bioRxiv : the preprint server for biology最新文献

{"title":"An origin for a eukaryotic lipid transfer protein fold in Asgard archaea.","authors":"Nicolas-Frédéric Lipp, Itay Budin","doi":"10.1101/2025.05.16.653879","DOIUrl":"10.1101/2025.05.16.653879","url":null,"abstract":"<p><p>The evolution of eukaryotic cells necessitated the advent of machinery to transport molecular building blocks for organelles to proliferate. Extant eukaryotes share several classes of highly conserved lipid transfer proteins (LTP) that associate with donor membranes, bind individual lipid molecules, and shuttle them to acceptor membranes. Because cells lacking organelles do not require extensive lipid transport networks, it is not known if this machinery pre-dated eukaryotic organelles or had to evolve alongside them. Here we describe a class of putative lipid transporters in the Asgard archaeota superphylum that share a common ancestry with eukaryotic LTPs in the START domain superfamily. We identified three classes of Asgard START proteins, StarAsg1-3, which are conserved across most Asgard phyla. Of these, StarAsg1 family proteins contain the predicted structural features necessary for lipid transfer: large, hydrophobic binding pockets lined with amphipathic motifs for membrane docking. In contrast, StarAsg2 and StarAsg3 family proteins contain smaller binding cavities and minimal predicted membrane interactions. We found that StarAsg1 from Lokiarcheota interacts with anionic membranes both in vitro and in yeast cells and can transfer sterols between liposomes. Phylogenetic analysis of START domains across the tree of life indicates that eukaryotic LTPs share a common ancestry with StarAsg1 homologs, while StarAsg2 and StarAsg3 form a monophyletic group with eukaryotic heat shock protein cochaperones. We propose that the emergence of proteins for inter-membrane lipid transporters in the ancestors of eukaryotic cells could have facilitated the subsequent development of intracellular organelles.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12139934/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144236414","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrated Histology and Molecular Profiling of Postmortem Human Auditory and Vestibular Organs via a Poly(Methyl Methacrylate)-Based Workflow.","authors":"David Bächinger, Brock Peyton, Jacqueline Neubauer, Anbuselvan Dharmarajan, MengYu Zhu, Jennifer T O'Malley, Venus Kallupurackal, Steven Senese, Alison Brown, Sabina Wunderlin, Susanne Kreutzer, Nora M Weiss, Heiko Richter, Adrian Dalbert, Christof Röösli, Anja Kipar, Zsuzsanna Varga, Brigitte von Rechenberg, Sami S Amr, Andreas H Eckhard","doi":"10.1101/2025.06.04.657716","DOIUrl":"10.1101/2025.06.04.657716","url":null,"abstract":"<p><p>Hearing and balance disorders are the most prevalent sensory impairments, affecting hundreds of millions worldwide, yet their underlying cellular and molecular pathologies remain poorly understood. This knowledge gap stems from the inaccessibility of the ear's sensory organs-embedded within the temporal bone (TB), the hardest bone in the body-which cannot be biopsied in living patients without causing irreversible damage. Conventional histopathology workflows rely on postmortem en bloc extraction of TBs, followed by lengthy decalcification, celloidin embedding, and manual serial sectioning of these large specimens-a process that takes one to two years, is labor- and cost-intensive, and lacks compatibility with most modern protein, DNA, and RNA assays. Here, we present a rapid, reversible polymethyl methacrylate (rPMMA) workflow that enables advanced molecular histopathology studies on formalin-fixed, calcified TBs. Our protocol uses low-temperature (-40 °C to +4 °C) resin embedding, precision near-serial sectioning (10-50 µm) via femtosecond laser microtomy or precision diamond wire sawing, and subsequent deacrylation to fully restore tissue accessibility for high-fidelity histomorphology, multiplexed immunofluorescence, whole-genome sequencing, and in situ mRNA detection (RNAscope^™) assays. Compared to the gold-standard celloidin workflow, our method reduces processing time and costs by approximately 90% while integrating equivalent histomorphology with advanced molecular assays, providing a new benchmark for multidimensional studies in human hearing and balance pathologies.