Biology subjects

Rodrigues, A.

Publications and source records attributed to Rodrigues, A..

At least 19 recordsLinked to original sources

bioRxiv2026

StaBLE: digital metrics capture balance performance across a wide spectrum of balance tasks and abilities

Balance is fundamental to mobility and independence, yet it remains difficult to measure in a way that is both clinically meaningful and feasible for widespread adoption. There is a need for more quantitative, objective, and accessible balance assessments. To this end, we created the Stanford Balance Level Evaluation (StaBLE), a battery of 18 balance-challenging tasks and accompanying digital performance metrics computed from video that aimed to capture widely ranging balance abilities across 180 participants. We validated the StaBLE score against existing clinical scales and found that it correlated with age ({rho} = -0.76, p<0.001), the Activities-Specific Balance Confidence Scale ({rho} = 0.60, p<0.001), a Mini-Balance Evaluation Systems Test task ({tau}=0.55, p<0.001), 4-Stage Balance Test ({tau} = 0.55, p<0.001), and Short Physical Performance Battery ({tau} =0.57, p<0.001). StaBLE overcame ceiling effects in these existing scales and had a different underlying distribution for those identified at risk of falling and not (KS: 0.67, p<0.001). To reduce the time it takes to perform the test, we identified subsets of balance tasks that best predicted the StaBLE score across different populations. Together, our dataset, balance assessment protocol, and performance-based score demonstrate the validity of digital measures of balance.

bioengineering↗

bioRxiv2026

MicroRNA correlates of resilience to Alzheimer's disease identify candidate therapeutic targets for neuroprotection

Many aged individuals accumulate advanced Alzheimers disease (AD) neuropathology without cognitive decline, signifying potent endogenous resilience mechanisms. Mimicking resilience to AD may offer new therapeutic opportunities by emulating endogenous neuroprotection, but the molecular pathways that underpin resilience remain largely unknown. MicroRNAs (miRNAs)--small noncoding RNA molecules that post-transcriptionally regulate gene expression--influence neuronal and glial processes associated with development, aging, and AD neurodegeneration but their role in resilience has not been characterized. We investigated resilience-associated miRNAs and their targeted programs in the dorsolateral prefrontal cortex of post-mortem human brains. We analyzed matched miRNA and messenger RNA (mRNA) expression across resilient, AD, and control subjects from the Religious Order Study and Memory and Aging Project (ROSMAP) cohort. Nine miRNAs were differentially expressed in resilience compared with AD, including previously unreported miR-362-3p and miR-433-3p and new resilience-associated roles for known AD-related miRNAs, including neuronal miR-132-3p and miR-129-5p. By analyzing miRNA activity across AD progression, we found additional cognitive-associated miRNAs (e.g., miR-335-5p and miR-19b) and one plaque-restricted miRNA (miR-199a-5p). Sex-specific miRNA dysregulation was observed: miR-7-5p showed male-specific upregulation in AD versus resilience and suggested sex-specific differences in AD patients. Integrated co-expression and target-enrichment analyses linked resilience-associated miRNAs to pathways that were associated with cognitive decline, including transforming growth factor {beta}, Rho guanosine triphosphatases, and neurotransmitter receptor regulation. We also report restricted co-activity in AD subjects for miR-362-3p with inflammatory pathways, including tumor necrosis factor and Toll-like receptor signaling. These results demonstrate systematic involvement of miRNAs across neuronal and glial programs of AD resilience, cognitive decline, and sex-specific regulation. Our study provides an important resource for discovery of actionable regulatory programs that could lead to new therapies, based on endogenous molecules, that emulate natural resilience to AD. Key pointsO_LINine distinct cortical microRNA (miRNA) signatures correlate with resilience to Alzheimers disease (AD)-related cognitive decline, pointing to endogenous regulatory programs that may help preserve cognitive function. C_LIO_LIAt the miRNA level, resilient brains show minimal or no detectable differences to healthy individuals, despite the profound pathological differences, supporting the idea that resilience reflects preserved molecular homeostasis rather than a separate disease state. C_LIO_LISome miRNA signatures are specific to a certain pathology (e.g., one linked to amyloid-{beta} plaques independently of tau pathology and cognitive decline), helping to disentangle the regulation of these processes. C_LIO_LISex-specific miRNA dysregulation in AD suggests that resilience-linked pathways may be regulated differently in males and females, reinforcing the value of sex-stratified analyses and the potential for differential therapeutic strategies. C_LIO_LIIntegrative miRNA-pathway analysis highlights candidate regulatory networks involved in inflammation, matrisome, and cellular stress responses, providing a framework for resilience-linked therapeutic targets. C_LI

neuroscience↗

bioRxiv2026

Atta Leafcutter Ants are Fine-Scale Bioindicators of Geographic and Seasonal Climate Changes Across the Americas

