Biology subjects

Kulkarni, M.

Publications and source records attributed to Kulkarni, M..

At least 19 recordsLinked to original sources

bioRxiv2026

Human caspase-4 and caspase-5 assemble into non-canonical inflammasomes in response to overlapping and distinct LPS structures

Innate immune sensing of bacterial lipopolysaccharide (LPS) is critical to host defense, but a dysregulated response can lead to sepsis, a life-threatening condition associated with multiple organ failure and shock. LPS is sensed at the cell surface by Toll-Like Receptor 4 (TLR4) and in the cytosol by mouse caspase-11 (Casp11) or its human orthologs, caspase-4 and 5 (CASP4/5). In response to cytosolic LPS, Casp11 oligomerizes in a catalytically dependent manner into supramolecular organizing centers (SMOCs). We demonstrate that CASP4 and CASP5 also form catalytically dependent SMOCs in response to bacterial LPS in macrophages. Furthermore, we find that CASP4 and CASP5 differentially respond to distinct LPS structural variants that fail to activate CASP11 and that only activating LPS structures induce SMOC formation by the corresponding caspase. Notably, transgenic mice expressing CASP4 in lieu of Casp11 exert more effective control of Francisella novicida, a rodent pathogen whose LPS evades TLR4 and Casp11 detection. Conversely, CASP4-expressing mice are more susceptible to sepsis when exposed to E. coli expressing a lipid A variant that fails to activate TLR4 and Casp11. Together, these findings reveal human CASP4 and CASP5 have evolved to assemble into SMOCs in response to a broader range of LPS structures than mouse Casp11 and highlight the potential tradeoff between systemic inflammation and control of bacterial infection that may underline these differences. Our findings may inform the development of targeted therapeutic strategies for gram-negative bacterial infection and sepsis that are effective in humans.

immunology↗

bioRxiv2026

DIPTAR: A synthetic biology platform for functional interrogation of protein degradation

Protein degradation regulates cellular homeostasis, yet many degradation events are difficult to study because they lack a readily selectable phenotype. Here, we develop Degradation-Induced Pyroptosis TArgeting Receptors (DIPTAR), a modular synthetic biology platform that couples protein degradation to CARD8-mediated pyroptosis. Using HIF-1 as a model substrate, we show that DIPTAR faithfully reports oxygen-dependent VHL-mediated degradation and enables pooled CRISPR screening to identify established and previously unrecognized regulators of HIF-1 stability. DIPTAR is functional across multiple cell types and can be programmed with diverse proteins, including BRD4, I{kappa}B, and p53, to convert distinct degradation stimuli into a common pyroptotic output. DIPTAR also detects pathogen-mediated perturbations of host degradation pathways, including both inhibition and induction of degradation-dependent signaling. By converting protein degradation into a robust selectable phenotype, DIPTAR provides a scalable platform for functional genetic discovery, interrogation of degradation pathways, degrader characterization, and investigation of host-pathogen interactions.

synthetic biology↗

bioRxiv2026

Rod pathway blockade improves visual function in a mouse model of photoreceptor degeneration

Retinitis pigmentosa (RP) refers to a group of inherited photoreceptor degenerations characterized by the initial death of rods and secondary loss of cones. In animal models of RP, the ganglion cells (GCs) exhibit increased spontaneous activity that can mask residual photoreceptor signals and reduce the efficacy of treatments for vision restoration. The A2 amacrine cells (ACs) are inhibitory interneurons that propagate aberrant activity to GCs, but it remains unclear whether rod or cone pathways drive altered A2-AC signaling or whether the relative contributions of these pathways change with disease progression. We examined the circuit mechanisms underlying aberrant activity in the rd10 mouse model and found that blocking rod pathway input to A2-ACs suppressed aberrant activity and improved the signal-to-noise ratio of residual cone-driven responses in On- GCs. The results also indicate that the On- but not the Off-pathway exhibits circuit-level compensation during the loss of photoreceptor input. The results suggest that pharmacological blockade of the rod pathway may improve residual cone-mediated function and improve the efficacy of vision restoration in rod-cone photoreceptor degenerations.

systems biology↗

bioRxiv2026

Characterization and Validation of Compressed Sensing for Time-of-Flight MRI Angiography of the Human Brain at 3T and 7T

