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Hernandez, S.

Publications and source records attributed to Hernandez, S..

At least 19 recordsLinked to original sources
bioRxiv2026

A foundation model for brain organoid network dynamics uncovers intrinsic computation and convergent disease phenotypes

Brain organoids develop self-organizing neural networks that can be used to model neurodevelopment and disease. Here we introduce PULSE, a foundation model developed using over 900 hours of high-density microelectrode array recordings of in vitro and ex vivo neural tissue. We show that this single model can address two open questions: how disease-associated variants alter network development, and how naive circuits encode information. PULSE revealed accelerated network maturation in held-out human 16p11.2-deletion organoids. It also revealed a change in network coordination shared with SHANK3 deficiency. Because organoids develop without sensory input, they let us ask how circuits encode information before any representation is learned. Closed-loop control of a virtual inverted pendulum engaged existing spontaneous activity patterns rather than creating new ones. PULSE provides a common reference frame for neural circuits across genotypes, protocols and tasks.

neuroscience↗
bioRxiv2026

Soluble pathogenic tau transmission to astrocytes drives acute oxidative damage, cellular senescence, and neurovascular uncoupling in a model of Alzheimers tauopathy

We previously found that soluble pathogenic tau aggregates (tau oligomers) enter brain microvascular endothelial cells and induce cellular senescence and microvascular dysfunction in a tauopathy mouse model. This study shows that soluble pathogenic tau is also transmitted to astrocytes, where it induces mitochondrial dysfunction, mediates senescence, and impairs neurovascular coupling responses. Single-cell RNA sequencing of hTau mouse cortex revealed astrocytes as one of the most transcriptionally altered cell type, showing coordinated downregulation of electron transport chain genes and upregulation of stress-induced and inflammatory markers, also elevated in hTau mouse brain and astrocyte-enriched fractions. Similar to neuron-to-neuron transmission, soluble tau aggregates entered primary human astrocytes via a heparin-sensitive process, causing microtubule destabilization, ATP depletion, and mitochondrial ROS accumulation before induction of cell-cycle-arrest-associated markers. Mitochondrial ROS scavenging with Mito TEMPO reduced tau-induced SASP cytokine activation in astrocytes in vitro and in vivo. Coculturing neurons with astrocytes undergoing tau induced senescence decreased dendritic spine density, branch level, and dendritic area in a non-cell-autonomous manner. Tau and the SASP-associated cytokines IL-1{beta} and IL-6 contributed to distinct aspects of the neuronal structural phenotype. Astrocyte-targeted SOD2 overexpression attenuated the hTau-related deficit in evoked cerebral blood flow responses; however, this partial improvement in response magnitude and duration did not achieve statistical significance compared to hTau mice expressing GFP. These results identify astrocyte senescence as a potential mechanism connecting astrocyte tau uptake, mitochondrial stress, and neuronal structural impairment. The findings motivate further study of mitochondrial antioxidant defense in tau-associated astrocyte dysfunction.

neuroscience↗
bioRxiv2026

ATG-9-Induced Lysosomal Membrane Permeabilization and Cell Death in a Caenorhabditis elegans model of Mucolipidosis type IV

Mucolipidosis type IV is a lysosomal storage disease that is characterized by delayed psychomotor development and retinal degeneration due to cell death, in addition to other symptoms that are due to aberrant functions of live tissues. Caenorhabditis elegans CUP-5 is the orthologue of human TRPML1, the protein that is dysfunctional in Mucolipidosis type IV patients. Mirroring Mucolipidosis type IV pathology, loss of C. elegans CUP-5 results in developing intestinal cell death in embryos leading to embryonic lethality, while other tissues in adults lacking CUP-5 are alive but dysfunctional. We had previously shown that ESCRT-Associated proteins and the ATP-Binding Cassette Transporter MRP-4 are necessary for acquiring aberrant and poorly functional lysosomes in the absence of CUP-5. In this study, we show that the aberrant lysosomes permeabilize or rupture, thus releasing lysosomal degradative enzymes that kill cells in the absence of CUP-5. We also show that the autophagy-related protein ATG-9 mediates, in an autophagy-independent manner, this lysosomal permeabilization. We finally propose phenotypic and biochemical models linking CUP-5 to lysosomal defects and cell death.

