Biology subjects

Robert, G.

Publications and source records attributed to Robert, G..

11 recordsLinked to original sources

bioRxiv2025

Non-canonical caspase-8 activation by cathepsin B drives anti-inflammatory human macrophage polarization

Anti-inflammatory monocyte-derived macrophages are essential to maintain tissue homeostasis but can also contribute to disease progression, notably in cancer and fibrosis. Deciphering the signaling pathways that govern their generation could therefore unlock new therapeutic opportunities. Here we uncover a previously unrecognized, non-apoptotic function of caspase-8 in driving both monocyte-to-macrophage differentiation and anti-inflammatory macrophages polarization. We identified cathepsin B as a novel upstream activator of caspase-8 activation through a non-canonical cleavage mechanism, conferring to caspase-8 an original activity profile distinct from its apoptotic role. Disruption of this cathepsin-B-caspase-8 axis, either genetically or pharmacologically, not only impairs the generation of anti-inflammatory macrophages but also reprograms these cells towards a pro-inflammatory phenotype. Our findings position the cathepsin-B-caspase-8 axis as a critical regulatory node in macrophage fate decisions and a promising target for therapeutic reprogramming of human macrophages in cancer, inflammation and fibrotic diseases.

immunology↗

bioRxiv2025

Transdiagnostic electrophysiological subtypes reveal brain-behavior dimensions in youth psychiatry

Conventional psychiatric diagnoses often fail to reflect the underlying neurobiological and behavioral complexity of mental health conditions. Here, we propose a transdiagnostic, data-driven framework for stratifying youth based on large-scale multisite electroencephalography (EEG) data from 1,707 individuals aged 5-18 years, including healthy controls and individuals diagnosed with attention-deficit/hyperactivity disorder (ADHD), autism spectrum disorder (ASD), anxiety disorders (ANX), and learning disabilities (LD), along with their common comorbidities. By applying normative modeling to quantify individual deviations from typical brain functional maturation, and integrating multidimensional EEG features across spectral, temporal, complexity, and dynamical domains via similarity network fusion clustering, we identified three robust neurophysiological biotypes. These biotypes showed distinct electrophysiological and behavioral profiles, and captured meaningful brain-behavior relationships. Our findings suggest that biologically informed subtypes capture meaningful neuropsychiatric heterogeneity in youth, challenging conventional diagnostic boundaries in psychiatric nosology.

neuroscience↗

bioRxiv2024

Dual targeting of GPX4 and TXNRD1 triggers eradication of AML cells through induction of apoptosis and ferroptosis

MyeloDysplastic Syndromes (MDS) are a group of heterogeneous hematological disorders characterized by bone marrow failure and abnormal hematopoietic cell expansion, often progressing to acute myeloid leukemia (AML). Current treatments for AML and high-risk MDS have limited efficacy, requiring the exploration of new therapeutic approaches. Recent research highlighted the potential of inducing cell death through ferroptosis, either independently or alongside traditional chemotherapy, as promising approaches for treating MDS/AML cells. We described here two novel compounds, HA344 and #231, capable of targeting both ferroptosis and apoptosis, leading to the effective eradication of cell lines and primary blasts from MDS/AML patients, while sparing normal hematopoietic cells. RNASeq analysis identified oxidation reduction and apoptotic processes as highly significant induced pathways in two different AML cell lines. Using click-chemistry approaches coupled to mass spectrometry, we identified glutathione peroxidase 4 (GPX4) and thioredoxin reductase 1 (TXNRD1) as the main targets of HA344 and #231 in a large panel of AML cells. Accordingly, both compounds inhibited GPX4 and TXNRD1 activity in the micromolar range and triggered GPX4 degradation. Moreover, using recombinant GPX4 carrying or not a selenium (GPX4-Se and GPX4-S), we confirmed by mass spectrometry that HA344 and #231 bind more efficiently GPX4-Se than GPX4-S. In conclusion, these compounds might represent a new pharmacological approach in the treatment of MDS and AML, offering a potential avenue for future therapies.

