We previously showed that ubiquitous overexpression of the chromatin remodeling factor SWItch3-related gene (SRG3) promotes M2 macrophage differentiation, resulting in anti-inflammatory responses in the experimental autoimmune encephalomyelitis model of multiple sclerosis. Since hepatic macrophages are responsible for sepsis-induced liver injury, we investigated herein the capacity of transgenic SRG3 overexpression (SRG3^β-actin mice) to modulate sepsis in mice exposed to lipopolysaccharide (LPS) plus d-galactosamine (d-GalN). Our results demonstrated that ubiquitous SRG3 overexpression significantly protects mice from LPS/d-GalN-induced lethality mediated by hepatic M1 macrophages. These protective effects of SRG3 overexpression correlated with the phenotypic conversion of hepatic macrophages from an M1 toward an M2 phenotype. Furthermore, SRG3^β-actin mice had decreased numbers and activation of natural killer (NK) cells but not natural killer T (NKT) cells in the liver during sepsis, indicating that SRG3 overexpression might contribute to cross-talk between NK cells and macrophages in the liver. Finally, we demonstrated that NKT cell-deficient CD1d KO/SRG3^β-actin mice are protected from LPS/d-GalN-induced sepsis, indicating that NKT cells are dispensable for SRG3-mediated sepsis suppression. Taken together, our findings provide strong evidence that SRG3 overexpression may serve as a therapeutic approach to control overwhelming inflammatory diseases such as sepsis.

We have previously shown that Vα14 TCR Tg (Vα14^Tg) NC/Nga (NC) mice contain increased numbers of double-negative (DN) invariant natural killer T (iNKT) cells that protect against spontaneous development of atopic dermatitis (AD). iNKT cells can regulate immune responses by producing various cytokines such as IFNγ and IL4 rapidly upon stimulation with α-galactosylceramide (α-GalCer), a prototypical iNKT cell agonist. However, the precise role of α-GalCer-activated iNKT cells in AD development remains unclear. Therefore, we examined whether repeated activation of iNKT cells with α-GalCer can regulate the pathogenesis of AD in Vα14^Tg NC mice. We found that Vα14^Tg NC mice injected repeatedly with α-GalCer display exacerbated AD symptoms (e.g., a higher clinical score, IgE hyperproduction, and increased numbers of splenic mast cells and neutrophils) compared with vehicle-injected Vα14^Tg NC mice. Moreover, the severity of AD pathogenesis in α-GalCer-injected Vα14^Tg NC mice correlated with increased Th2 cells but reduced Th1 and Foxp3⁺ Treg cells. Furthermore, the resulting alterations in the Th1/Th2 and Treg/Th2 balance were strongly associated with a biased expansion of type 2 cytokine-deviated iNKT cells in α-GalCer-treated Vα14^Tg NC mice. Collectively, our results have demonstrated the adverse effect of repeated α-GalCer treatment on skin inflammation mediated by type 2 immunity.

The SWItch (SWI)3-related gene (SRG3) product, a SWI/Sucrose Non-Fermenting (SNF) chromatin remodeling subunit, plays a critical role in regulating immune responses. We have previously shown that ubiquitous SRG3 overexpression attenuates the progression of Th1/Th17-mediated experimental autoimmune encephalomyelitis. However, it is unclear whether SRG3 overexpression can affect the pathogenesis of inflammatory skin diseases such as atopic dermatitis (AD), a Th2-type immune disorder. Thus, to elucidate the effects of SRG3 overexpression in AD development, we bred NC/Nga (NC) mice with transgenic mice where SRG3 expression is driven by the β-actin promoter (SRG3^β-actin mice). We found that SRG3^β-actin NC mice exhibit increased AD development (e.g., a higher clinical score, immunoglobulin E (IgE) hyperproduction, and an increased number of infiltrated mast cells and basophils in skin lesions) compared with wild-type NC mice. Moreover, the severity of AD pathogenesis in SRG3^β-actin NC mice correlated with expansion of interleukin 4 (IL4)-producing basophils and mast cells, and M2 macrophages. Furthermore, this accelerated AD development is strongly associated with Treg cell suppression. Collectively, our results have identified that modulation of SRG3 function can be applied as one of the options to control AD pathogenesis.

