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Das, A.

Publications and source records attributed to Das, A..

At least 37 records · Page 2Linked to original sources

Functional genomic map of local adaptation in sorghum to guide allele mining

Genomic data from genebanks could be exploited to find alleles adapted to target environments for resilience breeding, but it can be difficult to prioritize among the thousands of accessions and millions of genomic variants. There are competing hypotheses for the molecular basis and architecture of local adaptations: e.g. whether cis-regulatory versus amino acid changing variants are more important; or whether small-effect, low pleiotropy versus large-effect, high pleiotropy variants are more important. Here, we compare a range of variant types and genomic contexts thought to influence effect size, pleiotropy, and selection for their role in local adaptation in 443 whole genome resequenced African sorghum landraces. We used genotype-environment associations (GEAs) as evidence of local adaptation. We found that GEA were particularly enriched in the vicinity of genes and depleted elsewhere. However, enrichment was strongest in likely cis-regulatory contexts: accessible chromatin, unmethylated regions, and in transposable elements close to genes. Near genes, there were clear peaks in GEAs at the transcription start site, where mutations are demonstrated to have the largest expression effects. Additionally, GEAs in accessible chromatin and unmethylated regions were better predictors of genetic variation in response to experimental drought than comparable loci. Having tested hypotheses about the variants underlying local adaptation, we can now use this knowledge of the importance of cis-regulatory variation in the search for new environmentally-adaptive alleles for plant improvement.

evolutionary biology↗

Principles of Local and Global Grouping that Underlie Segmentation of Natural Texture Images

The human visual system segments images using both high-level recognition mechanisms and low-level mechanisms that are largely independent of specific prior experience. The low-level mechanisms are essential for initiating recognition processes, and for learning to recognize new materials, objects, and contexts. Here we describe a hierarchical Bayesian observer (HBO) model of texture segmentation that is biologically plausible, takes into account the statistics of natural scenes, and does not depend on prior experience. The HBO model consists of five steps: local similarity grouping with local normalization, mutual similarity grouping (local grouping is strengthened if the neighboring regions are similar to the same set of other regions), transitive grouping (good continuation), confidence grouping (neighboring regions far from the same-different decision boundary guide grouping of regions near the decision boundary), and region grouping (similarity grouping of the regions from the initial segmentation). We find that a local similarity grouping process, trained to maximize accuracy, predicts human texture discrimination accuracy. We then find that the four additional steps accurately segment images with randomly shaped regions containing arbitrary natural textures. The success of the model depends on all the steps, but especially on local-similarity and transitive grouping. We also find that the transitive grouping allows correct segmentation of non-stationary texture regions (e.g., textures slanted in depth). Further, we find that when illumination varies across the image, local normalization enables both correct texture segmentation and estimation of illumination change. Finally, we find that unlike our model large state-of-the-art deep networks often fail on these stimuli.

animal behavior and cognition↗

Railway Catenary Sparking as a Source of Toxic Copper Ultrafine Particles: Evidence from Realistic In Vitro Inhalation Exposure

Railway catenary sparking generates airborne ultrafine particles (UFPs) that may pose health risks due to their metallic composition and ability to penetrate deep into the alveolar region of the lungs. Copper, widely used in wires and pantographs, is a major component of these emissions, making copper-rich particles common in railway environments such as subways. However, exposure levels and health impacts remain poorly characterized, and localized hotspots may represent an underrecognized risk in densely populated areas. This study investigated the toxicity of copper UFPs under realistic dosimetry and deposition conditions. Copper UFPs were generated using a spark discharge generator and applied to two in vitro lung models: a 3D co-culture of Calu-3 epithelial cells, THP-1-derived macrophages, and EA.hy926 endothelial cells, and a monoculture of A549 alveolar epithelial cells. Cells were exposed at the air-liquid interface (ALI) using an automated platform to mimic inhalation exposure and UFPs deposition. Copper deposition ranged from 6.5 to 41 ng/cm2, within occupationally relevant levels. A549 cells showed cytotoxic responses consistent with previous studies, whereas the 3D co-culture model revealed broader adverse effects, including inflammation, impaired epithelial barrier integrity, oxidative stress, and early DNA damage. Inflammatory activation also differed between models: A549 cells mainly exhibited transcriptional responses, while the 3D model showed significant secretion of IL-6 and IL-8, associated with interferon signaling. These findings highlight the potential health risks of copper UFPs from railway systems and emphasize the need for improved characterization of UFP exposure in environmental and occupational railway settings.