biochemistry↗

bioRxiv2024

Beyond homogeneity: Charting the landscape of heterogeneity in psychiatric electroencephalography

Electroencephalography (EEG) has been thoroughly studied for decades in psychiatry research. Yet its integration into clinical practice as a diagnostic/prognostic tool remains unachieved. We hypothesize that a key reason is the underlying patients heterogeneity, overlooked in psychiatric EEG research relying on a case-control approach. We combine HD-EEG with normative modeling to quantify this heterogeneity using two well-established and extensively investigated EEG characteristics -spectral power and functional connectivity-across a cohort of 1674 patients with attention-deficit/hyperactivity disorder, autism spectrum disorder, learning disorder, or anxiety, and 560 matched controls. Normative models showed that deviations from population norms among patients were highly heterogeneous and frequency-dependent. Deviation spatial overlap across patients did not exceed 40% and 24% for spectral and connectivity, respectively. Considering individual deviations in patients has significantly enhanced comparative analysis, and the identification of patient-specific markers has demonstrated a correlation with clinical assessments, representing a crucial step towards attaining precision psychiatry through EEG.

neuroscience↗

bioRxiv2023

HIF-1 inactivation empowers HIF-2 to drive hypoxia adaptation in aggressive forms of medulloblastoma

Medulloblastoma (MB) is the most prevalent brain cancer in children. Four subgroups of MB have been identified; of these, Group 3 is the most metastatic. Its genetics and biology remain less clear than the other groups, and it has a poor prognosis and few effective treatments available. Tumor hypoxia and the resulting metabolism are known to be important in the growth and survival of tumors but, to date, have been only minimally explored in MB. Here we show that Group 3 MB tumors do not depend on the canonical transcription factor hypoxia-inducible factor-1 (HIF-1) to mount an adaptive response to hypoxia. We discovered that HIF-1 is rendered inactive either through post-translational methylation, preventing its nuclear localization specifically in Group 3 MB, or by a low expression that prevents modulation of HIF-target genes. Strikingly, we found that HIF-2 takes over the role of HIF-1 in the nucleus and promotes the activation of hypoxia-dependent anabolic pathways. The exclusion of HIF-1 from the nucleus in Group 3 MB cells enhances the reliance on HIF-2s transcriptional role, making it a viable target for potential anticancer strategies. By combining pharmacological inhibition of HIF-2 with the use of metformin, a mitochondrial complex I inhibitor to block respiration, we effectively induced Group 3 MB cell death, surpassing the effectiveness observed in Non-Group 3 MB cells. Overall, the unique dependence of MB cells, but not normal cells, on HIF-2-mediated anabolic metabolism presents an appealing therapeutic opportunity for treating Group 3 MB patients with minimal toxicity.

cancer biology↗

bioRxiv2023

DIFFERENTIAL EFFECTS OF REACTIVE OXYGEN SPECIES ON AUTOPHAGY IN ARABIDOPSIS

Autophagy plays a key role in the responses to different stress condition in plants. Reactive oxygen species (ROS) are common modulators of stress responses, having both toxic and signaling functions. In this context, the relationship between ROS and autophagy regulation remains unclear, and in some aspects, contradictory. In this study, we employed pharmacological and genetic approaches to investigate the effects of different ROS on the cytoplastic redox state and autophagic flux in Arabidopsis thaliana. Ours results demonstrated that oxidative treatments with H2O2 and MV, which drastically increased the oxidized state of the cytoplasm, reduced the autophagic flux. Conversely, singlet oxygen, which did not have significant effects on the cytoplasmic redox state, increased the autophagic flux. Additionally, our findings indicated that after H2O2 and high light treatments and during the recovery period, the cytoplasm returned to its reduced state, while autophagy was markedly induced. In summary, our study unveils the differential effects of ROS on the autophagic flux, establishing a correlation with the redox state of the cytoplasm. Moreover, it emphasizes the dynamic nature of autophagy in response to oxidative stress and the subsequent recovery period. HIGHLIGHTSO_LIThis research shows the differential effects of reactive oxygen species on autophagic modulation, highlighting their impact on the cytoplasmic redox state. The relationship between ROS and autophagy regulation remains unclear, and in some aspects contradictory. Here, we present a comprehensive investigation characterizing the effects of different ROS, such as hydrogen peroxide and singlet oxygen, on the modulation of autophagy in Arabidopsis. In brief, our findings reveal differential impacts on cytoplasmic redox states and autophagic flux, providing insight into the dynamic nature of autophagy, especially in stress and post-stress conditions. C_LIO_LITo the best of our knowledge, our work is the first to evaluate autophagic flux both during and after oxidative stress in plants. Our results indicate that this differentiation is crucial when analyzing the effects of oxidative stress on autophagy. C_LI