AimWe develop Atta leafcutter-ants as bioindicators that respond at fine scales to geographic and seasonal climate changes in the Americas, thereby addressing the paucity of versatile insect bioindicator systems capable of monitoring climate in both Southern and Northern Hemispheres. LocationAmerican tropics and sub-tropics from latitudes S33.6{degrees} to N33.2{degrees}, with case studies from Colombia, Mexico, and southern USA. Time Period2012-2024. Taxon StudiedAtta leafcutter-ants. MethodsWe elucidate biogeographic patterns of mating-flight phenology of Atta leafcutter-ants across the entire Atta range from Uruguay/Argentina to the USA, using 2335 records of Atta reproductives from the community database iNaturalist, then ground-truth these patterns by comparison with (i) mating-flight records (n=806) from the Atta literature; and (ii) mating-flight observations (n=836) accumulated by a consortium of experts who have researched Atta for a combined 1000+ work-years. Onset of mating flights can be timed with great precision in Atta populations because mass-mating flights are synchronized and triggered by the first major rainfall of a rainy season. ResultsBiogeographic patterns in climate-dependent mating-flight phenologies recorded at iNaturalist are corroborated by observations accumulated in the literature and by Atta experts. Analyses reveal so-far unknown gradients in mating-flight phenology (e.g., Colombia to USA) that are correlated to geographic climate gradients, and season switches of mating flights from early to late in the year between proximate Atta populations (e.g., 200 kilometers apart), for example in the climatically complex Andean regions of Colombia. Regional differences in Atta mating-flight phenology correspond to temporal differences in rainfall between ecoregions of Colombia. Main ConclusionsAtta ants are tractable insect bioindicators to monitor climate impacts with detailed spatial and temporal resolution across a 9200-kilometer trans-equatorial transect in the Americas. We outline future research directions to explore climate-dependent biology using the continuously growing, and now ground-truthed, information at iNaturalist on Atta mating behavior.

ecology↗

bioRxiv2026

High residual pesticide contamination despite contrasted feeding treatments in semi-captive bred grey partridges

Food security is one of the major ongoing global anthropic challenges, driving the intensification of agricultural systems notably through the widespread use of phyto-pharmaceutical products (PPPs). Although effective for maximizing production yields through pest control, PPP contamination is pervasive across the entire agroecosystem, raising concerns about human and non-target species exposure and health effects. Organic farming has therefore been promoted as a sustainable alternative to ensure human food security and mitigate these impacts. Yet, the pathways of PPP non-target organisms contamination are insufficiently understood. While most research has focused on ingestion as the primary route of exposure, increasing evidence suggests more complex contamination dynamics. In this study, we investigated the role of food intake in shaping PPP contamination profiles in semi-captive grey partridges (Perdix perdix). To do so, 80 birds were housed in open aviaries within an agricultural landscape and fed either conventional or organic grains over a five-month period. After drawing their PPP blood profiles at the end of the exposure period, we assessed their loads and compared the pesticide prevalence and abundance between food treatments. We found similar quantity and diversity of PPP residues in blood from conventionally- and organically-fed partridges, with few differences in specific PPP compounds. Overall, these findings suggest that food intake is not necessarily the only source of contamination for non-target organisms, with alternative pathways (e.g., inhalation, dermal contact, soil and water exposure) likely playing a substantial role. Integrating multi-pathway contamination monitoring across trophic levels within a One Health framework would now be essential to better understand and mitigate PPP contamination risks.

ecology↗

bioRxiv2026

Scalable microvascular networks-on-chip enable 8-week unidirectional perfusion for long-term vascular toxicity screening

