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Previous research has established a link between the genus Corynebacterium in the skin microbiota and skin health; however, studies investigating the specific effects of Corynebacterium-derived substances on skin cells remain limited. In this study, we isolated eight strains of Corynebacterium from the facial skin of female participant. To investigate the potential skin improvement effects of these isolates, three cultured skin cell lines (Hs68, HaCaT, and B16-F10) were treated with the supernatants of each strain. We then evaluated and compared the gene expression levels (ELN, COL1A1, IL-1{beta}, and HAS3) alongside their inhibitory effects on melanin biosynthesis. Corynebacterium amycolatum SB-1 was selected as the final candidate for subsequent analysis, primarily based on its rapid growth rate and high cell yield. Using LC-TOF/MS, we screened the postbiotic candidates from the SB-1 strain and identified ellagic acid as a known polyphenolic antioxidant with potent anti-melanogenic effects beneficial for skin improvement. To validate the effects of ellagic acid on skin cells, we assessed FLG gene expression and its inhibition of melanin biosynthesis. Additionally, RNA sequencing was performed to elucidate the underlying molecular mechanisms, revealing that ellagic acid is significantly involved in inhibiting melanogenesis and promoting collagen synthesis. Taken together, our results highlight ellagic acid derived from strain SB-1 as a highly effective bioactive compound for improving skin health, specifically targeting hyperpigmentation and skin aging.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.748710v1?rss=1

Kim, S., Mun, S., Kim, M., Kim, M.-J., Lee, D.-G., Kang, S., Jo, H., Han, K.

During chronic infection, the persistence of antigen and inflammation leads to the differentiation of CD8+ T cells into an exhausted state characterised by expression of inhibitory receptors (IRs) and the progressive loss of T cell functions. Among the different subsets of exhausted CD8+ T (Tex) cells, Tex progenitors expressing SLAMF6 and the transcription factor TCF-1 (TCF-1+) give rise to Tex effector-like cells expressing CX3CR1 and Tex terminal cells expressing CD101. PD-1/PD-L1 blockade acts on TCF-1+ Tex progenitor cells and promotes their differentiation into Tex effector-like cells. The molecular events controlling CD8+ Tex cell differentiation are still poorly defined. As Notch signaling may be sustained during chronic infection by persistent TCR stimulation and inflammation, we tested whether Notch signaling influences CD8+ T cell exhaustion. Using mice lacking (N1N2{triangleup}/{triangleup}) or not (N1N2fl/fl) Notch1/2 expression only in mature CD8+ T cells, we showed that the absence of Notch signal causes severe CD8+ T cell exhaustion during chronic LCMV infection. N1N2{triangleup}/{triangleup} Tex cells express higher levels of IRs and are less functional when compared to their wild-typee counterpart. In the absence of N1N2 receptors, Tex progenitor and Tex terminal cells accumulate and Tex cells cannot be reinvigorated by PD-1/PD-L1 blockade. We further demonstrated that Notch signaling is essential to promote the differentiation of Tex progenitors into Tex effector-like cells. Moreover, Notch signals, provided by stromal cells expressing the ligands Delta-like 1 and 4, are necessary during all stages of the infection to prevent severe exhaustion. Single-nucleus RNA and ATAC multiome profiling identifies Notch signaling as a critical role on effector transcriptional programming in exhausted CD8 T cells. Loss of Notch signaling impairs transcriptional program associated with migration and perception of CD4+ T cell help. Together, these alterations drive the differentiation of Tex progenitor cells toward a terminally exhausted Tex fate.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748651v1?rss=1

Maurice De Sousa, D., Boulet, S., Duval, F., Perkey, E., Daudelin, J.-F., Le Corre, L., Lebel, M.-E., Lamarre, A., Radtke, F., Ludewig, B., Siebel, C. W., Maillard, I., Labrecque, N.

SNRNP70 is a core spliceosome RNA-binding protein best known for its essential role in nuclear pre-mRNA splicing. Although traditionally associated with nuclear RNA processing, previous studies have identified important extranuclear functions for SNRNP70 in neurons, including roles in mRNA stability, localisation, and axonal transport. Yet, much of our understanding of SNRNP70 localisation has relied on overexpression or transgenic approaches, leaving a critical gap in our knowledge of where endogenous SNRNP70 resides and how it behaves in living neurons under physiological expression conditions. Here, we address this limitation by establishing and validating a novel zebrafish SNRNP70-eGFP CRISPR knock-in line, enabling direct visualisation of the endogenous protein. eGFP was fused to the C-terminus of endogenous SNRNP70 while retaining the native 3' untranslated region, preserving key regulatory features of the endogenous locus. We demonstrate that the knock-in faithfully reports endogenous SNRNP70 expression and reveals widespread physiological localisation throughout the developing nervous system, including prominent enrichment within axonal and synaptic compartments. Crucially, live in vivo imaging reveals that endogenous SNRNP70 is dynamically localised within neuronal mRNP granules, providing direct evidence of its physiological behaviour in these structures without the confounding effects of protein overexpression. Proximity ligation analyses further demonstrates associations between endogenous SNRNP70 and PABPC1B, FUS, and UPF1, which are established neuronal mRNP granule components. Together, our work provides a validated genetic and imaging resource for investigating SNRNP70 at endogenous levels in the living nervous system. By overcoming key limitations of conventional transgenic and overexpression-based approaches, the SNRNP70-eGFP knock-in enables physiological analysis of SNRNP70 localisation and dynamics and reveals its prominent and dynamic organisation within neuronal mRNP granules. More broadly, this work highlights how endogenous fluorescent tagging can provide a versatile platform for resolving the spatial organisation of RNA-binding proteins in living neurons under physiological expression conditions and provide new insight into the regulation of neuronal mRNA fate.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748523v1?rss=1

Lloyd-Jones, J., Sartorius, S., Gurevich, D., Nikolaou, N.

