Biology subjects

Kelly, C.

Publications and source records attributed to Kelly, C..

At least 19 recordsLinked to original sources

bioRxiv2026

Cross-state stability and maturational similarity of the social brain connectome in children and adolescents

The intra-individual stability of functional connectivity (FC) across two or more scans and the inter-individual similarity of FC patterns relative to peers capture developmental variability in functional brain organization. Despite their different referents, these measures are strongly coupled at the whole brain level; however, it remains unclear whether this relationship emerges in development, whether it extends to the social brain, and whether it differs by sex. Based on the importance of social development in this age range, we hypothesized that known developmental relationships and sex-specific effects would be more pronounced in the social brain relative to the whole brain. Using a large psychiatrically enriched sample of 769 youth ages 6-21 (female N = 327, mean age = 12.27 years), we computed FC during rest and movie-watching scans and quantified stability across scans as well as similarity to mature subjects (n=36) within each condition. We then tested whether sex-dependent age relationships were captured by these measures or by the strength of their coupling. Our results showed that cross-state stability and maturational similarity are strongly coupled in development, and that this coupling is enhanced in the social brain relative to the whole brain. Furthermore, the strength of stability-similarity coupling increased as a function of age in females, while males showed the opposite effect. When assessing relationships between age and each FC measure independently, no individual effects were significant after FDR correction. However, our multivariate age-prediction models fared much better, demonstrating that edgewise stability and maturational similarity measures can capture spatially heterogeneous developmental effects that are lost when collapsing across edges. Together, these results support the emerging idea that the degree of FC stability across scans is related to how closely an individuals functional architecture aligns to a shared template. Here, we newly show that this stability-similarity coupling is stronger in the social brain and varies with age in sex-specific ways. Future longitudinal work is needed to establish whether these cross-sectional relationships also characterize within-person developmental change.

neuroscience↗

bioRxiv2026

Reply to: Structural polymorphism of amyloid fibrils in ATTR amyloidosis revealed by cryo-electron microscopy

In the paper by Nguyen et al, published in Nature Communications in 2024, we describe structural polymorphism in fibrils extracted from patients with ATTR amyloidosis. ATTR amyloidosis is characterized by deposition of transthyretin amyloid fibrils across multiple organs, resulting in marked phenotypic variability. Previous studies on ATTR suggest its phenotypic variability may be linked to differences in fibril structure, as demonstrated in amyloid diseases such as AA amyloidosis, tauopathies, or synucleinopathies. Thus far, most ATTR fibrils adopt a strikingly similar fold across patients and affected organs. The most notable exception is described in the study by Nguyen et al., where we report the identification of two co-existing polymorphs in cardiac fibrils from three patients with the ATTRv-I84S variant. These cardiac polymorphs differ in conformation of the segment Gly57 to Gly67, termed the gate, which regulates access to a central polar channel within the fibril core. In one polymorph, the gate adopts an open conformation, exposing the channel, whereas in the other it is closed, shielding it. Whether these polymorphs extend beyond cardiac tissue remains unknown. Here, we address this question by extending our analysis to additional organs from a fourth patient with ATTRv-I84S amyloidosis.