Cardiovascular toxicity is a leading cause of late-stage drug attrition, yet current in vitro vascular models lack the longevity and scalability needed to capture clinically relevant responses in mature vasculature. We developed a vascular microphysiological system comprising 32 parallel, self-assembled microvascular networks from primary human endothelial cells and stromal fibroblasts. Networks were unidirectionally perfused by gravity-driven, pump-free flow. They remained functionally perfusable for at least 57 days, the longest duration reported for self-assembled microvascular networks, and underwent progressive maturation characterised by perivascular fibroblast organisation, basement membrane deposition, and matrix remodelling. Variance decomposition confirmed high reproducibility, with intra-plate variability of 3-7% and no operator-dependent effects on network morphometry. Exploiting this extended culture window, we reveal maturation-dependent shifts in endothelial inflammatory responsiveness and cytotoxic susceptibility. Distinct acute and chronic toxicity profiles of clinically used tyrosine kinase inhibitors are resolved, and a concentration- and time-dependent spectrum of sorafenib-induced microvascular responses is produced. These results show that vascular longevity and progressive maturation are key requirements for assessing vascular toxicity, and provide a scalable platform for evaluating acute and chronic drug responses in mature human microvascular tissues.

cell biology↗

bioRxiv2026

Host cell traversal by Plasmodium parasites is essential for sterilizing hepatic cellular immunity.

An effective vaccine capable of inducing sterile protection against Plasmodium, the causative agent of malaria, is critical to aid global eradication. Whole-organism vaccines using liver-infective sporozoites provide high levels of sterile protection against pre-erythrocytic infection. Yet, determinants of sporozoite immunogenicity remain poorly characterized. Using rodent models of vaccination, we demonstrate that the ability of Plasmodium sporozoites to actively migrate through multiple host cells prior to infecting hepatocytes is required for sterilizing immunity, regardless of the time of intrahepatic development of immunizing parasites. We further establish that host cell traversal is sufficient to trigger robust protection against Plasmodium hepatic infection. Impaired cell traversal precludes protective liver-resident memory CD8 T cell responses following vaccination, but not the production of anti-plasmodial antibodies. Our findings challenge the prevailing notion that intrahepatic parasite development is the sole determinant of whole-sporozoite vaccination-induced protection, and highlight parasite behavior traits as critical immunogenic events shaping sterilizing cellular immunity against Plasmodium liver stages.

immunology↗

bioRxiv2025

Synergistic non-neutralizing plasma antibodies to PfRH5 drive potent malaria parasite growth inhibition

An effective blood-stage vaccine is needed to protect against malaria pre-erythrocytic stage breakthrough. P. falciparum reticulocyte-binding protein homolog 5 (PfRH5) has emerged as a promising blood-stage vaccine antigen candidate, reducing parasite growth in humans during malaria challenge and showing field efficacy in children. Here, we characterize the human plasma IgG response to the RH5.1 vaccine candidate at monoclonal resolution, revealing that plasma repertoires are dominated by abundant, non-neutralizing antibodies. Using oligoclonal reconstitution experiments, in which defined pools of recombinant plasma mAbs are reassembled and functionally tested, we map how individual antibody interactions shape parasite growth inhibition activity. This approach allows us to discern which antibodies, within a polyclonal setting, act additively or synergistically, thereby revealing the emergent properties of anti-PfRH5 IgG. We further show that IgG lineages targeting linear epitopes lack neutralizing activity, while non-neutralizing IgG lineages that bind conformational epitopes can exhibit potent, interdependent synergy with each other and with neutralizing mAbs. These synergistic antibodies were identified in the plasma IgG compartments of five volunteers and highlight non-neutralizing PfRH5 epitopes that are critical for polyclonal-mediated growth inhibition. Our findings have broad implications for PfRH5 vaccine immunogen engineering and the role of non-neutralizing antibodies in infectious disease immunity.

immunology↗

bioRxiv2025

Structural basis for synergistic antibody protection against the essential malaria invasion complex protein RIPR

