysis is labor-intensive, costly, and potentially detrimental to cell viability. To streamline experimental workflows and reduce expenses, it is imperative to identify the most relevant channels for downstream analysis. In this study, we present PXPermute, a user-friendly and powerful method that assesses the significance of IFC channels for a given task, such as cell profiling. Our approach evaluates channel importance by permuting pixel values within each channel and analyzing the resulting impact on the performance of machine learning or deep learning models. Through rigorous evaluation on three multi-channel IFC image datasets, we demonstrate the superiority of PXPermute in accurately identifying the most informative channels, aligning with established biological knowledge. To facilitate systematic investigations of channel importance and aid biologists in optimizing their experimental designs, we have released PXPermute as an easy-to-use open-source Python package.

bioinformatics↗

Spatial proteomics in neurons at single-protein resolution

To fully understand biological processes and functions, it is necessary to reveal the molecular heterogeneity of cells and even subcellular assemblies by gaining access to the location and interaction of all biomolecules. The study of protein arrangements has seen significant advancements through super-resolution microscopy, but such methods are still far from reaching the multiplexing capacity of spatial proteomics. Here, we introduce Secondary label-based Unlimited Multiplexed DNA-PAINT (SUM-PAINT), a high-throughput imaging method capable of achieving virtually unlimited multiplexing at better than 15 nm spatial resolution. Using SUM-PAINT, we generated the most extensive multiprotein dataset to date at single-protein spatial resolution, comprising up to 30 distinct protein targets in parallel and adapted omics-inspired analysis workflows to explore these feature-rich datasets. Remarkably, by examining the multiplexed protein content of almost 900 individual synapses at single-protein resolution, we revealed the complexity of synaptic heterogeneity, ultimately leading to the discovery of a new synapse type. This work provides not only a feature-rich resource for researchers, but also an integrated data acquisition and analysis workflow for comprehensive spatial proteomics at single-protein resolution, paving the way for Localizomics.

neuroscience↗

scifAI: Explainable machine learning for profiling the immunological synapse and functional characterization of therapeutic antibodies

Therapeutic antibodies are widely used to treat severe diseases. Most of them alter immune cells and act within the immunological synapse; an essential cell-to-cell interaction to direct the humoral immune response. Although many antibody designs are generated and evaluated, a high-throughput tool for systematic antibody characterization and prediction of function is lacking. Here, we introduce the first comprehensive open-source framework, scifAI (single-cell imaging flow cytometry AI), for preprocessing, feature engineering and explainable, predictive machine learning on imaging flow cytometry (IFC) data. Additionally, we generate the largest publicly available IFC data set of the human immunological synapse containing over 2.8 million images. Using scifAI, we analyze class frequency- and morphological changes under different immune stimulation. T cell cytokine production across multiple donors and therapeutic antibodies is quantitatively predicted in vitro, linking morphological features with function and demonstrating the potential to significantly impact antibody design. scifAI is universally applicable to IFC data. Given its modular architecture it is straightforward to incorporate into existing workflows and analysis pipelines, e.g. for rapid antibody screening and functional characterization.

bioinformatics↗

Label-free imaging of 3D pluripotent stem cell differentiation dynamics on chip

The dynamic chemical and architectural microenvironments of 3D stem cell cultures can be controlled by integration into a microfluidic chip. Massive parallelized 3D stem cell cultures for engineering in vitro human cell types require new imaging methods with high time and spatial resolution to fully exploit technological advances in cell culture. Here, we introduce a label-free deep learning method called Bright2Nuc to predict in silico nuclear staining in 3D from bright-field images obtained using traditional confocal microscopy. Bright2Nuc was trained and applied to several hundred 3D human induced pluripotent stem cell cultures differentiating towards definitive endoderm on a microfluidic platform. Combined with existing image analysis tools, Bright2Nuc segmented individual nuclei from bright-field images, quantified their morphological properties, predicted stem cell differentiation state, and tracked the cells over time. Our methods are available in an open-source pipeline that enables researchers to upscale 3D cell phenotyping in stem cell culture.

bioinformatics↗

BEN: a generalizable Brain Extraction Net for multimodal MRI data from rodents, nonhuman primates, and humans