Knockout of the UDP-glucose-activated P2Y14 receptor (P2Y14R) in adipocytes or whole-body has been reported to provide metabolic benefits in obese mice. We hypothesized that selective P2Y14R activation would lead to metabolic impairments, whereas pharmacological antagonism would improve metabolic deficits in diet-induced obese (DIO) mice. Here, we investigated the metabolic effects of a synthetic P2Y14R agonist, UDP-like methylene-bridged MRS2905. Acute P2Y14R activation with MRS2905 triggered a robust and prolonged hyperglycemic effect in lean and obese mice with impaired glucose homeostasis. Moreover, MRS2905 treatment of obese mice lowered fasting plasma insulin and increased glucagon levels, along with upregulation of liver JNK phosphorylation and the expression of rate-limiting gluconeogenic genes. The MRS2905-induced hyperglycemic effect was blunted in whole-body P2Y14R knockout mice compared with wild-type control. In contrast, a potent P2Y14R antagonist mono-ester prodrug (MRS4779) partially reversed the agonist-induced hyperglycemia and restored proper glucose homeostasis after acute treatment. Chronic MRS4779 administration in DIO mice reduced fat mass and improved various metabolic parameters including liver steatosis. Additionally, we examined the roles of P2Y14R in hepatocytes of DIO mice. Here, we report that P2Y14R was upregulated in liver and hepatocytes from obese mice compared to lean mice. Overnight fasting also upregulated hepatic P2Y14R expression. P2Y14R deletion from hepatocytes in DIO mice improved fasting blood glucose level and lipid metabolism without improving glucose homeostasis. These results suggest a novel P2Y14R function in hepatic lipid metabolism, and P2Y14R antagonists may prove useful for the treatment of obesity and obesity-related metabolic disorders.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748648v1?rss=1

Pramanik, A., Hariprasad Kurma, S., Wen, Z., Cai, H., Violet, P.-C., Walter, P. J., Demby, T., Gavrilova, O., Pittala, S., Sajiir, H., Umarova, R., Wing, A., Rotman, Y., Wess, J., Jacobson, K. A.

Large-scale chemical screens are important tools for hazard identification and chemical prioritization, but they less commonly progress from identifying a phenotype to identifying its mechanism. Here, we used zebrafish embryos to screen 4,657 chemicals from the U.S. EPA ToxCast Phase III library for disruption of embryonic development and advanced selected hits through sequential validation using original library stocks, independently sourced chemicals, concentration-response analysis, transcriptomics, and functional experiments. Of 61 primary hits subjected to repeat testing, 33 reproduced the original phenotype, and four of eight compounds subsequently tested using independently sourced chemicals exhibited reproducible concentration-dependent developmental toxicity. We then investigated purpurin, an understudied anthraquinone pigment that caused pericardial edema, circulation defects, and body-axis abnormalities. Transcriptomic analysis of purpurin-exposed embryos revealed coordinated suppression of pathways involved in calcium regulation, ion transport, and neuronal signaling. Increasing extracellular calcium produced a concentration-dependent rescue of purpurin-induced developmental abnormalities, whereas equivalent magnesium supplementation did not, supporting a role for calcium availability or homeostasis in purpurin developmental toxicity. These results demonstrate that large-scale in vivo toxicity screening can be integrated with independent chemical validation and functional follow-up to move beyond hazard identification toward mechanistic understanding of how environmental chemicals disrupt embryonic development.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748647v1?rss=1

Shahriar, S., Gorelick, D. A.

DNA nanostructures can be tailored to perform a wide variety of functions, with continued interest in biological applications. Some aspects of DNA nanostructure assembly can hinder the ability of nanostructures to be useful in physiological environments. Typical assembly methods employ magnesium ions to stabilize the structure, which leave the structure susceptible to damage by nucleases in body fluids. Further, DNA nanostructure assembly typically involves a thermal annealing protocol in which DNA strands are heated in a specific buffer to a high temperature and cooled slowly at specific rates, preventing convenient encapsulation of temperature-sensitive guest molecules. In this work, we demonstrate the assembly of a wide variety of DNA nanostructures and 3D crystals in a hydrated ionic liquid (choline dihydrogen phosphate, CDHP) instead of magnesium at constant moderate temperatures, thus avoiding thermal annealing. CDHP-assembled structures show enhanced biostability against a variety of nucleases. Molecular dynamics simulations show that choline ions stabilize DNA nanostructures by a direct and close-range interaction in contrast to the predominantly water-mediated interactions of Mg2+, leading to enhanced nuclease resistance in CDHP-containing environments. CDHP-assembled structures do not affect the viability of HepG2 cells and show higher cell internalization. Overall, this work develops a potential method to construct more biostable DNA nanostructures and 3D crystals in a simple one pot process. Assembly of DNA nanostructures under isothermal conditions is desirable for scaffolding biomolecules and to reduce the need for thermal annealing instruments, allowing nanostructure preparation in low-resource settings.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.744742v1?rss=1

Talbot, H., Nguyen, L., Gandavadi, D., Nowzari, Z., Mathur, C., Gallagher, G., Mongin, K., Vangaveti, S., Begley, T. J., Wang, X., Chandrasekaran, A. R.

The Wnt/{beta}-catenin pathway is constitutively active in most colorectal and other cancers. Tankyrase (TNKS) promotes Wnt signalling by PARylating AXIN, a rate-limiting scaffold subunit of the {beta}-catenin destruction complex, and its inhibition is a validated strategy for pathway downregulation. Existing small-molecule TNKS inhibitors that target the catalytic PARP domain suffer from off-target effects across the PARP family. Here, we present an alternative approach that targets the substrate binding domains of TNKS using a short TNKS-binding peptide (TBP) presented multivalently on DNA nanostructures. Such a strategy may be required to effectively disrupt the function of targets such as TNKS, which is known to form high-order assemblies in the cell. We engineered DNA nanostructures with two distinct geometries: a 2D triangle (~100 nm) displaying 27 copies of TBP, and a compact 3D tetrahedron (~10 nm) displaying 2 TBP copies. We show that both nanostructures assemble efficiently, can be functionalised with TBP in high yields, and retain binding to TNKS protein in vitro. DNA nanostructures are efficiently internalised to the cytoplasm by HeLa Kyoto and colorectal cancer cells, with TBP functionalisation enhancing rather than hindering uptake. In HeLa cells, both triangle-TBP and tetrahedron-TBP downregulated Wnt signalling to a similar extent despite an order-of-magnitude difference in TBP copy number, while the same concentration of free TBP had no effect. This finding likely reflects the two functions of the DNA nanostructures - intracellular delivery and multivalent display - whereby the smaller nanostructures more efficiently internalise the TBP ligand but have lower valency. These results establish DNA nanostructures as a modular, tuneable platform for multivalent inhibition of intracellular clustered targets such as TNKS.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.03.749096v1?rss=1

Zacharopoulou, M., Cassidy, Z., Qin, S., Grannum, S., Sridhar, A., Chambers, J. E., de la Roche, M. A., Itzhaki, L. S., Mela, I.

