Search bioRxiv⌕ Search

Biology subjects

Mayo, A.

Publications and source records attributed to Mayo, A..

At least 19 recordsLinked to original sources

Automated behavioral segmentation and markerless pose tracking of mice during spaceflight

The NASA Rodent Habitat aboard the International Space Station enabled long-duration studies of behavioral responses to spaceflight, but video-based behavioral analysis has relied on laborious manual annotation. No study has tested whether deep learning tools can automate this analysis under the demanding imaging conditions of orbital vivaria. We applied pose estimation (SLEAP) and behavioral segmentation (DeepEthogram) to archival footage from the Rodent Research-1 mission. Nine labelers annotated 3,249 pose labels across 2,063 frames, and three behaviorists labeled 411,194 frames across 66 videos. Pose tracking accuracy approximated human inter-annotator variability despite progressive lens soiling, grid occlusions, and spherical aberration. Behavioral classification across eight categories achieved accuracy of 0.86-0.90 and suggests progressive behavioral adaptations to microgravity. Kinematic reconstruction of circling estimated centripetal accelerations periodically approaching 1g. This is the first application of deep learning-based pose estimation and behavioral segmentation to rodents in spaceflight, establishing benchmarks for future monitoring systems. One-Sentence SummaryDeep learning-based pose estimation and behavioral segmentation of ISS video reveal progressive behavioral adaptations of mice to microgravity

neuroscience↗

Cell surface ATP6V1B2 marks a subset of persistent senescent cells with increased resistance to apoptosis

Accumulation of senescent cells promotes ageing and age-related diseases. While senescent cells are heterogenous and increasingly persistent in vivo with age, the mechanisms underlying their heterogeneity, resistance to apoptosis, and tissue accumulation remain insufficiently understood. Here we report that in response to DNA damage, a subset of senescent cells upregulates the v-type ATPase subunit, ATP6V1B2 (V1B2) on the cell surface. This upregulation is associated with altered lysosomal activity and changes in intracellular pH. Heterogeneity of senescent cells marked by cell surface V1B2 (csV1B2) is present in naturally occurring senescent cells within both ageing and fibrotic lungs. Senescent cells expressing csV1B2 show an age-independent transcriptional signature associated with DNA repair and resistance to apoptosis. Consistent with this, we show that csV1B2 expression correlates with senescent cell resistance to ABT-737-induced apoptosis in culture. Our study identifies a subset of senescent cells, marked by csV1B2, with a distinct signature of apoptosis resistance. Understanding the functional heterogeneity of senescent cells and the mechanisms accountable for persistence of specific subpopulations in tissues may facilitate the development of improved senotherapeutic strategies for age-related diseases.

cell biology↗

Single-cell quantification of senescence burden reveals cell type-specific ageing dynamics across organs

Cellular senescence, a hallmark of ageing, drives tissue dysfunction by promoting inflammation and fuelling disease. Yet, the dynamics of senescent cell accumulation across tissues and their cell type identity remain poorly understood. Here, we introduce the first, single-cell, protein-level approach, combining multiple senescence markers for the identification and quantification of senescent cells across multiple tissues in mice and in human PBMCs. Applying this method, we reveal widespread but heterogeneous changes in senescence marker expression across cell types and tissues. The cells we identify as senescent displayed transcriptomic senescence signatures, providing a direct molecular link between protein- and mRNA-level detection of senescence. Importantly, senescence accumulation was strongly coordinated within organs but showed little correlation across them, supporting the idea of a tissue specific progression of ageing. These findings refine our understanding of the tissue-specific dynamics of senescence accumulation with age, and provide a framework for evaluating diverse therapeutic interventions.