Plasmodium falciparum RH5-interacting protein (RIPR) is central to the essential PTRAMP-CSS-RIPR-CyRPA-RH5 (PCRCR)-complex; a leading target of blood-stage malaria vaccines. However, mechanisms whereby anti-RIPR antibodies inhibit parasite invasion are poorly understood. Here, we characterise 83 human IgG mAbs from RIPR-vaccinated Kymouse platform mice. Single mAbs have minimal neutralising activity, however, high-level synergistic inhibition is observed with pools of mAbs targeting the RIPR-Tail region. Structural characterisation and molecular dynamics simulations of RIPR-Tail show that mAbs targeting EGF-like domains 6-8 (RIPREGF (6-8)), but not EGF-like domains 9-10 or the C-terminal domain (RIPREGF (9-10)-CTD), synergise to constrain the RIPR-Tail conformation. The same antibodies dissociate PTRAMP-CSS from RIPR, thereby enabling anti-RIPREGF (9-10)-CTD mAbs or anti-CSS sdAbs to bind and potentiate anti-RIPREGF (6-8) IgG. Addition of these mAbs to IgG from humans immunised with the R78C (RIPREGF (7-8)-CyRPA) candidate vaccine enhances malaria growth inhibition. These data provide a framework to guide next-generation blood-stage malaria vaccine design.

immunology↗

bioRxiv2025

Comparative genomics of Sympodiorosea identifies genome evolution mediated through selective pressure on the metabolic gene repertoire

Biological interactions involving host-associated fungi are manifestations of chemistry shaped over evolutionary time. For antagonistic fungi, specialization to host fungi is often facilitated through the acquisition of genes encoding novel secretions, including proteins and specialized metabolites that shape interactions with host defenses, as well the acquisition of nutrients. Alternatively, novel function may arise from diversifying selection eliciting evolutionary innovation in inert secretions. Through specialization, fungal metabolic capacity can serve as an imprint of the selective forces imposed on the antagonist and provide insights into its natural history and occupied niche. Here, we conduct a phylogenomic investigation of antagonistic ascomycetes in the genus Sympodiorosea, which are commonly found in basal fungus-growing ant gardens. Sympodiorosea and closely related genera of Escovopsis, Luteomyces, and Escovopsioides are canonically referenced as virulent mycoparasites, however, recent work has illuminated the possibility for diverse, non-virulent species to emerge within these attine-associated fungi. We explored genomic variation in Sympodiorosea to gain insight into the genomic potential for alternative lifestyles, focusing on diversity and evolution of metabolic genes. Our study revealed a constrained selective landscape across the Sympodiorosea genome. However, outcomes of in vitro interactions with host fungi are diverse and predictable based on the antagonists ant-species-of-origin, suggesting functional diversification. Phylogenomics reveals that gain/loss events of genes involved in secretions (secretome) are potential drivers of diversification. Additionally, we demonstrate that purifying selection acts more intensely on secretome-related genes relative to other genes. In contrast, few genes from the secretome experience diversifying selection, suggesting a mechanistic role for driving both functional differences between strains and host-specialization. Comparative genomics including other fungi within the family Hypocreales reveals that Sympodiorosea has experienced expansions and contractions in proteases genes that are discordant from expectations under a strictly mycoparasitic lifestyle, indicating either the potential for an alternative lifestyle within ant gardens, or that Sympodiorosea recently evolved from other niches. These results provide novel insight into genomic evolution of these fungi and inform future experimental studies of the ecology and chemistry of interactions within the complex fungus-growing ant symbiosis.

evolutionary biology↗

bioRxiv2025

Spatiotemporal data integration for marine megafauna SDMs in dynamic environments: A point process approach