BackgroundCerebral vasculature is a key biomarker of brain health, and time-of-flight (TOF) magnetic resonance angiography (MRA) provides noninvasive assessment of vascular anatomy. However, conventional TOF-MRA requires long scan times, increasing patient burden and susceptibility to motion artifacts. Compressed sensing (CS) offers a feasible acceleration strategy. PurposeTo quantitatively evaluate CS acceleration in TOF-MRA at 3T and 7T using automated whole-FOV vascular segmentation and semi-automatic segmentation of representative vessels. Study typeProspective Population23 healthy human participants (3T) and 8 healthy human participants (7T). Field Strength/SequenceCS TOF-MRA (CS factors 4 and 8 at 3T; 8 at 7T) was compared against non-accelerated (CS0) TOF-MRA. AssessmentVisual comparison and vascular segmentation were performed using automated whole-FOV methods and semi-automatic segmentation of the posterior cerebral artery and anterior choroidal artery. Statistical TestsContrast-to-noise ratio (CNR), voxel count, and vessel diameter were assessed using two-tailed paired t-tests. ResultsWhole-FOV CNR differed significantly across CS factors at 3T (CS0 > CS4: p < 0.001, d = 0.77; CS0 < CS8: p = 0.008, d = 0.36; CS4 < CS8: p < 0.001, d = 1.11) and 7T (CS0 < CS8: p = 0.002, d = 0.54), with semi-automatic segmentation yielding consistent findings (p < 0.01 for all comparisons). The diameter measurements for segmented vessels are also higher with high CS-factors (PCA 7T: left: p = 0.006, d = 0.93, right: p = 0.045, d = 0.43; AChA 7T: left: p < 0.001, d = 0.66, right: p = 0.009, d = 1.06; PCA 3T: p < 0.001 for all comparison dLeft = 0.52 (CS0 vs. CS4), 0.56 (CS4 vs. CS8), 1.11 (CS0 vs. CS8) and dRight = 0.78 (CS0 vs. CS4), 0.57 (CS4 vs. CS8), 1.17 (CS0 vs. CS8)). Data ConclusionCS shows promise for enhancing clinical applicability of TOF-MRA, with advantages most pronounced at 7T.

bioengineering↗

bioRxiv2026

Functional reorganization of motor cortex connectivity during learning

Learning new tasks requires the brain to reshape the flow of neural activity, but how these changes arise from dynamic neural connectivity remains unclear. Here, we used two-photon photostimulation and calcium imaging to map learning-related changes in connectivity in layer 2/3 of mouse motor cortex, induced by learning of an optical brain-computer interface (BCI) task. Mice rapidly (within minutes) learned to change activity in a conditioned neuron to earn rewards. Activity changes were sparse; the conditioned neuron increased activity more than surrounding neurons. Mapping connectivity before and after learning revealed changes in motor cortex connectivity, enriched in neurons that were active before trial initiation, analogous to motor cortex populations that are active preceding movement. Motor cortex plasticity reroutes preparatory activity to neurons that are active later and control the conditioned neuron. Our findings show how rapid learning can be achieved through structured changes in motor cortex connectivity.

neuroscience↗

bioRxiv2026

Early electrical stimulation promotes functional recovery after volumetric muscle loss

Volumetric muscle loss (VML) injuries overwhelm the inherent regenerative capacity of skeletal muscle, causing persistent functional deficits with no routinely effective therapies. Electrical stimulation (ES) has been shown to preserve muscle structure in other injury models, but technical barriers have prevented daily delivery during the acute post-injury window when critical regenerative programs are established. Here, we developed a fully implantable bioelectronic system with nanoporous platinum-modified electrodes enabling daily therapeutic stimulation and electromyographic recording without repeated anesthesia in a rat tibialis anterior VML model. Animals receiving ES during the acute post-injury period (10 sessions over days 0-14) showed sustained functional improvement, reaching 90% of baseline torque at 8 weeks compared to 71% in unstimulated controls. This recovery reflected enhanced remodeling of injured muscle rather than synergistic muscle compensation. Histological analysis revealed coordinated early increases in vascularization, pro-regenerative macrophages, and satellite cells. These findings establish early ES as a promising intervention for promoting muscle regeneration after catastrophic injury.

bioengineering↗

bioRxiv2026

The heat-ramp method to study regulated cell death in a pathogenic yeastCryptococcus neoformans