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12157654/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277628","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Engineering gene expression dynamics via self-amplifying RNA with drug-responsive non-structural proteins.","authors":"Parisa Yousefpour, Justin R Gregory, Kristen Si, Jan Lonzaric, Yingzhong Li, Junmin Wang, Kashif Qureshi, Amir Ledbetter, Ashley A Lemnios, Jonathan Dye, Tanaka K Remba, Rachel Yeung, Melissa Güereca, Linette Rodriguez, Yuebao Zhang, Shengwei Wu, Yizhou Dong, Ron Weiss, Darrell J Irvine","doi":"10.1101/2025.06.08.658495","DOIUrl":"10.1101/2025.06.08.658495","url":null,"abstract":"<p><p>The design of gene therapies with drug-regulatable expression of therapeutic payloads is of interest for diverse applications. We hypothesized that a regulated expression system based on alphavirus-derived self-amplifying RNAs (saRNAs), which encode 4 non-structural proteins (nsPs) that copy the RNA backbone to enable sustained expression, would have advantages in safety and simplicity of delivery. Here we designed saRNAs where payload expression is regulated by the FDA-approved drug trimethoprim (TMP), by fusing TMP-responsive degradation domains (DDs) to nsPs to regulate RNA self-amplification. Screening a library of nsP-DD fusions, we identified an optimal design with DDs fused to nsP2, nsP3, and the payload, achieving a high fold-change in expression level in response to TMP and low expression in the off state. In mice, this saRNA circuit enabled diverse dynamic expression patterns in response to oral TMP. Implementing this circuit for controlled expression of an HIV antigen, an escalating TMP regimen significantly enhanced germinal center responses critical for B cell affinity maturation. This drug-regulated RNA technology holds potential for vaccines, immunotherapies, and gene therapies.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12157543/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277539","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Pathological Role and Therapeutic potential of ALDH2 in acrolein detoxification Following Spinal Cord Injury in Mice.","authors":"Siyuan Sun, Seth Herr, Anna Alford, Rachel Stingle, Zaiyang Zhang, Timothy Dong, Riyi Shi","doi":"10.1101/2025.06.08.658505","DOIUrl":"10.1101/2025.06.08.658505","url":null,"abstract":"<p><p>Oxidative stress and lipid peroxidation-derived aldehydes, such as acrolein, play a central role in the pathology of spinal cord injury (SCI) and have emerged as promising therapeutic targets. Mitochondrial aldehyde dehydrogenase-2 (ALDH2) is a key oxidoreductase responsible for detoxifying reactive aldehydes. Pharmacological activation of ALDH2 using Alda-1, a selective agonist, has been shown to reduce aldehyde accumulation, alleviate inflammation, and enhance functional recovery in experimental SCI models. However, approximately 8% of the global population carries the ALDH2*2 mutation, which severely impairs this detoxification pathway. In this study, we used a transgenic ALDH2*2 mouse model to investigate the role of ALDH2 in SCI pathology. This model mimics the human ALDH2*2 condition, allowing us to examine the impact of impaired aldehyde clearance on acrolein accumulation and its pathological consequences. We modulated endogenous aldehyde detoxification through both genetic deficiency and pharmacological activation with Alda-1. Our results showed that ALDH2 deficiency led to significantly elevated acrolein levels, which were associated with increased microglial activation, cytokine storm, neuronal loss, demyelination, and tissue damage compared to wild-type (WT) mice. Treatment with Alda-1 enhanced ALDH2 activity and significantly reduced acrolein levels in both ALDH2*2 and WT mice from 2 to 28 days post-SCI. This was accompanied by reduced inflammation, improved preservation of myelin, and marked improvements in locomotor and sensory function, especially in ALDH2*2 mice. Notably, even beyond the traditionally ideal treatment window, Alda-1 treatment remained effective in promoting recovery, particularly in motor function and to a greater extent in ALDH2*2 mice. Our study comprehensively evaluated ALDH2's role in SCI by both genetically impairing and pharmacologically enhancing its activity, highlighting ALDH2 as a critical modulator of acrolein-mediated damage and suggesting its potential as a therapeutic target, especially for individuals with the ALDH2*2 mutation.