Species distribution models (SDMs) are widely used to predict where and when species occur, but the data available to fit them often resolve space and time unevenly. Designed surveys, such as line-transect distance-sampling surveys, provide structured spatial coverage and the detection information needed to estimate detectability, but logistical and funding constraints often limit them to short, infrequent periods. Opportunistic or citizen-science records are collected more frequently across the year, but from areas shaped by observer access and interest rather than survey design, and usually without comparable detection information. As a result, models built from either source alone tend to capture spatial structure or temporal dynamics well, but not both, limiting predictions for mobile species in dynamic environments. Here, we present a general framework, based on a log-Gaussian Cox process (LGCP) fitted using integrated nested Laplace approximations (INLA), that links multiple data sources to a shared spatiotemporal measure of species density through source-specific observation models: a detection function for survey transects, and spatial constraints reflecting the footprint of opportunistic effort. We apply the framework to cetacean line-transect surveys and whale-watching records. In a simulation with one month of survey data and twelve months of opportunistic data, the integrated model recovered spatiotemporal distribution patterns well (median correlation with the true pattern = 0.91) and estimated monthly abundance with a mean error of +10.7%, outperforming single-source models. In a case study of common dolphins (Delphinus delphis) off mainland Portugal, integration improved the spatial accuracy of predictions, although weak covariate effects limited the temporal variation the model could resolve. These results show that integration adds the most value when the two data sources resolve complementary aspects of distribution, in this case broad spatial structure from the survey and temporal replication from whale-watching, while temporal predictions remain limited when covariates carry weak seasonal signal. The framework is general and can be extended to other taxa, systems, and combinations of structured and opportunistic data by substituting the relevant observation models.

ecology↗

bioRxiv2025

A Comprehensive Structural and Functional Analysis of Saccharomyces Killer Toxins.

Antifungal killer toxins are cytotoxic proteins that have the potential to combat the growing threat of fungi to human health and agriculture. A lack of empirical tertiary structures has placed limitations on understanding their mechanisms of action and targeting of pathogens. AlphaFold and molecular dynamics simulations were used to create tertiary structure models of all canonical Saccharomyces killer toxins. These models have enabled the prediction of the functional domains of killer toxins and postranslational modifications, including sites of proteolytic cleavage and disulfide bonds. They have also revealed unexpected homology between Saccharomyces killer toxins, suggesting that all but K28 are likely ionophores. Structural homology with the well-studied killer toxins K1 and K2 enabled prior empirical data to predict the antifungal and immunity mechanisms of the K1L, K21, K45, K74, and KHS toxins. The understudied killer toxins Klus, KHR, and K62 were found to have homology to bacterial and plant toxins, including members of the aerolysin family and antifungal lectins. These structural similarities provided clues for the mechanisms of killer toxin carbohydrate binding, oligomerization, and membrane attack. Given the hundreds of sequence homologs of the Saccharomyces killer toxin identified across fungi, modeling studies offer an exciting opportunity to characterize novel toxin-like proteins. This approach is strengthened by the continued use of the model yeast S. cerevisiae to study killer toxins and the wealth of functional data gathered in the decades since their first discovery.

molecular biology↗

bioRxiv2025

Genomics insights reveal multi-year maintenance of a new Deltacoronavirus infecting Seabirds from the Cagarras Island Archipelago Natural Monument, Brazil

Previous studies have identified various pathogens in seabirds, notably coronaviruses (CoVs) and influenza A viruses (IAVs), due to their potential to cause significant morbidity and mortality. The Cagarras Island Archipelago Natural Monument, located near Rio de Janeiro, Brazil, serves as nesting site for two species, the magnificent frigatebird (Fregata magnificens) and the brown booby (Sula leucogaster). Despite its ecological importance, no prior studies have investigated viral infections in these species, which share habitat interfaces with densely populated human areas. To address this gap, we sampled and tested seabirds for CoVs and IAVs from January 2022 to April 2024. Birds were captured and identified by species, age, and sex. Oropharyngeal and cloacal swabs, as well as blood samples, were collected. Viral RNA was extracted using the QIAamp Viral RNA Mini Kit, and the presence of IAVs was screened via real-time RT-PCR, while CoVs were screened using semi-nested RT-PCR. Sanger and metatranscriptomic sequencing were performed to identify viral strains and assess phylogenetic relationships. Of the 153 seabirds sampled, CoVs were detected in 6 individuals (9.1%) of F. magnificens and 16 individuals (18.4%) of S. leucogaster. No IAVs were found in either oropharyngeal or cloacal swabs, and all serum samples were negative for the presence of antibodies against the virus. We recovered two full deltacoronavirus genomes and eight additional draft genomes from S. leucogaster samples obtained from distinct sampling expeditions and additional enteroviruses, passeriviruses, and picornaviruses. Phylogenetic analysis revealed that the detected CoVs are closely related to avian deltacoronaviruses from environmental samples of S. leucogaster in the Sao Pedro and Sao Paulo Archipelago, indicating potential viral exchange between these seabird populations living at these distant islands. Moreover, multiple detections in different individuals at different time points are associated with specific Spike NTD deletions that have been shown to accumulate in immune escape lineages, supporting the long-term maintenance through new infections and reinfection of this virus in these bird populations. This is the first detection of CoVs in F. magnificens, highlighting their circulation in marine ecosystems. Further research is needed to understand the ecological and epidemiological implications, including potential cross-species transmission.

