AXL kinase-mediated astrocytic phagocytosis modulates outcomes of traumatic brain injury

Abstract

Background Complex changes in the brain microenvironment following traumatic brain injury (TBI) can cause neurological impairments for which there are few efficacious therapeutic interventions. The reactivity of astrocytes is one of the keys to microenvironmental changes, such as neuroinflammation, but its role and the molecular mechanisms that underpin it remain unclear. Methods Male C57BL/6J mice were subjected to the controlled cortical impact (CCI) to develop a TBI model. The specific ligand of AXL receptor tyrosine kinase (AXL), recombinant mouse growth arrest-specific 6 (rmGas6) was intracerebroventricularly administered, and selective AXL antagonist R428 was intraperitoneally applied at 30 min post-modeling separately. Post-TBI assessments included neurobehavioral assessments, transmission electron microscopy, immunohistochemistry, and western blotting. Real-time polymerase chain reaction (RT-PCR), siRNA transfection, and flow cytometry were performed for mechanism assessments in primary cultured astrocytes. Results AXL is upregulated mainly in astrocytes after TBI and promotes astrocytes switching to a phenotype that exhibits the capability of ingesting degenerated neurons or debris. As a result, this astrocytic transformation promotes the limitation of neuroinflammation and recovery of neurological dysfunction. Pharmacological inhibition of AXL in astrocytes significantly decreased astrocytic phagocytosis both in vivo and in primary astrocyte cultures, in contrast to the effect of treatment with the rmGas6. AXL activates the signal transducer and activator of the transcription 1 (STAT1) pathway thereby further upregulating ATP-binding cassette transporter 1 (ABCA1). Moreover, the supernatant from GAS6-depleted BV2 cells induced limited enhancement of astrocytic phagocytosis in vitro. Conclusion Our work establishes the role of AXL in the transformation of astrocytes to a phagocytic phenotype via the AXL/STAT1/ABCA1 pathway which contributes to the separation of healthy brain tissue from injury-induced cell debris, further ameliorating neuroinflammation and neurological impairments after TBI. Collectively, our findings provide a potential therapeutic target for TBI.

Recombinant growth arrest-specific protein 6 protects the blood–brain barrier by regulating microglia polarization via the GAS6/Axl/SOCS pathway in post-stroke hemorrhagic transformation model

“…Our study had some limitations. First, it is important to acknowledge that the GAS6/TAM system exerts multiple actions in the CNS [21,53,54]. As previously reported, GAS6 e ciently recognized phosphatidylserine on the surface of apoptotic cells, leading to enhanced engulfment by macrophages and reduced in ammation [20,21].…”

Identification of Key Genes and Pathways in the Hippocampus after Traumatic Brain Injury: Bioinformatics Analysis and Experimental Validation

“…Microglia act as important phagocytes in the brain and can rapidly remove cellular and myelin debris after brain injury [47]. Moderate microglial phagocytosis is beneficial for remodeling the brain microenvironment and alleviating secondary injury [47,48], whereas excessive phagocytosis by microglia of neurons, synapses, or myelin may impede tissue and functional recovery [49,50]. Current evidence indicates extensive crosstalk between the complement and coagulation systems, which have fundamental clinical implications for inflammation, immunity, and tissue damage [51].…”

Neuroinflammation of traumatic brain injury: Roles of extracellular vesicles

“…During CNS infections, injuries, and diseases, astrocytes switch to an inflammatory phenotype and may contribute to subsequent inflammatory responses ( 47 ). In contrast, reactive astrocytes contribute to removing debris from the injured brain and forming glial scars to ameliorate neuroinflammation ( 167 ). Additionally, TGF-β, nerve growth factor, and brain-derived neurotrophic factor, which exert anti-inflammatory roles in TBI, are released by astrocytes ( 168 ).…”