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12157631/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277920","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cyclo-stationary distributions of mRNA and Protein counts for random cell division times.","authors":"Syed Yunus Ali, Aditya Saran, Ashok Prasad, Abhyudai Singh, Dibyendu Das","doi":"10.1101/2025.06.06.658238","DOIUrl":"10.1101/2025.06.06.658238","url":null,"abstract":"<p><p>There is a long history of using experimental and computational approaches to study noise in single-cell levels of mRNA and proteins. The noise originates from a myriad of factors: intrinsic processes of gene expression, partitioning errors during division, and extrinsic effects, such as, random cell-cycle times. Although theoretical methods are well developed to analytically understand full statistics of copy numbers for fixed or Erlang distributed cell cycle times, the general problem of random division times is still open. For any random (but uncorrelated) division time distribution, we present a method to address this challenging problem and obtain exact series representations of the copy number distributions in the cyclo-stationary state. We provide explicit cell age-specific and age-averaged results, and analyze the relative contribution to noise from intrinsic and extrinsic sources. Our analytical approach will aid the analysis of single-cell expression data and help in disentangling the impact of variability in division times.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12157499/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277468","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Short chain fatty acids regulate the chromatin landscape and distinct gene expression changes in human colorectal cancer cells.","authors":"Tohfa Kabir, Charlotte A Connamacher, Zara Nadeem, Matthew R Paul, Matthew R Smutny, Zoe K Lawler, Annaelle M Djomo, Thomas S Carroll, Leah A Gates","doi":"10.1101/2025.05.07.652677","DOIUrl":"10.1101/2025.05.07.652677","url":null,"abstract":"<p><p>Short chain fatty acids (SCFAs) are small metabolites that are produced through the activity of microbes and have important roles in human physiology. These metabolites have varied mechanisms in interacting with the host, of which one such mode is decorating the chromatin landscape. We previously detected specific histone modifications in the mouse gut that can be derived from SCFAs and are regulated by the microbiota. However, the roles of these SCFAs on chromatin and how they impact gene regulation in human cells is largely unknown. Now, our studies demonstrate these microbiota-dependent histone posttranslational modifications (PTMs) are associated with alterations in gene regulation in human cells. We show that histone butyrylation on H3K27 is detected in human colon samples. Furthermore, histone acetylation, butyrylation, and propionylation on H3K9 and H3K27 are responsive to levels of SCFAs in human colon cancer cell lines and are associated with active gene regulatory elements. In addition, treatment of human cancer cell lines with individual metabolites or combinations of SCFAs replicating the intestinal lumen environment result in distinct and overlapping gene program changes, with butyrate largely driving gene regulatory effects of SCFA combinations. Lastly, we define butyrate effects on gene expression that are independent of HDAC inhibition and are dependent on p300/CBP, suggesting potential gene programs regulated by histone butyrylation. Together, these results demonstrate that SCFAs are key regulators of the chromatin landscape and gene programs in human colorectal cancer cells.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12132577/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144218442","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Genomic structural equation modeling of impulsivity and risk-taking traits reveals three latent factors distinctly associated with brain structure and development.","authors":"Mari Shishikura, Lang Liu, Eric Yu, Rui Zhu, Laura Vilar-Ribó, Renata B Cupertino, Abraham A Palmer, Sandra Sanchez-Roige, Uku Vainik, Filip Morys, Ziv Gan-Or, Bratislav Misic, Alain Dagher","doi":"10.1101/2025.06.05.657840","DOIUrl":"https://doi.org/10.1101/2025.06.05.657840","url":null,"abstract":"<p><strong>Background: </strong>Impulsivity is a multifaceted trait that emerges in childhood and is linked to several psychiatric disorders, such as attention-deficit/hyperactivity disorder and substance use disorders. Recent genomic and neuroimaging studies have identified genetic loci and brain systems associated with impulsivity and risk-taking behaviors. However, how these genetic underpinnings overlap across different facets of impulsivity and risk-taking, and how they are associated with brain morphology during early development remain unknown.