Biology subjects

Hasan, M.

Publications and source records attributed to Hasan, M..

At least 19 recordsLinked to original sources

bioRxiv2026

Hemodynamic-Structural Multi-Fidelity Assessment of Endovascular Flow Diversion Strategies for Cerebral Aneurysm Treatment: An Integrated Computational and Experimental Framework

Subarachnoid hemorrhage after cerebral aneurysm rupture has a 50% first-month fatality rate. Endovascular flow diversion is the primary treatment modality. However, no prior study has compared stents, coils, and the Woven Endo-Bridge hemodynamically integrating hemodynamic results with structural device behavior in one framework. An aneurysm model was reconstructed from computed tomography imaging of a patient, and computational fluid dynamics evaluated multiple hemodynamic metrics across three flow-diversion strategies. Finite element analysis assessed stent geometries with 50%, 55%, and 60% porosity in 316L stainless steel, poly-L-lactic acid, and nitinol. Universal testing machine tests validated the models with printed polylactic acid and thermoplastic polyurethane analogues. Dimensional and elastic-tube analyses tested scale equivalence between the FEA and UTM geometries and the rigid-wall CFD assumption. Patient-specific CFD confirmed a high-risk hemodynamic profile. The stent yielded the lowest peak intra-sac velocity, greatest pressure reduction, near-zero wall shear stress, the only reduction in time-averaged wall shear stress, cycle-averaged TAWSS, and the largest peak-systolic reductions in velocity, WSS, and vorticity. OSI and RRT rose for all three treated configurations, consistent with the hemodynamic signature of flow diversion. FEA reproduced the expected stiffness hierarchy (Steel > Nitinol > PLLA), and TPU was the closest surrogate for Nitinol's elastic recovery. Scaled specimens matched native-scale stiffness within 2.3%. Predicted radial distension was orders of magnitude below native wall values, supporting rigid-wall CFD. Stent-assisted diversion demonstrated the most favorable hemodynamic profile. A 50%-porosity Nitinol stent best matched target elastic mechanics, while 316L stainless steel provided the greatest resistance to radial compression.

neuroscience↗

bioRxiv2026

Joint recording of past and present transcriptomes reconstructs single-cell trajectories

Tracing molecular cell state transitions through development, homeostasis and disease is still beyond reach at single-cell resolution. We present scReMeMber-Seq, a methylation-based cell-memory recorder connecting past transcriptional states with present transcriptome and DNA methylome in individual cells. scReMeMber-Seq leverages pulse induction of DCM-Polr2b (TimeMachine) and single-cell methylome-transcriptome (scMT) sequencing for endpoint readout. In mouse neural stem cells (NSCs) and pancreatic organoids, single-cell DCM methylation profiles (scDCMome) accurately discriminate cell types. Over time, scDCMome also record NSC quiescence-activation state changes across multiple cell divisions. In zebrafish embryos, scDCMome resolve individual transcriptional trajectories from progenitor to neuron. Together, scReMeMber-Seq enables real time reconstruction of cellular trajectories, connecting past and present molecular states in single cells, at genome scale, across experimental systems, in vitro and in vivo.

neuroscience↗

bioRxiv2026

Shared phylogeographic structuring and recurrent hybridization underlie the diversification of Microhyla frog complexes in the Indochinese biodiversity hotspot

Comparative phylogeography provides a powerful framework to identify the historical processes shaping biodiversity hotspots by testing whether co-distributed species exhibit shared patterns of diversification. Southeast Asia harbors exceptional biodiversity, yet the extent to which common paleogeographic and climatic drivers have structured diversification across taxa remains poorly understood. Here, we investigated the evolutionary history of five widespread Microhyla species complexes distributed across the Indochinese Peninsula and adjacent regions using dense mitochondrial sampling (1,388 individuals) combined with genome-scale ddRAD sequencing (280 individuals). Across all complexes, both approaches recovered multiple lineages and remarkably congruent phylogeographic breaks and contact/intergradation zones associated with major Indochinese regions, including Myanmar, the Tenasserim-Malay Peninsula, southern Vietnam, and northern Vietnam-southern China, supporting the hypothesis that Indochina functions as a mosaic of stable biogeographic units. However, lineage divergence varied substantially among complexes, suggesting that common biogeographic drivers interacted with species-specific demographic histories. Phylogenomic analyses and cyto-nuclear discordance further revealed historical introgression in every complex, indicating that diversification involved both long-term allopatric isolation and reticulate evolution. These findings portray the Indochinese biodiversity hotspot as a dynamic evolutionary system where cycles of fragmentation and reconnection have repeatedly reshaped lineage boundaries. Moreover, the complex phylogeographic structure recovered exemplifies the urgent need for taxonomic revisions, for which genome-scale data provide an essential framework to validate mitochondrial hypotheses and assess admixture patterns for species delimitation. Finally, regions such as the southern Annamites and Tenasserim Hills emerged as recurrent hotspots of genetic diversity across independent lineages, highlighting their importance for conserving not only species/lineage richness but also the evolutionary processes and adaptive potential that sustain biodiversity.