cell biology↗

A Mechanistic Model for the HPA Axis Cortisol Paradox in PTSD

Post-traumatic Stress Disorder (PTSD) is a debilitating psychiatric condition characterized by intrusive memories, hyperarousal, avoidance, and cognitive and mood disturbances. A longstanding biological paradox in PTSD is the observation of low basal cortisol levels, despite the expectation of elevated cortisol under chronic stress. This "low cortisol paradox" challenges traditional hypothalamic-pituitary-adrenal (HPA) axis regulation models. Individuals with PTSD also exhibit normal or near-normal adrenocorticotropic hormone (ACTH) levels despite reduced cortisol and blunted hormonal responses to acute stress. In this paper, we provide further evidence of reduced cortisol levels using a large medical database from thousands of individuals diagnosed with PTSD. To understand this dysregulation, we apply a systems-level mathematical model of HPA axis regulation that incorporates dynamic changes in gland functional mass, the pituitary corticotrophs and adrenal cortex, over weeks to months. Using this model, we demonstrate that enhanced glucocorticoid receptor (GR) sensitivity, a known risk factor for PTSD, can lead to a steady state with low cortisol and normal ACTH, reconciling key clinical observations. The model also recapitulates the blunted cortisol and ACTH responses to acute stress and the dexamethasone/ corticotropin-releasing hormone (DEX/CRH) test, reported in PTSD cohorts. Importantly, despite low cortisol levels, basal glucocorticoid receptor (GR) activity is higher than normal due to the reduced receptor affinity. Thus, individuals with PTSD effectively experience elevated cortisol signaling relative to their GR sensitivity. These findings provide a unified mechanistic explanation for HPA axis dysregulation in PTSD, grounded in the slow time scale of changes in gland functional mass and supported by literature and clinical data.

systems biology↗

OSDR2.0 infers microenvironment-driven cell-state transitions and population dynamics from a single spatial biopsy

Cell populations in human tissues change over time by cell division, death and transitions between functional states. In the tumor microenvironment (TME), such dynamics are central to immune evasion, stromal remodeling and therapeutic response. However, it is difficult to measure such dynamics in vivo because usually only a single biopsy is available providing a static snapshot. To obtain cell population dynamics from a snapshot we previously developed One Shot Dynamic Reconstruction (OSDR1.0), an algorithm that reconstructs cell population dynamics over weeks to months from a spatial biopsy using cell-type information and a proliferation marker. OSDR1.0 however does not include transitions between cell states. Here we present OSDR2.0, an extension that incorporates transitions between cell states inferred from the local cellular neighborhood. The algorithm OSDR2.0 models the probability of a cell being in a given state (e.g., PD1+ vs. PD1- T cell, or cancer-associated vs. resting fibroblast) as a function of its surrounding cell types. These state-transition rules are then integrated into simulations of population dynamics. After the cell population is advanced by a timestep, the cell states are adjusted according to the new neighborhoods, using the fact that cell state transitions, which take hours, are typically much more rapid than cell population changes on the scale of weeks, and can thus be considered at quasi-steady-state. Applying OSDR2.0 to spatial proteomics data from triple-negative breast cancer (TNBC), we find that cell state is strongly associated with local microenvironment composition. Incorporating state transitions significantly improves the ability to predict treatment response - OSDR2.0 accurately separates responders from nonresponders to chemotherapy and immunotherapy based on early post-treatment biopsies, outperforming OSDR1.0 that does not include cell state transitions. This work highlights the importance of cell state plasticity in shaping tumor response and opens a way to infer both population dynamics and cell state transitions from static clinical samples.

cancer biology↗

A fibroblast-centric network drives cold fibrosis in the tumor microenvironment of lung squamous cell carcinoma

