Biology subjects

Coon, J.

Publications and source records attributed to Coon, J..

At least 19 recordsLinked to original sources

bioRxiv2026

The Aspergillus fumigatus C2-Domain Protein SppA is required for septal integrity and alters susceptibility to echinocandins and neutrophil killing during infection

Aspergillus fumigatus is a major opportunistic fungal pathogen whose ability to maintain hyphal integrity and withstand host defenses is critical for virulence. Septal pores, which connect hyphal compartments, are dynamically regulated to preserve cellular integrity under stress, yet the molecular components governing this process remain incompletely defined. Here, we identify and characterize a septal pore-associated protein, SppA, and demonstrate its essential role in maintaining septal integrity in A. fumigatus. We show that expression of SppA is positively regulated by the transcription factor ZfpA and is induced in response to the cell wall-targeting antifungal caspofungin. Deletion of sppA resulted in defective septal organization and increased susceptibility to hyphal damage. The mutant exhibited heightened sensitivity to cell wall-targeting antifungal agents, indicating a role in cell wall stress tolerance. In a zebrafish model of invasive aspergillosis, loss of SppA significantly attenuated virulence which was abrogated in neutrophil-deficient zebrafish. Further, the mutant strain displayed increased susceptibility to killing by primary human neutrophils, suggesting that proper septal pore formation contributes to fungal survival during host immune attack. Together, our findings establish SppA as a critical determinant of septal integrity, antifungal tolerance, and pathogenicity in A. fumigatus, and position it as part of a ZfpA-regulated, caspofungin-responsive pathway that supports fungal survival during stress and infection. Author SummaryAspergillus fumigatus is a common environmental mold that can cause life-threatening infections in people with weakened immune systems. For successful invasion of host tissue, the fungus requires the ability to protection sections of its hyphae from cell wall targeting antifungals and host immune cell attack by closing septal (cross wall) pores distributed throughout hyphal strands. We have identified an A. fumigatus protein, SppA, required for proper septal pore closure. Loss of SppA reduces the ability of hyphae to withstand treatment with antifungals and the ability of A. fumigatus to cause disease in a zebrafish infection model. The SppA mutant was particularly susceptible to killing by neutrophils, key immune cells that help control fungal infections. Our findings reveal an important mechanism that helps A. fumigatus survive environmental and host-imposed stresses and highlight septal pore regulation as a potential target for future antifungal strategies.

microbiology↗

bioRxiv2026

Performance Evaluation of a Quantitative Metabolomics Workflow Incorporating Microchip Capillary Electrophoresis, Indexed Migration Time, and Single-Point External Calibration

Capillary electrophoresis-mass spectrometry (CE-MS) offers unique analytical advantages for polar metabolite profiling but has remained underutilized in metabolomics relative to liquid chromatography-MS (LC-MS), in part due to challenges in managing migration time drift during data analysis. Here we introduce the use of indexed migration time (iMT) for easily managing this aspect of CE-MS data for metabolomics. Migration time indexing using a panel of stable isotope-labeled (SIL) amino acid reference standards, stored as an iRT database in Skyline, outperformed both uncorrected migration time and relative migration time (RMT) correction across three independent analytical batches spanning 90 samples from four biological matrices. The indexed migration time approach achieved sub-1% relative standard deviation (RSD) in migration index across batches, compared to up to ~15% RSD for uncorrected migration times. Additionally, we evaluate the use of single-point external calibration in Skyline for the purposes of metabolite quantification from complex matrices in order to ease the burden of translational metabolite quantification from metabolomics using high-resolution mass spectrometry (HRMS). Single-point external calibration using a biological matrix-based calibrator was benchmarked against a 13-point linear calibration curve across a panel of amino acids; above 1 M, greater than 95% of back-calculated concentrations fell within {+/-}20% of multi-point calibration. Application of the complete workflow to plasma, serum, urine, and NIST Standard Reference Material (SRM)-1950 demonstrated low inter-batch variability by principal components analysis, broad metabolite coverage across 126 quantifiable analytes, and strong quantitative concordance (Deming slope = 0.862, pseudo-R2 = 0.994, n = 64 analytes) with an independent comprehensive reference dataset for NIST SRM-1950. Together, these results establish a practical mCE-HRMS metabolomics workflow that bridges targeted and discovery metabolomics paradigms and lays the groundwork for single-point external calibration as a powerful tool for translational metabolomics.