microbiology↗

bioRxiv2024

A polyketide-based biosynthetic platform for diols, amino alcohols and hydroxyacids

Medium- and branched-chain diols and amino alcohols are important industrial solvents, polymer building blocks, cosmetics and pharmaceutical ingredients, yet biosynthetically challenging to produce. Here, we present a novel approach utilising a modular polyketide synthase (PKS) platform for the efficient production of these compounds. This platform takes advantage of a versatile loading module from the rimocidin PKS and NADPH-dependent terminal thioreductases (TRs), previously untapped in engineered PKSs. Reduction of the terminal aldehyde with specific alcohol dehydrogenases enables production of diols, oxidation enables production of hydroxy acids, and transamination with specific transaminases enables production of various amino alcohols. Furthermore, replacement of the malonyl-coenzyme A (CoA)-specific acyltransferase (AT) in the extension module with methyl- or ethylmalonyl- CoA-specific ATs enables production of branched-chain diols and amino alcohols. In total, we demonstrated production of nine 1,3-diols (including the difficult-to-produce insect repellent and cosmetic ingredient 2-ethyl-1,3-hexanediol), six amino alcohols, and two carboxylic acids using our PKS platform in Streptomyces albus. Finally, tuning production of the PKS acyl-CoA substrates enabled production of high titers of specific diols and amino alcohols (1 g/L diol titer in shake flasks), demonstrating high tunability and efficiency of the platform.

bioengineering↗

bioRxiv2024

The Prevalence of Killer Yeasts in the Gardens of Fungus-Growing Ants and the Discovery of Novel Killer Toxin named Ksino.

Killer toxins are proteinaceous antifungal molecules produced by yeasts, with activity against a wide range of human and plant pathogenic fungi. Fungus gardens of attine ants in Brazil were surveyed to determine the presence of killer toxin-producing yeasts and to define their antifungal activities and ecological importance. Our results indicate that 10 out of 59 yeasts species isolated from fungal gardens are killer yeasts. Killer yeasts were less likely to inhibit the growth of yeasts isolated from the same environment but more effective at inhibiting yeast isolated from other environments, supporting a role for killer yeasts in shaping community composition. All killer yeasts harbored genome-encoded killer toxins lacked cytoplasmic toxin-encoding elements (i.e., double-stranded RNA satellites and linear double-stranded DNAs). Of all the killer yeasts associated with attine ants, Candida sinolaborantium (strain LESF 1467) showed a broad spectrum of antifungal activities against 39 out of 69 57% of yeast strains tested for toxin susceptibility. The complete genome sequence of C. sinolaborantium LESF 1467 identified a new killer toxin, Ksino, with similarities in primary sequence and tertiary structure to the Saccharomyces cerevisiae killer toxin named Klus. Surveys of publicly available genome databases identified homologs of Ksino in the genomes of yeast strains of Saccharomycetes and Pichiomycetes, as well as other species of Ascomycota and Basidiomycota filamentous fungi. This demonstrates that killer yeasts can be widespread in attine ant fungus gardens, possibly influencing fungal community composition and the importance of these complex microbial communities for discovering novel antifungal molecules. ImportanceAttine ants perform essential ecosystem services through the harvesting of substrates for fungiculture. The cultured fungi are a food source for attine ants. Characterizing antifungal toxin-producing yeasts (killer yeasts) is vital to understanding how they might protect gardens from invasion by unwanted fungal species. This study describes a new toxin named Ksino from the yeast Candida sinolaborantium, a member of a new group of toxins found across many different species of fungi. This work supports the role of killer yeasts in the ecology of fungicultures and competition between fungi. The observed high prevalence of killer yeasts in fungal gardens also enables the discovery of novel antifungal molecules with the potential to be applied against disease-causing fungi.