cell biology↗

bioRxiv2023

MCB-613 exploits a collateral sensitivity in drug-resistant EGFR-mutant non-small cell lung cancer through covalent inhibition of KEAP1

Targeted therapies have revolutionized cancer chemotherapy. Unfortunately, most patients develop multifocal resistance to these drugs within a matter of months. Here, we used a high-throughput phenotypic small molecule screen to identify MCB-613 as a compound that selectively targets EGFR-mutant, EGFR inhibitor-resistant non-small cell lung cancer (NSCLC) cells harboring diverse resistance mechanisms. Subsequent proteomic and functional genomic screens involving MCB-613 identified its target in this context to be KEAP1, revealing that this gene is selectively essential in the setting of EGFR inhibitor resistance. In-depth molecular characterization demonstrated that (1) MCB-613 binds KEAP1 covalently; (2) a single molecule of MCB-613 is capable of bridging two KEAP1 monomers together; and, (3) this modification interferes with the degradation of canonical KEAP1 substrates such as NRF2. Surprisingly, NRF2 knockout sensitizes cells to MCB-613, suggesting that the drug functions through modulation of an alternative KEAP1 substrate. Together, these findings advance MCB-613 as a new tool for exploiting the selective essentiality of KEAP1 in drug-resistant, EGFR-mutant NSCLC cells.

cancer biology↗

bioRxiv2022

HSPA8 chaperone complex drives chaperone-mediated autophagy regulation in acute promyelocytic leukemia differentiation

Acute myeloid leukemia (AML) is a cancer of the hematopoietic system characterized by the hyperproliferation of undifferentiated cells of the myeloid lineage. While most of AML therapy are focusing on tumor debulking, all-trans retinoic acid (ATRA) induces differentiation in acute promeylocytic leukemia (APL) a particular subtype. Macroautophagy has been extensively investigated in the context of various cancers and is often dysregulated in AML where it can have context-dependent pro- or anti-leukemogenic effects. On the contrary, the implications of chaperone-mediated autophagy (CMA) on the pathophysiology of diseases are still being explored and its role in AML has remained elusive. To answer our questions we took advantages of human AML primary samples and databases. Furthermore, we used ATRA-sensitive (NB4) and -resistant (NB4-R1) cells to further dissect a potential function for CMA in ATRA-mediated neutrophil differentiation. NB4-R1 cells are unique in that they do respond to retinoic acid transcriptionally, but do not mature in response to retinoid signaling alone unless maturation is triggered by adding cAMP. Here, we report that CMA related mRNA transcripts are higher expressed in immature hematopoietic cells as compared to neutrophils. In line, lysosomal degradation of mCherry-KFERQ CMA reporter decreases during ATRA-induced differentiation of APL cells. On the other hand, using NB4-R1 cells we found that macroautophagy flux primed the ATRA resistant NB4-R1 cells to differentiate upon ATRA treatment, but reduced association of LAMP-2A and HSPA8 is necessary for complete neutrophil maturation. Accordingly, depletion of HSPA8 attenuated CMA activity and facilitated APL differentiation. In contrast, maintaining high CMA activity by ectopic expression of LAMP-2A impeded APL differentiation. Overall, our findings demonstrate that both normal and APL neutrophil differentiation require CMA downregulation and this pathway is predominantly dependent on HSPA8 assisted by other co-chaperones.