molecular biology↗

bioRxiv2026

UFMylation of Pyruvate Dehydrogenase Regulates Mitochondrial Metabolism

The ubiquitin-fold modifier 1 (UFM1) post-translational modification (PTM), or UFMylation, regulates protein homeostasis and is essential for human development. Yet the roles of the de-UFMylase, UFM1-specific peptidase 2 (UFSP2), which removes UFM1 from UFMylated proteins, remain poorly characterized. Here, we demonstrate that UFMylation and UFSP2 regulate mitochondrial metabolism. Quantitative proteomics in UFSP2-deficient cells revealed the accumulation of many proteins previously unknown to be impacted by UFMylation. These included components of the mitochondrial ribosome, electron transport chain (ETC), and pyruvate dehydrogenase (PDH) complex. Functional analyses demonstrated that excessive UFMylation in UFSP2-deficient cells increases mitochondrial respiration, glucose oxidation in the tricarboxylic acid (TCA) cycle, and PDH enzymatic activity. We identified dihydrolipoamide S-acetyltransferase (DLAT), the E2 component of PDH, as a direct UFMylation substrate, with lysine 118 (K118) as the primary conjugation site. Mutating K118 to arginine (K118R) abolished DLAT UFMylation and reduced pyruvate oxidation, identifying this modification as an activator of PDH. These findings reveal a UFMylation-based regulatory mechanism that controls mitochondrial function by inducing utilization of pyruvate as a TCA cycle fuel.

biochemistry↗

bioRxiv2026

A Multi-tissue Transcriptomic-Metabolomic Map Linking Maternal High-Fiber Diet to Reduced Offspring Type 2 Diabetes

BackgroundEarly-life nutritional exposures are increasingly recognized as critical determinants of long-term metabolic health, yet the molecular mechanisms linking maternal diet to offspring type 2 diabetes susceptibility remain incompletely understood. Experimental models are essential to disentangle maternal dietary effects from later-life metabolic influences. MethodsUsing the Nile rat, a genetically heterogeneous model of diet-induced diabetes, we quantified the impact of maternal high-fiber diet on offspring diabetes incidence using sex-stratified time-to-event analyses in 762 offspring. To identify molecular mediators, we performed transcriptomic profiling across 13 offspring tissues, independently contrasting maternal diet exposure and early-onset diabetes status. Overlapping differentially expressed genes were prioritized and evaluated for cardiometabolic associations in human whole-blood transcriptomic data from the GENE-FORECAST cohort. Untargeted plasma metabolomics was integrated to identify circulating metabolites associated with candidate genes. ResultsOffspring born to dams maintained on a high-fiber diet exhibited a markedly reduced risk of developing type 2 diabetes, with approximately 70% lower hazard of diabetes onset in both males and females compared with offspring from regular chow-fed dams. Multi-tissue transcriptomic analyses identified 147 genes differentially expressed in association with both maternal diet and early-onset diabetes, with most effects being tissue-specific. Asnsd1 uniquely showed consistent regulation across the aorta, brown adipose tissue, and skeletal muscle, with higher expression in offspring exposed to a high-fiber maternal diet and lower expression in offspring with early-onset diabetes. In human whole-blood transcriptomic data, ASNSD1 expression was significantly associated with blood pressure-related cardiometabolic traits, including hypertension, systolic blood pressure, and mean arterial pressure. In the animal model, circulating succinic acid was positively correlated with Asnsd1 expression in the aorta but not in other tissues. ConclusionsThis study provides a multi-tissue transcriptomic-metabolomic framework linking maternal high-fiber diet to reduced offspring type 2 diabetes risk. The findings identify ASNSD1 as a maternal diet-sensitive gene associated with diabetes susceptibility across multiple tissues and with cardiometabolic traits in humans, while highlighting tissue-specific relationships between gene expression and circulating metabolites. Together, these results offer mechanistic insight into how early-life nutrition can durably influence diabetes risk across the life course.