pharmacology and toxicology↗

Herbivory-induced alterations in cytosolic proteins of pigeon pea (Cajanus cajan) leaves

Insect herbivory triggers cytosolic proteome reprogramming by activating defense pathways and modulating key metabolic processes. We found that simulated herbivory in pigeon pea (Cajanus cajan) induced reactive oxygen species (ROS) production and molecular alterations within 12 hours (h) of post treatment. We compared the leaf proteome profiles of two cultivated genotypes, ICPL 332 (moderately resistant) and ICPL 87 (susceptible), using two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) coupled with mass spectrometry (MS). More than 220 protein spots were detected in ICPL 332 and over 200 in ICPL 87. Comparative analysis revealed 75 differentially accumulated proteins (DAPs), of which 40 were consistently reproducible across biological replicates. These included 11 unique to ICPL 87, 9 unique to ICPL 332, and 10 common to both genotypes. Among the shared DAPs, ICPL 332 showed five upregulated and five downregulated, whereas ICPL 87 exhibited only two upregulated and eight downregulated. Functional categorization grouped DAPs into primary metabolism, stress response, and growth and development. Proteins related to primary metabolism were largely downregulated in both genotypes, while stress-associated proteins exhibited substantial downregulation in ICPL 87 compared to ICPL 332. Overall, the results demonstrate proteomic adjustments underlying defense responses in pigeon pea genotypes.

plant biology↗

spatiAlytica: Viewer-Grounded Multimodal Agentic System for Interactive Spatial Omics Analysis

Spatial transcriptomics and proteomics map tissue architecture and cellular interactions, but analysis remains limited by programming demands and text-centered AI agents that lack viewer grounding and cross-turn context. We present spatiAlytica, a viewer-centric multimodal interactive agentic system embedded in the Napari viewer that enables non-programmer biologists to perform iterative, hypothesis-driven spatial omics analysis via natural language. spatiAlytica couples viewer-state serialization, agentic memory, biological concept-to-data-field mapping, code generation and debugging, Spatial VQA, and grounded interpretation to support an exploratory analysis and interpretive reasoning workflow. We introduce spatiAlyticaBench, a comprehensive benchmark spanning 222 single-turn spatial analytical coding questions, 178 multi-turn sequential workflow questions, and 7,350 image-grounded reasoning questions. spatiAlytica outperformed strong agentic baselines, while using less time and tokens. Case studies across Kaposis sarcoma, colorectal cancer, and ovarian cancer recapitulated known spatial patterns and uncovered progressive CD8 T-cell dysfunction during KS progression.

bioinformatics↗

MRI-Based Blood Clot Phenotyping: An In Vitro Study

Background and PurposeNeurointerventional outcomes depend on clot composition and may be influenced by clot contraction. Thus, a priori identification of clot composition and contraction could inform procedural strategies and improve outcomes. The goal of our work is to conduct an in vitro test to determine whether MRI can reliably predict both clot composition and contractile state. Materials and MethodsTo this end, we prepared blood clots spanning clinically observed compositions (0-80% red blood cells (RBCs)) in both contracted and uncontracted states. Contraction was controlled by coagulating blood with or without thrombin. We imaged these clots using quantitative, clinical, and investigational MRI sequences. Using these data, we then determined whether MRI signal intensities, quantitative parameters, and radiomic features capturing intensity and texture patterns can (i) predict clot hematocrit and (ii) classify clots by composition (RBC-rich vs. fibrin-rich) and contraction state. ResultsQuantitative MRI parameters (T1, T2, ADC) decreased with increasing hematocrit (R2 = 0.56-0.85, p < 0.001), while signal intensities from clinical sequences showed weaker correlations (R2 = 0.46-0.62, p < 0.001). Radiomic models predicted hematocrit with performance comparable to MRI parameters. When applied to classification, radiomic features accurately discriminated RBC-versus fibrin-rich clots, with AUCs exceeding 0.90 across nearly all sequences. In contrast, classification of contraction state showed greater variability in AUCs across sequences but remained high for quantitative T1 and T2 values (AUCs up to 0.88). Trends were consistent across clots coagulated with and without thrombin. Pooling features across sequences did not outperform the best individual sequence for either regression or classification. ConclusionsWe demonstrate that MRI-based radiomic analysis quantitatively characterizes clot composition and contraction in vitro. These findings support the feasibility of using MRI for pre-interventional clot phenotyping, with potential to inform thrombolytic and mechanical thrombectomy strategies. Thus, in vivo studies validating these results are warranted.