The tumor microenvironment (TME) of chronic inflammation-associated cancers (CIACs) is shaped by cycles of injury and maladaptive repair, yet the principles organizing fibrotic stroma in these tumors remain unclear. Here, we applied the concept of hot versus cold fibrosis, originally credentialed in non-cancerous fibrosis of heart and kidney, to lung squamous cell carcinoma (LUSC), a prototypical CIAC. Single-cell transcriptomics of matched tumor and adjacent-normal tissue from 16 treatment-naive LUSC patients identified a cold fibrotic architecture in the LUSC TME: cancer-associated fibroblasts (CAFs) expanded and adopted myofibroblast and stress-response states, while macrophages were depleted. This macrophage-poor, CAF-rich stroma was maintained by CAF autocrine growth factor loops, including TIMP1, INHBA, TGFB1, and GMFB. In parallel, the immune compartment exhibited a hot tumor phenotype with abundant T and B cells, forming spatially distinct but molecularly engaged networks with CAFs. CAF gene programs typifying cold fibrosis in LUSC were conserved in other CIACs, including esophageal and gastric adenocarcinomas. These results redefine desmoplastic regions of tumors through the lens of a non-cancer fibrosis model, demonstrating that conserved stromal circuits constitute therapeutic vulnerabilities in CIACs.

cancer biology↗

Dynamic growth trajectories distinguish bacteriostatic and bactericidal antibiotics at subinhibitory concentrations

Subinhibitory antibiotic exposures are common in clinical and environmental contexts, yet their effects on bacterial growth dynamics remain incompletely understood. We studied the temporal response of Escherichia coli to a panel of bactericidal ("cidal") and bacteriostatic ("static") antibiotics at sub-minimum inhibitory concentrations (sub-MIC). We uncover a sharp dynamical distinction between the two classes. Bacteriostatic antibiotics reduce the initial growth rate in a dose-dependent manner, similar to a nutrient starvation response. In contrast, bactericidal antibiotics do not alter initial growth rates - cells continue to grow as fast as untreated cells - until an abrupt slowdown in growth rate. The onset of slowdown occurs earlier with increasing dose, suggesting a damage accumulation mechanism leading to a lethal threshold. Cidals also show a steeper dose-response curve. We propose that bacteria respond to cidal antibiotics with a "grow-fast-then-crash" strategy that is adaptive for transient lethal threats, whereas static antibiotics trigger stress adaptation and slower growth. While clinical outcomes of statics and cidals may be similar at full inhibitory doses, these sub-MIC dynamical signatures could influence resistance evolution and treatment outcomes in biofilms or partially resistant strains. Our findings offer a dynamic framework for antibiotic classification and raise new questions about how bacteria respond to sublethal antibiotic stress.

systems biology↗

Unifying regulatory motifs in endocrine circuits

In this study we identify unifying design principles in human endocrine systems. We find that 43 hormone systems, controlling diverse physiological functions, fall into 5 classes of circuits with shared structure - thus only a small number of the possible circuits actually occur. Each class uses a different regulatory logic to perform specific dynamical functions, such as homeostasis, acute input-output response or adjustable set points. The circuits employ interactions on two timescales: hormone secretion on the scale of minutes-hours and growth and shrinkage of endocrine gland mass on the scale of months, which impacts the amount of hormone the glands secrete. This two-timescale principle recurs in several classes of circuits, including the most complex class, which has an intermediate gland, the pituitary. We analyze the pituitary circuit in detail and find tradeoffs between endocrine amplification, buffering of hypersecreting tumors, and rapid response times. These unifying principles of regulation build a foundation for systems endocrinology.

systems biology↗

Hormone circuit analysis explains why most HPA drugs fail for mood disorders and predicts the few that work

Elevated cortisol causes morbidity in chronic stress and mood disorders, including metabolic and cardiovascular diseases. There is therefore interest in developing drugs that lower cortisol by targeting its endocrine pathway, the hypothalamic-pituitary-adrenal (HPA) axis. Several promising HPA-modulating drugs have, however, failed to lower long-term cortisol in mood disorders such as major depressive disorder despite their effectiveness in situations where high cortisol is caused by a tumor (Cushings syndrome). Why these drugs failed is not well understood. Here we use a mathematical model of the HPA axis to show that the pituitary and the adrenal glands compensate for the effect of drugs by adjusting their functional mass, a feedback compensation that is absent in Cushing tumors. To find potential drug targets, we carried out a systematic in silico analysis of points of intervention in the HPA axis. We find that only two interventions that target corticotropin-releasing hormone (CRH) can lower long-term cortisol. Other drug targets either fail to lower cortisol due to gland-mass compensation or lower cortisol but harm other aspects of the HPA axis. Thus, we identify potential drug targets, including CRH-neutralizing antibodies and CRH-synthesis inhibitors, for lowering long-term cortisol in mood disorders and in those suffering from chronic stress. More generally, this study indicates that understanding the slow compensatory mechanisms in endocrine axes can be crucial in order to prioritize drug targets.

