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Klonjkowski, B.

Publications and source records attributed to Klonjkowski, B..

5 recordsLinked to original sources

Microglial Foxo3 shapes dopaminergic vulnerability in Parkinson disease

Neurodegenerative diseases such as Parkinsons disease (PD) result from complex interactions between neuronal stress and the surrounding tissue environment, yet the determinants that govern this interplay remain incompletely understood. While neuronal responses to mitochondrial dysfunction and proteotoxic stress have been extensively characterized, the contribution of the microglial state to disease progression remains unclear. Here, we identify the transcription factor Foxo3 as a key regulator of dopaminergic vulnerability acting predominantly through microglia-, rather than neuron-intrinsic, mechanisms. Foxo3 was rapidly induced and translocated to the nucleus in dopaminergic-like cells in response to mitochondrial complex I inhibition and -synuclein aggregation, indicating activation of a conserved neuronal stress response. However, neuron-specific deletion of Foxo3 attenuated early Parkinson-like transcriptional signatures but did not confer sustained in vivo neuroprotection. In contrast, microglia-specific deletion of Foxo3 provided robust and durable protection against dopaminergic degeneration across complementary mouse models, including both MPTP intoxication and -synuclein-driven pathology. Translatomic profiling revealed that Foxo3 deficiency induces extensive transcriptional remodeling of microglia at baseline, establishing a distinct state enriched in immune-related and phagocytic pathways that remains compatible with tissue homeostasis. Strikingly, neuroprotection occurred despite minimal transcriptional reprogramming following neurotoxic insult, suggesting that disease outcome is dictated by the baseline microglial state rather than by the magnitude of transcriptional responses to neurotoxic insult. Mechanistically, neuroprotection correlated with selective modulation of discrete signaling nodes, including reinforcement of the TREM2-TYROBP axis and attenuation of pathways involved in intracellular signal amplification, rather than with broad suppression of inflammatory programs. These findings indicate that Foxo3 does not primarily regulate the magnitude of microglial activation, but instead defines the baseline configuration and response thresholds that shape subsequent neuroimmune dynamics. Together, our results identify baseline microglial state as a determinant of neurodegenerative trajectory and position Foxo3 as a central regulator of this process. More broadly, these findings provide a framework in which microglial state-setting, rather than the magnitude of reactive responses alone, governs dopaminergic vulnerability, with potential implications for therapeutic strategies targeting neuroimmune interactions in Parkinsons disease.

neuroscience↗

SARS-CoV-2 Delta variant induces severe damages in the nasal cavity from the first day post-infection in the Syrian hamster model.

SARS-CoV-2 replication initiates in the nasal cavity and can spread to the lower respiratory tract. However, the early physiopathological events that occur in the nasal cavity after infection remain poorly understood. In this study, we investigated the initial steps of viral infection from 1 day post-infection (dpi) in Syrian hamsters infected with SARS-CoV-2 D614G, Delta and Omicron (BA.1) variants and compared them with animals sacrificed at 4 dpi. While the level of viral replication in the nasal turbinates of the three groups of hamsters was equivalent at 4dpi, the amount of viral RNA at 1dpi was higher in D614G- and Delta-infected animals than in the Omicron group. No difference in viral RNA levels or inflammatory markers in the nasal turbinates was observed between D614G- and Delta-infected animals, except for a significantly higher level of IFN-{lambda} in the Delta group at 1dpi. Additionally, histological analysis revealed a more rapid diffusion of the Delta virus reaching the posterior zone of the nasal cavity at 1dpi inducing significant damage to the olfactory epithelium. At the same time, the D614G and Omicron infections were essentially restricted to the anterior part of the nasal cavity with less damage observed. Consistently, viral replication was already effective in the lungs of all Delta- infected hamsters at 1 dpi, but only in two of the six D614G animals. Our results highlight the importance of studying viral infection in the nasal cavity very early after infection with a spatial approach to better understand the physiopathology of the different SARS-CoV-2 variants.