cell biology↗
bioRxiv2026

The differentiation and functional landscape of tumor-resident CD8⁺ T cells shapes immunity and clinical outcomes in clear cell renal cell carcinoma

Clear cell renal cell carcinoma (ccRCC) is a common and lethal kidney cancer in which CD8+ T cell-targeted immunotherapies are standard of care. However, tumor CD8+ T cell infiltration correlates with divergent clinical outcomes, reflecting functional heterogeneity. We comprehensively characterized tumor-specific memory CD8+ T cells in ccRCC using phenotypic, transcriptional and functional analyses. Three major subsets were identified based on tissue-residency markers: circulating (Tcirc), tissue-resident (Trm) and CD69+CD103- Trm-like cells, all recognizing autologous RCC cells in an HLA class I-dependent manner. Tcirc cells exhibited greater stemness and cytotoxic potential, whereas Trm populations displayed tissue-residency programs, enhanced tumor reactivity and exhaustion features. Single-cell transcriptomics analyses uncovered substantial heterogeneity within Trm cells, including progenitor, activated, transitional, type-17, interferon-responsive, regulatory and terminally exhausted states. Trajectory and TCR analyses indicated progressive differentiation from central-memory Tcirc cells toward tissue residency while losing circulation, cytotoxicity and stemness potentials and then progress toward terminal exhaustion. Enrichment of progenitor Trm cells associated with improved survival and favorable immunotherapy response, whereas regulatory or exhausted Trm cells correlated with poor outcomes. These findings define the differentiation and functional landscape of tumor-specific CD8+ T cells that shapes immunity and clinical outcomes in ccRCC.

immunology↗
bioRxiv2026

Cloud-Connected Pluripotent Stem Cell Platform Enhances Scientific Identity in Underrepresented Students

Stem cell research offers unique opportunities for authentic scientific engagement, yet infrastructure requirements have confined participation to elite institutions, perpetuating workforce disparities. We developed an integrated framework combining engineered biology, cloud-connected microscopy, and validated psychometric assessment to make pluripotent stem cell (PSC) experimentation widely accessible. The framework comprises three components: a doxycycline-inducible NGN2 mouse embryonic stem cell line for rapid neuronal specification, low-cost cloud microscopy for remote observation, and the validated Stem Cell Research Identity Scale (SCRIS) for quantifying educational outcomes. Implementation across a Title I high school and urban community college demonstrated significant increases in scientific identity. Students using differentiating PSCs showed broader science identity development than those using neuroblastoma cells, particularly in competence, research readiness, and recognition. High school students showed enhanced research competence gains compared to community college students despite equivalent intervention duration. Demographic analyses revealed enhanced effectiveness for Hispanic and first-generation college students. This framework provides a scalable model for broadening participation in advanced biomedical research.

scientific communication and education↗
bioRxiv2025

Characterisation of Two Plasmodium Virulence Factors Important for Lipid Metabolism and Disease Progression In Vivo

Malaria is caused by Plasmodium parasites and claims 610,000 deaths annually. Parasite invasion of red blood cells (RBCs) involves secretion of multiple proteins from specialised apical organelles (micronemes, rhoptries, and dense granules). These collectively facilitate host cell entry, establish a protective parasitophorous vacuole (PV), and directly and indirectly mediate interactions between the infected RBC (iRBC) and the endothelial wall. The cytoadherence of iRBCs results in sequestration within organs and plays a critical role in pathogenesis and virulence, but the function of many secreted parasite proteins remains poorly characterised. Here, we leverage spatial proteomics from Plasmodium falciparum to identify two novel Plasmodium berghei orthologue proteins containing hydrolase domains (MAP1, PBANKA_1425900, and RhoSH, PBANKA_1001500). Ultrastructure expansion microscopy reveals their localisation to the rhoptries in late schizogony, where co-immunoprecipitation assays show they interact with each other. In vivo studies demonstrate that these proteins help the parasite evade spleen-mediated clearance and contribute to disease progression. Deletion of both genes disrupts lipid metabolism and PV membrane morphology, suggesting a role in establishing or maintaining the protective PV membrane. Loss of map1 reduces sequestration of P. berghei iRBCs to adipose tissue, and conditional knockdown of the P. falciparum map1 orthologue results in reduced CD36-mediated cytoadhesion, suggesting a cross-species role in sequestration and cytoadherence. Our findings identify MAP1 and RhoSH as key mediators of Plasmodium virulence.