zoology↗

bioRxiv2026

CyFj11: FlowJo v11 Workspace Import and Legacy Format Export for R-Based Flow Cytometry Analysis

AO_SCPLOWBSTRACTC_SCPLOWHigh-dimensional flow cytometry measures immune cells at single-cell resolution, enabling systematic characterization of cell populations at scale. But harnessing this potential requires seamless interoperability between the interactive gating tools used by biologists and the statistical environments used for detailed downstream analysis. FlowJo, one of the most widely used commercial cytometry analysis software packages, now stores workspaces in a format that existing R tools cannot read, leaving researchers unable to import their gating strategies into R, or to return R-based results to FlowJo for visual review or collaborative sharing, without manual reconstruction. We present CyFj11, an R package that closes this gap, enabling import of FlowJo v11 gating hierarchies into R and export of R-defined gates back to FlowJo (throughout this paper, "import" refers to bringing a FlowJo v11 workspace into R, and "export" to writing an R-derived GatingSet back out to FlowJo). Using a combination of synthetic test scenarios and a real-world immunophenotyping dataset, we show that population counts in FlowJo 10 and 11 matched R-derived values with Pearson correlation coefficients exceeding 0.99. CyFj11 is platform-independent, requires no additional software infrastructure, and is freely available at https://github.com/C3BI-pasteur-fr/CyFj11.

bioinformatics↗

bioRxiv2026

Cross-Sectional Physiological and Neuromuscular Profiling of Elite and Recreational University Badminton Athletes: Preliminary Benchmarks for Exercise-Based Injury Risk Stratification

Background and PurposeBadminton imposes considerable cardiovascular and musculoskeletal stress. Physiological profiling can identify modifiable injury risk factors and inform exercise-based prevention and rehabilitation. This study compared cardiovascular recovery, neuromuscular activation, and limb strength between elite and recreational male university badminton players to derive preliminary physiological benchmarks for injury risk stratification and exercise rehabilitation guidance. MethodsForty male athletes (20 elite: national/university representatives with [≥]5 years of competitive experience; 20 recreational: <3 years of experience) completed assessments of heart rate recovery (HRR), biceps brachii surface electromyography (sEMG; SENIAM protocol), handgrip strength (JAMAR dynamometry), and maximal bodyweight squat repetitions. Independent-sample t-tests with Cohens d ( = 0.05) and Pearson correlations were applied. ResultsElite players demonstrated significantly greater handgrip strength (49.00{+/-}6.12 vs. 39.00{+/-}5.45 kg, p = 0.001, d = 1.72) and lower-limb (LL) strength (60.35{+/-}11.29 vs. 41.75{+/-}6.72 repetitions, p < 0.001, d = 1.96). Normalized sEMG root mean square (RMS) was higher in elite athletes during flexion (11.56{+/-}4.16% vs. 7.26{+/-}5.15%, p = 0.004, d = 0.94) and extension (12.67{+/-}4.56% vs. 7.85{+/-}5.73%, p = 0.003, d = 0.94). HRR did not differ significantly between groups (p = 0.17, d = 0.43, observed power = 0.34). Elite players nonetheless showed a more favorable recovery distribution. sEMG -HRR correlations were weak and non-significant in both groups. ConclusionsElite badminton players exhibit a distinct physiological profile of greater strength and more efficient neuromuscular activation. These preliminary cross-sectional findings may support the design of exercise-based injury-prevention and rehabilitation in university badminton.

biophysics↗

bioRxiv2026

Structure-Based and Stability-Validated Prioritization of BACE1 Inhibitors Integrating Meta-Ensemble QSAR and Molecular Dynamics