microbiology↗

Biosynthetic proteins targeting the SARS-CoV-2 spike as anti-virals

The binding of the SARS-CoV-2 spike to angiotensin-converting enzyme 2 (ACE2) promotes virus entry into the cell. Targeting this interaction represents a promising strategy to generate antivirals. By screening a phage-display library of biosynthetic protein sequences build on a rigid alpha-helicoidal HEAT-like scaffold (named Reps), we selected candidates recognizing the spike receptor binding domain (RBD). Two of them (F9 and C2) bind the RBD with affinities in the nM range, displaying neutralisation activity in vitro and recognizing distinct sites, F9 overlapping the ACE2 binding motif. The F9-C2 fusion protein and a trivalent Rep form (C2-foldon) display 0.1 nM affinities and EC50 of 8-18 nM for neutralization of SARS-CoV-2. In hamsters, F9-C2 instillation in the nasal cavity before or during infections effectively reduced the replication of a SARS-CoV-2 strain harbouring the D614G mutation in the nasal epithelium. Furthermore, F9-C2 and/or C2-foldon effectively neutralized SARS-CoV-2 variants (including delta and omicron variants) with EC50 values ranging from 13 to 32 nM. With their high stability and their high potency against SARS-CoV-2 variants, Reps provide a promising tool for SARS-CoV-2 therapeutics to target the nasal cavity and mitigate virus dissemination in the proximal environment. Author SummaryThe entry of SARS-CoV-2 in permissive cells is mediated by the binding of its spike to angiotensin-converting enzyme 2 (ACE2) on the cell surface. To select ligands able to block this interaction, we screened a library of phages encoding artificial proteins (named Reps) for binding to its receptor binding domain (RBD). Two of them were able to bind the RBD with high affinity and block efficiently the virus entry in cultured cells. Assembled Reps through covalent or non-covalent linkages blocked virus entry at lower concentration than their precursors (with around 20-fold activity increase for a trimeric Rep). These Reps derivates neutralize efficiently SARS-CoV-2 {beta}, {gamma}, {delta} and Omicron virus variants. Instillation of an Rep dimer in the nasal cavity effectively reduced virus replication in the hamster model of SARS-CoV-2 and pathogenicity.

microbiology↗

Neutrophils initiate the destruction of the olfactory epithelium during SARS-CoV-2 infection in hamsters

The loss of smell related to SARS-CoV-2 infection is one of the most prevalent symptoms of COVID-19. It is now clear that this symptom is related to the massive infection by SARS-CoV-2 of the olfactory epithelium leading to its desquamation. However, the molecular mechanism behind the destabilization of the olfactory epithelium is less clear. Using golden Syrian hamster, we show here that while apoptosis remains at a low level in damaged infected epithelium, the latter is invaded by innate immunity cells. By depleting the neutrophil population or blocking the activity of neutrophil elastase-like proteinases, we reduced the damage induced by the SARS-CoV-2 infection. Surprisingly, the impairment of neutrophil activity led to a decrease of SARS-CoV-2 infection levels in the nasal cavity. Our results indicate a counterproductive role of neutrophils leading to the release of infected cells in the lumen of the nasal cavity and thereby enhanced spreading of the virus.

immunology↗

Melatonin drugs inhibit SARS-CoV-2 entry into the brain and virus-induced damage of cerebral small vessels

COVID-19 is a complex disease with short- and long-term respiratory, inflammatory and neurological symptoms that are triggered by the infection with SARS-CoV-2. Invasion of the brain by SARS-CoV-2 has been observed in humans and is postulated to be involved in post COVID condition. Brain infection is particularly pronounced in the K18-hACE2 mouse model of COVID-19. Here, we show that treatment of K18-hACE2 mice with melatonin and two melatonin-derived marketed drugs, agomelatine and ramelteon, prevent SARS-CoV-2 entry in the brain thereby reducing virus-induced damage of small cerebral vessels, immune cell infiltration and brain inflammation. Brain entry of SARS-CoV-2 through endothelial cells is prevented by melatonin through allosteric binding to human angiotensin-converting enzyme 2 (ACE2), which interferes with the cell entry receptor function of ACE2 for SARS-CoV-2. Our findings open new perspectives for the repurposing of melatonergic drugs in the prevention of brain infection by SARS-CoV-2 and COVID-19-related long-term neurological symptoms.

pharmacology and toxicology↗