microbiology↗
bioRxiv2025

Fronto-Parietal Oscillatory Dynamics of Emotion Regulation as a Function of Adult Attachment Orientations

Emotion regulation enables individuals to modulate emotional experiences and behaviors according to situational demands. Within attachment theory, individual differences in attachment anxiety and avoidance are conceived as interpersonal dispositions that influence the quality and efficiency of emotion regulation strategies, potentially shaping the underlying neural dynamics. This study examined cortical oscillatory activity during two regulation strategies--cognitive reappraisal and expressive suppression--focusing on theta (4-8 Hz) and beta (15-30 Hz) bands at the source level. Forty adults (21 male, 18 female, 1 gender-unspecified; M = 27.58 years, SD = 8.71) performed an emotion regulation task involving emotionally evocative images from the International Affective Picture System (IAPS) while EEG was recorded. Cortical sources were reconstructed using standardized low-resolution brain electromagnetic tomography (sLORETA). Linear mixed-effects models (LMMs) assessed the effects of condition, region of interest (ROI), and attachment orientations on oscillatory power. Results showed that higher attachment anxiety predicted reduced theta power in the right dorsolateral prefrontal cortex (dlPFC) during reappraisal, indicating attenuated recruitment of cognitive control mechanisms. In the beta band, suppression reduced activity in the right parietal lobe/precuneus -- a region involved in self-referential and attentional processes -- across participants, while during reappraisal, attachment anxiety was linked to lower beta power and attachment avoidance to higher beta power in the left dlPFC. Together, these findings indicate that cognitive reappraisal and expressive suppression engage distinct cortical oscillatory systems, and that interpersonal dispositions modulate their neural implementation. Frontal theta appears to index top-down control during reappraisal, whereas beta activity shows a dual pattern: frontal beta variations reflect differences in control stability associated with attachment orientations, and parietal beta decreases during suppression suggest attentional disengagement from self-referential processing.

neuroscience↗
bioRxiv2025

Incubator-Free Organoid Culture in a Sealed Recirculatory System

Discovery in human biology is pivoting toward high-dimensional computational analysis of 3D in vitro models, but this progress is limited by reliance on conventional cell culture techniques. Realism and data collection are hindered by the environmental instabilities and accessibility constraints of standard incubators. We introduce an automated, sealed recirculatory system that eliminates these barriers, enabling unconstrained instrument integration and infrastructure-independent scalability. By employing gas-tight sealing, a liquid-phase gas buffer and a non-porous plastic gas exchanger, our technology maintains biological stability without the compromises of open-air vessels. This design eliminates the need for CO2 incubators and prevents the evaporative drift that typically plagues conventional open-culture vessels. Operating on the benchtop outside the cell culture suite, we demonstrate that our system supports continuous, multi-week live imaging of vascular organoids while maintaining metabolic viability, structural fidelity and electrophysiological activity in brain organoids comparable to traditional in-incubator cultures. TeaserA sealed benchtop device enables data-rich organoid culture beyond the constraints of the cell culture laboratory.

bioengineering↗
bioRxiv2025

United States stakeholder insights on genetic testing for equine health and breeding