biophysics↗

bioRxiv2026

Decoupled evolution of antimicrobial repertoires in lichen-forming fungi

Lichens are stable multipartite symbioses in which a fungal mycobiont associates with one or more photosynthetic partners while hosting complex microbiomes. Secreted antimicrobial proteins (AMPs) have recently emerged as important mediators of fungal interactions with surrounding microbial communities, yet their evolutionary distribution and diversity in lichen-forming fungi remain largely unexplored. Here, we predicted AMP repertoires across 80 phylogenetically diverse lichen-forming fungi representing three photobiont association types. Contrary to our expectation, average repertoire size did not differ between chlorolichens, cyanolichens and tripartite lichens (363 AMPs per species overall), even though repertoire size varied almost five-fold among species and 23 AMP families showed photobiont-associated enrichment or depletion. This quantitative conservation masks extensive compositional turnover, with families occurring in at least 90% of species coexisting alongside numerous lineage-specific groups. Pooled population sequencing of 11 Umbilicaria phaea populations along two elevational gradients further revealed recurrent AMP presence-absence variation within a single species. In contrast to AMPs, biosynthetic gene clusters (BGCs) and carbohydrate-active enzymes (CAZymes), which include known antimicrobials, were markedly reduced in cyanolichens relative to chlorolichens. Lichen-forming fungi thus combine a conserved baseline antimicrobial protein capacity, whose composition nonetheless turns over between lineages and populations, with chemical and enzymatic systems that differentiate along photobiont association and may support ecological and lifestyle-associated adaptation.

microbiology↗

bioRxiv2026

A chromosome-scale super-pangenome of the lichen genus Peltigera reveals genome architecture and expanded interaction repertoires shared across pathogenic and mutualistic fungi

Fungi engage in associations with other organisms across a continuum from pathogenic to mutualistic lifestyles. Hence, they require a compendium of molecular capacities, including partner recognition, extracellular signaling, nutrient exchange, immune modulation, and control of microbial competitors. In filamentous pathogens such traits are frequently associated with compartmentalized genomes, including rapidly evolving secreted proteins known as effectors and expanded receptor families, but it remains unclear whether similar genomic principles shape mutualistic fungal symbioses. Here, we generated a chromosome-scale super-pangenome for the lichen-forming genus Peltigera, comprising 41 mycobiont assemblies representing eleven species, together with genomes of associated Nostoc and, in tripartite species, Coccomyxa photobionts. The mycobiont genomes revealed extensive variation in genome size, transposable element content, biosynthetic gene clusters, and lineage-specific gene content, with pronounced expansions in tripartite species. Across Peltigera, secreted protein encoding genes were preferentially located in TE-rich regions. We further identified Starship-like transposon elements, expanded antimicrobial protein repertoires, and a large, previously underestimated repertoire of fungal GPCRs dominated by Pth11-like receptors. Layer-specific transcriptomics of a P. rufescens thallus showed differential expression of several interaction-associated gene families, e.g. lectins, antimicrobial proteins and Pth11-like GPCRs. These data indicate that pathogenic and mutualistic fungi might exhibit shared genomic principles, including genome compartmentalization, mobile-element-associated diversification, and the expansion of molecular repertoires involved in recognition, extracellular control and signaling.

genomics↗

bioRxiv2026

Individualised mapping of living human brain mitochondria by MRI reveals signatures of bioenergetic defects.

Mitochondria support the bioenergetic processes that enable brain function and cognition, but we have lacked a label-free, non-invasive approach to explore how brain mitochondria are linked to ageing, disease, and cognition in humans. A recently introduced MitoBrainMap neuroimaging framework predicts mitochondrial features from magnetic resonance data alone, potentially bridging cellular biology with macroscale brain organization. Here, we tested whether this framework captures meaningful age- and pathology-related mitochondrial variation. Consistent with existing literature, we find that MR-predicted mitochondrial density and tissue respiratory capacity consistently declined with age, whereas mitochondrial respiratory capacity-an index of mitochondrial quality-was relatively preserved across the lifespan. Moreover, the relations among specific mitochondrial features predicted from our algorithm were consistent with their biological organization, supporting preliminary construct validity for MR-predicted mitochondrial features. In patients with rare mitochondrial diseases, predicted maps revealed region-specific alterations in mitochondrial density and respiratory chain components, particularly the expected compensatory upregulation of complex II, but not of other mitochondrial genome-encoded components. Finally, the MR-based mitochondrial features were associated with the energetic stress marker GDF15 measured in blood, as well as with cognitive performance measures, linking the novel predictions of brain mitochondria to systemic stress and behavior. These findings introduce a first-generation, label-free, neuroimaging-based mitochondrial mapping as a non-invasive window into living human brain mitochondria.