systems biology↗

Quantification of beta cell carrying capacity in prediabetes

Prediabetes, a subclinical state of high glucose, carries a risk of transition to diabetes. One cause of prediabetes is insulin resistance, which impairs the ability of insulin to control blood glucose. However, many individuals with high insulin resistance retain normal glucose due to compensation by enhanced insulin secretion by beta cells. Individuals seem to differ in their maximum compensation level, termed beta cell carrying capacity, such that low carrying capacity is associated with a higher risk of prediabetes and diabetes. Carrying capacity has not been quantified using a mathematical model and cannot be estimated directly from measured glucose and insulin levels in patients, unlike insulin resistance and beta cell function which can be estimated using HOMA-IR and HOMA-B formula. Here we present a mathematical model of beta cell compensation and carrying capacity, and develop a new formula called HOMA-C to estimate it from glucose and insulin measurements. HOMA-C estimates the maximal potential beta cell function of an individual, rather than the current beta cell function. We test this approach using longitudinal cohorts of prediabetic people, finding 10-fold variation in carrying capacity. Low carrying capacity is associated with higher risk of transitioning to diabetes. We estimate the timescales of beta cell compensation and insulin resistance using large datasets, showing that, unlike previous mathematical models, the new model can explain the slow rise in glucose over decades. Our mathematical understanding of beta cell carrying capacity may help to assess the risk of prediabetes in each individual.

systems biology↗

Compression of sickspan by interventions that steepen the survival curve

Longevity research aims to extend the period of healthy life, known as the healthspan, and to minimize the duration of disability and morbidity, known as the sickspan. Most longevity interventions in model organisms extend healthspan, but it is not known whether they extend healthspan relative to the lifespan - that is, whether they compress relative sickspan. Here, we present a theory that predicts which interventions effectively compress relative sickspan. The theory is based on the shape of the survival curve - the fraction of organisms surviving as a function of age. Interventions that extend mean lifespan while preserving the shape of the survival curve, known as scaling, are predicted to extend the sickspan proportionally, without compressing it. Such interventions include caloric restriction and many other longevity interventions. Conversely, a small subset of interventions that extend lifespan and steepen the shape of the survival curve, are predicted to compress the relative sickspan. We explain this based on the saturating removal mathematical model of aging, and present evidence from longitudinal health data in mice, Caenorhabditis elegans and Drosophila melanogaster. We discuss longevity interventions in mice that steepen the survival curves, including senolytics, ketogenic diet, and agents that reduce glucose spikes and protect blood vessels, as potential candidates for compressing the sickspan. We apply the theory to combinations of longevity interventions and discuss human healthspan data. This approach offers potential strategies for compressing morbidity and extending healthspan.

systems biology↗

Longitudinal hair cortisol in bipolar disorder and a mechanism based on HPA dynamics