Accurate brain tissue extraction on magnetic resonance imaging (MRI) data is crucial for analyzing brain structure and function. While several conventional tools have been optimized to handle human brain data, there have been no generalizable methods to extract brain tissues for multimodal MRI data from rodents, nonhuman primates, and humans. Therefore, developing a flexible and generalizable method for extracting whole brain tissue across species would allow researchers to analyze and compare experiment results more efficiently. Here, we propose a domain-adaptive and semi-supervised deep neural network, named the Brain Extraction Net (BEN), to extract brain tissues across species, MRI modalities, and MR scanners. We have evaluated BEN on 18 independent datasets, including 783 rodent MRI scans, 246 nonhuman primate MRI scans, and 4,601 human MRI scans, covering five species, four modalities, and six MR scanners with various magnetic field strengths. Compared to conventional toolboxes, the superiority of BEN is illustrated by its robustness, accuracy, and generalizability. Our proposed method not only provides a generalized solution for extracting brain tissue across species but also significantly improves the accuracy of atlas registration, thereby benefiting the downstream processing tasks. As a novel fully automated deep-learning method, BEN is designed as an open-source software to enable high-throughput processing of neuroimaging data across species in preclinical and clinical applications.

neuroscience↗

Gal1 repression memory in budding yeast

Cells must continuously adapt to changing environments and, thus, have evolved mechanisms allowing them to respond to repeated stimuli. For example, faster gene induction upon a repeated stimulus aids adaptation - a process known as reinduction memory. However, whether such a memory exists for gene repression is unclear. Here, we studied gene repression across repeated carbon source shifts in over 2,500 single Saccharomyces cerevisiae cells. By monitoring the expression of a carbon source-responsive gene, galactokinase 1 (Gal1), and mathematical modeling, we discovered repression memory at the population and single-cell level. Using a repressor model to estimate single-cell repression parameters, we show that repression memory is due to a shortened repression delay, the estimated time gap between carbon source shift and Gal1 expression termination, upon the repeated carbon source shift. Additionally, we show that cells lacking Elp6 display a gain-of-repression-memory phenotype characterized by a stronger decrease in repression delay between two consecutive carbon source shifts. Collectively, our study provides the first quantitative description of repression memory in single cells.

systems biology↗

SHAPR - An AI approach to predict 3D cell shapes from 2D microscopic images

Reconstruction of shapes and sizes of three-dimensional (3D) objects from two-dimensional (2D) information is an intensely studied subject in computer vision. We here consider the level of single cells and nuclei and present a neural network-based SHApe PRediction autoencoder. For proof-of-concept, SHAPR reconstructs 3D shapes of red blood cells from single view 2D confocal microscopy images more accurately than naive stereological models and significantly increases the feature-based prediction of red blood cell types from F1 = 79.0% to F1 = 87.4%. Applied to 2D images containing spheroidal aggregates of densely grown human induced pluripotent stem cells, we find that SHAPR learns fundamental shape properties of cell nuclei and allows for prediction-based morphometry. Reducing imaging time and data storage, SHAPR will help to optimize and up-scale image-based high-throughput applications for biomedicine.

bioinformatics↗

Annotation-efficient classification combining active learning, pre-training and semi-supervised learning for biomedical images

Deep learning based classification of biomedical images requires manual annotation by experts, which is time-consuming and expensive. Incomplete-supervision approaches including active learning, pre-training and semi-supervised learning address this issue and aim to increase classification performance with a limited number of annotated images. Up to now, these approaches have been mostly benchmarked on natural image datasets, where image complexity and class balance typically differ considerably from biomedical classification tasks. In addition, it is not clear how to combine them to improve classification performance on biomedical image data. We thus performed an extensive grid search combining seven active learning algorithms, three pre-training methods and two training strategies as well as respective baselines (random sampling, random initialization, and supervised learning). For four biomedical datasets, we started training with 1% of labeled data and increased it by 5% iteratively, using 4-fold cross-validation in each cycle. We found that the contribution of pre-training and semi-supervised learning can reach up to 25% macro F1-score in each cycle. In contrast, the state-of-the-art active learning algorithms contribute less than 5% to macro F1-score in each cycle. Based on performance, implementation ease and computation requirements, we recommend the combination of BADGE active learning, ImageNet-weights pre-training, and pseudo-labeling as training strategy, which reached over 90% of fully supervised results with only 25% of annotated data for three out of four datasets. We believe that our study is an important step towards annotation and resource efficient model training for biomedical classification challenges.

bioinformatics↗