bioengineering↗

Engineering a Glucose-Inducible Whole-Cell Biosensor via CRISPRi-Based Promoter Reprogramming

Precise monitoring of intracellular glucose dynamics is essential for understanding carbon flux, optimizing microbial bioprocesses, and enabling responsive control of engineered metabolic pathways. Here, we develop a modular whole-cell biosensor in Escherichia coli that converts the native glucose repression phenotype of a CAP-sensitive promoter into a tunable, glucose-inducible output using CRISPR interference (CRISPRi). By placing a guide RNA (gRNA) under the control of the CAP promoter and positioning dCas9 to target the -10 region of a constitutive promoter driving sfGFP, we created an inversion circuit in which glucose suppresses gRNA expression, thereby relieving dCas9-mediated repression and activating fluorescence. Systematic evaluation of gRNA strand orientation and target site selection revealed that template-strand targeting yielded strong repression ([~]90 %) but reduced sensing range, whereas moderately repressive non-template gRNAs ([~]27-35 % repression) enabled optimal signal inversion. The resulting biosensor demonstrated a robust, linear fluorescence response across 200 M -50 mM glucose (R{superscript 2} > 0.97), with high specificity against other sugars and a strong correlation between glucose consumption and fluorescence accumulation (R{superscript 2} {approx} 0.996). To extend the functionality of the platform, we integrated the sensor with a secreted {beta}-glucosidase module that hydrolyzes cellobiose to glucose. The biosensor accurately quantified glucose released during cellobiose degradation, with engineered strains producing up to [~]33 mM glucose from 50 mM cellobiose in a two-plasmid system. This coupling of enzymatic conversion with intracellular sensing enabled real-time, non-destructive monitoring of metabolic transitions. Together, this work establishes a programmable CRISPRi-based strategy for inverting native promoter logic and provides a sensitive, specific, and modular platform for metabolite sensing in bacteria. The approach is broadly applicable for dynamic pathway regulation, monitoring carbon fluxes, and building responsive genetic circuits in metabolic engineering and synthetic microbial ecosystems. TOC O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=116 SRC="FIGDIR/small/717388v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@158eb53org.highwire.dtl.DTLVardef@c1a21eorg.highwire.dtl.DTLVardef@7a87f7org.highwire.dtl.DTLVardef@746e80_HPS_FORMAT_FIGEXP M_FIG C_FIG

bioengineering↗

A domain-general neural signature of serial order memory across action and perception

Remembering events in the correct order, and generating ordered sequences of actions, are fundamental abilities across species. Behavioral studies, and theoretical work, raise the possibility that the brain represents serial order by a domain-general neural code, following the principle of Competitive Queuing. However, direct neurophysiological evidence for Competitive Queuing exists only in the motor domain. When humans and non-human primates prepare for a series of movements, several of the upcoming movements are represented in parallel, with their representational strength reflecting ordinal position in the sequence. We test the generalizability of this so-called primacy gradient across motor sequences and memorized auditory sequences. Using a multivariate decoding approach, Experiment 1 replicated the presence of a Competitive Queuing primacy gradient in magnetoencephalography (MEG) data of young healthy adults (n = 23) when they prepared a sequence of finger movements from memory. Importantly, we observed a similar primacy gradient when participants anticipated a sequence of tones they had learned before, in the absence of any movement. In Experiment 2 (n = 23; naive cohort), we rule out the possibility that this primacy gradient in auditory memory is explained by any learnt association between tones and movements, or by MEG signal fluctuations that are unrelated to discrete sequential events. In sum, we find a similar neural signature of serial order coding when humans prepare a sequence of movements, and when they anticipate a sequence of sounds. This lends support to the generalizability of Competitive Queuing.