neuroscience↗

bioRxiv2026

Longitudinal consensus clustering reveals the functional architecture of the developing rat brain

Resting-state fMRI-based mapping of functional networks in developing human populations holds promise for enhancing the diagnosis and treatment of psychiatric disorders, which often emerge during adolescence. Preclinical models offer experimentally tractable lifespans to investigate these trajectories longitudinally, while also enabling direct experimental manipulations that can reveal causal influences on brain and behavioural outcomes. However, unlike in the human neuroimaging field, preclinical neuroimaging often lacks standardisation, making comparison across studies and translation of findings more difficult. In particular, we lack developmentally informed functional brain atlases. Working with a longitudinal sex-balanced resting fMRI dataset collected using a standardised protocol, we aimed to identify robust functional networks in the rat brain and investigate how they change over development (P28, P35, P49, P70 and P91). Employing a consensus clustering approach informed by cluster quality metrics, we mapped functional network development of the rat brain across multiple spatial scales (k=3, k=5 and k=7) from juvenile (pre-puberty) to early adulthood. Force-directed spring embeddings and graph metrics showed global shifts in brain organisation; with increased global system segregation over time, while global network integration (participation coefficient) decreased. At the network-level, the rat brain was characterised by functionally heterogenous network maturation that recapitulated sensorimotor - higher cognitive gradients of development observed in humans. Longitudinal mapping over the adolescent period revealed both linear and non-linear developmental trajectories, and shows that maturation of the rat brain is characterised by network fractionation and delayed cortical specialisation, with the brain transitioning from globally interconnected juvenile networks into functionally segregated adult networks. The functional architecture we describe offers a generalisable structure and nomenclature for the preclinical imaging community, analogous to the Yeo 7-network parcellation in humans. By openly sharing all data, analytic resources, and outputs with the community, this work provides a developmentally informed functional atlas developed under a standardised protocol for anaesthesia and data acquisition and offers resources that will enable preclinical researchers to further advance our understanding of brain development during this critical period.

neuroscience↗

bioRxiv2026

A sex-balanced longitudinal developmental task-free fMRI rat dataset.

Here we present a preclinical longitudinal developmental dataset spanning pre-puberty (juvenile) to early adulthood in Wistar rats. Thirty-six rats (18 female) were scanned during five sessions at approximately postnatal day 28, 35, 49, 70 and 91. A standardised consensus protocol was used for animal sedation and MRI data acquisition (structural MRI and resting-state fMRI). The dataset also includes daily body weight measurements, day of puberty onset, and daily cytological smear images for oestrous tracking. This openly available dataset addresses the need for sex-balanced developmental preclinical datasets spanning puberty and enables researchers to investigate longitudinal brain functional developmental trajectories and Sex As a Biological Variable (SABV).

neuroscience↗

bioRxiv2026

Transthyretin amyloid fibrils adopt distinct folds in the brain

Amyloid deposition in the central nervous system is increasingly recognized in transthyretin (ATTR) amyloidosis, particularly in patients with prolonged survival following liver transplantation or disease-modifying therapies. However, the structural basis of transthyretin aggregation in the brain remains unknown. Here we determine cryo-electron microscopy (cryo-EM) structures of ex vivo brain-derived ATTR fibrils from patients carrying the ATTRv-V30M and ATTRv-V30G variants. Both fibrils adopt folds distinct from those previously reported in peripheral tissues and the vitreous humor. V30M fibrils exhibit a continuous ordered core spanning residues Pro11-Asn124, whereas V30G fibrils consist of a substantially reduced ordered core, revealing pronounced structural divergence even within the same tissue environment. Despite this diversity, comparative analyses identify conserved regions across ATTR fibrils, including a segment implicated in transthyretin aggregation and targeted for diagnostic and therapeutic development. These results provide direct evidence that local tissue context can shape amyloid fibril architecture in human disease.