developmental biology↗

bioRxiv2026

Proteotyping reactivant toxoplasmic encephalitis reveals virulence-associated dense granule protein GRA5 polymorphisms

A fatal case of toxoplasmic encephalitis, and others like it, has caused microbiologists and clinicians to question whether different strains of T. gondii have more pathogenic potential than others. This raises significant concern, as T. gondii is a widely spread parasitic organism that is presumed to lie dormant in a third of the worlds general population. In this study, we expand on a previously published proteomicanalysis reactivated toxoplasmic encephalitis and have been able to identify T. gondii-specific peptides in the cerebrospinal fluid (CSF) of this patient and two additional cases of toxoplasmic reactivation. Multilocus PCR-restriction fragment-length polymorphism (PCR-RFLP) was used to genetically identify the T. gondii strain that resulted in this fatal case, which belonged to ToxoDB PCR-RFLP genotype #7. Using other T. gondii strains of the same genotype, we performed bioassays to compare the pathogenicity of this genotype with that of a clinically relevant strain, ME49. Both of the tested genotype #7 strains appear to have a greater pathogenic potential, although through likely different mechanisms. Of the most abundant T. gondii-specific CSF peptides across multiple patients, we identified a polymorphic region of the dense granule protein GRA5 that appears to have strain specificity. This approach could represent a "proteotype" that allows for T. gondii strain risk stratification within clinical samples. Ultimately spinal fluid could be a valuable tool in distinguishing between T. gondii exposed individuals with no cyst forms in the brain versus those exposed individuals that harbor "clinically silent" but viable brain cysts. SIGNIFICANCEA third of the worlds population is exposed to the parasite Toxoplasma gondii, residing in a dormant, encysted stage within neurons. T. gondii is a diverse microorganism with some strains having greater reactivation potential. There is no means of identifying which individuals are at risk of reactivation. Proteomic analysis of cerebrospinal fluid from patients with reactivated toxoplasmosis demonstrated a consistent pattern of T. gondii peptides, including GRA5, a secreted virulence factor. Postmortem analysis identified a ToxoDB PCR-RFLP genotype #7 strain associated with a fatality. Cerebrospinal fluid could provide clues to persistent brain infection and may identify strains able to reactivate.

microbiology↗

bioRxiv2025

Regarding Emitter Positioning for Nanoflow Electrospray Ionization

Nanoflow electrospray ionization is commonly used for proteomics due to its high sensitivity. Signal intensity, however, is dependent on optimal emitter positioning relative to the mass spectrometer inlet. Here, we characterize the effect of varied emitter positions on peptide signal intensity in all three dimensions using emitters and flows consistent with standard proteomic analyses. We observe improved signal robustness to x/y variations at increasing z distances and demonstrate that positioning within 1 to 2 mm of the optimal location will maintain consistent signal. Signal intensity behavior is consistent across the m/z range, suggesting a certain level of analyte-independence for proteomics analyses. These results provide insight for proteomics researchers using nanoflow LC-MS/MS. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=93 SRC="FIGDIR/small/695161v1_ufig1.gif" ALT="Figure 1"> View larger version (18K): org.highwire.dtl.DTLVardef@1a976ccorg.highwire.dtl.DTLVardef@b6627forg.highwire.dtl.DTLVardef@1fca93eorg.highwire.dtl.DTLVardef@cb61f6_HPS_FORMAT_FIGEXP M_FIG C_FIG

biochemistry↗

bioRxiv2025

Developmentally-specific physiological and metabolic responses support drought resilience in switchgrass and constrains biofuel yield