The S1094 United States Department of Agriculture (USDA) Multistate Research Project is a community of scientists working together to utilize equine genetics and genomics to improve the health and well-being of horses while also providing educational and genetic testing resources to those in the equine industry. To assess the knowledge and priorities of stakeholders regarding genetic testing, we conducted an online survey targeting horse owners and enthusiasts in the United States. Data were collected from March to October 2024 with 412 respondents from 44 states completing the survey. Most of the survey participants were horse owners, and the two most common age ranges for respondents were 31-50 (33.6%) and 51-70 (33.3%) years old. The two most common use categories for horses reported by respondents were sport and stock horses. The survey revealed that a majority of respondents were confident in their knowledge of genetics and that they relied primarily on breed and discipline organizations, as well as universities for their genetic information. Secondarily they relied upon social media and their veterinarians. Respondents expressed significant interest in genetic testing for diversity and performance traits. Across all horse use categories, arthritis, colic and intestinal disease, laminitis, metabolic disorders, and tendon and ligament issues had the greatest level of concern - all of which are complex genetic traits and will require collaborative efforts to discern levels of genetic involvement. Results from the survey underscore the need for continued outreach and education of horse owners, as well as further development of genetic testing tools to support informed decision-making among stakeholders in the equine industry. Lay SummaryGenetic testing helps horse owners, breeders, and breed registries and associations verify parentage, evaluate health, and improve performance. The S1094 USDA Multistate Research Project has a mission of improving genomics and genetics tools, as well as providing educational resources for those in the equine industry. To better understand the knowledge and priorities of stakeholders in the equine industry, we conducted an online survey netting responses from 412 individuals in the United States. Respondents were primarily horse owners who self-identified as confident in their knowledge of horse genetics. Furthermore, those responding indicated that their primary resources for information about genetics were breed and discipline association websites and universities. They also expressed interest in genetic testing for diversity and performance traits. Across all horse use categories, arthritis, colic and intestinal disease, laminitis, metabolic disorders, and tendon and ligament issues had the greatest level of concern. Given the complex nature of these conditions, collaborative efforts will be required to discern levels of genetic involvement. Respondents felt that genetic testing and education efforts have significant impacts on the equine industry because they aid owners in making breeding decisions and in the management of horse health and well-being. Teaser TextThis nationwide survey of U.S. horse owners discerned strong stakeholder interest in equine genetic testing, determined sources for knowledge acquisition, and highlighted key priorities for the development of genetic tools and educational materials.

genetics↗
bioRxiv2025

Epigenetic Subtypes of High-Grade T1 Bladder Cancer Reveal Intra-Tumor Heterogeneity and Distinct Interactions with Tumor Microenvironment.

High-Grade T1 (HGT1) Non-Muscle Invasive Bladder Cancer (NMIBC) is a clinically heterogeneous disease, characterized by unpredictable treatment responses and limited tools for recurrence prediction. Although molecular classification efforts have been made, patient stratification still primarily relies on clinicopathological features, which offer limited clinical precision. In this study, we integrated chromatin profiling in bulk with single-nuclei(sn) RNA-seq, immunohistochemistry and spatial transcriptomics to define epigenetic subtypes of HGT1, characterize their heterogeneity, and investigate tumor-microenvironment interactions. Our findings reveal distinct chromatin profiles differentiating urothelial (URO) and micropapillary (MP) histological variants of high-risk HGT1. We identified three epigenetic states: two within the URO group, luminal-like inflammatory (LLI) and basal-like (BL), and a separate signature unique to MP tumors. Single-cell and spatial resolution approaches validated intratumoral heterogeneity and provided insight into subtype-specific microenvironmental contexts. Notably, [~]40% of URO tumors exhibited spatially distinct coexisting LLI and BL components. BL regions showed enrichment for angiogenesis and hypoxia pathways and were preferentially located near vascular stroma, while LLI regions showed to be located at the core of the tumor. MP tumors featured a markedly different microenvironment, characterized by diverse populations of cancer-associated fibroblasts (CAFs) and M2-polarized macrophages intermingled with tumor cells, suggesting a more immunosuppressive niche which could account for their worse clinical outcome. URO tumors, by comparison, showed a more immune-excluded phenotype. These findings provide a detailed molecular and spatial map of bladder cancer heterogeneity, highlighting how distinct epigenetic subtypes and histological variants are associated with tumor architecture and microenvironmental interactions. Our data underscore the need for subtype-specific therapeutic strategies to more effectively address the complexity already existing at HGT1 bladder cancer.