biophysics↗

bioRxiv2026

REVOLUTA regulates cell fate and wall patterning in the fruit endocarp

Explosive seed dispersal distinguishes Cardamine species from Arabidopsis and depends on polarized secondary cell wall (SCW) deposition in fruit endocarp b (endb) cells. How this SCW pattern is specified and environmentally modulated remains unclear. The polyploid Cardamine chenopodiifolia produces explosive aerial fruit and non-explosive subterranean fruit, creating a tractable system to address this problem. We show light triggers underground fruit to explode by reprogramming endb SCW patterning from uniform to polar. We identify the HD-ZIPIII transcription factor REVOLUTA as a central regulator of endb cell fate, SCW formation, and organ polarity in Arabidopsis and Cardamine hirsuta. In C. hirsuta, duplicated REVOLUTA paralogs are required for endb SCW deposition, while other HD-ZIPIII genes contribute redundantly to cell fate and organ polarity. REVOLUTA over-expression converts polar endb SCWs to uniform, producing non-explosive fruit. Together, these findings reveal a tunable developmental module underlying evolutionary transitions between explosive and non-explosive seed dispersal strategies.

plant biology↗

bioRxiv2026

Germ-free piglets display variable neuroinflammatory-like perturbations in prefrontal cortical microglia

Communication between gut microbiota and immune cells within the brain is essential for neurotypical development. Specifically, microglia are known to play a key role in regulating and supporting neural progenitor stem cell production during brain development, and are sensitive to changes in the maternal gut microbial composition during perinatal development. Here, we employed a germ-free (GF) porcine paradigm to examine how the absence of the microbiome affects microglial dynamics during a key epoch of brain development. We utilized automated software to evaluate microglial density and morphology across three developmentally significant regions: the ventricular/subventricular zone (VZ/SVZ), the prefrontal subcortical white matter (PFCSWM), and layers II/III of the prefrontal cortex (PFCII-III). We found no significant differences in microglial morphology or density in the VZ/SVZ or PFCSWM. In contrast, the PFCII-III of P16 piglets exhibited an increase in microglia density paired with morphologies indicative of an activated/reactive functional state. Notably, these effects were identified with no overall changes in microglial density in any of the regions assessed. Transcriptomics on RNA isolated from the PFCII-III revealed a significant upregulation of genes related to neuroinflammation, in agreement with a region-specific microglial and immune response in the absence of microbial colonization during postnatal development. Together, these findings build on the limited knowledge available on how microbiota influence brain development in large animal model organisms with high similarities to human brain anatomy and developmental trajectories. Significance StatementThe prefrontal cortex of porcine display unique, ramified microglia which are sensitive to germ-free conditions whereby they display alterations in morphology with a more transcriptionally reactive signature. These findings indicate that microglia are regionally sensitive to stimuli in the periphery, and studies in lissencephalic mammalian models may not be directly correlative to other higher-order species. The neuroanatomical heterogeneity of microglia across species is informative and understudied, but necessary, to draw conclusions on the array of perturbations spanning neurodevelopmental trajectories in health and disease.

neuroscience↗

bioRxiv2026

Historical ecology and stakeholder perspectives can inform peatland fire management

Climate change is increasing wildfire risk globally and peatlands have become increasingly vulnerable to fire in recent decades. We combine social research methods with analysis of historical ecological (palaeoecological) records to understand links between fire, climate, vegetation and land use. Stakeholders in the Peak District (UK) highlight the need for scientific research and local knowledge to be more frequently embedded into policy. Analyses of historical ecological datasets reveal regime shifts in moorland vegetation following periods of fire activity and managed burning. This type of disturbance can lead to dominance of grasses, which may have a negative impact on peatland carbon balance under warmer climatic conditions. Recent fires are contributing to the loss of Sphagnum moss and greater dominance of heath and grass. Restoring peatlands, through re-establishing native woodland and improving peat bog hydrological conditions, alongside careful planning around controlled burning, are key measures to enhance resilience to future fire events.

ecology↗

bioRxiv2026

Conserved sequence elements in the final exon of MDM2-eight-exon skipping event reveal a cassette regulon model of alternative splicing controlled by a distal regulatory element.