</p><p><strong>Methods: </strong>We applied genomic structural equation modeling to 17 impulsivity and risk-taking GWAS datasets to explore the overlapping genetic architecture underlying these phenotypes. We then computed polygenic scores (PGSs) for each genetic latent factor in 4,142 participants from the Adolescent Brain Cognitive Development Study and examined how each factor was associated with the brain structure during early development. We further tested whether socioeconomic status modulated the association between the PGSs and brain structures.</p><p><strong>Results: </strong>We identified three genetic latent factors, which we labeled as lack of self-control, reward drive, and sensation seeking. These showed distinct associations with brain structure in late childhood and early adolescence, relating to the frontal cortex, subcortical cellularity, and white matter structure in different ways. The association between PGS of lack of self-control and white matter mean diffusivity was modulated by socioeconomic status.</p><p><strong>Conclusions: </strong>Our findings revealed that genetic predisposition for impulsivity and risk-taking is associated with morphological brain differences as early as ages 9-10. We also highlighted the importance of capturing the multidimensional nature of these traits to better understand their neurodevelopmental basis.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12157643/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277591","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"PTP1B inhibition promotes microglial phagocytosis in Alzheimer's disease models by enhancing SYK signaling.","authors":"Yuxin Cen, Steven R Alves, Dongyan Song, Jonathan Preall, Linda Van Aelst, Nicholas K Tonks","doi":"10.1101/2025.05.30.657101","DOIUrl":"https://doi.org/10.1101/2025.05.30.657101","url":null,"abstract":"<p><p>Amyloid-β (Aβ) accumulation is a hallmark of Alzheimer's disease (AD). Emerging evidence suggests that impaired microglial Aβ phagocytosis is a key feature in AD, highlighting the therapeutic potential of enhancing this innate immune function. Here, we demonstrate that genetic deletion or pharmacological inhibition of protein tyrosine phosphatase 1B (PTP1B) ameliorated memory deficits and reduced Aβ burden in APP/PS1 mice. Moreover, we show that PTP1B was highly expressed in microglia, and its deficiency promoted a transcriptional shift toward immune activation and phagocytosis. Consistently, PTP1B deletion in microglia enhanced phagocytosis and metabolic fitness, supported by increased AKT-mTOR signaling, a pathway essential for meeting the energy demands of activation. Mechanistically, we identified spleen tyrosine kinase (SYK), a key regulator of microglial phagocytosis, as a direct substrate of PTP1B. Inhibition of SYK showed that PTP1B modulates microglial activation in a SYK-dependent manner. These findings established PTP1B as a critical modulator of microglial activation and a potential therapeutic target for AD.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12154680/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277829","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The Role of Paraventricular Nucleus of Thalamus in Sleep Disturbance Induced by Withdrawal from Repeated Ethanol Exposure.","authors":"Aubrey Bennett, Jeyeon Lee, Hyunjung Kim, Vatsala Kapoor, David Jury, Seungwoo Kang","doi":"10.1101/2025.06.04.657945","DOIUrl":"https://doi.org/10.1101/2025.06.04.657945","url":null,"abstract":"<p><p>Sleep disturbance is known to be comorbid with withdrawal from repeated ethanol exposure and could be a negative reinforcement for the majority of people with alcohol use disorder (AUD). The paraventricular nucleus of the thalamus (PVT) has been highlighted for its function in integrating arousal states and associated modulation in sleep homeostasis. However, there is limited understanding of the involvement