Kelly B. Menees

The NLRP3 inflammasome is a central mediator of metabolic inflammation, yet the upstream mechanisms remain poorly defined. We identified RGS10 as a critical regulator of NLRP3-driven immunometabolic responses. In macrophages, loss of RGS10 enhanced calcium-dependent NLRP3 inflammasome activity during both priming and activation. In high-fat diet-fed male mice, Nlrp3 deletion improved glucose tolerance, adipose remodeling, and hepatic and hypothalamic pathology, whereas Rgs10 deletion induced the opposite phenotype. These benefits required RGS10: in Nlrp3 ^-/-/ Rgs10 ^-/- (dKO) males, the improvements in glucose tolerance, adipose remodeling, and hypothalamic microgliosis were abolished. This requirement was selective; RGS10 acted dominantly to restrain myeloid and hepatic inflammation independent of NLRP3, and the interaction was sex-dependent. Finally, RGS10 declined in aged and Parkinson’s disease human microglia, and aged Rgs10 ^-/- mice developed glucose intolerance. Together, these findings establish RGS10 as a dominant, sex-dependent upstream regulator of NLPR3-driven inflammation, suggesting that age- or inflammation-associated RGS10 loss may limit the efficacy of NLRP3-targeted therapies.

The inflammatory response mounted by the immune system is a protective process necessary for survival following exposure to insults. However, it must be ti

Regulator of G-protein signaling 10 (RGS10) has been shown to regulate multiple inflammatory pathways relevant to disease pathogenesis. Of particular importance is the ability of RGS10 to negatively regulate the NFkB pathway, a prominent pro-inflammatory pathway implicated in multiple inflammatory disease phenotypes. However, the exact mechanism by which RGS10 regulates NFkB is unknown. Considering that RGS10 translocates into the nucleus upon stimulation, we hypothesize that RGS10 may regulate NFKB through transcription. To determine whether RGS10 mediates NFkB transcription, we stimulated RGS10 KO and B6 peritoneal macrophages and collected cell lysate over 24 hours to assess transcript levels of NFkB and related proinflammatory cytokines. Here we found that RGS10 differentially regulates the transcription of NκKB subunits and NFκB-dependent cytokines. Further studies are warranted to understand the potential role of RGS10 in transcriptional regulation of inflammatory states.

SOP for cell lysis, BCA protein assay, sample preparation and western immunoblotting

SOP for Immunocytochemistry. SOP for immunocytochemistry.. Includes step-by-step instructions and materials - an experimental workflow on protocols.io

Parkinson’s disease (PD) is the fastest-growing neurodegenerative disease in the world ¹ . Gastrointestinal (GI) dysfunction can occur decades before motor impairments and in up to 80% of individuals living with PD ^2–4 . We investigated peripheral relationships that may underlie mechanisms along the gut–blood axis that contribute to PD progression. Single-cell multiomic spatial molecular imaging (SMI) of colonic tissue localized and identified inflammatory injury within epithelial cells that appear to be associated with iron mishandling in both inflammatory bowel disease (IBD) and PD biosamples. We found that both the single-cell SMI of RNA and protein revealed parallel cross-modal dysregulation in the gut epithelium, in both IBD and PD biosamples. These data are accompanied by plasma (PD) and stool (IBD) protein depletion of CCL22. Our findings suggest iron mishandling along the gut barrier likely contributes to systemic inflammation, which may be one catalyst that primes circulating immune cells to body-first PD progression.

Parkinson’s disease (PD) is the fastest growing neurodegenerative disease in the world ¹ . Gastrointestinal (GI) dysfunction can occur decades before motor impairments and in up to 80% of individuals living with PD ^2,3,4 . We investigated peripheral relationships that may underlie mechanisms along the gut-blood axis that contribute to PD pathogenesis. Single-cell multiomic spatial molecular imaging (SMI) of colonic tissue localized inflammatory injury within epithelial cells that appear to be associated with iron mishandling in both inflammatory bowel disease (IBD) and PD biosamples. We found that both the single-cell SMI of RNA and protein revealed parallel cross-modal dysregulation in the gut epithelium, in both IBD and PD biosamples. These data are accompanied by plasma (PD) and stool (IBD) protein depletion of CCL22. Our findings suggest iron mishandling along the gut barrier likely contributes to systemic inflammation, which may be the catalyst that primes circulating immune cells to body-first PD pathogenesis.

Regulator of G-protein signaling 10 (RGS10), a key homeostatic regulator of immune cells, has been implicated in multiple diseases associated with aging and chronic inflammation including Parkinson’s Disease (PD). Interestingly, subjects with idiopathic PD display reduced levels of RGS10 in subsets of peripheral immune cells. Additionally, individuals with PD have been shown to have increased activated peripheral immune cells in cerebrospinal fluid (CSF) compared to age-matched healthy controls. However, it is unknown whether peripheral immune cells in the CSF of individuals with PD also exhibit decreased levels of RGS10. Utilizing the Michael J. Fox Foundation Parkinson’s Progression Markers Initiative (PPMI) study we found that RGS10 levels are decreased in the CSF of individuals with PD compared to healthy controls and prodromal individuals. As RGS10 levels are decreased in the CSF and circulating peripheral immune cells of individuals with PD, we hypothesized that RGS10 regulates peripheral immune cell responses to chronic systemic inflammation (CSI) prior to the onset of neurodegeneration. To test this, we induced CSI for 6 weeks in C57BL6/J mice and RGS10 KO mice to assess circulating and CNS-associated immune cell responses. We found that RGS10 deficiency synergizes with CSI to induce a bias for inflammatory and cytotoxic cell populations, a reduction in antigen presentation machinery in peripheral blood immune cells, as well as in and around the brain that is most notable in males. These results highlight RGS10 as an important regulator of the systemic immune response to CSI and implicate RGS10 as a potential contributor to the development of immune dysregulation in PD.