Modern sequencing technologies have revealed that cancers exhibit widespread dysregulation of alternative splicing, which plays a significant role in driving tumor hallmarks. Alternative splicing enables a single gene to generate multiple mRNA variants, a fundamental mechanism that shapes the distinctive characteristics of normal cells as well as cancer cells. These splicing events are governed by complex interactions between cis- and trans-acting factors and understanding these mechanisms is crucial for effectively targeting cancer cells. This study highlights a unique splicing event in which genotoxic stress induces the skipping of eight-exons in the proto-oncogene MDM2, resulting in the formation of MDM2-ALT1, an isoform that is overexpressed in various cancers. To effectively target this splicing event, it is important to unfold the regulatory mechanism behind it. We hypothesize that the event occurs either by the autonomous regulation of individual exons (Exon autonomous model), where each exon is regulated independently or by the coordinated exclusion of an eight-exon-regulon unit (Exon Regulon model). Utilizing in-silico tools, a comprehensive modular minigene system, CRISPR-mutated cell line, and murine models, we demonstrate that this complex MDM2 splicing event is regulated by sequences within a distal terminal exon, which emerge as critical regulators, orchestrating the alternative splicing of MDM2. Constitutive expression of the Mdm2-MS2 isoform (the mouse ortholog of the human MDM2-ALT1 splice variant), achieved by mutating the Srsf2 binding-site on exon 11, modulates proliferation and apoptosis dynamics in NIH3T3 cells. Moreover, in a p53-wildtype in-vivo setting, this isoform confers a protective effect against age-induced neoplasia. Our findings support the Exon Regulon model of MDM2 splicing, regulated by distal elements analogous to distal enhancer elements that control transcription. These finding sheds light on intricacies in the splicing code that could have significant implications for developing splice variant targeting cancer therapies.

molecular biology↗

bioRxiv2026

A motif-vocabulary model of CAR T-cell intracellular domains identifies determinants of immunophenotype differentiation

Chimeric antigen receptor (CAR) T-cell efficacy depends critically on the costimulatory domain, which shapes downstream signaling and the immunophenotype of manufactured products. Despite mechanistic evidence that immune receptors function as motif-based signaling scaffolds, CAR engineering has focused on a narrow set of costimulatory domains--principally CD28 and 4-1BB--leaving much of the available signaling design space unexplored. Here, we screened 1,243 naturally occurring intracellular domains as costimulatory modules in an anti-CD20 CAR backbone in primary human CD8+ T cells and quantified construct enrichment across memory-differentiation and PD-1-defined immunophenotypic compartments. Using Eukaryotic Linear Motif (ELM) annotations, we analyzed motif-phenotype associations via complementary statistical approaches: Mann-Whitney screening and negative binomial regression identified ELM features associated with differential construct representation, while Dirichlet-Multinomial modeling--which properly accounts for the compositional structure of FACS-partitioned data--revealed that individual ELMs do not significantly alter phenotype distributions. This discrepancy indicates that single motifs primarily affect proliferation or survival rather than differentiation fate. In contrast, construct-level analysis using a leave-one-out compositional test identified specific costimulatory domains with significant phenotype-shifting effects, demonstrating that particular combinations of ELMs--rather than individual motifs--determine immunophenotype. These results suggest that CAR T-cell differentiation state is governed by the integrated output of multiple signaling motifs and provide a combinatorial framework for rational costimulatory domain engineering.