The SWItch3-related gene (SRG3) is a core component of ATP-dependent SWI/SNF complexes, which are crucial for regulating immune cell development and function (e.g., macrophages and CD4⁺ T cells), embryonic development, and non-immune cell differentiation. Notably, SRG3 overexpression has been shown to polarize macrophages in the central nervous system toward an anti-inflammatory M2 phenotype, thereby protecting against the development of experimental autoimmune encephalomyelitis in mice. However, the effect of SRG3 on immune responses in adipose tissues remains unclear. To address this issue, we examined the cellularity and inflammatory status of adipose tissue in B10.PL mice overexpressing the SRG3 gene under the ubiquitous β-actin promoter (SRG3^β-actin). Interestingly, SRG3 overexpression significantly reduced adipocyte size in both white and brown adipose tissues, without affecting the overall adipose tissue weight. Such phenotypic effects might be associated with the improved glucose tolerance observed in SRG3^β-actin B10.PL mice. Moreover, we found that SRG3 overexpression down-regulates IL1β-expressing M1 macrophages, leading to a significant decrease in the M1/M2 macrophage ratio. Additionally, SRG3^β-actin B10.PL mice showed a dramatic reduction in neutrophils as well as IL1β- and IL17-producing T cells in adipose tissues. Taken together, our results indicate that SRG3 plays a vital role in maintaining immune homeostasis within adipose tissues.

Background: Ovarian and gastric cancers are associated with poor prognosis and derive limited benefit from current therapies, highlighting the need for novel immunotherapeutic approaches. The L1 cell adhesion molecule (L1CAM) is aberrantly expressed in several types of solid tumors and is associated with aggressive clinical behavior. This study aimed to develop and characterize optimized L1CAM-targeted chimeric antigen receptor-T (CAR-T) cells with improved efficacy and safety, using patient-derived preclinical models of ovarian and gastric cancer. Methods: L1CAM expression was assessed in tumor tissues from patients with ovarian cancer (OC;n=227) and gastric cancer (GC;n=132), followed by an analysis of its clinical impact. A panel of L1CAM-targeted CAR constructs incorporating distinct hinge and co-stimulatory domains was systematically evaluated. Functional activity was assessed by cytotoxicity and cytokine release assays using cancer cell lines and patient-derived tumor cells. Antitumor efficacy, persistence, and safety were evaluated in xenograft and patient-derived xenograft (PDX) models, including NanoLuc-based in vivo tracking of CAR-T cell distribution. Results: High L1CAM expression correlated with aggressive clinicopathological features and adverse prognosis in patients with OC. Among the constructs tested, CAR-T cells incorporating an IgD/IgG1 hybrid hinge and dual CD28-ICOS co-stimulatory domains demonstrated the most potent antigen-specific activity. These cells exhibited enhanced cytotoxicity, robust interferon-γ production, and sustained functional activity. In vivo, optimized L1CAM CAR-T cells achieved complete tumor regression in OC PDX models and significant tumor suppression in GC models, with durable tumor control and CAR-T persistence. Intraperitoneal delivery further improved therapeutic efficacy and CAR-T cell localization compared with intravenous administration. Importantly, no evidence of neurotoxicity or systemic toxicity was observed, supporting a favorable safety profile. Conclusions: Optimized L1CAM-targeted CAR-T cells demonstrate potent, durable, and antigen-specific antitumor activity with a favorable safety profile in preclinical models of OC and GC. The combination of rational CAR design (CD28-ICOS signaling) and optimized delivery strategy provides a strong rationale for the clinical development of L1CAM-targeted CAR-T therapy, particularly in patients with L1CAM-positive solid tumors.