neuroscience↗

Structure-Activity Mapping of Intraperitoneal mRNA-LNPs: Decoupling Tumor and Liver Biodistribution in Pancreatic Cancer

Pancreatic ductal adenocarcinoma (PDAC) remains difficult to treat with nucleic acid therapeutics because efficient intratumoral delivery is limited and off-target liver accumulation is common. Here, we developed a structure-activity map for intraperitoneally administered mRNA lipid nanoparticles (mRNA-LNPs) to identify formulation features that improve delivery to pancreatic tumors while reducing liver expression. A full-factorial library of 48 mRNA-LNP formulations was generated by varying ionizable lipid, sterol, phospholipid, and PEG-lipid components. Formulations were characterized for size, polydispersity, zeta potential, and encapsulation, then evaluated in an orthotopic KPC8060 pancreatic tumor model after intraperitoneal administration of firefly luciferase mRNA-loaded LNPs. Biodistribution was assessed by Rhodamine B fluorescence and functional delivery by luciferase expression 12 h after dosing. Lipid composition strongly influenced both physicochemical properties and in vivo performance. G0-C14-based formulations produced the smallest and most homogeneous particles, whereas FTT5-containing formulations were generally larger. Across the 48-formulation library, mRNA expression and nanoparticle biodistribution varied significantly among tumor, pancreas, liver, and spleen. Statistical, decision-tree, and predictive modeling analyses identified composition rules associated with organ-selective delivery. High tumor expression was associated primarily with G0-C14 combined with DSPC and {beta}-sitosterol, whereas liver expression was favored by C12-200 or DLin-MC3-DMA with DOPE and DSPE-PEG. Notably, a G0-C14/DSPC/DSPE-PEG formulation emerged as a lead candidate, producing a greater than 6-fold increase in tumor luciferase signal relative to the library median while reducing liver exposure by approximately 60%. Histopathology showed no treatment-related liver or lung toxicity. These findings define actionable formulation rules for tuning intraperitoneal mRNA-LNP delivery in PDAC and support further development of tumor-selective mRNA therapeutics for pancreatic cancer.

bioengineering↗

STiLE: Automated Tissue Microarray Dearraying for Spatial Transcriptomics

Tissue microarrays (TMAs) enable high-throughput spatial transcriptomic profiling of dozens of tissue cores on a single slide. However, existing dearraying methods operate on histological images and do not support the coordinate-based outputs of spatial transcriptomics platforms. Therefore, task of assigning cells to their respective cores (dearraying) remains a manual bottleneck. We present STiLE, a tool for automated TMA dearraying that operates solely on cell centroid coordinates. By eliminating dependence on image data, STiLE is robust to artifacts such as variable staining quality and uneven illumination. The algorithm combines connectivity-based component detection, density-based clustering (HDBSCAN), component-guided cluster merging, and optional grid-based peak detection. Validation on eleven public TMA samples (50-150 cores, three platforms) achieved ARI > 0.99, while systematic benchmarking on 396 synthetic datasets with realistic artifacts demonstrated consistently robust performance (mean ARI = 0.992). STiLE accepts standard formats (AnnData, CSV) and is platform-agnostic, supporting diverse platforms including Vizgen MERSCOPE, 10x Xenium, and NanoString CosMx. An interactive Streamlit interface enables parameter tuning, visual inspection, and region-based processing for large slides. Availability and Implementationhttps://pypi.org/project/stile; source at https://github.com/Huang-AI4Medicine-Lab/stile Contactyu Supplementary InformationSupplementary data are available at Bioinformatics online