bioengineering↗

bioRxiv2026

Overmaturation as a major trajectory of naive CD4 T cell aging

Aging is associated with progressive changes of cellular functional states, but whether these changes arise from novel programs or from distortion of normal developmental trajectories often remains unclear. Here, we investigate this question in the context of human immune aging by analyzing age-associated changes in CD4 T cells at single-cell resolution. We show that CD4 T cell subsets accumulate senescence-associated features at different rates and that the naive CD4 T cell compartment is highly heterogeneous with respect to senescence markers. Strikingly, transcriptional changes associated with aging of naive CD4 T cells parallel those observed during normal post-thymic maturation from recent thymic emigrants (RTE) to mature naive cells. This similarity defines a transcriptional aging trajectory that we term overmaturation, characterized by exaggerated execution of a physiological maturation program. This overmaturation is consequential for T cell function, as we demonstrate through perturbation of transcription factor TOX, which is predominantly expressed in RTE and whose expression decreases with age. Together, our findings identify transcriptional overmaturation as a major trajectory of naive CD4 T cell aging and suggest that aging-associated distortion of developmental programs can strongly contribute to immunosenescence.

cell biology↗

bioRxiv2025

Translation control by altered start codon usage as a means of modulating the general stress response and virulence in Listeria monocytogenes.

In the food-borne pathogen Listeria monocytogenes, SigB is the central regulator of general stress response (GSR) and it mediates host entry by promoting acid resistance and epithelial cell attachment. The post-translational regulation of SigB activity is complex and is controlled by multiple genes. There is significant evidence that mutations can readily arise in these regulatory genes, or in sigB itself, leading to reduced SigB activity, which suggests that there is considerable genetic plasticity In the GSR. To further investigate this, we defined the complete genome sequence of a clinical isolate with attenuated SigB activity and investigated how it adapts to lethal acidic challenge (mimicking the selective pressure encountered during entry into the host). Acid resistance developed rapidly and 6 acid resistant derivatives (ARDs) were selected for further investigation. All 6 ARDs restored SigB activity due to mutations acquired in rsbW, which encodes an antagonist of SigB. These mutations resulted in non-canonical start codons (rsbWATG to rsbWATA or rsbWATT) or premature translation termination (rsbW-). A translational reporter assay demonstrated distinct differences in translation efficiency between three start codons: ATG>ATA>ATT, suggesting that a perturbation of RsbW:SigB stoichiometry alters SigB activity. We then analysed start codon usage for all conserved genes in 60,692 L. monocytogenes genomes available in the NCBI database. This analysis revealed flexible usage of start codons associated with genetic clades in 39 conserved genes, 13 of which are involved in virulence and stress response. Further, we show that flexible use of canonical start codons (ATG and GTG) also mediates different levels of expression of virulence and stress response genes. Taken together, we show the genetic plasticity of GSR regulation in a model pathogen, and highlight the importance of translational control as a means of fine-tuning gene expression during short-term adaptation and long-term evolution for optimal fitness. Author SummaryThe general stress response (GSR) in foodborne pathogen Listeria monocytogenes is important for environmental stress response and for host entry, but GSR is also highly variable across wild isolates. In this study, we analysed the evolutionary trajectory of a clinical isolate with attenuated GSR and characterized the adaption by this strain to a host-mimicking stress (acidic condition). Under extreme selective pressure, mutations disabling a negative regulator of the GSR were enriched. Interestingly, several independently occurring mutations negatively affect the translation initiation by using non-canonical start codons. Prompted by this, we analysed the population-wide start codon usage by examining all available L. monocytogenes genomes available (n = 60,690). This analysis revealed differential start codon usage in 39 conserved genes that are associated with different genetic clades. Furthermore, we demonstrated differential translation efficiencies between the different canonical start codons. This work highlights the genetic plasticity of GSR in the important food-borne pathogen L. monocytogenes and shows that altered translational start codon can be used as means of regulatory control. Together the data suggest that genetic changes in the regulation of the GSR might confer niche-specific fitness advantages.