Alzheimers disease remains a major therapeutic challenge, and no {beta}-secretase (BACE1) inhibitor has achieved clinical approval. A key limitation of prior discovery efforts is reliance on single-parameter optimization, often resulting in candidates with limited translational potential. In this study, we developed a biology-informed computational framework integrating meta-ensemble QSAR modeling, molecular docking, Protein Language Model (ESM-1b)-guided residue interaction weighting, and ADMET profiling within a normalized multi-parameter ranking scheme. Model performance was validated using cross-validation, external validation, and Y-randomization (n = 100; p = 0.009), while applicability domain analysis based on Tanimoto similarity highlighted reduced reliability for extrapolative predictions. Sensitivity analysis showed high ranking stability under moderate perturbations (Spearman {rho} = 0.998 for {+/-}10%; 0.963 for {+/-}25%), with reduced agreement under randomized weighting ({rho} = 0.821), indicating that prioritization is robust but influenced by weight selection. Screening of 16,196 compounds identified 153 predicted actives (accuracy = 0.852; ROC-AUC = 0.920), which were refined to 111 candidates and seven prioritized leads. Molecular dynamics simulations (200 ns) indicated stable binding and persistent catalytic interactions, with Mol-2 showing favorable dynamic stability and ADMET characteristics. Overall, this study presents an interpretable and quantitatively evaluated framework for multi-parameter compound prioritization, supporting more reliable virtual screening in early-stage CNS drug discovery.

bioinformatics↗

bioRxiv2026

Genetically Programmed Shape-morphing of Engineered Living Materials

Engineered living materials (ELMs) promise genetically programmable functions by coupling biological regulation to synthetic material responses. Here, we introduce genetically encoded, reversible shape-morphing in a peptide-crosslinked polyethylene glycol (PEG) hydrogel whose network density is modulated by opposing enzymatic pairs that induce crosslinking or hydrolysis. This molecular programmability alternates the hydrogel between deswelling and swelling/disintegration and produces 2 - 5-fold changes in mechanical properties. By fabricating a bilayer hydrogel with an inert layer, these molecular modulations are translated into a reversible and directional motion with angular bending motions exceeding 80{degrees}. Further, by embedding genetically engineered bacteria or interfacing mammalian cells, producing the relevant enzymatic cues, the reversible shape-morphing of these ELMs is programmed at the genetic level. We further demonstrate genetically programmed, autonomous reversible bending in a bilayer hydrogel controlled by out-of-equilibrium counteracting biochemical reactions with dynamically changing respective reaction rates. This work establishes a concept where coordinated polymer/peptide material engineering and synthetic biology yield autonomous shape-morphing ELMs, opening avenues toward biohybrid soft robotics, adaptive microfluidic systems, and dynamic biomedical interfaces.

synthetic biology↗

bioRxiv2026

A selective Cullin 3 RING E3 ligase inhibitor attenuates hyperglycemia via dual insulin sensitizing and insulinotropic action

Hyperglycemia is a hallmark of type-2 diabetes and a key pathogenic driver of diabetic complications. Cullin RING E3 ligases (CRLs) are multi-subunit E3 ubiquitin ligases that mediate cellular protein turnover. The activity of CRLs requires cullin neddylation, a post-translational modification that can be pharmacologically targeted with therapeutic potentials. By using hyperinsulinemic-euglycemic clamp analysis, we discover that pan neddylation inhibitor exerts both insulin sensitization effect in liver and muscle and insulinotropic effect in pancreatic {beta} cells. This dual action is mediated by Cullin 3 (Cul3), a member of the 7 canonical cullin family proteins. DI-1859, a selective Cul3 neddylation inhibitor, effectively protects against hyperglycemia in obese mice. DI-1859 enhances insulin signaling by preventing Cul3-mediated insulin receptor substrate degradation in liver and muscle cells. DI-1859 increases insulin secretion in a glucagon-like peptide-1-independent manner in mice and directly potentiates glucose-stimulated insulin secretion in INS-1 832/13 {beta} cells and human islets. Mechanistic studies reveal that DI-1859 does not promote glycolytic flux or bioenergetics function but potentiates glucose-stimulated insulin secretion via mechanisms involving RhoA activation and cytoskeleton remodeling in {beta} cells. This study shows that a single agent targeting Cul3 neddylation simultaneously promotes insulin sensitization and insulin secretion to attenuate hyperglycemia in mice. Article Highlightsa. Pan cullin neddylation inhibitors exhibit potent hypoglycemic effect. b. The target organs and mechanisms underlying the hypoglycemia effect of cullin pan neddylation inhibitors are incompletely understood. c. We found that inhibition of Cul3 leads to a dual insulin sensitization and insulinotropic effect. d. Selective inhibition of Cul3 neddylation is a feasible approach to lower hyperglycemia.