of PVT neurons in regulating sleep patterns, especially during withdrawal from chronic ethanol exposure. In this study, we investigated the potential function of the PVT in sleep disturbance during ethanol withdrawal using electrophysiology, in vivo calcium imaging, biochemical, and chemogenetic approaches. At 24 hours post-withdrawal from chronic intermittent ethanol exposure (CIE) for four weeks, there is an increase in wake time and a decrease in non-rapid eye movement (NREM) sleep. The calcium transient levels in the PVT neurons are positively correlated with the transition from sleep to wakefulness. CIE elevates the PVT neuronal activity in a subregion-specific manner, resulting in a significant rise in cFos levels in the anterior PVT (aPVT). Temporal suppression of aPVT excitatory neurons via chemogenetics ameliorates the disturbance in sleep patterns generated by CIE. The aPVT has a notable distinction in the expression of the m-type potassium channel subunit, KCNQ2, with a higher expression level compared to the posterior PVT (pPVT). While the expression of KCNQ2 in the aPVT is reduced in CIE mice, the restoration of KCNQ2 expression using viral gene transfer within the aPVT alleviates the sleep disturbances produced by CIE. This data indicates a significant role of the PVT in sleep disturbance during ethanol withdrawal, which may partially be due to the downregulation of M-channels, hence underscoring M-channels in the PVT as a potential therapeutic target for sleep disturbance in alcohol use disorder.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12157549/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277906","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrative Chemical Genetics Platform Identifies Condensate Modulators Linked to Neurological Disorders.","authors":"Dylan Poch, Chandrayee Mukherjee, Sunanda Mallik, Vanessa Todorow, E F Elsiena Kuiper, Nalini Dhingra, Yulia V Surovtseva, Christian Schlieker","doi":"10.1101/2025.06.07.658469","DOIUrl":"https://doi.org/10.1101/2025.06.07.658469","url":null,"abstract":"<p><p>Aberrant biomolecular condensates are implicated in multiple incurable neurological disorders, including Amyotrophic Lateral Sclerosis (ALS), Frontotemporal Dementia (FTD), and DYT1 dystonia. However, the role of condensates in driving disease etiology remains poorly understood. Here, we identify myeloid leukemia factor 2 (MLF2) as a disease-agnostic biomarker for phase transitions, including stress granules and nuclear condensates associated with dystonia. Exploiting fluorophore-derivatized MLF2 constructs, we developed a high-content platform and computational pipeline to screen modulators of NE condensates across chemical and genetic space. We identified RNF26 and ZNF335 as protective factors that prevent the buildup of nuclear condensates sequestering K48-linked polyubiquitinated proteins. Chemical screening identified four FDA-approved drugs that potently modulate condensates by resolving polyubiquitinated cargo and MLF2 accumulation. Our exploratory integrated chemical-genetics approach suggests that modulation of zinc, and potentially autophagy and oxidative stress, is critical for condensate modulation and nuclear proteostasis, offering potential therapeutic strategies for neurological disorders. Application of our platform to a genome-wide CRISPR KO screen identified strong enrichment of candidate genes linked to primary microcephaly and related neurodevelopmental disorders. Two hypomorphic microcephaly-associated alleles of ZNF335 failed to rescue nuclear condensate accumulation in ZNF335 KO cells, suggesting that aberrant condensates and impaired nuclear proteostasis may contribute to the pathogenesis of microcephaly.</p><p><strong>Highlights: </strong>MLF2 emerges as a disease-agnostic condensate biomarker co-localizing with TDP-43 and G3BP1FDA-approved drugs target condensates linked to perturbed proteostasis.RNF26 and ZNF335 are identified as modulators of nuclear phase transitions.Microcephaly patient disease alleles fail to counteract aberrant condensates.</p>","PeriodicalId":519960,"journal":{"name":"bioRxiv : the preprint server for biology","volume":" ","pages":""},"PeriodicalIF":0.0,"publicationDate":"2025-06-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12157464/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144277631","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}