Barberry is an ecologically and medicinally important perennial shrub and an alternate host of Puccinia striiformis f. sp. tritici (Pst), the causal agent of wheat stripe rust. However, its genome architecture and molecular responses to Pst infection remain poorly understood. Here, we generated a haplotype-resolved, chromosome-level genome assembly of Berberis aggregata using PacBio HiFi and Hi-C sequencing. The two haplotypes, Ba5A and Ba5B, were assembled into 14 pseudochromosomes each, with genome sizes of approximately 1.13 and 1.16 Gb, respectively, and showed high continuity and completeness. Comparative analyses revealed extensive divergence between the haplotypes, including widespread structural variation, substantial hemizygosity, and TE-rich, gene-poor non-alignable regions. Haplotype-specific hemizygous genes also exhibited distinct functional biases, with Ba5A enriched in immune-related processes and Ba5B associated with transport and cell-envelope functions. Dual RNA-seq across infection stages revealed stage-dependent host transcriptional reprogramming, characterized by early pathogen perception followed by stronger defense activation and photosynthesis repression, whereas Pst displayed a coordinated colonization program consistent with its biotrophic lifestyle. Allele-specific expression analysis showed that ~25-28% of genes exhibited significant haplotype-biased expression, and a subset transitioned from unbiased expression at 0 dpi to Ba5A- or Ba5B-biased expression during infection. Integration with differential expression analysis further identified infection-responsive induced-biased genes associated with defense-related processes, including responses to fungal pathogens. These findings show that haplotype divergence and dynamic allele-specific regulation jointly contribute to transcriptional responses during Pst infection, providing new insights into the genomic basis of barberry-rust pathogen interactions.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748723v1?rss=1

Wang, J., Xu, Y., Duan, Y., Kang, Z., Zhao, J.

In female meiosis, each of the four chromatids in a tetrad has a 25% chance to be selected for transmission through the pronucleus, realizing Mendel's second law. Here, we characterize a cheating but unselfish behaviour that has documented cases across diverse animal taxa including humans and flies, whereby chromatids with crossovers (COs) have a transmission advantage, resulting in increased production of recombinant offspring. Taking advantage of Drosophila ovarian physiology, we show that this form of meiotic cheating which we call recombinant drive occurs on the autosomes when females are under nutrient stress, potentially as a mechanism for recombination plasticity, while curiously, the effect is suppressed on the X by the distributive system. We explored recombinant drive using simulations and identified unique quantitative signatures that are at odds with several intrinsic properties of linkage. One violation is the production of more offspring with recombinant versus parental allele combinations, which we empirically demonstrated to be possible. Further, we show that recombinant drive can appear to modify CO patterning effectively acting as assurance and interference mechanisms, even when it has no influence on and acts downstream of CO spacing. We discuss the potential benefits and consequences of having a conserved method that can rapidly increase recombination in response to stress, and speculate on a mechanism for the preferential transmission of COs at meiosis II. Overall, our study revealed distinct properties and behaviours of a poorly understood, but potentially widespread, conserved phenomenon and offers avenues for broad detection and mechanistic dissection.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748656v1?rss=1

Chauhan, A., Yeung, N., Kim, H., Wei, K.

Depletion of B cells in multiple sclerosis (MS) is beneficial yet there is no defined pathogenic B cell subset in MS. Here, we demonstrate that T-bet+ memory B cells are rare in control human brain specimens but are found in MS lesions. Transfer of murine T-bet+ memory B cells into mice with ongoing experimental autoimmune encephalomyelitis (EAE), a model of MS, worsened their disability and demyelination. Microglia/macrophage density increased in central nervous system parenchyma even though B cells mostly remained in barriers. In culture, secreted factors from T-bet+ memory B cells promoted macrophage migration. IFN-{gamma} produced by T-bet+ memory B cells activated microglia to secrete chemokines that further enabled macrophage migration. Consistent with their age-associated elevation, conditional deletion of T-bet in B cells lowered EAE severity in aged but not young mice. We define T-bet+ memory B cells as pathogenic in EAE and MS through their IFN-{gamma}-facilitated interactions with microglia/macrophages.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748682v1?rss=1

Jain, R. W., Goiko, M., Mendes, A., Xue, S., Zein, S., Gorter, R. P., Morch, M. T., Prat, A., Li, R., Yong, V. W.

Retrotransposons are mobile genetic elements that can impair genome integrity, and their insertions and dysregulation have been implicated in various diseases. However, growing evidence suggests that retrotransposons may play important physiological roles. At the Drosophila larval neuromuscular junction (NMJ), the ViSyToR (Viral Synaptic Transfer of RNA) pathway requires the active retrotransposon copia as a key negative regulator of synaptogenesis. Here, we identify a long non-coding RNA, Statera (Stae), as a regulator of copia, initially discovered through its physical association with the Copia protein. Stae is highly expressed in the central nervous system and body wall muscles of Drosophila larvae. CRISPR knockout or RNAi knockdown of Stae leads to a general decrease of bouton number at the NMJ, indicating its role in promoting synaptogenesis. Furthermore, copia is highly upregulated in Stae mutants, suggesting that Stae functions to repress copia. Mechanistically, Stae reduces copia RNA stability and limits copia DNA copy number. Additional knockdown of copia in motor neuron-specific Stae RNAi animals rescues the abnormal NMJ morphology. Together, our findings reveal an unexpected role for a long non-coding RNA in promoting synaptogenesis by repressing a retrotransposon.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748680v1?rss=1

Wang, S., M'Angale, P., Xiao, C., Alegre, G., Simkin, A., Malinkevich, A., Thomson, T.