Human fungal pathogens cause a significant public health burden. While no reliable surveilence data are available, estimations suggest that 1 billion infections and over 2 million deaths are attributable to fungal infections annually worldwide. This drove the World Health Organization to generate a priority list of fungal pathogens for reearch, which includes the yeast Cryptococcus neoformans in a top critical priority. With the rise of drug-resistance and emerging fungal pathogens, new conceptual strategies for antifungal therapies are needed in addition to existing antibiotic development pipelines to meet clinical needs. Intrinsic cell death pathways encoded by pathogenic fungi are largely unstudied but could be leveraged for antifungal therapy analogous to anti-cancer therapeutics that activate apoptosis or other cell death mechanisms. Thus far, molecularly defined fungal cell death mechanisms are best characterized for only a few, predominantly model filamentous species. To extend these studies to pathogenic yeast, here we describe and demonstrate a tunable heat-ramp stimulus that when applied to small volumes of yeast cell suspensions reveals a protracted cell death process in the pathogenic yeast Cryptococcus neoformans. This low cost protocol induces robust and reproducible phenotypes to study gene-dependent mechanisms in laboratory strains and clinical isolates.

microbiology↗

bioRxiv2025

NELFA-mediated promoter-proximal pausing restrains YAP-driven transcription and shapes context-dependent outcomes in breast cancer

Overexpression of YAP is associated with tumor progression in multiple malignancies. YAP is the key transcriptional effector of Hippo signaling: a conserved pathway from Drosophila (Yki) to humans (YAP). In a Drosophila screen, we identified NELFA, a core component of the promoter-proximal pausing (PPP) complex, as a suppressor of Yki-driven hyperproliferation. We investigated whether PPP-YAP regulatory interaction is conserved in mammalian system. In HEK293T and MDA-MB-231 cells, we demonstrate that NELFA depletion amplifies YAP-driven transcription. Whole-transcriptome analysis of MDA-MB-231 cells revealed widespread reprogramming upon NELFA loss, with strong enrichment of YAP signatures, EMT and TGF-{beta} transcriptional networks--indicating that NELFA regulates a broad YAP-centered gene regulatory module. Clinical analysis of two independent breast cancer cohorts revealed a context-dependent role for NELFA. Low NELFA expression predicted poor disease-free survival in YAP-high tumors--particularly in triple-negative breast cancer (TNBC)--suggesting a tumor-suppressive role. While High NELFA expression correlated with poorer overall survival in all the subtypes. These findings confirm that NELFAs role in YAP-driven transcription is conserved in mammalian system and it manifests in a context-dependent manner in the breast cancer.

cancer biology↗

bioRxiv2025

Human non-canonical inflammasomes activate CASP3 to limit intracellular Salmonella replication in macrophages.

Inflammasomes are multiprotein signaling platforms that activate inflammatory caspases to initiate innate immune signaling. In humans, canonical inflammasomes activate CASP1, which cleaves the pore-forming protein gasdermin D (GSDMD) and the cytokines IL-1{beta} and IL-18. In contrast, the non-canonical inflammasome detects bacterial lipopolysaccharide (LPS) through CASP4/5, which cleave GSDMD to drive pyroptosis. While CASP1 substrates are well characterized, CASP4/5 substrates remain less defined. Here, we show that in response to intracellular LPS and gram-negative bacterial infection, CASP4/5 directly cleave and activate the executioner caspases CASP3/7. CASP3 in turn cleaves and activates gasdermin E (GSDME). Surprisingly, CASP3, but not GSDME, was required for restricting intracellular Salmonella replication, suggesting that CASP4/5-induced apoptosis contributes to host defense. We further show that most GSDMD cleavage during non-canonical inflammasome activation is mediated by CASP1, and that GSDMD is the primary driver of pyroptosis. Finally, we confirm that CASP4/5 activate CASP3/7 and GSDME in human primary macrophages. These findings establish CASP4/5 as dual apoptotic initiator and inflammatory caspases and reveal a central role for the apoptotic signaling cascade in non-canonical inflammasome-mediated immunity.

cell biology↗

bioRxiv2025

HomRank: Homogeneous RNA Ranking for 3D Structure Evaluation

Leveraging rigorously curated RNA sequences and a novel ranking-based evaluation paradigm, we propose an improved pipeline for RNA 3D structure assessment. To enhance generalization, we construct a dataset comprising non-redundant single-chain RNA sequences from the PDB and apply unsupervised clustering to minimize data leakage. Based on this, we retrain the previous state-of-the-art method, ARES, to develop ARES+, which improves the top-1 success rate by 21% on the RNA Puzzles benchmark. To further boost near-native identification, we introduce HomRank, a homogeneous RNA ranking method that directly optimizes the selection of near-native conformations from candidate sets, resembling expert evaluation strategies. HomRank achieves 95% and 100% top-1 and top-5 success rates, respectively, significantly outperforming ARES+. These results demonstrate that carefully designed datasets and the expert-like selection paradigm can substantially improve the accuracy and robustness of RNA 3D structure assessment, offering a promising direction for deep learning-based RNA evaluation and near-native conformation selection.