Switchgrass (Panicum virgatum) is a promising bioenergy crop due in part to its resilience to drought stress. However, the significance of drought timing remains poorly understood, both from a plant biology perspective and its impact on downstream biofuel production. This study determines the developmental stage-specific physiological and metabolic responses of switchgrass to drought stress and its implications for biofuel production using a custom-built programmable irrigation system. Vegetative, flowering, and senescence-stage drought significantly reduced CO2 assimilation, and stomatal conductance without affecting biomass yield. Metabolic profiling revealed significant accumulation of glucose, fructose, quinic acid, shikimate and GABA during vegetative-stage drought, while flowering and senescence stages exhibited limited metabolic changes. Similarly, specialized metabolites also displayed distinct developmental patterns, with vegetative-stage drought driving the most pronounced metabolic alterations. Thermochemically-treated and hydrolyzed switchgrass biomass from vegetative-stage drought showed elevated lignocellulose-derived compounds and saponins with the latter most positively correlating with fermentation lag times. Conversely, senescence-stage drought enhanced ethanol yields while lowering saponin levels in the hydrolysates. While vegetative-stage drought enhanced physiological resilience, it compromises downstream biofuel production by introducing fermentation inhibitors, particularly saponins.

plant biology↗

bioRxiv2025

SLX4IP acts in parallel to FANCM to limit BLM-dependent replication stress at ALT telomeres

Alternative Lengthening of Telomeres (ALT) is a telomerase-independent telomere maintenance mechanism that enables cancer cells to gain unlimited replicative capacity. ALT relies on recombination-mediated telomere elongation and is promoted by telomeric replication stress. However, ALT requires strict regulation, as excessive replication stress or recombination are cytotoxic. Central to ALT is the RecQ helicase BLM, which regulates telomeric replication stress and promotes telomere recombination and DNA synthesis. Despite its key role in the ALT pathway, BLM must be tightly regulated to prevent deleterious outcomes. Here, we identify SLX4IP as a key suppressor of BLM-driven replication stress at ALT telomeres. Loss of SLX4IP in ALT-positive cells leads to BLM-dependent telomeric replication stress and impaired replication fork progression. Mechanistically, SLX4IP limits the unwinding of unligated Okazaki fragments by BLM on the lagging strand during telomere replication. This reduces the formation of toxic 5' DNA flaps and prevents hyperactivation of ATR signalling and deleterious recombination levels. We also uncover a synthetic lethal interaction between SLX4IP and FANCM, an ATPase/translocase that is a known regulator of BLM at telomeric replication forks in ALT cells. We demonstrate that SLX4IP and FANCM act in parallel to restrain BLM activity, thereby maintaining the balance of replication stress and recombination that is necessary for productive ALT. These findings reveal a vulnerability in ALT-positive cancers lacking SLX4IP and establish SLX4IP as a potential biomarker for therapeutic strategies targeting FANCM. HIGHLIGHTSO_LISLX4IP depletion activates the replication stress response at ALT telomeres C_LIO_LISLX4IP acts in parallel to FANCM to limit replication stress at ALT telomeres C_LIO_LIThe synthetic lethal interaction between SLX4IP and FANCM is dependent on BLM C_LIO_LISLX4IP depletion causes BLM-dependent lagging-strand replication stress C_LI

cell biology↗

bioRxiv2024

Boosting the Sensitivity of Quantitative Single-Cell Proteomics with Activated Ion-Tandem Mass Tags (AI-TMT)