Objective: The original goal was to map neuronal circuits activated by spontaneous seizures in models of temporal lobe epilepsy. Studies used the c-fos driven TRAP2 system, which has been used successfully to label neurons after seizures. Unexpectedly, astrocytes were also labeled, then shown to express c-fos in a sustained manner after seizures. The role of these so-called fostrocytes in spontaneous seizures was studied using novel Cre-dependent AAVs. Methods: Studies used a homozygous mouse line produced by crossing TRAP2 Cre-driver mice with the Ai9 Cre-reporter line. Seizures were induced using electrical stimulation, kainic acid, or pilocarpine. Cre-dependent AAVs used the GFAP promoter to drive expression of either the catalytic A chain of diphtheria toxin (DTA), GFP, or empty vector. Spontaneous seizures were continuously recorded using EEG. Results: Discrete seizures in naive mice evoked transient c-fos expression in all astrocytes, while status epilepticus evoked a higher and sustained level of c-fos expression. Timing the activation of the TRAP2 system allowed specific labeling of the subset of astrocytes with high c-fos expression. These cells were colocalized with established astrocyte markers, showed reactive astrocyte morphology, and were selectively labeled by GFAP-driven Cre-dependent AAVs. Ablation of fostrocytes in spontaneously seizing mice increased seizure frequency. Significance: Next-generation sequencing studies have revealed a great diversity of astrocyte subtypes, identifying clusters with up-regulated c-fos expression in patients with neurological disorders. In conclusion, these studies suggest that therapies that augment the activity of c-fos expressing astrocytes would have anti-seizure activity.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748637v1?rss=1

Failor, M. J., Bork, B. P., San Pietro, J. M., Maciejczuk, A., Elmer, K., Lile-King, C., Oliinyk, M., Choi, J. L., Huang, A. H., Tabor, D. J., Gaykema, R. P., Perez-Reyes, E.

Objective: T cells are increased in the brain in Alzheimer's disease (AD) and primary tauopathies where they correlate with tau pathology. In mouse models, T cells exacerbate tau-related neuroinflammation and neurodegeneration, and depletion of T cells is neuroprotective. Here, we investigate whether the multiple sclerosis drug fingolimod may inhibit infiltration of T cells into the brain in the setting of tau pathology and thereby attenuate tau-mediated neurodegeneration. Methods: P301S-tau transgenic mice expressing human APOE4 (TE4), which develop tau pathology, neurodegeneration, and neuroinflammation with T cell accumulation, were randomly assigned to receive 1 mg/kg/d fingolimod in drinking water or standard mouse drinking water beginning just prior to the accumulation of neurofibrillary tangle pathology at 6 months of age. Mice were then assessed with a battery of cognitive behavioral assays at 9 months of age and euthanized at 9.5 months of age to analyze the blood and brain using flow cytometry, single molecule array (SIMOA) technology, stereotactic brain measurements, and immunohistochemistry. Results: Fingolimod produced the expected lymphopenia in peripheral blood but unexpectedly exacerbated brain T cell accumulation, microgliosis, astrogliosis, and tau pathology, with no significant effect on neurodegeneration or behavioral impairment in TE4 mice. Interpretation: These results indicate fundamental differences in the nature of the adaptive immune response to tau pathology versus other neuroinflammatory diseases such as multiple sclerosis and underscores the importance of understanding the impact of immune modulating therapies on all aspects of AD-related pathology in preclinical models prior to consideration and initiation of clinical trials.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748583v1?rss=1

Rudman, M. D., Litvinchuk, A., Smith, C., Remolina Serrano, J., Manis, M., Bao, X., Yuede, C. M., Ulrich, J. D., Holtzman, D. M.

Individuals with autism spectrum disorder (ASD) often show aversion to affective touch (AT). However, the neural mechanism of this abnormality in cortices remains poorly understood probably due to the lack of effective animal models. Here, we used a canine model to address this issue by leveraging the intimate dog-human interactions. In a newly-designed heterospecific AT paradigm, we found that dogs carrying mutations in Shank3, a high-risk gene for ASD, avoided human AT. In vivo single-unit recording analysis showed that AT-evoked oscillations in the dorsal medial prefrontal cortex (dmPFC) were significantly altered in Shank3 mutant dogs. Shank3 mutation also reduced the number of neurons encoding AT in the dmPFC. Importantly, the aversion to AT and altered neural processing in Shank3 mutants were largely rescued by a GABAA receptor antagonist pentylenetetrazole. Together, these findings provide neural mechanisms for abnormal AT processing in ASD and suggest potential biomarkers for therapeutic strategies.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748311v1?rss=1

Zhou, Y., He, Y., Dan, M., Jia, L., Guo, K., Fang, Y., Hu, L., Yu, X., Wang, J., Xing, D., Zhang, Y. Q.

TAR DNA-binding protein 43 (TDP-43) aggregation is the defining pathological hallmark of nearly all cases of amyotrophic lateral sclerosis (ALS), yet physiologically relevant human models that faithfully recapitulate disease-associated TDP-43 proteinopathy and dysfunction remain limited. To cover this gap, we generated a novel human-based model of cortical neurons carrying the endogenous ALS-linked TDP-43 A315T mutation together with an in-frame Dendra2 fluorescent reporter, enabling temporal and spatial monitoring of the protein. Neurons expressing TDP-43 A315T exhibited progressive neurite degeneration, altered neuronal activity, and impaired mitochondrial respiration, recapitulating several ALS-associated phenotypes. Our model also displays autophagy-dependent accumulation of cytoplasmic aggregates of mutant TDP-43 without overt loss of nuclear function, maintaining normal processing of canonical cryptic exon targets. In contrast, experimental induction of TDP-43 nuclear exclusion readily triggered cryptic exon incorporation, demonstrating that the model faithfully reproduces loss-of-function phenotypes under stress conditions. In line with perturbed protein solubility, mutant TDP-43 neurons show increased stress granule (SG) formation at baseline and under oxidative stress condition. Finally, treatment with the RNA chaperone Clip34 significantly reduced TDP-43 aggregation under both basal and oxidative stress conditions as well as its localization to SGs. Taken together, these findings establish a physiologically relevant human model that separates early TDP-43 toxic gain-of-function from basal loss-of-function while providing a robust platform for investigating TDP-43 biology and accelerating therapeutic discovery in ALS.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.745749v1?rss=1

Cicardi, M. E., Antonini, S., Martorell Serra, I., Muench, M., Kinney, N., Girdhar, A., Cristofani, R., Poletti, A., Sterneckert, J., Crippa, V., Bordi, M., Pasinelli, P., Guo, L., Tigano, M., Trotti, D.