Bipolar disorder (BD) is a dynamic disease in which mania, depression and mixed states vary on a timescale of months to years. BD patients characteristically exhibit elevated levels of the hormone cortisol. Persistently elevated cortisol can also cause mood episodes in a substantial fraction of the general population. Although BD is a dynamic disease that is related to cortisol, longitudinal cortisol dynamics in BD have rarely been studied. Here we use hair to measure past cortisol where each cm of hair corresponds to a month of growth. Cortisol was measured in 12 cm hair samples from people with BD (n=26) and controls (n=59), corresponding to one year of cortisol data. We found that hair cortisol exhibited a frequency spectrum with enhanced year-scale fluctuations whose amplitude was about 4-fold higher on average in BD compared to controls. Cortisol in the proximal 2 cm hair segment correlated with mood scales that report on mood in the past two months. In line with the notion that cortisol correlates with mood, we find that the mean frequency spectrum of depression (n=266) and mania (n=273) scores from a large longitudinal study of BD is similar to the hair cortisol spectrum from the present cohort. Taken together, these results suggest a mechanism for BD as the intersection of two neuropsychological traits: cortisol-induced mood episodes (CIM) and high emotional reactivity (ER). High ER causes fluctuations in which cortisol is elevated for months, as shown by a mathematical model of the hypothalamic-pituitary-adrenal (HPA) axis that regulates cortisol. In individuals with CIM, the magnitude of these persistent cortisol fluctuations can be high enough to trigger mood episodes. Thus, this study combines longitudinal cortisol measurements and mathematical modeling to provide a potential mechanistic link between the timescales of cortisol and moods in BD.

systems biology↗

Unifying design principles of endocrine gland mass and its regulatory circuits

Hormones are regulatory molecules that impact physiological functions. Much is known about individual hormones, but general rules that connect the regulatory logic of different hormone systems are limited. In this study, we analyzed a range of human hormone systems using a mathematical approach to integrate knowledge on endocrine cells, target tissues and regulation, to uncover unifying principles and regulatory circuits. We find that the number of cells in an endocrine gland is proportional to the number of cells in its target tissues, as one single endocrine cell serves approximately 2000 target cells. We identified five classes of regulatory circuits, each has specific regulatory functions such as homeostasis or allostasis. The most complex class includes an intermediate gland, the pituitary, which can otherwise be considered redundant and exposes to fragilities. We suggest a tradeoff: with the price of fragilities comes advantages -amplification, buffering of hypersecreting tumors, and faster response times. By elucidating these unifying principles and circuits, this study deepens our understanding of the control of endocrine processes and builds the foundation for systems endocrinology.

physiology↗

Pregnancy and postpartum dynamics revealed by an atlas of millions of lab tests

Pregnancy and delivery involve dynamic alterations in many physiological systems. However, the physiological dynamics during pregnancy and after delivery have not been systematically analyzed at high temporal resolution in a large human population. Here we present the dynamics of 76 lab tests based on a cross-sectional analysis of roughly 41 million measurements from over 300,000 pregnancies. We analyzed each test at weekly intervals from 20 weeks preconception to 80 weeks postpartum, providing detailed temporal profiles. About half of the tests take three months to a year to return to baseline during postpartum, highlighting the physiological load of childbirth. The precision of the data revealed effects of preconception supplements, overshoots after delivery and intricate temporal responses to changes in blood volume and renal filtration rate. Pregnancy complications - gestational diabetes, pre-eclampsia and postpartum hemorrhage - showed distinct dynamical changes. These results provide a comprehensive dynamic portrait of the systems physiology of pregnancy.

physiology↗

Excitable dynamics of flares and relapses in autoimmune diseases

Many autoimmune diseases show flares in which symptoms erupt and then decline. A prominent example is multiple sclerosis (MS) in its relapsing-remitting phase. Mathematical models attempting to capture the flares in multiple sclerosis have often been oscillatory in nature, assuming a regular pattern of symptom flare-ups and remissions. However, this fails to account for the non-periodic nature of flares, which can appear at seemingly random intervals. Here we propose that flares resemble excitable dynamics triggered by stochastic events and show that a minimal mathematical model of autoimmune cells and inhibitory regulatory cells can provide such excitability. In our model, autoimmune response releases antigens that cause autoimmune cells to expand in a positive feedback loop, while regulatory cells inhibit the autoimmune cells in a negative feedback loop. The model can quantitatively explain the decline of MS relapses during pregnancy and their postpartum surge based on lymphocyte dynamics, as well as the decline in MS relapses with age. The model also points to potential therapeutic targets and predicts that even small modulation of regulatory T cell production, removal or activity can have a large effect on relapse rate. Excitable dynamics may underlie flares and relapses found across autoimmune diseases, thus providing an understanding that may help improve treatment strategies.