bioinformatics↗

The extracellular matrix gene mec-9 regulates C. elegans sensory cilia

Cilia are critical sensory organelles that project from the cell surface into the tissue environment, where they are surrounded by extracellular matrix (ECM). Abnormal ECM and fibrosis are two hallmarks of ciliopathies, yet the relationship between cilia and ECM is not well understood. Using the sense organs of C. elegans as a model, we found that a neomorphic mutation in the ECM gene mec-9 impacts sensory cilia function, ciliary protein localization, microtubule ultrastructure, and shedding of ciliary extracellular vesicles (EVs). We show that mec-9 is not expressed in EV releasing neurons, but rather by companion neurons in the sense organs, and may act cell non-autonomously. Our studies reveal pleiotropic roles for mec-9 in the C. elegans ciliated nervous system and provide an in vivo model to study the relationship between cilia and ECM.

cell biology↗

Dynamics of single cell-cell junctions as an indicator of cell state switch

Cell-cell junctions (CCJs) are dynamic biopolymeric systems essential for adherence of biological cells organizing into a tissue or organ. In vertebrate organisms, CCJs present in stable epithelial tissues are maintained primarily by cell-to-cell protein bridges made of an adhesion receptor E-cadherin. CCJs are destabilized during Epithelial (E) to mesenchymal (M) transformation (EMT), an essential step in cancer metastasis, where cells switch states and acquire migratory features. An essential trigger for EMT is a cadherin switch process from E-cadherins with N-cadherin, another cadherin isoform. EMT proceeds through several intermediate states referred to as hybrid E/M cells. These states are characterized by mixed levels of E- and N-cadherins at the junctions and exhibit versatile cancerous traits that are more aggressive than cancerous fully mesenchymal cells. As a result, many such states have emerged as key targets in cancer therapy. However, development of a therapeutic design to counter the hybrid E/M cells has been limited by the absence of a comprehensive understanding of the mechanics and dynamics of hybrid E/M states. Here, we develop a physical model of CCJs as a non-equilibrium system composed of a variable ratio of E- and N-cadherins, considered as coarse-grained molecules driven by ATP-powered machinery observed at CCJs. Our model predicts a robust measure of strength of junctions that captures previous experimental observations, and reveals a minimal mechanochemical landscape of hybrid E/M states. We show the emergence of several groups of CCJ states in this landscape with variable adhesion strengths, many of which resemble different hybrid E/M-characteristics observed experimentally. Finally, we identify that a difference in mechanosenstivity of the two cadherin isoforms towards cytoskeletal forces could be why the hybrid E/M states come into existence.

biophysics↗

Mechanism of Gating and Isoform-Specific Inhibition in Renal CLC Chloride Channels

Hyponatremia is a prevalent disorder marked by excess water retention and substantial morbidity, motivating interest in the CLC-Ka chloride channel as a therapeutic target. Selectively inhibiting CLC-Ka without affecting the closely related CLC-Kb is essential for preventing serious side effects. However, developing isoform-selective inhibitors has been challenging because most small molecules do not distinguish between CLC-Ka and CLC-Kb, and the basis for selectivity in the few known exceptions remains unclear. The small molecule BIM1 preferentially inhibits CLC-Ka over CLC-Kb, providing an opportunity to dissect isoform-specific pharmacology. To investigate this mechanism, we determined cryo-EM structures of BIM1 and BIM15, a related nonselective analog, bound to a CLC-K variant engineered to match the human CLC-Ka binding pocket. Structural and computational analyses reveal that inhibition and isoform selectivity are anchored by interactions with a conserved lysine, with surrounding binding-site residues subtly tuning the local electrostatic environment to promote or disfavor these contacts. These analyses further identify a dynamic extracellular loop that intermittently occludes the shared pathway accessing the inhibitor-binding site and pore. Bound BIM15 engages this gating loop more extensively than BIM1, suggesting that differential loop engagement contributes to inhibitor selectivity, a prediction validated by mutagenesis. Because loop dynamics block the pore, we examined the structural impact of Ca{superscript 2}, which favors opening, and found the gating loop ordered and withdrawn from the pathway. Together, these findings define how binding-site microenvironments and gating-loop dynamics shape isoform-specific inhibition and pore access in CLC-K channels. Significance StatementHyponatremia is a major clinical problem with limited therapeutic options. The kidney chloride channel CLC-Ka is an attractive drug target, but its high sequence identity to CLC-Kb has hindered the development of isoform-selective inhibitors needed for safe therapy. A low-micromolar CLC-Ka-selective inhibitor had been identified, providing a foothold for drug development, but the structural basis of its selectivity was unknown. Here, by integrating cryo-EM structures with molecular dynamics simulations, we define the inhibitor-binding site and reveal the mechanism that enables preferential CLC-Ka inhibition. We further show that a dynamic extracellular loop functions as a gating element shaping inhibitor access and engagement. These findings establish a mechanistic foundation for developing improved treatments for hyponatremia.

