Biology subjects

Guergues, J.

Publications and source records attributed to Guergues, J..

4 recordsLinked to original sources

bioRxiv2026

Dendrimer Delivered shRNA Targeting the CCL20-CCR6 Axis Suppresses Complement-Mediated Microglial Synaptic Pruning and Ameliorates Chronic Neuroinflammation After Repetitive Traumatic Brain Injury

Repetitive traumatic brain injury (rTBI) induces persistent microglial activation and chronic neuroinflammation, yet the upstream signals driving long-term synaptic injury remain unclear. In this study, we identify the CCL20-CCR6 chemokine axis as a critical regulator of sustained microglial activation and complement-dependent synaptic loss after rTBI. Proteomic profiling at 30 days post-injury (dpi) showed broad normalization of complement-linked inflammatory and synaptic pathways in the cortex and hippocampus, underscoring a mechanistic link between chemokine signaling, microglial activation, and synaptic vulnerability. To therapeutically target this axis, we developed a dendrimer-based shRNA platform (shCombo-DPX) that simultaneously silences CCL20 and CCR6. Intranasal and intravenous delivery in rTBI mice effectively reduced CCL20-CCR6 expression, attenuated chronic microgliosis and astrogliosis, and suppressed complement activation. Treatment limited microglial synaptic engulfment, preserved synaptic proteins, restored BDNF levels, and improved motor, anxiety-related, and cognitive outcomes. In microglia-neuron coculture systems, CCL20 silencing reduced LPS-induced complement signaling and prevented synaptic loss, neuronal apoptosis, and BDNF depletion. Conversely, exposure to recombinant CCL20 induced dendritic degeneration, caspase-3 activation, microglial reactivity, complement dysregulation, and synaptic injury both in vitro and in vivo. Collectively, these findings establish CCL20-CCR6 as a key upstream driver of chronic complement-mediated synaptic degeneration after rTBI and support dendrimer-delivered shRNA therapy as a targeted strategy to mitigate long-term neurodegeneration.

neuroscience↗

bioRxiv2026

CCL20-CCR6 Signaling as a Prognostic Biomarker and Therapeutic Target in Temozolomide-Resistant Glioblastoma

Glioblastoma remains highly lethal, with median survival of ~15 months. Resistance to temozolomide is ubiquitous, yet its mechanisms are incompletely understood. Here, we identify the CCL20-CCR6 chemokine axis as a stress-responsive survival pathway limiting therapeutic efficacy. Targeting CCL20-CCR6 in combination with temozolomide and cannabidiol was evaluated using clinical datasets, GBM cell lines, tumor organoids, and a syngeneic CT-2A mouse model integrating proteomic and lipidomic profiling. Low CCL20 expression was associated with improved survival, supporting its prognostic relevance. Across models, TMZ alone or with CBD induced CCL20 expression while exerting limited antitumor activity. Targeted disruption of CCL20-CCR6 signaling using dendrimer-delivered shRNA enhanced therapeutic response in murine models and GBM organoids. Multi-omic analyses revealed that CCL20 inhibition reprograms the tumor microenvironment and induces mitochondrial dysfunction, resulting in elevated reactive oxygen species (ROS) and tumor cell death. This effect was accompanied by accumulation of 17-hydroxydocosahexaenoic acid and activation of oxidative stress-associated cytotoxic pathways. Functional assays confirmed that CCL20 blockade selectively amplifies mitochondrial ROS beyond levels induced by TMZ alone potentiating TMZ efficacy by promoting mitochondrial oxidative stress. Targeting this axis represents a promising strategy to overcome chemoresistance and positions CCL20 as both a prognostic biomarker and a therapeutic vulnerability in GBM.

cancer biology↗

bioRxiv2025

Proteomic characterization of the Alzheimer's disease risk factor BIN1 interactome

The gene BIN1 is the second-largest genetic risk factor for late-onset Alzheimer's disease (LOAD). It is expressed in neurons and glia in the brain as cell-type specific and ubiquitous isoforms. BIN1 is an adaptor protein that regulates membrane dynamics in many cell types. Previously, we reported that BIN1 predominantly localizes to presynaptic terminals in neurons and regulates presynaptic vesicular release. However, the function of neuronal BIN1 in relation to LOAD is not yet fully understood. A significant gap in the field is the unbiased characterization of neuronal BIN1-interacting proteins and proximal neighbors. To address this gap and help define the functions of neuronal BIN1 in the brain, we employed TurboID-based proximity labeling to identify proteins biotinylated by the neuronal BIN1 isoform 1-TurboID fusion protein (BIN1iso1-TID) in cultured mouse neuroblastoma (N2a) cells in vitro and in adult mouse brain neurons in vivo. Label-free quantification-based proteomic analysis of the BIN1iso1-TID biotinylated proteins led to the discovery of 361 proteins in N2a cells and 897 proteins in mouse brain neurons, identified as BIN1iso1-associated (proximal) or interacting proteins. A total of 92 proteins were common in both datasets, indicating that these are high-confidence BIN1-interacting or proximity proteins. SynapticGO analysis of the mouse brain dataset revealed that BIN1iso1-TurboID labeled 159 synaptic proteins, with 60 corresponding to the synaptic vesicle cycle. Based on phosphorylation site analysis of the neuronal BIN1iso1-TID interactome and related kinase prediction, we selected AAK1, CDK16, SYNJ1, PP2BA, and RANG for validation through immunostaining and proximity ligation assays as members of the BIN1 interactome in the mouse brain. By identifying several previously unknown proximal and potential interacting proteins of BIN1, this study establishes a foundation for further investigations into the function of neuronal BIN1.

neuroscience↗

bioRxiv2024

A Simplified Method for Comprehensive Capture of the Staphylococcus aureus Proteome

Staphylococcus aureus is a major human pathogen causing myriad infections in both community and healthcare settings. Although well studied, a comprehensive exploration of its dynamic and adaptive proteome is still somewhat lacking. Herein, we employed streamlined liquid- and gas-phase fractionation with PASEF analysis on a TIMS-TOF instrument to expand coverage and explore the S. aureus dark proteome. In so doing, we captured the most comprehensive S. aureus proteome to date, totaling 2,231 proteins (85.6% coverage), using a significantly simplified process that demonstrated high reproducibility with minimal input material. We then showcase application of this library for differential expression profiling by investigating temporal dynamics of the S. aureus proteome. This revealed alterations in metabolic processes, ATP production, RNA processing, and stress-response proteins as cultures progressed to stationary growth. Notably, a significant portion of the library (94%) and proteome (80.5%) was identified by this single-shot, DIA-based analysis. Overall, our study shines new light on the hidden S. aureus proteome, generating a valuable new resource to facilitate further study of this dangerous pathogen.

microbiology↗