Single-cell proteomics is a powerful approach to precisely profile protein landscapes within individual cells toward a comprehensive understanding of proteomic functions and tissue and cellular states. The inherent challenges associated with limited starting material in single-cell analyses demands heightened analytical sensitivity. Just as advances in sample preparation maximize the amount of material that makes it from the cell to the mass spectrometer, we strive to maximize the number of ions that make it from ion source to the detector. In isobaric tagging experiments, limited reporter ion generation limits quantitative accuracy and precision. The combination of infrared photoactivation and ion parking circumvents the m/z dependence inherent in HCD, maximizing reporter generation and avoiding unintended degradation of TMT reporter molecules in a method we term activated ion-tandem mass tags (AI-TMT). The method was applied to single-cell human proteomes using 18-plex TMTpro, resulting in a 4-5-fold increase in reporter ion signal on average compared to conventional SPS-MS3 approaches. AI-TMT enables faster duty cycles, higher throughput, and increased peptide identification and quantification. Comparative experiments showcase 4-5-fold lower injection times for AI-TMT, providing superior sensitivity without compromising accuracy. In all, AI-TMT enhances the sensitivity and dynamic range of proteomic experiments and is compatible with other techniques, including gas-phase fractionation and real-time searching, promising increased gains in the study of cellular heterogeneity and disease mechanisms.

biochemistry↗

bioRxiv2024

High-content phenotypic analysis of a C. elegans recombinant inbred population identifies genetic and molecular regulators of lifespan

Lifespan is influenced by complex interactions between genetic and environmental factors. Studying those factors in model organisms of a single genetic background limits their translational value for humans. Here, we mapped lifespan determinants in 85 genetically diverse C. elegans recombinant intercross advanced inbred lines (RIAILs). We assessed molecular profiles - transcriptome, proteome, and lipidome - and life-history traits, including lifespan, development, growth dynamics, and reproduction. RIAILs exhibited large variations in lifespan, which positively correlated with developmental time. Among the top candidates obtained from multi-omics data integration and QTL mapping, we validated known and novel longevity modulators, including rict-1, gfm-1 and mltn-1. We translated their relevance to humans using UK Biobank data and showed that variants in RICTOR and GFM1 are associated with an elevated risk of age-related heart disease, dementia, diabetes, kidney, and liver diseases. We organized our dataset as a resource (https://lisp-lms.shinyapps.io/RIAILs/) that allows interactive explorations for new longevity targets.

systems biology↗

bioRxiv2023

Heme biosynthesis regulates BCAA catabolism and thermogenesis in brown adipose tissue

With age, people tend to accumulate body fat and reduce energy expenditure1. Brown (BAT) and beige adipose tissue dissipate heat and increase energy expenditure via the activity of the uncoupling protein UCP1 and other thermogenic futile cycles2,3. The activity of brown and beige depots inversely correlates with BMI and age4-11, suggesting that promoting thermogenesis may be an effective approach for combating age-related metabolic disease12-15. Heme is an enzyme cofactor and signaling molecule that we recently showed to regulate BAT function16. Here, we show that heme biosynthesis is the primary contributor to intracellular heme levels in brown adipocytes. Inhibition of heme biosynthesis leads to mitochondrial dysfunction and reduction in UCP1. Although supplementing heme can restore mitochondrial function in heme-synthesis-deficient cells, the downregulation of UCP1 persists due to the accumulation of the heme precursors, particularly propionyl-CoA, which is a product of branched-chain amino acids (BCAA) catabolism. Cold exposure promotes BCAA uptake in BAT, and defects in BCAA catabolism in this tissue hinder thermogenesis17. However, BCAAs contribution to the TCA cycle in BAT and WAT never exceeds 2% of total TCA flux18. Our work offers a way to integrate current literature by describing heme biosynthesis as an important metabolic sink for BCAAs.

cell biology↗

bioRxiv2023

SCD1 and monounsaturated lipids are required for autophagy and survival of adipocytes