biophysics↗

Myeloid DRP1 Sulfenylation Drives Reparative Macrophage Polarization and Neovascularization in Ischemic Muscle

Reparative macrophage polarization and macrophage-derived reactive oxygen species (ROS) are required for ischemia-induced revascularization in peripheral artery disease (PAD). Our previous study showed that mitochondrial fission protein DRP1 promotes reparative polarization and metabolic reprogramming in macrophages and post-ischemic neovascularization. However, the redox-dependent mechanism governing DRP1 activation in this context remains elusive. Here, using a mouse hindlimb ischemia (HLI) model of PAD, we identify cysteine sulfenylation (CysOH) of DRP1 as a critical redox modification induced in ischemic bone marrow (BM)-derived cells. BM chimeric mice reconstituted with CRISPR/Cas9-generated "redox-dead" DRP1-C631A knock-in mutant (Drp1C/A) BM exhibited markedly reduced limb perfusion recovery and CD31 capillary density in ischemic muscles following HLI. These defects were associated with enhanced Ly6G neutrophil accumulation, pro-inflammatory F4/80CD80 M1 macrophages and reduced anti-inflammatory F4/80CD206 M2 macrophages in ischemic muscle. Mechanistically, using an in vitro PAD model, hypoxia-serum starvation (HSS) rapidly induced cytosolic ROS production and DRP1-CysOH formation in wild type macrophages. In contrast, Drp1C/A macrophages failed to undergo DRP1-CysOH-dependent mitochondrial fission under HSS, resulting in aberrant metabolic reprogramming characterized by enhanced glycolysis and mitochondrial ROS, pro-inflammatory p-NF-{kappa}B and M1-genes, and suppressed anti-inflammatory p-AMPK and M2-genes. Thus, our findings establish DRP1 sulfenylation as a previously unrecognized redox-sensing mechanism that links ischemia-induced ROS to reparative macrophage reprogramming and revascularization, identifying a novel therapeutic target for PAD.

cell biology↗

Acute Sublethal Heat Stress Impairs Blood Feeding and Trypanosome Infection in the Kissing Bug, Rhodnius prolixus

Kissing bugs are the primary vectors of Trypanosoma cruzi, the causative agent of Chagas disease. Kissing bugs are exposed to thermal variability, including short periods of heat stress, which can induce mortality or exert sublethal effects. This study investigated Rhodnius prolixus following brief periods of high thermal stress with respect to survival, blood feeding, developmental processes, and T. cruzi infection, with a focus on sublethal effects. Our results demonstrated a significant decrease in survival for R. prolixus at 42 {degrees}C for 8 hours. When exposed to sub-lethal thermal stress (40{degrees}C for 8 hours), blood ingestion (amount and proportion) was reduced after 24 hours of recovery from thermal stress. Among the bugs that fed after 24 hours, molting was not impacted by temperature exposure. The infection rate decreased after heat exposure, likely due to reduced blood volume ingested when feeding 24 hours after heat stress. A week of recovery after exposure to higher temperatures improved feeding and increased infection rates to levels comparable to those of kissing bugs not exposed to thermal stress. Our findings offer insights into how extreme temperature events may influence Chagas disease. Specifically, these studies highlight the need to clarify how temperature, particularly at sublethal levels, interacts with vector biology to alter parasite transmission.

physiology↗

Non-Additive Transcriptional Regulation Underpins Additive Survival Trait In Populations Of Drosophila Selected For Increased Post Infection Survival

