DeBerry, J. J. | bioRxiv Science

bioRxiv2024

Inhibition of the RNA Regulator HuR mitigates spinal cord injury by potently suppressing post-injury neuroinflammation

BackgroundNeuroinflammation plays a significant role in promoting secondary tissue injury after spinal cord trauma. Within minutes after spinal cord injury (SCI), microglia and astrocytes become activated and produce inflammatory mediators such as TNF-, IL-6, iNOS and COX-2 which induce tissue injury through cytotoxicity, vascular hyperpermeability, and secondary ischemia. The inflammatory cascade is amplified by chemokines such as CCL2 and CXCL1 that promote recruitment of peripheral inflammatory cells into the injured spinal cord. HuR is a key post-transcriptional RNA regulator that controls glial expression of many pro-inflammatory factors by binding to adenylate- and uridylate-rich elements in 3 untranslated regions of the mRNA. SRI-42127 is a small molecule inhibitor that blocks HuR nucleocytoplasmic translocation, a process critical for its regulatory function. The goal of this study was to assess the potential of SRI-42127 for suppressing neuroinflammation after SCI and improving functional outcome. MethodsAdult female mice underwent a contusion injury at the T10 level. SRI-42127 or vehicle was administered intraperitoneally starting 1 h after injury and up to 5 days. Locomotor function was assessed by open field testing, balance beam and rotarod. Immunohistochemistry was used to assess lesion size, neuronal loss, myelin sparing, microglial activation and HuR localization. Molecular analyses of spinal cord and peripheral tissues for expression of inflammatory mediators included qPCR, immunohistochemistry, ELISA, or western blot. Post-SCI pain was assessed by the mouse grimace scale. ResultsSRI-42127 significantly attenuated loss of locomotor function and post-SCI pain. Histologic correlates to these beneficial effects included reduced lesion size, neuronal loss, and an increase in myelin sparing. There was reduced microglial activation at the epicenter with concomitant attenuation of HuR nucleocytoplasmic translocation. Molecular analysis revealed a striking reduction of pro-inflammatory mediators at the epicenter including IL-6, MMP-12, IL-1{beta}, TNF-, iNOS, COX-2, and chemokines CCL2, CXCL1, and CXCL2. Suppression of inflammatory responses extended peripherally including serum, liver, and spleen. ConclusionTargeting HuR after SCI is a viable therapeutic approach for suppressing neuroinflammatory responses after tissue injury and improving functional outcome.

neuroscience↗

bioRxiv2020

Heme Attenuates Endogenous Opioid Levels in Leukocytes of HIV positive individuals with Chronic Widespread Pain

ABSTRACTThe prevalence of chronic widespread pain (CWP) in people with HIV (PWH) is high, yet the underlying mechanisms are elusive. Leukocytes synthesize the endogenous opioid, β-endorphin (β-END), within their endoplasmic reticulum (ER). When released into plasma, β-END dampens nociceptive transmission by binding to opioid receptors on sensory neurons. In the present study, we hypothesized that heme-induced ER stress attenuates leukocyte levels/release of β-END, thereby increasing pain sensitivity in PWH. Results demonstrate that PWH with CWP have fragile erythrocytes, high plasma levels of cell-free heme, and impaired heme metabolism. Leukocytes from PWH with CWP also had high ER stress and low β-END compared to PWH without CWP and HIV-negative individuals with or without pain. In vitro heme exposure decreased β-END levels/secretion in murine monocytes/macrophages, which was prevented by treatment with sodium 4-phenylbutyrate, an ER stress inhibitor. To mimic hemolytic effects in a preclinical model in vivo, C57BL/6 mice were injected with phenylhydrazine hydrochloride (PHZ). PHZ increased cell-free heme and ER stress, decreased leukocyte β-END levels and hindpaw mechanical sensitivity thresholds. Treatment of PHZ-injected mice with the heme scavenger, hemopexin, blocked these effects, suggesting that heme-induced ER stress and a subsequent decrease in leukocyte β-END may contribute to CWP in PWH.Competing Interest StatementThe authors have declared no competing interest.View Full Text

pathology↗