microbiology↗

bioRxiv2024

TBX15 regulates a network of immune response genes in adipose tissue and alters fat mass and depot weight in heterozygous knockout mice

Adipose tissue distribution in the body is an indicator of metabolic disease risk, independent of body mass index (BMI), and is indirectly measured by waist-hip-ratio (WHR). T-Box transcription factor-15 (TBX15) has been implicated in regulation of adipose distribution in multiple human and mouse studies, and the TBX15-WARS2 genome-wide association study locus has been associated with BMI-adjusted-WHR signals in multiple investigations. As a potential mediator of this signal, we investigated the role of Tbx15 using heterozygous and homozygous mouse knockout models to determine if loss of this gene alters adipose physiology, and to identify the transcriptional network regulated by Tbx15 in adipose tissue and preadipocyte cells. In a metabolic phenotyping experiment we provided either low fat diet (LFD) or high fat diet (HFD) to male and female heterozygous Tbx15+/- and wildtype Tbx15+/+mice from weaning and maintained for 24 weeks. Only Tbx15+/-mice maintained on LFD weighed less than wildtype LFD controls, and female LFD Tbx15+/- mice had lower fat mass overall. We found that in LFD Tbx15+/- mice, multiple visceral fat depots weighed less than wildtype controls, and this was maintained when corrected for body mass for both gonadal and mesenteric visceral adipose depots. When comparing adipocyte size in multiple adipose depots, some reduction in number of larger adipocytes was detected in the perirenal adipose tissue of female HFD Tbx15+/- vs Tbx15+/+ mice, mesenteric adipose tissue from female LFD Tbx15+/- vs Tbx15+/+ mice and male HFD Tbx15+/- vs Tbx15+/+mice. RNA-sequencing of subcutaneous (inguinal) adipose tissues from 12-week old male and female knockout Tbx15-/-, Tbx15+/- and Tbx15+/+ mice raised on a standard chow diet identified 897 upregulated genes and 2328 downregulated genes in female Tbx15-/- mice compared to Tbx15+/+mice. We then combined this dataset with TBX15 ChIP-sequencing data from mouse preadipocyte 3T3-L1 cells overexpressing TBX15 to identify a credible set of genes directly regulated by TBX15. These 52 genes were enriched for B- and T-cell receptor signalling, JAK-STAT signalling and haematopoietic cell lineage pathways; suggesting a direct regulatory role for TBX15 in these pathways in adipose tissue. Together, these data highlight a role for TBX15 in regulation of differential adipose tissue expansion, particularly under low caloric conditions. Further, we identify a potentially important role for TBX15 in the well described adipocyte-immune cell crosstalk associated with obesity and type 2 diabetes mellitus.

physiology↗

bioRxiv2024

VTA μ-opioidergic neurons facilitate low sociability in protracted opioid withdrawal

Opioids initiate dynamic maladaptation in brain reward and affect circuits that occur throughout chronic exposure and withdrawal that persist beyond cessation. Protracted withdrawal is characterized by negative affective behaviors such as heightened anxiety, irritability, dysphoria, and anhedonia, which pose a significant risk factor for relapse. While the ventral tegmental area (VTA) and mu-opioid receptors (MORs) are critical for opioid reinforcement, the specific contributions of VTAMOR neurons in mediating protracted withdrawal-induced negative affect is not fully understood. In our study, we elucidate the role of VTAMOR neurons in mediating negative affect and altered brain-wide neuronal activities following opioid exposure and withdrawal in male and female mice. Utilizing a chronic oral morphine administration model, we observe increased social deficit, anxiety-related, and despair-like behaviors during protracted withdrawal. VTAMOR neurons show heightened neuronal FOS activation at the onset of withdrawal and connect to an array of brain regions that mediate reward and affective processes. Viral re-expression of MORs selectively within the VTA of MOR knockout mice demonstrates that the disrupted social interaction observed during protracted withdrawal is facilitated by this neural population, without affecting other protracted withdrawal behaviors. Lastly, VTAMORs contribute to heightened neuronal FOS activation in the anterior cingulate cortex (ACC) in response to an acute morphine challenge, suggesting their unique role in modulating ACC-specific neuronal activity. These findings identify VTAMOR neurons as critical modulators of low sociability during protracted withdrawal and highlight their potential as a mechanistic target to alleviate negative affective behaviors associated with opioid withdrawal. SIGNFICANCEThe compelling urge for relief from negative affective states during long-term opioid withdrawal presents a crucial challenge for maintaining abstinence. The ventral tegmental area (VTA) and its mu-opioid receptor-expressing (VTAMOR) neurons represent a critical target of opioidergic action that underlie dependence and withdrawal. Chronic activation of VTAMOR neurons during opioid exposure induces maladaptations within these neurons and their structurally connected circuitries, which alter reward processing and contribute to negative affect. Using an oral morphine drinking paradigm to induce dependence, we demonstrate that withdrawal engages VTAMOR neurons and identify this neuronal population as key mediators of opioid withdrawal-induced social deficits. These findings hold promise to inform development of targeted therapies aimed at alleviating negative affective states associated with protracted opioid withdrawal.