Stress is a common trigger of headache and widespread somatic pain. Repeated exposure to stress establishes latent sensitization that is tonically suppressed by endogenous inhibitory systems. Using brain-penetrating and peripherally restricted receptor antagonists in a mouse model of repetitive restraint stress, we systematically identified these protective pathways. Peripheral kappa opioid receptors as well as GABA type A and type B (GABAA and GABAB) receptors, alongside central opioid and cannabinoid signaling, actively suppress headache-related facial mechanical hypersensitivity in stressed mice. Blocking any pathway quickly reinstated cephalic allodynia. Peripheral opioid and GABAA receptor signaling also inhibited stress-induced latent sensitization on hindpaw. In contrast, depletion of anti-inflammatory regulatory T (Treg) cells prolonged facial but not hindpaw sensitization, suggesting that Tregs preferentially limit stress-induced chronic headache. Next, we administered low-dose interleukin-2 (LD-IL-2) to stressed mice to preferentially expand and activate Treg cells. Following LD-IL-2 treatment, neither subthreshold pain triggers nor blockade of endogenous inhibitory pathways reinstated cephalic or hindpaw allodynia in either sex, indicating elimination of stress-induced sensitization. Mechanistically, LD-IL-2 acted through Treg cells to recruit multiple peripheral cytokine pathways without engaging endogenous opioid, GABA, or cannabinoid receptor signaling. Notably, LD-IL-2 was more effective than anti-CGRP (calcitonin gene-related peptide) therapy in preventing headache-related chronic sensitization in stressed mice. Collectively, these findings reveal multiple central and peripheral pathways that act concertedly to mask stress-induced latent sensitization and strongly support further evaluation of LD-IL-2 as a novel treatment for stress-induced headache and widespread pain with a distinct mechanism of action.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748528v1?rss=1

Simoes, R. R., Kraus, S., Zhang, J., Do Nascimento, L. F., Tosh, D. K., Unsinger, J., Hotchkiss, R. S., Che, T., Jacobson, K. A., Cao, Y.-Q.

Background: Diabetic kidney disease (DKD) is the leading cause of chronic kidney disease (CKD). However, most proteomic studies of DKD rely on bulk kidney tissue, which cannot distinguish the contribution or response of individual nephron compartments to injury. Methods: Diabetes was induced in male mice by streptozotocin (STZ) injections. After 16-weeks, the mice and vehicle-treated controls were characterized physiologically, biochemically, and histologically. A deep learning-powered Deep Visual Proteomics (DVP) pipeline, validated against manual annotation, was adapted to isolate proximal tubule (PT) and glomeruli from Megalin stained kidney sections by automated laser microdissection. Bulk kidney, PT, and glomerular proteomes were generated by data independent acquisition mass spectrometry. PT-enriched candidates were prioritized using a composite scoring approach and compared with human tubulointerstitial proteomic data from the Kidney Precision Medicine Project. Results: STZ mice developed sustained hyperglycaemia and albuminuria, alongside elevated markers of tubular injury and interstitial fibrosis. Segmentation models isolated PT and glomeruli with high fidelity (Dice coefficients 0.878 and 0.914; area correlations r=0.993 and r=0.996). Compartment-resolved proteomics determined that PT and glomeruli underwent largely distinct, non-overlapping remodelling: PT exhibited loss of proteostatic, cell cycle, and structural programs with compensatory mitochondrial and lipid metabolic upregulation, whereas glomeruli showed broad loss of oxidative metabolic capacity without any compensatory metabolic program. Fourteen of the top twenty prioritized PT candidates, including LARS2 and ANXA2, changed in the same direction in human CKD tubulointerstitial proteomic data. The STZ PT proteome correlated significantly with this human dataset, while the glomerular comparison did not. Conclusions: Compartment-resolved and deep learning-guided visual proteomics can uncover divergent, biologically coherent PT and glomerular injury programs in DKD that are masked in bulk tissue analysis. A PT injury signature was uncovered that is partially conserved in human CKD, identifying novel candidate mechanisms and biomarkers for future exploration.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748754v1?rss=1

Zheng, X., d'Acierno, M., Rosenbaek, L. K., Rinschen, M., Wu, Q., Fenton, R. A.

Pseudomonas aeruginosa relies on heme acquisition to sustain growth and virulence in the iron-limited environment of the host, particularly in the acidic airways of patients with cystic fibrosis where antibiotic efficacy is markedly reduced. The Has heme assimilation system is initiated by the secreted hemophore HasA, which binds extracellular heme and delivers it to the outer-membrane receptor HasR. During infection, HasA is proteolytically processed, generating a truncated form that constitutes the biologically relevant species. Here, we characterise the structural and functional properties of truncated HasA under disease-associated acidic conditions (pH 6.5) and perform a rationally designed Systemic Evolution of Ligands by EXponential enrichment (SELEX) method to identify DNA aptamers capable of binding this hemophore. Biophysical analyses revealed that truncated HasA is folded, displays a mixed /{beta} secondary structure, and exists as a concentration-dependent mixture of monomers and domain-swapped dimers. Circular dichroism and nano-differential scanning fluorimetry identified two thermal transitions, consistent with the coexistence of apo/holo and monomer/dimer species. The apo protein bound heme with high affinity (KD = 113 nM) and a 1:1 stoichiometry, confirming preservation of its functional binding mechanism. Twelve rounds of SELEX, incorporating target switching and platform switching, yielded a highly enriched aptamer pool dominated by two sequences, HasA_1 and HasA_2. ELONA (Enzyme-Linked Oligonucleotide Assay) assays demonstrated that both aptamers bind specifically to truncated HasA, and biolayer interferometry revealed nanomolar dissociation constants (KD = 0.59 M and 0.32 M, respectively). These aptamers did not bind full-length HasA, BSA, or control sequences, confirming that selection under acidic conditions drove specificity toward the physiologically relevant form of the hemophore. Our findings identify HasA-binding DNA aptamers that retain function in acidic environments where antibiotic potency is compromised, highlighting their potential as molecular tools to disrupt heme acquisition in P. aeruginosa. This work establishes a foundation for developing aptamer-based antimicrobial strategies targeting the Has system, particularly under acidic conditions where conventional antibiotics are compromised.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.03.749098v1?rss=1

Bosetto, F., Wei, T., Stott, K., McLaughlin, S. H., Zacharopoulou, M., Itzhaki, L. S., Mela, I.