cancer biology↗
bioRxiv2025

The Green Band: Using Production and Catch to Judge Distortive Pressure on an Ecosystem

Ecosystem based fisheries management is an increasingly accepted approach for many fisheries management agencies. Indicators and methods that can usefully support the aspiration of making ecosystem based fisheries management decisions on tactical time frames are slowly accumulating, but have primarily focused on biomass status and trends. We first verify that fishing can distort the cumulative distribution of connections within an ecosystem, and then show (via theoretical testing and application to four contrasting case study locations) how relating catch with production by defining an acceptable band of exploitation pressure - the "Green Band" - can help inform fisheries decision makers as to whether an ecosystem is under distortive patterns of fishing pressure that will ultimately undermine the structure of the ecosystem.

ecology↗
bioRxiv2025

Self-Organizing Neural Networks in Organoids Reveal Principles of Forebrain Circuit Assembly

The mouse cortex is a canonical model for studying how functional neural networks emerge, yet it remains unclear which topological features arise from intrinsic cellular organization versus external regional cues. Mouse forebrain organoids provide a powerful system to investigate these intrinsic mechanisms. We generated dorsal (DF) and ventral (VF) forebrain organoids from mouse pluripotent stem cells and tracked their development using longitudinal electrophysiology. DF organoids showed progressively stronger network-wide correlations, while VF organoids developed more refined activity patterns, enhanced small-world topology, and increased modular organization. These differences emerged without extrinsic inputs and may be driven by the increased generation of Pvalb+ interneurons in VF organoids. Our findings demonstrate how variations in cellular composition influence the self-organization of neural circuits, establishing mouse forebrain organoids as a tractable platform to study how neuronal populations shape cortical network architecture.

neuroscience↗
bioRxiv2025

HIPPIE: A Multimodal Deep Learning Model for Electrophysiological Classification of Neurons

Extracellular electrophysiological recordings present unique computational challenges for neuronal classification due to noise, technical variability, and batch effects across experimental systems. We introduce HIPPIE (High-dimensional Interpretation of Physiological Patterns In Extracellular recordings), a deep learning framework that combines self-supervised pretraining on unlabeled datasets with supervised fine-tuning to classify neurons from extracellular recordings. Using conditional convolutional joint autoencoders, HIPPIE learns robust, technology-adjusted representations of waveforms and spiking dynamics. This model can be applied to electrophysiological classification and clustering across diverse biological cultures and technologies. We validated HIPPIE on both in vivo mouse recordings and in vitro brain slices, where it demonstrated superior performance over other unsupervised methods in cell-type discrimination and aligned closely with anatomically defined classes. Its latent space organizes neurons along electrophysiological gradients, while enabling batch and individual corrected alignment of recordings across experiments. HIPPIE establishes a general framework for systematically decoding neuronal diversity in native and engineered systems.

neuroscience↗
bioRxiv2025

Protein Secretion From Liver Ex Vivo Reveals Regulation of Hepatokines by Sex and Circadian Rhythm

Here, we use an ex vivo approach compatible with the circadian timescale to interrogate protein secretion from liver, revealing several findings. Proteomic analyses in male and female mice identify hundreds of proteins that exhibit time-of-day-dependent or clock-dependent secretion involved in extracellular matrix, immune, redox, xenobiotic, and fatty acid functions. Among these, the liver secretes more endostatin, a cleavage product of collagen type XVIII alpha 1 (COL18A1), during the inactive, fasting phase of the diurnal cycle. Temporal regulation of COL18A1/endostatin is dysregulated upon loss of Bmal1 through combined effects on Col18a1 transcriptional repression and proteolytic processing. Functional experiments in vivo and in vitro reveal that endostatin suppresses mitochondrial gene expression in white adipose tissue in a time-dependent manner and reduces mitochondrial respiration in adipocytes, while enhancing lipolysis. These results support a mechanism of inter-organ crosstalk whereby hepatically derived, temporally-restricted endostatin tunes adipocytes toward metabolic activities required during the fasting phase.