bioinformatics↗

bioRxiv2025

Understudied and underprotected: biodiversity and conservation challenges in the Konkan region of Maharashtra

In biodiversity hotspots, such as the Western Ghats, periodic synthesis of existing ecological research can help identify knowledge gaps and address critical threats to biodiversity. The Maharashtra part of the Konkan region, situated between the Sahyadri foothills and the west coast, with diverse tropical vegetation, harbours unique open ecosystems, such as lateritic plateaus, and supports a multitude of threatened species, including hornbills and tigers. However, the region remains relatively understudied and has not received adequate protection through state or national conservation policies and laws. The area is undergoing rapid human-driven land-use change. These activities can impact the regions biodiversity, necessitating an effort to identify knowledge gaps and critical threats. Through a combination of a literature review and focus group discussions with 44 participants from various institutions and non-governmental organisations, we synthesised existing published information on the Konkan region of Maharashtra and identified key research gaps and conservation challenges. Our review of 138 studies found that while agroforestry and human-wildlife interactions have received some research attention, the effects of climate change on the regions biodiversity remain poorly understood. Focus group discussions highlighted major threats, including land-use changes due to expanding monoculture plantations, clear-felling of forests, forest fires, environmental pollution, and rapid infrastructure development, which are leading to habitat loss and fragmentation. Some of these concerns were validated by publicly available regional data, which revealed a 30% increase in roads, a 14% expansion of cashew plantations, and associated forest loss. This synthesis offers valuable insights for government and non-governmental organisations to inform future research and conservation efforts in the region.

ecology↗

bioRxiv2025

Chemical Tools Based on the Tetrapeptide Sequence of IL-18 Reveals Shared Specificities between Inflammatory and Apoptotic Initiator Caspases

Caspases are a family of cysteine proteases that act as molecular scissors to cleave substrates and regulate biological processes such as programmed cell death and inflammation. Extensive efforts have been made to identify caspase substrates and to determine factors that dictate substrate specificity. We recently discovered that that the human inflammatory caspases (caspases-1, -4, and -5) cleave the cytokines IL-1{beta} and IL-18 in a sequence-dependent manner. Here, we report the development of a new peptide-based probe and inhibitor based on the tetrapeptide sequence of IL-18 (LESD). We found that this inhibitor was most selective and potent at inhibiting caspase-8 activity (IC50 = 50 nM). We also discovered that our LESD-based inhibitor is more potent than the currently used z-IETD-FMK inhibitor that is thought to be the most selective and potent inhibitor of caspase-8. Accordingly, we demonstrate that the LESD based inhibitor prevents caspase-8 activation during Yersinia pseudotuberculosis infection in primary bone-marrow derived macrophages. Furthermore, we characterize the selectivity and potency of currently known substrates and inhibitors for the apoptotic and inflammatory caspases using the same activity units of each caspase. Our findings reveal that VX-765, a known caspase-1 inhibitor, also inhibits caspase-8 (IC50 = 1 {micro}M) and even when specificities are shared, the caspases have different efficiencies and potencies for shared substrates and inhibitors. Altogether, we report the development of new tools that will facilitate the study of caspases and their roles in biology.

biochemistry↗

bioRxiv2025

Development and Implementation of a Minority Health International Infectious Diseases Research Training Program

From 2014-2019, the University of Hawai`i (UH) at Manoa offered a National Institutes of Health funded Minority Health International Research Training (MHIRT) Program for undergraduate research experiences in infectious diseases. The goal of the program was to immerse undergraduate students in conducting global infectious diseases research to train a new generation of scientists to combat future global pandemics. The MHIRT program trained educationally underrepresented groups unique in Hawai`i: Native Hawai`ians and other Pacific Islanders, and underrepresented Asian Americans, e.g., Filipinos and Vietnamese. A 12-member Advisory Committee guided the program development, implementation, recruitment strategies, recruitment, retention, and program evaluation. The program provided an eight-month classroom and field based international research training. Students learned about "8 steps of research" and cultural competency, experienced a hands-on research project abroad, and presented their results to the research community in the host country and at UH. Students research topics focused on ongoing projects of UH faculty including those with international collaborations in infectious disease and tropical medicine. Students spent two months on an international research mentors project at an international site under the mentorship of a Hawai`i based research faculty. After eight months, students graduated from the program by presenting their research to family, friends, mentors, and UH faculty and administrators. Trainees in this program pursued academic graduate or professional health degrees. The majority of students pursued further studies related to infectious diseases contributing to workforce development in infectious disease health research. The UH-MHIRT program provides an example of student immersion in international infectious disease research to foster future interest in global health research providing potential benefits for the student and faculty and their research efforts.