Complete ammonia oxidizers (comammox) are widespread nitrifiers that can dominate ammonia oxidation in diverse environments by efficiently converting ammonia to nitrate within a single cell, yet the molecular basis distinguishing their ammonia monooxygenase (AMO) from those of canonical bacterial AMO remains unresolved. Here we report cryo-electron microscopy (cryo-EM) structures of AMO from the comammox Nitrospira inopinata (NiAMO) in inhibitor-free and allylthiourea (ATU)-bound states at 2.47 and 2.68 angstrom, respectively. NiAMO displays distinctive auxiliary-subunit organization, copper-site configuration and hydrophobic-channel architecture. Integrative molecular dynamics (MD) and quantum mechanics/molecular mechanics (QM/MM) calculations support a methyl-plastoquinol (methyl-PQH2)-coupled, CuD-centric catalytic model, with CuC potentially facilitating quinone redox cycling. N. inopinata exhibited broad susceptibility to several known nitrification inhibitors, and ATU-bound NiAMO structure localized the inhibitor to the CuC-CuD region, accompanied by constriction of the hydrophobic channel, which is consistent with the competitive role of ATU demonstrated in recovery assays. Multi-omics analyses further revealed an energy-limited stress response to ATU, including induction of urea transport and utilization systems. Collectively, these findings define a methyl-PQ-linked catalytic and inhibitor-responsive architecture of comammox AMO and establish a mechanistic framework for lineage-aware management of nitrification in natural and engineered ecosystems.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.748776v1?rss=1

Mao, T.-Q., Yang, X., He, Z.-C., Yang, J., Wu, R., Leung, K. M. Y., Li, S., Han, P., Peng, W., Li, Z.

The chromatoid body (CB) is a hallmark of haploid male germ cells, but how it selects and regulates RNAs has remained unclear. Here we define the RNA landscape of the CB by transcriptome and small RNA profiling of isolated CBs throughout mouse round spermatid differentiation. We find that the CB selectively concentrates pachytene piRNAs, retrotransposon transcripts, specific mRNA isoforms and intron-retaining mRNAs, revealing extensive RNA sorting within this cytoplasmic germline condensate. The enrichment of transposable element transcripts together with PIWI-piRNA complexes identifies the CB as a surveillance center that may safeguard genome integrity in post-meiotic haploid cells. Unexpectedly, mRNA recruitment is not determined by predicted piRNA targeting. Instead, transcript localization is encoded by a combinatorial set of intrinsic sequence and structural features that accurately predict CB enrichment at isoform resolution. These findings establish the CB as a dynamic post-transcriptional regulatory compartment that integrates genome surveillance, RNA quality control and selective transcript sorting, uncovering general principles through which biomolecular condensates shape cell-specific transcriptomes.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.744273v1?rss=1

Ma, L., Ahmedani, A., Charmpramary, S., Laasanen, S., Olotu, O., Kotaja, N.

Motivation: Comparative studies of trajectory inference (TI) methods evaluate complete computational pipelines, making it impossible to isolate how much distortion is introduced specifically by the dimensionality reduction (DR) step. To our knowledge, no study has directly and systematically evaluated how well DR methods alone preserve a known reference path when projecting high-dimensional single-cell data to two dimensions, and no current study has introduced a dedicated set of metrics to quantify the degree of path-preservation quality after dimensionality reduction. This gap matters because DR is a universal preprocessing choice that shapes all downstream trajectory analysis, yet its independent geometric effect on path structure remains uncharacterized, and practitioners have no principled way to quantify it. Methods: We tested a panel of candidate path-preservation metrics on two single-cell datasets with known reference trajectories, one linear and one cyclic, to determine whether the resulting metric values, DR-method rankings, and overall conclusions are sensitive to the number of points used to construct and display the path, and whether they remain stable once that choice is fixed. The primary linear dataset is a CD4+ T-cell surface-protein dataset (3,096 cells, 51 proteins); a ground-truth reference path was constructed from cells lying close to the first principal component (PC1) of a single cluster, providing a known linear trajectory in the high-dimensional space. Sixteen DR methods were applied and twelve geometric path-preservation metrics were computed, spanning log-ratio distortions of length, curvature, and spatial similarity; Spearman rank correlations of pairwise distances and segment lengths; and structural complexity measures including self-intersection frequency and coiling. To test the sensitivity of this evaluation framework to path density, we varied the fraction of cells used to define the reference path from 1% to 10% (31-310 path points) and tracked how method rankings responded. The same analysis was repeated on a topologically distinct reference, a closed B-cell cell-cycle loop detected by persistent homology in a separate CyTOF dataset, to test whether these conclusions about metric and method stability hold for cyclic as well as linear trajectories. Results: The central sensitivity question, whether the number of points used to construct the reference path changes the evaluation's conclusions, was answered negatively on both datasets. On the linear PC1 trajectory, absolute values of all twelve metrics shifted smoothly as the path-density threshold was varied from 1% to 10% (31-310 points), reflecting the broadening of the reference band, but each method's composite rank remained stable across every threshold: no method changed performance tier as the hyperparameter varied. A composite rank aggregating all twelve metrics identified the same consistently high-performing methods (UMAP, MDS, CNPE, TSNE, SPE, LPMIP) and consistently low-performing methods (SPMDS, LPP, DVE, LAPEIG, PHATE) at every density level tested. Considered on its own, `SpatDistSpear`, the single most discriminating metric, separated a high-fidelity group (LPMIP, DM, MDS, SPMDS, DVE, CISOMAP, CNPE; all r > 0.80) from a mid-range group (LAPEIG, SPE, PHATE, UMAP, TSNE, FOSMOD) and a low-fidelity group (PFA, NNP, LPP); global distance preservation and overall composite performance therefore do not always agree on the same "top tier" of methods, but this disagreement in which metric identifies the best methods is itself density-independent rather than an artifact of the specific threshold chosen. The cyclic loop reproduced the same density-independence: absolute metric values drifted with the per-segment band width, yet each method's composite rank again held constant across all eleven density levels. The identity of the best and worst performers was largely, though not entirely, conserved between the two topologies, with CNPE, LPMIP, SPE, and MDS as top performers and LAPEIG, DVE, and SPMDS as poor performers on both the linear path and the closed loop. UMAP and TSNE were exceptions, dropping from top performers on the linear path to the middle of the sixteen-method panel, rather than the worst tier, on the closed loop. This topology-dependence is a property of the reference geometry rather than of path density: it holds consistently regardless of how many points are used to define the path. Significance: This work introduces a direct, pipeline-independent evaluation of how DR methods distort trajectory geometry, a benchmarking dimension absent from existing TI comparisons. The within-dataset rank stability result, demonstrated on both a linear and a cyclic reference trajectory, validates the use of a fixed reference-path threshold as a robust operating point for large-scale DR benchmarking; however, the partial reordering of top performers between topologies shows that a method's DR benchmark ranking is trajectory-shape-dependent and should not be assumed to transfer from a linear to a cyclic reference.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748633v1?rss=1

Bombina, P., Coombes, K. R.