physiology↗

bioRxiv2026

Single-cell spatially resolved transcriptomic characterization of the developing mouse cochlea

The cochlea, the sensory organ of hearing, functions as a frequency analyzer, analogous to a musical instrument. During development, while the medio-lateral axis supports differentiation of sensory cells and their surrounding supporting cells, the longitudinal axis underlies frequency-dependent properties of the cochlea. The combination of these two gene expression gradients defines unique physiological attributes of each cell intimately linked to its position within the cochlea. To determine which cochlear cell-types have a transcriptomic signature sensitive to these two gradients and identify the underlying gene regulatory networks, we took advantage of the advent of spatial single cell transcriptomics methodologies. We therefore generated a spatial transcriptomic atlas reaching single cell resolution based on the Visium HD technique, a sequencing-based technology that employ arrays of spatially barcoded probes to capture RNA molecules unbiasedly from histological tissue sections. Spatial transcriptional changes during embryonic stages, E14 and E16, as well as during postnatal development, P1 and P8, were investigated. Based on this dataset, not only cell-type assignment in single cell RNA-seq experiments could be validated, but the classification for some cell-types could be refined. Gradients of gene expression along the medio-lateral and longitudinal axes in multiple cell-types together with their temporal dynamics across development were also uncovered. Altogether, this atlas paves the way for deciphering gene regulatory networks controlling gene expression as a function of position in the cochlear cell types, providing a valuable resource for the design of efficient, robust and safe gene therapy strategies.

cell biology↗

bioRxiv2025

KCNQ1 regulates human neuronal development through mitochondrial and insulin signalling pathways

KCNQ1 encodes a voltage-gated potassium channel implicated in various peripheral and neurological disorders, yet its role during human neuronal development remains unclear. To investigate this, we generated KCNQ1 knockouts (KO) in human induced pluripotent stem cell lines and differentiated them into neural stem cells (NSCs) and cortical neurons. KCNQ1-deficient NSCs showed impaired neurite outgrowth, linked to reduced cell adhesion and disrupted neural cell adhesion molecule (NCAM) signalling. This phenotype was reproduced in wild-type and heterozygous lines by pharmacological KCNQ1 inhibition. Whole transcriptome, proteome, and follow-up analyses revealed mitochondrial dysfunction in KO NSCs, including reduced mitochondrial copy number and ATP synthase expression. Additionally, evidence was obtained for an impairment of insulin signalling in NSCs and neurons, with diminished insulin receptor gene expression and perturbation of key downstream signalling pathways (RAS-MAPK, PI3K-AKT). In neurons, KCNQ1 loss resulted in decreased synaptic activity and a more immature gene expression profile. Overall, our work reveals a novel role for KCNQ1 in human neurodevelopment by regulating cell adhesion, mitochondrial function, and insulin signalling. This work increases our understanding of KCNQ1 function in neurons and its contribution to neurological phenotypes observed in patients with KCNQ1-related diseases.

molecular biology↗

bioRxiv2025

Linking Genomic Landscape to Disease Mechanism: Core Genetic Factors Underlying Pathogenesis and Antimicrobial Resistance in Diarrheal Pathogens

BackgroundDiarrheal diseases remain a major global health burden, as they severely affect children, particularly in Bangladesh. After decades of research, the molecular mechanisms of diarrheal pathogens for disease pathogenesis and antibiotic resistance are still unknown, notably in Gram-negative bacteria. This pilot study fills the gap by employing whole genome sequencing and pan-genome analysis to analyze Bangladeshi diarrheal pathogens to identify genetic variables that cause disease pathogenesis and antibiotic resistance. ResultsHence, we investigated the genetic diversity of bacterial isolates from 31 clinical stool samples by a combination of whole-genome sequencing (WGS) and pan-genomic analysis. A core group of 50 genes, conserved across a significant number of strains, was identified via pan-genomic analysis, with considerable variation in accessory genes. This signifies a significant degree of genetic flexibility. Gene ontology analysis yielded substantial insights into prospective therapeutic targets by emphasizing the critical function of these core genes in bacterial survival and pathogenicity. Furthermore, the findings of the antimicrobial susceptibility test (AST) revealed concerning resistance trends, particularly to fluoroquinolones and beta-lactams, underscoring the necessity for enhanced surveillance and alternative therapeutic approaches. ConclusionThis study provides a comprehensive genetic framework to improve understanding of the complexity of diarrheal infections and the mechanisms underlying their resistance, fostering opportunities for potential therapeutic advancements.