molecular biology↗
bioRxiv2025

The liver clock tunes transcriptional rhythms in skeletal muscle to regulate mitochondrial function

Circadian clocks present throughout the brain and body coordinate diverse physiological processes to support daily homeostasis and respond to changing environmental conditions. The local dependencies within the mammalian clock network are not well defined. We previously demonstrated that the skeletal muscle clock controls transcript oscillations of genes involved in fatty acid metabolism in the liver, yet whether the liver clock also regulates the muscle was unknown. Here, we use hepatocyte-specific Bmal1 KO mice (Bmal1hep-/-) and reveal that approximately one third of transcriptional rhythms in skeletal muscle are regulated by the liver clock vivo. Treatment of myotubes with serum harvested from Bmal1hep-/- mice inhibited expression of genes involved in metabolic pathways, including oxidative phosphorylation. Overall, the transcriptional changes induced by liver clock-driven endocrine-communication revealed from our in vitro system were small in magnitude, leading us to surmise that the liver clock acts to fine-tune metabolic gene expression in muscle. Strikingly, treatment of myotubes with serum from Bmal1hep-/- mice inhibited mitochondrial ATP production compared to WT and this effect was only observed with serum harvested during the active phase. Overall, our results reveal communication between the liver clock and skeletal muscle-uncovering a bidirectional endocrine communication pathway dependent on clocks in these two key metabolic tissues. Targeting liver and muscle circadian clocks may represent a potential avenue for exploration for diseases associated with dysregulation of metabolism in these tissues.

cell biology↗
bioRxiv2024

Comparison of imaging-based single-cell resolution spatial transcriptomics profiling platforms using formalin-fixed, paraffin-embedded tumor samples

Imaging-based spatial transcriptomics (ST) is evolving rapidly as a pivotal technology in studying the biology of tumors and their associated microenvironments. However, the strengths of the commercially available ST platforms in studying spatial biology have not been systematically evaluated using rigorously controlled experiments. In this study, we used serial 5-m sections of formalin-fixed, paraffin-embedded surgically resected lung adenocarcinoma and pleural mesothelioma tumor samples in tissue microarrays to compare the performance of the single cell ST platforms CosMx, MERFISH, and Xenium (uni/multi-modal) platforms in reference to bulk RNA sequencing, multiplex immunofluorescence, GeoMx Digital Spatial Profiler, and hematoxylin and eosin staining data for the same samples. In addition to objective assessment of automatic cell segmentation and phenotyping, we performed pixel-resolution manual evaluation of phenotyping to carry out pathologically meaningful comparison between ST platforms. Our study detailed the intricate differences between the ST platforms, revealed the importance of parameters such as tissue age and probe design in determining the data quality, and suggested reliable workflows for accurate spatial profiling and molecular discovery.

bioinformatics↗
bioRxiv2024

Goal-Directed Learning in Cortical Organoids

Experimental neuroscience techniques are advancing rapidly, with major recent developments in high-density electrophysiology and targeted electrical stimulation. In combination with these techniques, cortical organoids derived from pluripotent stem cells show great promise as in vitro models of brain development and function. Although sensory input is vital to neurodevelopment in vivo, few studies have explored the effect of meaningful input to in vitro neural cultures over time. In this work, we demonstrate the first example of goal-directed learning in brain organoids. We developed a closed-loop electrophysiology framework to embody mouse cortical organoids into a simulated dynamical task (the inverted pendulum problem known as Cartpole) and evaluate learning through high-frequency training signals. Longitudinal experiments enabled by this framework illuminate how different methods of selecting training signals enable improvement on the tasks. We found that for most organoids, training signals chosen by artificial reinforcement learning yield better performance on the task than randomly chosen training signals or the absence of a training signal. This systematic approach to studying learning mechanisms in vitro opens new possibilities for therapeutic interventions and biological computation.

neuroscience↗