DNA tetrahedra (DNA Td) are promising nanocarriers for drug delivery, but how hypoxia affects their cellular internalisation remains poorly understood. We synthesised and characterised Cy5-labelled DNA Td and established chemical hypoxia; in HeLa, MDA-MB-231, and MCF-7 cells. Hypoxia was confirmed by HIF-1; nuclear translocation. Confocal microscopy revealed significantly reduced DNA Td uptake under hypoxia, whereas transferrin and cholera toxin B uptake increased, indicating cargo-selective regulation. Temperature-arrest experiments confirmed reduced energy-dependent internalisation. Pharmacological profiling showed a shift from clathrin-mediated and galectin/glycan-dependent pathways toward lipid raft/cholesterol-dependent uptake. Hypoxia increased plasma membrane electronegativity, suggesting a biophysical barrier to DNA Td uptake. Importantly, DOTMA complexation restored uptake to normoxic levels, identifying electrostatic repulsion as a key determinant. In zebrafish larvae, hypoxia significantly enhanced whole-larva DNA Td accumulation. These findings highlight surface charge engineering as a strategy for improving DNA nanostructure delivery under hypoxic conditions.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.748847v1?rss=1

Kosara, S., Prakash, G., Dave, H., Dhanasekaran, S., Bhatia, D. D.

Sensitive human data underpin discoveries across medicine, biology and the social sciences, yet privacy regulation often prevents sharing them with collaborators or artificial intelligence (AI) systems. We introduce SPHERE, a model-free method that makes sensitive datasets directly usable by AI and shareable for open science as a synthetic twin, while the original records never leave the local environment. Across 33 datasets spanning five scientific domains, SPHERE protects individual privacy against adversarial re-identification attacks while preserving the data's statistical structure: means, variances and correlations are reproduced exactly, effect size and P value in linear statistical analysis is numerically identical, nonlinear machine-learning utility is retained, and each twin is generated in seconds on a laptop. Frontier AI agents running on the twin reach the same scientific conclusions as on the original records. Analyses of the twin reproduce genome- and proteome-wide results at UK Biobank scale and recover the findings of landmark studies across three independent cohorts and consortia. The approach also extends to deep-learning embeddings across language, vision and time-series, with minimal utility loss. We make the Stanford Alzheimer's Disease Research Center cohort openly available for the first time, as a SPHERE twin spanning nine modalities that any registered researcher can analyze without an approval process. We release SPHERE with certification of each twin's privacy and fidelity, and an AI agent that autonomously executes research tasks on sensitive data without ever accessing it. Sensitive datasets that are currently closed to research could thus become routine inputs to open science and AI to enable key discoveries.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748580v1?rss=1

He, Z., Park, J., Pulgrossi, R. C., Lee, J., Butler, R. R., Weber, A., Tian, L., Zhang, X., Wang, J., Sha, S., Mormino, E. C., Wyss-Coray, T., Henderson, V. W., Longo, F. M., Zou, J., Desai, M., Altman, R.

In biomedical research, multiple types of high-dimensional data, such as genomic, transcriptomic, proteomic, and metabolomic data, are increasingly collected from the same subjects. Integrating these multiple data views can improve prediction by exploiting shared or complementary information across the views. Cooperative learning provides an agreement-based framework for multiview supervised learning by encouraging predictions obtained from individual views to be similar. However, the original framework assumes independent observations and therefore does not account for clustered structures, such as repeated measurements obtained from the same subject. To address this limitation, we propose Cooperative Learning with a penalized Linear Mixed Model (CL-pLMM) for high-dimensional multiview data with a clustered structure. CL-pLMM replaces the ordinary prediction loss in cooperative learning with a covariance-weighted loss that accounts for within-cluster dependence, while retaining the agreement penalty between views and a Lasso penalty for variable selection. We further show that its objective function can be represented as a penalized linear mixed-effects model applied to augmented data, allowing existing estimation procedures to be used. The performance of CL-pLMM is evaluated through simulation studies under various signal and dependence settings and an application to longitudinal proteomic and metabolomic data for predicting the time to spontaneous labor.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748742v1?rss=1

Yoshimura, S., Takagishi, M., Tanioka, K.

Sleep expression in wild animals is regulated by their ecological context. Yet, how populations of a species differ in their sleep in response to environmental challenges such as urbanization, remains to be fully understood. Robust quantification of sleep in small animals, such as lizards, has so far been methodologically difficult, limiting population-level assessments. Using recently developed miniature loggers, we recorded sleep in 19 wild-caught individuals of the Peninsular rock agama lizard (Psammophilus dorsalis). We compared individuals from urban and rural environments and measured electro-oculogram (EOG)-derived sleep parameters: sleep duration, bout frequency, bout duration, and interval between bouts. We then subjected all individuals to an acute stressor (handling-restraint) and examined changes in sleep expression. We found that this species slept mostly at night, with an average total sleep time of 11h 10min. EOG-derived sleep characteristics did not differ substantially between populations, although daytime sleep bouts were more consolidated (shorter interval between bouts) in urban lizards. Contrary to evidence from mammals, acute restraint stress did not impact sleep, overall or for either population. To complement these measures, we also measured the latency to arousal after exposure to vibration stimuli in a separate set of urban and rural lizards (n = 12 each), and found that urban individuals responded marginally quicker at night compared rural individuals, demonstrating greater vigilance and/or more fragmented sleep. Overall, we show that sleep characteristics of wild animals in a common garden condition can be conserved within a species, with limited context-dependent variation between populations.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748758v1?rss=1

Mohanty, N. P., Libourel, P.-A. P., Bharath, D., Joshi, M., Thaker, M.

Methylation-associated deamination removes CpG from vertebrate genomes, but how it affects the dinucleotide profile remains unresolved. We analysed 753 vertebrate and 481 invertebrate genomes to test whether CpG loss defines a compositional axis and to identify its strongest signature. CpG depletion was the dominant axis of vertebrate dinucleotide variation. Unexpectedly, its strongest between-genome correlate was AG/CT rather than the direct mutational product TpG/CpA, which showed the expected dataset-wide mass balance but varied little among genomes. A forward-evolution model based on measured seven-nucleotide human germline substitution rates produced neither CpG depletion nor AG/CT enrichment when methylated-CpG mutability was excluded. Adding one CpG-specific mutability term, calibrated only to the mammalian CpG ratio, reproduced both features, identifying AG/CT as a second-order consequence of the context-dependent mutation network. Within genomes, CpG depletion was strongest in transposable elements and weakened with distance from them. Across vertebrates, the axis followed Amniota more closely than endothermy and was associated with an expanded GC-rich isochore compartment. A Machine Learning analysis shows that CpG depletion and AG/CT were the principal features separating vertebrates from invertebrates, in which both were markedly attenuated. Thus, a methylation-associated axis organises vertebrate dinucleotide composition, and its strongest marker is not the immediate product of CpG deamination.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.747768v1?rss=1

Bobbo, T., Waththe Liyanage, W. W., Boattini, A., Lio, P., Taccioli, C.