microbiology↗

bioRxiv2024

Host and rabies virus gene expression is shaped by human brain cell type and reveals a preexisting pro-viral transcriptional state in astrocytes

How virus-host cell interactions and innate immune antagonism shape neurotropic infection dynamics across diverse brain cell types are largely unknown. To "unmask" and study how innate immune inhibition affects cell type-specific transcriptional regulation of the human and viral genome, we performed single-cell RNA sequencing of human brain cell co-cultures, comparing an isolate of rabies virus (RABV) to its mutant incapable of antagonizing interferon- and NF-{kappa}B-dependent responses. RABV gene expression was shaped by host cell type. RABV induced small-scale, cell-type conserved transcriptional programs that likely support infection by 1) hijacking negative transcriptional feedback of pro-viral factors while 2) reducing anti-viral RNAs. Unexpectedly, disinhibited innate immune signaling increased RABV transcription, most strikingly in an infection-independent "pro-viral" astrocyte subpopulation. Further analysis suggested that "pro-viral"-like astrocytes are a rare subtype in the human brain and are primed to protect the brain during viral infection in concert with interferon-sensitive microglia recalcitrant to infection.

neuroscience↗

bioRxiv2024

IL-23 tunes inflammatory functions of human mucosal-associated invariant T (MAIT) cells

IL-23 signaling plays a key role in the pathogenesis of chronic inflammatory and infectious diseases, yet the cellular targets and signaling pathways affected by this cytokine remain poorly understood. We show that IL-23 receptors are expressed on the large majority of human MAIT, but not of conventional T cells. Protein and transcriptional profiling at the population and single cell level demonstrates that stimulation with IL-23 or the structurally related cytokine IL-12 drives distinct functional profiles, revealing a high level of plasticity of MAIT cells. IL-23, in particular, affects key molecules and pathways related to autoimmunity and cytotoxic functions. Integrated analysis of transcriptomic and chromatin accessibility, supported by CRISPR/Cas9 mediated deletion, shows that AP-1 transcription factors constitute a key regulatory node of the IL-23 pathway in MAIT cells. In conclusion, our findings indicate that MAIT cells are key mediators of IL-23 functions in immunity to infections and chronic inflammatory diseases.

immunology↗

bioRxiv2024

Differential predictive value of resident memory CD8+T cell subpopulations in non-small-cell lung cancer patients treated by immunotherapy

A high density of resident memory T cells (TRM) in tumors correlates with improved clinical outcomes in immunotherapy-treated patients. However, in preclinical models, only some subpopulations of TRM are associated with cancer vaccine efficacy. We identified two main TRM subpopulations in tumor-infiltrating lymphocytes derived from non-small cell lung cancer (NSCLC) patients: one co-expressing CD103 and CD49a (DP), and the other expressing only CD49a (MP); both exhibiting additional TRM surface markers like CD69. DP TRM exhibited greater functionality compared to MP TRM. Analysis of T-cell receptor (TCR) repertoire and of the stemness marker TCF-1 revealed shared TCRs between populations, with the MP subset appearing more progenitor-like phenotype. In two NSCLC patient cohorts, only DP TRM predicted PD-1 blockade response. Multivariate analysis, including various biomarkers (CD8, TCF1+CD8+T cells, and PD-L1) associated with responses to anti-PD(L)1, showed that only intra-tumoral infiltration by DP TRM remained significant. This study highlights the non-equivalence of TRM populations and emphasizes the importance of distinguishing between them to better define their role in antitumor immunity and as a biomarker of response to immunotherapy.

immunology↗

bioRxiv2023

Dysregulation of chromatin via H3K27 methylation underpins differentiation arrest in Isocitrate dehydrogenase-mutant Acute Myeloid Leukaemia.