scientific communication and education↗

bioRxiv2024

A method for authenticating the fidelity of Cryptococcus neoformans knockout collections

Gene knockout strain collections are important tools for discovery in microbiology. Cryptococcus neoformans is the only human pathogenic fungus with an available genome-wide deletion collection, and this resource is widely used by the research community. We uncovered mix-ups in the assembly of the commercially available C. neoformans deletion collection of [~]6,000 unique strains acquired by our lab. While pursuing the characterization of a gene-of-interest, the corresponding deletion strain from the C. neoformans KO collection displayed several interesting phenotypes associated with virulence. However, RNAseq analysis identified transcripts for the putative knockout gene, and the absence of transcripts for a different knockout strain found in the same plate position in an earlier partial knockout collection, raising the possibility that plates from one collection were substituted for the other. This was supported by determining the size of the nourseothricin (NAT)-resistance cassette used to generate the two separate knockout libraries and was confirmed by RNAseq and genome sequencing. Here we report that our KN99 collection is comprised of mixed plates from two independent KO libraries and present a simple authentication method that other investigators can use to distinguish the identities of these KO collections. ImportanceGene knockout strain collections are important tools for discovery in microbiology. Cryptococcus neoformans is the only human pathogenic fungus with an available genome-wide deletion collection, and this resource is widely used by the research community. Here we report that our KN99 collection is comprised of mixed plates from two independent KO libraries and present a simple authentication method that other investigators can use to distinguish the identities of these KO collections. Above all, this article serves as a reminder to 2015 library collection strains before undertaking large phenotyping experiments.

microbiology↗

bioRxiv2024

Wound state monitoring by multiplexed, electrochemical, real-time, localized, inflammation-tracking nitric oxide sensor (MERLIN)

Nitric oxide (NO) released endogenously by induced nitric oxide synthase (iNOS) in macrophages is a key regulatory biomarker for wound inflammation. Detecting NO directly on the wound bed is challenging due to its short half-life time (6 - 50 s), low physiological concentration (nM - {micro}M) and interferences in the complex wound environment. Existing NO sensors suffer from limitations such as rigid substrates, single point of detection and complex measurement setup. Here we present a compliant, multiplexed, electrochemical, real-time, localized, inflammation-tracking nitric oxide sensor (MERLIN) array for in vivo spatiotemporal measurement of NO, with high sensitivity (15.6 {+/-} 5.0 nA/M for 1.5 mm diameter electrodes), selectivity against nitrites (ca. 27900-fold), ascorbic acid (ca. 3800-fold), and uric acid (ca. 6900-fold), and low limit of detection (8.00 nM). We developed a robust device fabrication protocol, comprehensive and highly reproducible in vitro characterization yielding a consistent device performance deployed for wound healing diagnostics. MERLIN spatiotemporally tracked NO on rat skin wounds for 7 days and results indicated that NO peaks on day 3, in line with previously reported iNOS activity. MERLIN allows spatial mapping of the NO gradient across the wound bed, which can be used to provide diagnostic information to assist wound care.

bioengineering↗

bioRxiv2024

Discovery of free glycated amines and glycated urea as potential markers of diabetic nephropathy