Exposure of adipocytes to cool temperatures often found in the periphery of the body induces expression of Stearoyl-CoA Desaturase-1 (SCD1), an enzyme that converts saturated fatty acids to monounsaturated fatty acids. In this study, we employed Scd1 knockout cells and mouse models, along with pharmacological SCD1 inhibition, to investigate further the roles of SCD1 in adipocytes. Our study reveals that production of monounsaturated lipids by SCD1 is necessary for fusion of autophagosomes to lysosomes and that with a SCD1-deficiency, autophagosomes accumulate. In addition, SCD1-deficiency impairs lysosomal and autolysosomal acidification resulting in vacuole accumulation and eventual cell death. Blocking autophagosome formation or supplementation with monounsaturated fatty acids maintains vitality of SCD1-deficient adipocytes. Taken together, our results demonstrate that in vitro inhibition of SCD1 in adipocytes leads to autophagy-dependent cell death, and in vivo depletion leads to loss of bone marrow adipocytes.

molecular biology↗

bioRxiv2023

Cryogenic Soft Landing Improves Structural Preservation of Protein Complexes

We describe an apparatus for the cryogenic landing of particles from the ion beam of a mass spectrometer onto transmission electron microscope grids for cryo-electron microscopy. This system also allows for the controlled formation of thin films of amorphous ice on the grid surface. We demonstrate that as compared to room temperature landings, use of this cryogenic landing device greatly improves the structural preservation of deposited protein-protein complexes. Further, landing under cryogenic conditions can increase the diversity of particle orientations, allowing for improved 3D structural interpretation. Finally, we conclude that this approach allows for the direct coupling of mass spectrometry with cryo-electron microscopy.

biochemistry↗

bioRxiv2023

Nuclear cytoglobin associates with HMGB2 and regulates DNA damage and genome-wide transcriptional output in the vasculature

Identifying novel regulators of vascular smooth muscle cell function is necessary to further understand cardiovascular diseases. We previously identified cytoglobin, a hemoglobin homolog, with myogenic and cytoprotective roles in the vasculature. The specific mechanism of action of cytoglobin is unclear but does not seem to be related to oxygen transport or storage like hemoglobin. Herein, transcriptomic profiling of injured carotid arteries in cytoglobin global knockout mice revealed that cytoglobin deletion accelerated the loss of contractile genes and increased DNA damage. Overall, we show that cytoglobin is actively translocated into the nucleus of vascular smooth muscle cells through a redox signal driven by NOX4. We demonstrate that nuclear cytoglobin heterodimerizes with the non-histone chromatin structural protein HMGB2. Our results are consistent with a previously unknown function by which a non-erythrocytic hemoglobin inhibits DNA damage and regulates gene programs in the vasculature by modulating the genome-wide binding of HMGB2.

cell biology↗

bioRxiv2022

Early Endosome Capture Proteomics and its Application to Amyloid Precursor Protein Intramembrane Processing by β and γ-Secretases

Degradation and recycling of plasma membrane-localized proteins occurs via the endolysosomal system, wherein endosomes bud into the cytosol from the plasma membrane and subsequently mature into degradative lysosomal compartments. The endolysosomal system also participates in intracellular membrane protein trafficking, signaling, and protein degradation via autophagy. Our understanding of the endolysosomal system has been significantly advanced through the development of methods for selective capture of lysosomes (termed Lyso-IP), but methods for analogous isolation of early intermediates in the endolysosomal system are lacking. Here, we develop an approach for rapid isolation of early/sorting endosomes through affinity capture of the early endosome-associated protein EEA1 (Endo-IP) and provide proteomic and lipidomic snapshots of early endosomes in action. In addition to resident endosomal components, we identify numerous recycling, regulatory and membrane fusion complexes as well as candidate cargo, providing a proteomic landscape of early/sorting endosomes that we distinguished from lysosomes via comparative proteomics. The approach allows capture of internalized transferrin within minutes of its addition to cells, indicating the utility of the approach for dynamic analysis of early/sorting endosomes. Among the endocytic cargo identified by Endo-IP was the amyloid precursor protein (APP) genetically linked with Alzheimers disease. Processing of APP to amyloidogenic A{beta} peptides by {beta}- and {gamma}-Secretases can occur within the endolysosomal system among other compartments, but methods for spatial quantification of A{beta} products in individual organelles are lacking. We combined Endo- and Lyso-IP with targeted proteomics to provide a spatial digital snapshot of A{beta} products. This analysis revealed that products of A{beta} processing by {beta}- and {gamma}-Secretases, and alterations in the specificity of cleavage by small molecule {gamma}-Secretase modulators, can be quantified in both early/sorting endosomes and lysosomes. We anticipate that the Endo-IP approach will facilitate systematic interrogation of the many processes that are coordinated on early endosomes.

