Biology subjects

Servant, M. J.

Publications and source records attributed to Servant, M. J..

3 recordsLinked to original sources

bioRxiv2026

The orphan receptor IL-17RD is a negative regulator of RIG-I-like receptor-dependent antiviral innate immunity and restrains SARS-CoV-2-induced lung inflammation

Detection of viral RNA by the RIG-I-like receptors (RLRs) RIG-I and MDA5 triggers assembly of a MAVS-dependent signalosome that activates the TBK1-IRF3 and IKK{beta}-NF-{kappa}B axes together with the JNK and p38 MAPK modules, driving type I and type III interferon (IFN) and inflammatory cytokine production. Because unrestrained activity of this pathway is a major cause of immunopathology, host-encoded negative regulators are essential, yet the full complement of these brakes remains incompletely defined. Here we identify interleukin-17 receptor D (IL-17RD, also known as SEF), an orphan member of the IL-17 receptor family previously characterized as an antagonist of FGF and Toll-like receptor signaling, as a negative regulator of RLR-driven antiviral innate immunity. Using a CRISPR-engineered and shRNA-depleted human airway epithelial-derived lung carcinoma A549 cell line, we show that loss of IL-17RD amplifies and prolongs phosphorylation of TBK1 and IRF3 in response to poly I:C transfection and to infection with encephalomyocarditis virus (EMCV) or Sendai virus (SeV), and likewise potentiates the IKK{beta}-I{kappa}B module and the TAK1-JNK1/2 and p38 MAPK branches. This translates into increased nuclear accumulation of IRF3 and p65, and markedly elevated induction of IFNB1, IFNL1-3, CCL5, IL6, and NFKBIA transcripts, as well as secreted IFN-{beta} and IL-6. Silencing IL-17RD in ACE2-expressing A549 cells similarly derepresses the antiviral and inflammatory transcriptional programme following SARS-CoV-2 infection. Epistasis experiments place IL-17RD at the level of MAVS, downstream of the RLR sentinels. Mechanistically, IL-17RD localizes to the ER-to-Golgi intermediate compartment (ERGIC), the membrane platform on which the MAVS signalosome is present, and associates with RIG-I, MDA5, MAVS, TBK1 and IRF3. Its re-expression in depleted cells redistributes RLR effectors and TRAF proteins across low-molecular-weight signalosome fractions, reducing the amount of IRF3 recruited to the 670 kDa MAVS signalosome complex. Complementation of IL-17RD-deficient cells also indicates that the intracellular TIR subdomain is sufficient to confer this antagonistic activity. Finally, Il17rd-/- mice display a splenic transcriptome enriched for antiviral response signatures, and, following intranasal infection with a moderate dose of SARS-CoV-2, they mount an exaggerated pulmonary cytokine response and develop significantly greater lung inflammation and fibrosis than wild-type littermates. Together, these data establish IL-17RD as a bona fide brake on the RLR-MAVS axis that limits virus-induced immunopathology, and identify the SEFIR/TIR subdomain as the module responsible for this activity.

microbiology↗

bioRxiv2025

ARF6 controls VSMC cell phenotypic switching upon lipid stimulation to promote inflammatory signaling contributing to the progression of atherosclerosis

Vascular smooth muscle cells (VSMCs) play a pivotal role in the development and progression of atherosclerosis. Traditionally viewed as contractile cells that maintain vascular tone and structure, VSMCs undergo phenotypic switching in response to atherogenic stimuli, such as high circulating levels of LDL, thus adopting synthetic, osteogenic, or macrophage-like phenotypes. This plasticity contributes to plaque formation, extracellular matrix remodeling, and inflammatory signaling. We have previously shown that ADP-ribosylation factor 1 (ARF1), a small GTP-binding protein, regulates the expression and function of actin, which is important for maintaining the contractile phenotype of VSMCs. However, the role of ARF6 in phenotypic switching remains to be elucidated. Here, we demonstrate that ARF6 knockdown in human aortic smooth muscle cells (HASMCs) reduced lipid uptake through alterations in the expression of scavenger receptors (LOX-1, MSR1), cytokine production (IL-6) as well as modulation of inflammatory markers and pathways (adhesion molecules, PI3K, NFkB, p38). To confirm our findings in an in vivo setting, we engineered a novel conditional smooth muscle cell-specific ARF6 KO mouse in an atheroprone background (Acta2-Cre-ERT2+/-/ApoE-/-/Arf6f/f mice). Mice were fed a high-fat diet to accelerate plaque formation. ARF6 knockout resulted in a significant reduction of atherosclerotic lesions in the aortic arch, which was associated with a reduction of collagen and foam cells. Furthermore, we observed that ARF6 regulates the expression of inflammatory markers. These findings highlight the importance of ARF6 expression in VSMCs and its role in the pathogenesis of atherosclerosis.

biochemistry↗

bioRxiv2024

Early IKKb-dependent anabolic signature governs vascular smooth muscle cells fate and abdominal aortic aneurysm development

BackgroundAbdominal aortic aneurysm (AAA) is a detrimental disease with no effective pharmacological therapy. While inflammation is recognized as one of the key regulators of AAA, targeting inflammatory pathways once the disease is established does not impact the outcomes. However, understanding the earliest molecular indicators could shed light on the precise biological targets and prognostic markers for AAA. MethodsUsing apolipoprotein E (ApoE)-deficient mice fed with a standard diet and infused with Angiotensin II (Ang II), we conducted bulk RNA-sequencing (RNA-Seq) analysis on suprarenal (SRA) regions obtained from both unchallenged and challenged WT mice, specifically examining responses 24 hours after Ang II infusion to capture the initial phases of aortic stress response. We further created a unique model of hyperlipidemic mice in which the expression of the inhibitor of nuclear factor kappa B kinase subunit beta (IKK{beta}) can be conditionally (via tamoxifen injection) suppressed in vascular smooth muscle cells (VSMC). The development of AAA was evaluated using in situ examination and quantified using RT-qPCR, immunohistochemistry and fluorescence microscopy. Cultured VSMC were exposed to the selective IKK{beta} inhibitor MLN120b and the expression levels of phenotypic markers kruppel like factor 4 (KLF4), {beta}-Catenin and cellular communication network factor 2 (CCN2) were addressed using cellular extracts and immunoblot analysis. ResultsRNA-Seq data support the presence of early anabolic events in SRA regions detailing activation of the mammalian target of rapamycin complex 1 (mTORC1) pathway, which paralleled cellular anabolic processes including mitochondria, ribosome and sterol biosynthesis, the Unfolded Protein Response (UPR) and fibrogenesis. Conditional deletion of the Ikbkb gene in VSMC significantly reduces the incidence of SRA lesions as well as the rate of aneurysm ruptures in mice exposed to Ang II. In situ analysis further demonstrated that the protection conferred by the lack of IKK{beta} expression in VSMC is associated with reduced inflammatory response, the preservation of the contractile over the degradative VSMC phenotypes, and the absence of an anabolic signature. ConclusionOur results not only reinforce the major roles played by VSMC in the rapid adaptation leading to the deleterious remodeling of the vascular wall and aortic lesions but also support a paradigm aiming at repositioning the efforts focusing on anabolic rather than inflammatory events.

molecular biology↗