systems biology↗

Robust differentiation in a synthetic stem-cell circuit

Differentiation is a process fundamental to multicellularity. In its simplest form, differentiation converts self-renewing stem cells into non-proliferative cells with specified function. This process is inherently susceptible to mutant takeover -- mutant stem cells that never differentiate produce excess proliferative daughter cells, driving cancer-like expansion and decreasing the availability of differentiated cells to the organism. It has been proposed that coupling differentiation to an essential trait can select against these mutants by producing a biphasic fitness curve. This would provide mutant stem cells that do not differentiate with a selective disadvantage. However, this theory has yet to be tested experimentally. Here we use "fitness landscape engineering" to design and construct a synthetic biological model of stem cell differentiation in Escherichia coli with biphasic fitness. We find that this circuit is robust to mutations as predicted. Surprisingly, its optimal differentiation rate is robust to a wide range of environmental pressures. This environmental robustness is driven by transit-amplifying cells that differentiate and proliferate irrespective of environment. These results provide new interpretations for natural differentiation mechanisms and suggest strategies for engineering robust, complex multicellular consortia.

synthetic biology↗

p16-dependent upregulation of PD-L1 impairs immunosurveillance of senescent cells

The accumulation of senescent cells promotes aging, but a molecular mechanism that senescent cells use to evade immune clearance and accumulate remains to be elucidated. Here, we report that p16-positive senescent cells upregulate the immune checkpoint protein programmed death-ligand 1 (PD-L1) to accumulate in aging and chronic inflammation. p16-mediated inhibition of CDK4/6 promotes PD-L1 stability in senescent cells via the downregulation of ubiquitin-dependent degradation. p16 expression in infiltrating macrophages induces an immunosuppressive environment that can contribute to an increased burden of senescent cells. Treatment with immunostimulatory anti-PD-L1 antibody enhances the cytotoxic T cell activity and leads to the elimination of p16, PD-L1-positive cells. Our study uncovers a molecular mechanism of p16-dependent regulation of PD-L1 protein stability in senescent cells and reveals the potential of PD-L1 as a target for treating senescence-mediated age-associated diseases.

cell biology↗

Circuit to target approach defines an autocrine myofibroblast loop that drives cardiac fibrosis

Fibrosis is a broad pathology of excessive scarring with substantial medical implications. The fibrotic scar is produced by myofibroblasts that interact with macrophages. Fibrosis is a complex process involving thousands of factors, therefore, to better understand fibrosis and develop new therapeutic approaches, it is necessary to simplify and clarify the underlying concepts. Recently, we described a mathematical model for a macrophage-myofibroblast cell circuit, predicting two types of fibrosis - hot fibrosis with abundant macrophages and myofibroblasts, and cold fibrosis dominated by myofibroblasts alone. To test these concepts and intervention strategies in a medically relevant system, we use a widely studied in-vivo injury model for fibrosis, myocardial infarction (MI). We show that cold fibrosis is the final outcome of MI in both mice and pigs and demonstrate that fibrosis can shift toward healing in regenerative settings. MI begind with an increase of myofibroblasts and macrophages, followed by macrophage decline leading to persistent cold fibrosis (only myofibroblasts). During this process, fibroblasts, unlike macrophages, acquire distinct fate changes. Using mathematical modeling we predict that targeting of the autocrine signal for myofibroblast division could block cold fibrosis. We identify TIMP1 as an autocrine cardiac myofibroblast growth factor in-vitro. Treatment of adult mice after MI with anti-TIMP1 antibodies reduces fibrosis in-vivo. This study shows the utility of the concepts of hot and cold fibrosis and the feasibility of our circuit-to-target approach to reduce fibrosis after acute cardiac injury by inhibiting the myofibroblast autocrine loop.

systems biology↗