The role of protein glycation in the pathogenesis of diabetes is well established. Akin to proteins, free amino acids and other small-molecule amines are also susceptible to glycation in hyperglycemic conditions and may have a role in the pathogenesis of the disease. However, the information on glycation of free amino acids and other small- molecule amines is relatively obscure. In the quest to discover small molecule glycated amines in the plasma, we have synthesized glycated amino acids, glycated creatine, and glycated urea, and by using a high-resolution accurate mass spectrometer, a mass spectral library was developed comprising the precursor and predominant fragment masses of glycated amines. Using this information, we report the discovery of glycation of free lysine, arginine, and leucine/isoleucine from the plasma of diabetic patients. This has great physiological significance as glycation of these amino acids may create their deficiency and affect vital physiological processes such as protein synthesis, cell signaling and insulin secretion. Also, these glycated amino acids could serve as potential markers of diabetes and its complications. On the other hand, accumulation of creatinine and urea in the plasma act as biomarkers of diabetic nephropathy diagnosis. For the first time, we report the detection of glycated urea in diabetes, which is confirmed by matching the precursor and fragment masses with the in vitro synthesized glycated urea by using 12C6 and 13C6-glucose. Further, we quantified glycated urea detected in two forms, monoglycated urea (MGU) and diglycated urea (DGU), by a targeted mass spectrometric approach in the plasma of healthy, diabetes, and diabetic nephropathy subjects. Both MGU and DGU showed a positive correlation with clinical parameters such as blood glucose and HbA1c. Given that urea gets converted to glycated urea in hyperglycemic conditions, it is crucial to quantify MGU and DGU along with the urea for the diagnosis of diabetic nephropathy.

biochemistry↗

bioRxiv2023

Cryptococcus neoformans rapidly invades the murine brain by sequential breaching of airway and endothelial tissues barriers and engulfment by microglia

The fungus Cryptococcus neoformans causes lethal meningitis in humans with weakened immune systems and is estimated to account for 10-15% of AIDS-associated deaths worldwide. There are major gaps in our understanding of how this environmental fungus evades the immune system and invades the mammalian brain before the onset of overt symptoms. To investigate the dynamics of C. neoformans tissue invasion, we mapped early fungal localisation and host cell interactions at early times in infected brain, lung, and upper airways using mouse models of systemic and airway infection. To enable this, we developed an in situ imaging pipeline capable of measuring large volumes of tissue while preserving anatomical and cellular information by combining thick tissue sections, tissue clarification, and confocal imaging. Made possible by these techniques, we confirm high fungal burden in mouse upper airway turbinates after nasal inoculation. Surprisingly, most yeasts in turbinates were titan cells, indicating this microenvironment enables titan cell formation with faster kinetics than reported in mouse lungs. Importantly, we observed one instance of fungal cells enmeshed in lamina propria of upper airways, suggesting penetration of airway mucosa as a possible route of tissue invasion and dissemination to the bloodstream. We extend previous literature positing bloodstream dissemination of C. neoformans, via imaging C. neoformans within blood vessels of mouse lungs and finding viable fungi in the bloodstream of mice a few days after intranasal infection, suggesting that bloodstream access can occur via lung alveoli. In a model of systemic cryptococcosis, we show that as early as 24 h post infection, majority of C. neoformans cells traversed the blood-brain barrier, and are engulfed or in close proximity to microglia. Our work establishes that C. neoformans can breach multiple tissue barriers within the first days of infection. This work presents a new method for investigating cryptococcal invasion mechanisms and demonstrates microglia as the primary cells responding to C. neoformans invasion.

microbiology↗

bioRxiv2023

Bulky glycocalyx drives cancer invasiveness by modulating substrate-specific adhesion

Majority of the eukaryotic cell surface is decorated with a layer of membrane attached polysaccharides and glycoproteins collectively referred to as the glycocalyx. While formation of a bulky glycocalyx has been associated with cancer progression, the mechanisms by which the glycocalyx regulates cancer invasiveness is incompletely understood. We address this question by first documenting sub-type specific expression of the major glycocalyx glycoprotein Mucin-1 (MUC1) in breast cancer patient samples and breast cancer cell lines. Strikingly, glycocalyx disruption led to inhibition of 2D motility, loss of 3D invasion and reduction of clonal scattering of breast cancer cells at the population level. Tracking of 2D cell motility and 3D invasiveness of MUC1-based sorted sub-populations revealed fastest motility and invasiveness in intermediate MUC1-expressing cells, with glycocalyx disruption abolishing these effects. While differential sensitivity in 2D motility is attributed to a non-monotonic dependence of focal adhesion size on MUC1 levels, higher MUC1 levels enhance 3D invasiveness via increased traction generation. In contrast to inducing cell rounding on collagen-coated substrates, high MUC1 level promotes cell adhesion and confers resistance to shear flow on substrates coated with the endothelial surface protein E-selectin. Collectively, our findings illustrate how MUC1 drives cancer invasiveness by differentially regulating cell-substrate adhesion in a substrate-dependent manner.

cancer biology↗