cancer biology↗

bioRxiv2022

Autophagy modulates apical growth and development in the moss Physcomitrium patens

Different to root hairs and pollen tubes, Physcomitrium patens apical growing protonemal cells have the singularity that they continue to undergo cell divisions as the plant develops, allowing to study autophagy in the context of a multicellular apical growing tissue coupled to development. Herein, we showed that the core autophagy machinery is present in the moss P. patens, and deeply characterized the growth and development of wild-type, atg5 and atg7 loss-of-function mutants under optimal and nutrient-deprived conditions. Our results showed that the growth of the different morphological and functional protonemata apical growing cells, chloronema and caulonema, is differentially modulated by this process. These differences depend on the protonema cell type and position along the protonemal filament, and growth condition. As a global plant response, the absence of autophagy triggers the spread of the colony through protonemata growth at the expense of a reduction in buds and gametophore development, and thus the adult gametophytic and reproductive phases. Altogether this study provides valuable information indicating that autophagy has roles during apical growth with differential responses within the cell types of the same tissue and contributes to life cycle progression and thus the development of the 2D and 3D tissues of P. patens. HIGHLIGHTAutophagy is differentially induced in protonemal cells, and contributes to apical growth, life cycle progression, and thus the development of the 2D and 3D tissues of P. patens.

plant biology↗

bioRxiv2020

Low-frequency subthalamic neural oscillations are involved in explicit and implicit facial emotional processing - a local field potential study

In addition to the subthalamic nucleus (STN) role in motor control, STN deep brain stimulation (DBS) for Parkinsons disease (PD) has also uncovered its involvement in cognitive and limbic processing. STN neural oscillations analyzed through local field potential (LFP) recordings have been shown to contribute to emotional (mostly in the alpha band [8-12 Hz]) and cognitive processing (theta [4-7 Hz] and beta [13-30 Hz] bands). In this study, we aimed at testing the hypothesis that STN oscillatory activity is involved in explicit and implicit processing of emotions. To achieve this objective, we used a task that presented patients with fearful emotional facial expressions and asked them to identify the emotion (explicit task) or gender associated with the face (implicit task). We evaluated emotion and task effects on STN neural oscillations power and intertrial phase consistency. Our results revealed that accuracy was lower in the implicit task. Increased STN delta power and decreased alpha and beta power were observed after stimulus presentation. However, there was no influence of emotional facial expression, i.e. neutral versus fear, nor task demands. Intertrial phase consistency in the delta and theta band increased after stimulus onset, in the same time-period as delta power increased. However, similarly to oscillatory power, no changes related to emotional fear expression or task demand were found. These findings suggest that STN oscillatory activity is not specifically involved in explicit and/or implicit processing of emotions, and that power and phase synchronization changes might be more related to overall task-execution mechanisms. These conjectures remain to be confirmed. Highlights- STN LFPs were recorded during an emotional/gender recognition task in PD patients. - STN delta power increased, and alpha and beta power decreased after stimulus onset. - Power changes were not influenced by emotional fearful expression or task demands. - Delta/theta intertrial phase consistency increased after stimulus onset. - Intertrial phase consistency was not affected by emotional valence or task demands. - The observed STN activity was likely related to general task-execution mechanisms.

neuroscience↗

bioRxiv2020

Bilateral but not unilateral subthalamic stimulation promotes apathy: a translational study in rodents and Parkinson's disease patients

Apathy, depression, and anxiety represent the main neuropsychiatric symptoms of Parkinsons disease (PD). How subthalamic nucleus deep brain stimulation (STN-DBS) influences these symptoms, especially apathy, is a controversial topic. The present translational study investigates and compares the effect of bilateral or unilateral STN-DBS on this neuropsychiatric triad, combining a pre-clinical approach in rodents and a clinical follow-up of patients with PD. While depression and anxiety related behaviors remain unchanged, bilateral but not unilateral STN-DBS consistently induces apathy in patients and a reward seeking deficit in rodents. Together, these data substantiate the claim that STN-DBS may induce apathy by itself and suggest that bilateral but not the unilateral stimulation might be a critical factor.

neuroscience↗