microbiology↗

bioRxiv2025

Mitochondrial Energy Transformation Capacity Influences Brain Activation During Sensory, Affective, and Cognitive Tasks

Mitochondria supply the energy required for brain function, but how energetic capacity constrains human brain activity remains unclear. We compared patients with genetically defined mitochondrial disease to matched controls, testing whether bioenergetic stress selectively limits neural processes assessed with functional magnetic resonance imaging during rest and during tasks probing working memory, sensory perception, and affective responses (cold pain). We report impairment in working memory performance and reduced activation of associated brain networks in a subset of patients with the largest clinical and cognitive impairments. A resilient subgroup showed higher working-memory-related BOLD responses, which were associated with preserved task performance. Neural and behavioral effects scaled with the circulating markers of energetic stress growth differentiation factor 15 (GDF15). Brain activity related to sensory and affective responses and resting-state connectivity were comparable for patients and controls. These findings support an energy-hierarchy model in which complex cognition fails first, while sensory processes remain largely preserved under energetic stress.

neuroscience↗

bioRxiv2025

Impaired Complex I dysregulates neural/glial precursors and corpus callosum development revealing postnatal defects in Leigh Syndrome mice

Leigh syndrome (LS) is a complex, genetic mitochondrial disorder defined by neurodegenerative phenotypes with pediatric manifestation. However, recent clinical studies report behavioral phenotypes in human LS patients that are more reminiscent of neurodevelopmental delays. To determine if disruptions in epochs of rapid brain growth during infancy precede the hallmark brain lesions that arise during childhood, we evaluated neural and glial precursor cellular dynamics in a mouse model of LS. Single cell RNA sequencing along with histological and anatomical assessments were performed in NDUFS4 KO mice and compared with controls to determine the impact of Complex I deficiency on neural stem cells, their neuronal and oligodendroglial progeny, lineage progression, and overt differences in specific brain regions. Our findings show disruptions in all categories, specifically within the subventricular zone and corpus callosum. Given that LS is purely considered a neurodegenerative disease, we propose that mitochondrial dysfunction is a neurodevelopmental signature predating classic diagnosis in LS.

neuroscience↗

bioRxiv2025

Novel Lipophagy Inducers as Potential Therapeutics for Lipid Metabolism Disorders

Dysregulation of lipid homeostasis is associated with a wide range of pathologies encompassing neurological, metabolic, cardiovascular, oncological, and renal disorders. We previously showed that lipid droplet (LD) accumulation in podocytes contributes to the progression of diabetic kidney disease (DKD) and reducing LDs preserves podocyte function and prevents albuminuria. Here, we sought to identify compounds that treat pathological LD accumulation. We developed a phenotypic assay using human podocytes and deployed it to screen a combinatorial library comprising over 45 million unique small molecules. This led to the identification of a compound series that effectively reduce LD accumulation in stressed podocytes. Mechanistic studies revealed that these compounds activate lipophagy, reduce LD accumulation, and rescue podocytes from cell death. In contrast, compounds known to induce general autophagy failed to mimic these effects, indicating a novel lipophagy-specific mechanism of action (MoA), which was confirmed with unbiased phenotypic profiling. An advantage of this therapeutic strategy is its potential to not only halt the progression of pathological lipid accumulation but also reverse it. These compounds will serve as tools for uncovering novel drug targets and therapeutic MoAs for treating DKD and other diseases with similar etiologies. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=39 SRC="FIGDIR/small/642628v1_ufig1.gif" ALT="Figure 1"> View larger version (12K): org.highwire.dtl.DTLVardef@16039e2org.highwire.dtl.DTLVardef@7865c3org.highwire.dtl.DTLVardef@60e886org.highwire.dtl.DTLVardef@41db90_HPS_FORMAT_FIGEXP M_FIG C_FIG

cell biology↗