Understanding how immunity evolves requires integrating organismal resistance with the underlying genetic and regulatory mechanisms. Using Drosophila melanogaster populations experimentally selected for increased survival post infection with Enterococcus faecalis, we dissected the quantitative genetic basis of post-infection survival and the transcriptional architecture associated with it. F1 hybrid analyses revealed an additive genetic basis for survivorship in both sexes, consistent with a polygenic response to selection. In contrast, gene expression showed extensive sex-specific regulation and hybrid misexpression. Males strongly upregulated antimicrobial peptides yet suffered greater mortality, suggesting a costly or inefficient immune strategy. Females relied on distinct effector pathways and exhibited superior survivorship. Notably, transcriptional induction of the melanization precursor PPO1 did not correspond to increased phenoloxidase enzyme activity, revealing a post-transcriptional constraint on immune deployment. Together, these results show that while the genetic basis of survivorship is additive, the evolved immune phenotype is shaped by sex-specific regulatory divergence, hybrid incompatibilities, and functional bottlenecks. This emphasizes the need to integrate quantitative genetic, transcriptomic, and functional assays to understand the evolution of immune defense.

evolutionary biology↗

Coordinated inhibition of SOX9 and cell cycle progression by microRNA-200 restricts sebaceous gland fate specification

The microRNA-200 family (miR-200s) is widely recognized for their potent role in inhibiting epithelial-to-mesenchymal transition and cell cycle progression in cancer. However, their functional specificity in normal epithelial development remains poorly understood. Here we show that miR-200s are highly enriched in hair matrix progenitors but conspicuously absent from the upper hair follicle (HF), the anatomical location where sebaceous gland (SG) and HF stem cells are specified. We demonstrate that elevated miR-200 expression in this region abolishes SG fate specification while permitting hair morphogenesis. Genome-wide identification of miR-200 targets reveals that miR-200s regulate multiple negative regulators of WNT signaling, in addition to cell cycle regulators. Single-cell and spatial transcriptomic analyses uncover a mutually exclusive expression patterns between WNT activity and SOX9 in the upper HF, which is disrupted by miR-200 induction, resulting in compromised SOX9 function. Mechanistically, genome-wide identification of SOX9 targets uncovers a broad network of lipid and fatty acid metabolism genes critical for the transition of upper HF progenitors to the SG fate. The coordinated inhibition of the SOX9-dependent lipogenic program and the potent restriction of cell cycle progression, both mediated by miR-200s, collectively blocks SG specification. Taken together, this work reveals an unexpected specificity of miR-200 in restricting epithelial plasticity and elucidates a spatially defined SOX9 regulatory network essential for SG development.

developmental biology↗

3D genome architecture regulates the traffic of transcription factors throughout human chromosomes

Transcription factors (TFs) control the rate of transcription of genetic information by binding to specific DNA sequences. The time needed for a TF to find its specific target sites is a bottleneck to the genetic response mechanism. While TF target site search is a well-studied problem, the effect of genome 3D architecture on the TF target search times is poorly understood. Here, we use accurate and cell-specific 3D structural ensembles of human chromosomes to investigate how the spatial organization of binding sites on chromosomes influences the dynamics of TFs. We use Chromatin Immuno-Precipitation data to map the position of binding sites for several TFs on chromosomal structures and simulate the dynamics of individual TF within chromosomal territories. We find that the distribution of binding sites along chromosomes cooperates with the 3D folding of the chromatin fiber to induce dynamics in which TFs tend to visit sites distributed sequentially along the genome. In this way, genome 3D architecture appears to reduce the time each TF spends in the unbound state while commuting from one target site to the other. At the same time, genome 3D architecture further reduces the flux of TFs between binding sites already well separated along the genome, effectively isolating distant clusters of binding sites. We compare the TF traffic patterns generated by the 3D structures of human chromosomes with those generated by several alternative structural models characterized by increasing randomness. Finally, we study the effect of lengthwise compaction and phase separation, known architectural features of the human genome, in TFs target search. In short, our analysis demonstrates that genome architecture regulates the traffic of TFs within chromosomal territories and reduces the time each TF spends commuting between binding sites.

biophysics↗