Dysregulation of cellular differentiation is a hallmark of cancer. Isocitrate dehydrogenase (IDH) is commonly mutated multiple cancers including glioma, cholangiocarcinoma, lymphoma and Acute Myeloid Leukaemia (AML). Mutant IDH generates d-2-hydroxyglutarate that inhibits enzymes including Jumonji histone demethylases and TET2. Using primary human IDH2-mutant AML cells as a model, single cell RNA-seq and ATAC-seq, we demonstrated the continuum of cell states during restoration of neutrophilic differentiation to leukaemic progenitors by Enasidenib, a mutant IDH2 inhibitor. In cells which ultimately differentiate, there is co-expression of competing GATA2/RUNX3/SOX4-driven stem-progenitor and pro-differentiation EGR1/JUN/FOS programmes, followed by expression of cell cycle and terminal neutrophil programmes involving CEBP family and SPI1/PU.1. Genes upregulated during differentiation display loss of H3K27me3 in bivalent chromatin but not of H3K4me3, while downregulated genes are enriched for PRC2/EZH2 targets. In contrast to previous reports of a TET2-dependent mechanism for IDH-mutations, we observed only a modest link between promoter DNA CpG methylation and gene expression. For the first time in primary AML, we describe the lifting of differentiation block by de-repression of pro-differentiation genes through modulation of H3K27 demethylation in bivalent chromatin, and thus highlight a novel and important mechanism in how IDH mutations disrupt cell fates in cancer.

cancer biology↗

bioRxiv2023

Beyond 40 fluorescent probes for deep phenotyping of blood mononuclear cells, using spectral technology.

The analytical capability of flow cytometry is crucial for differentiating the growing number of cell subsets found in human blood. This is important for accurate immunophenotyping of patients with few cells and a large number of parameters to monitor. Here, we present a 43-parameter panel to analyze peripheral blood mononuclear cells from healthy individuals using 41 fluorescence-labelled monoclonal antibodies, an autofluorescent channel, and a viability dye. We demonstrate minimal population distortions that lead to optimized population identification and reproducible results. We have applied an advanced approach in panel design, in selection of sample acquisition parameters and in data analysis. Appropriate autofluorescence identification and integration in the unmixing matrix, allowed for resolution of unspecific signals and increased dimensionality. Addition of one laser without assigned fluorochrome resulted in decreased fluorescence spill over and improved discrimination of cell subsets. It also increased staining index when autofluorescence was integrated in the matrix. We conclude that spectral flow cytometry is highly valuable tool for high-end immunophenotyping, and that fine-tuning of major experimental steps is key for taking advantage of its full capacity.

cell biology↗

bioRxiv2023

High-dimensional spectral cytometry panels for whole blood immune phenotyping

The need to understand the mechanisms and pathways of immune responses in pathogenic conditions such as cancer and autoimmunity requires awareness of natural immune variability in healthy subjects. To this end, various systems immunology studies have been established. Among them, the Milieu Interieur (MI) study was established to define the boundaries of a healthy immune response and identify determinants of immune response variation. MI used immunophenotyping of a 1000 healthy donor cohort by flow cytometry as a principal outcome for immune variance at steady state. For the 10-year longitudinal MI study, we have developed two high-dimensional spectral flow cytometry panels that allow deep characterization of innate and adaptive whole blood immune cells (35 and 34 fluorescent markers, respectively) and standardized the protocol for sample handling, staining, acquisition, and data analysis. This permits the reproducible quantification of over 182 immune cell phenotypes through robust immunophenotyping at a single site. This highly standardized protocol was applied to samples from patients with autoimmune/inflammatory diseases. It is currently used for characterization of the impact of age and environmental factors on peripheral blood immune phenotypes of >400 donors from the initial MI cohort.

immunology↗

bioRxiv2023

UBL3 Interacts with Alpha-synuclein in Cells and the Interaction is Downregulated by the EGFR Pathway Inhibitor Osimertinib

Ubiquitin-like 3 (UBL3) acts as a post-translational modification (PTM) factor and regulates protein sorting into small extracellular vesicles (sEVs). sEVs have been reported as vectors for the pathology propagation of neurodegenerative diseases, such as -synucleinopathies. Alpha-synuclein (-syn) has been widely studied for its involvement in -synucleinopathies. However, it is still unknown whether UBL3 interacts with -syn, and is influenced by drugs or compounds. In this study, we investigated the interaction between UBL3 and -syn, and any ensuing possible functional and pathological implications. We found that UBL3 can interact with -syn by the Gaussia princeps based split luciferase complementation assay in cells and immunoprecipitation, while cysteine residues at its C-terminal, which are considered important as PTM factors for UBL3, were not essential for the interaction. The interaction was upregulated by 1-methyl-4-phenylpyridinium exposure. In drug screen results, the interaction was significantly downregulated by the treatment of osimertinib. These results suggest that UBL3 interacts with -syn in cells and be significantly downregulated by epidermal growth factor receptor (EGFR) pathway inhibitor osimertinib. Therefore, the UBL3 pathway may be a new therapeutic target for -synucleinopathies in the future.

molecular biology↗