cell biology↗

bioRxiv2021

3D Structure Determination of Protein Complexes using Matrix-Landing Mass Spectrometry

Native mass spectrometry (MS) is an emerging technology that can provide complementary data to electron microscopy (EM) for protein structure characterization. Beyond the ability to provide mass measurements of gas-phase biomolecular ions, MS instruments offer the ability to purify, select, and precisely control the spatial location of these ions. Here we present a modified Orbitrap MS system capable of depositing a native MS ion beam onto EM grids. We further describe use of a chemical landing matrix that both preserves and protects the structural integrity of the deposited particles. With this system we obtained the first 3D reconstructed structure of gas-phase, deposited biomolecular ions - the 800 KDa protein complex GroEL. These data provide direct evidence that non-covalent protein complexes can indeed retain their condensed-phase structures following ionization and vaporization. Finally, we describe how further developments of this technology could pave the way to an integrated MS-EM technology with promise to provide improved cryo-EM sample preparation over conventional plunge-freezing techniques.

molecular biology↗

bioRxiv2021

Multi-omics analysis identifies essential regulators of mitochondrial stress response in two wild-type C. elegans strains

The mitochondrial unfolded protein response (UPRmt) is a promising pharmacological target for aging and age-related diseases. However, the integrative analysis of the impact of UPRmt activation on different layers of signaling in animals with a different genetic background is lacking. In this study, we applied systems approaches to investigate the effect of UPRmt induced by administering doxycycline (Dox) on transcriptome, proteome, lipidome, and metabolome in two genetically divergent C. elegans strains. We found that Dox prolongs lifespan of both worm strains through pathways in both shared and strain-specific manners. From the integrated omics datasets, we observed a strong impact of Dox on mitochondrial functions, detected upregulated defense response and lipid metabolism, identified decreased triglycerides and lowered metabolome profiles in both strains. This conserved phenomic footprint has great translational value as it indicates that the beneficial effects of Dox-induced UPRmt on health and lifespan are consistent across different genetic backgrounds.

systems biology↗

bioRxiv2020

Nano-Emulsion Adjuvant Enhances Cross-presentation by Lipid Body Formation Independent of Glycolysis

Here, we report that a carbomer-based adjuvant, Adjuplex(R) (ADJ), stimulated robust CD8 T-cell responses to subunit antigens by modulating multiple steps in the cytosolic pathway of cross-presentation, and afforded effective immunity against virus and intracellular bacteria. Cross-presentation induced by TLR agonists requires a critical switch to anabolic metabolism, but ADJ enhanced cross presentation without this metabolic switch in DCs and NLRP3-driven caspase 1 activity. Instead, ADJ induced in DCs, an unique metabolic state, typified by dampened oxidative phosphorylation and basal levels of glycolysis. In the absence of increased glycolytic flux, induction of ROS and lipid bodies (LBs) and alterations in LB composition mediated by ADJ were critical for DC cross-presentation. These findings challenge the prevailing metabolic paradigm by suggesting that DCs can perform effective DC cross-presentation, independent of glycolysis to induce robust T cell-dependent protective immunity to intracellular pathogens. These findings have implications in the rational development of novel adjuvants.

immunology↗