Spatial transcriptomics has enabled the advancement of gene expression analysis, yet spatial co-expression remains understudied. We introduce spatial covariance regression (SCR), a scalable Bayesian factor-model-based framework for estimation of spatially-resolved gene co-expression networks across tissue domains. These networks provide the spatial map of gene-gene correlations and enable the identification of spatially co-expressed genes (SCGs), which serve as potential prognostic biomarkers and therapeutic targets.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748618v1?rss=1

Buker, I. E., Ni, Y., Hicks, S. C., Kang, J., Acharyya, S.

Dietary proteins induce antigen-specific immune responses, leading to oral tolerance or food allergies. While mouse models of allergic sensitization rely on antigen context and adjuvant signaling, exposure to purified proteins is usually sufficient for tolerance development in studies with soluble model antigens. In previous work, we discovered that the maize protein zein induces robust antigen-specific intestinal Tregs following normal chow exposure, but unexpectedly does not elicit a T cell response when delivered as a purified protein. Here, we investigated the mechanisms underlying these differential T cell responses, considering both biochemical properties of zein and its food matrix, as well as intestinal antigen processing. We focused on three zein preparations that induced different frequencies of antigen-specific intestinal T cells. While in vitro studies showed comparable presentation by dendritic cells, in vivo studies revealed stark differences in intestinal uptake. We identified differences in protein solubility and active sampling mechanisms as cooperative drivers of intestinal zein uptake. Different zein preparations could be sampled by either goblet cells, a route previously described for the model antigen OVA, or M cells, a route hypothesized for dietary proteins and established for transport of pathogenic bacteria. These findings also extend to peanuts, where the allergen Ara h 1 was selectively transported by goblet or M cells depending on food preparation, establishing examples of naturally occurring dietary M cell ligands. Finally, solubilizing zein increased intestinal uptake, and correspondingly feeding a diet with solubilized zein led to more intestinal zein-specific T cells compared to feeding the insoluble form. Overall, these findings suggest that intestinal uptake of dietary antigens is a regulated and context-dependent determinant of antigen-specific T cell induction, with implications for development of tolerance-restoring immunotherapies.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.01.748748v1?rss=1

Blum, J., Chang, Y.-T., Le, H., Morrison, A., Kong, R., Schulman, E., Chou, T.-W.

Oligodendrocytes play an essential role in axonal metabolic support and myelination, and their dysfunction is associated with amyotrophic lateral sclerosis (ALS). LINGO1 is a major inhibitor of oligodendrocyte differentiation and axonal regeneration, identified as a promising therapeutic targeted for inflammatory demyelinating diseases. Here, we investigated the expression pattern of LINGO1 in the well-established SOD1(G93A) transgenic mouse model of ALS. We show that LINGO1 protein expression is significantly elevated in both grey and white matter of the mutant spinal cord during both the symptomatic and end stages of the disease. LINGO1 is also notably co-expressed with astroglial and neuronal markers. However, neither the levels of LINGO1 transcripts, nor levels of microRNAs predicted to regulate its expression were altered in the mutant spinal cord, suggesting the involvement of alternative regulatory mechanisms. Our analysis of microRNAs revealed in the mutant a significant downregulation of miR-138, a microRNA known to promote myelination. The differential expression of LINGO1 and miR-138 in ALS mice thus may drive deleterious processes during disease progression in adulthood. Moreover, our data show that white-matter pathology arises earlier than initially thought, as evidenced by significant alterations of oligodendrogenesis that manifest as early as postnatal day 5. Study of the molecular determinants of this developmental defect shows increased miR-138 expression, which may promote oligodendrocyte progenitor cell maturation and involve compensatory mechanisms. Overall, our work identifies disrupted expression of LINGO1 and/or miR-138 in brain during early oligodendrogenesis and at symptomatic disease stages. These findings open the way to further investigations that may consider miR-138 as a potential presymptomatic biomarker and LINGO1 as a therapeutic target to promote remyelination in the context of ALS.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.03.749143v1?rss=1

Zahaf, A., Bourseguin, J., cobret, L., Kassoussi, A., Moussaed, M., Raoul, C., Traiffort, E., Morisset-Lopez, S.

The differentiation of naive CD4+ T cells into effector T cell subsets such as TH1, TH2, TH17 and TREG cells is governed by tightly coordinated activation programmes dependent on T cell receptor (TCR) engagement, co-stimulation, and cytokine signalling. This differentiation process involves regulation of stress-responsive signalling pathways such as the unfolded protein response (UPR) and the oxidative stress response (OSR) to adapt to the physiologic demands specific to distinct T cell subset functions. This suggests that pharmacologically targeting stress-responsive pathways offers a unique opportunity to selectively remodel the differentiation of these different T cell subsets. We previously identified compound AA147 as a metabolically activated proteostasis regulator that can induce both the ATF6 signalling arm of the UPR and, in certain cell types, the NRF2-regulated arm of the OSR. (Paxman et al., 2018; Plate et al., 2016) Here, we show that treatment with AA147 selectively reduces differentiation of pro-inflammatory TH17 cells by promoting degradation of the lineage-specifying transcription factor RORgammat, without impacting the transcription factors of other CD4+ T cell subsets. AA147-dependent reduction in TH17 differentiation is independent of ATF6 activation and involves activation of NRF2, which reduces intracellular reactive oxygen species (ROS) to hinder TH17 cell differentiation. Apart from RORgammat, we found that AA147 decreases expression of the TCR-responsive factor IRF4, thus suppressing production of select effector cytokines across effector T cells, demonstrating additional ways in which this compound reshapes the activities of these essential T cell subsets. Our results demonstrate the potential for metabolically activated proteostasis regulators such as AA147 to selectively reshape TH17 cell identity while broadly dampening effector cytokine responses across effector T cell subsets, through both NRF2-dependent and independent mechanisms during T cell differentiation.

Date: 2026-09-05
https://www.biorxiv.org/content/10.64898/2026.09.02.747634v1?rss=1

Chatterjee, P., Sanchez Ortiz, K., Dikiy, S., Bollong, M. J., Thaxton, J. E., Mendoza, A., Wiseman, L.