neuroscience↗

bioRxiv2024

Mimicking opioid analgesia in cortical pain circuits

The anterior cingulate cortex is a key brain region involved in the affective and motivational dimensions of pain, yet how opioid analgesics modulate this cortical circuit remains unclear. Uncovering how opioids alter nociceptive neural dynamics to produce pain relief is essential for developing safer and more targeted treatments for chronic pain. Here we show that a population of cingulate neurons encodes spontaneous pain-related behaviors and is selectively modulated by morphine. Using deep-learning behavioral analyses combined with longitudinal neural recordings in mice, we identified a persistent shift in cortical activity patterns following nerve injury that reflects the emergence of an unpleasant, affective chronic pain state. Morphine reversed these neuropathic neural dynamics and reduced affective-motivational behaviors without altering sensory detection or reflexive responses, mirroring how opioids alleviate pain unpleasantness in humans. Leveraging these findings, we built a biologically inspired gene therapy that targets opioid-sensitive neurons in the cingulate using a synthetic mu-opioid receptor promoter to drive chemogenetic inhibition. This opioid-mimetic gene therapy recapitulated the analgesic effects of morphine during chronic neuropathic pain, thereby offering a new strategy for precision pain management targeting a key nociceptive cortical opioid circuit with safe, on-demand analgesia. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=40 SRC="FIGDIR/small/591113v2_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@d294c0org.highwire.dtl.DTLVardef@123d4f4org.highwire.dtl.DTLVardef@1720deborg.highwire.dtl.DTLVardef@efc632_HPS_FORMAT_FIGEXP M_FIG C_FIG

neuroscience↗

bioRxiv2024

Preclinical Development of a Stabilized RH5 Virus-Like Particle Vaccine that Induces Improved Anti-Malarial Antibodies

The development of a highly effective vaccine against the pathogenic blood-stage infection of human malaria will require a delivery platform that can induce an antibody response of both maximal quantity and functional quality. One strategy to achieve this includes presenting antigens to the immune system on virus-like particles (VLPs). Here we sought to improve the design and delivery of the blood-stage Plasmodium falciparum reticulocyte-binding protein homolog 5 (RH5) antigen, which is currently in a Phase 2 clinical trial as a full-length soluble protein-in-adjuvant vaccine candidate called RH5.1/Matrix-M. We identify disordered regions of the full-length RH5 molecule induce non-growth inhibitory antibodies in human vaccinees, and a re-engineered and stabilized immunogen that includes just the alpha-helical core of RH5 induces a qualitatively superior growth-inhibitory antibody response in rats vaccinated with this protein formulated in Matrix-M adjuvant. In parallel, bioconjugation of this new immunogen, termed "RH5.2", to hepatitis B surface antigen VLPs using the "plug-and-display" SpyTag-SpyCatcher platform technology also enabled superior quantitative antibody immunogenicity over soluble antigen/adjuvant in vaccinated mice and rats. These studies identify a new blood-stage malaria vaccine candidate that may improve upon the current leading soluble protein vaccine candidate RH5.1/Matrix-M. The RH5.2-VLP/Matrix-M vaccine candidate is now under evaluation in Phase 1a/b clinical trials.

immunology↗