Biology subjects

Mansini, A. P.

Publications and source records attributed to Mansini, A. P..

3 recordsLinked to original sources

bioRxiv2026

Targeting neutrophil signaling networks in immune complex-mediated autoimmune disease

Fragment crystallizable gamma receptor (Fc{gamma}R)-induced signaling is a crucial process that determines the cellular response to immune complexes (IC) in autoimmune diseases. In several diseases including pemphigoid diseases (PD), such as epidermolysis bullosa acquisita (EBA), or rheumatoid arthritis (RA), neutrophils are prominently involved as effector cells, while others such as immune thrombocytopenia (ITP) are independent of neutrophils. At the same time, most diseases are commonly treated by broad-range immunosuppression accompanied by severe risk for adverse effects. Signal transduction inhibitors (STIs) have been successfully applied in cancer therapy. However, their use in autoimmune diseases is an emerging, but so far understudied potential treatment avenue. Therefore, we screened a target-selective compound library consisting of 155 STIs in a neutrophil-based assay and conducted a multiplex kinase activity profiling with IC-stimulated neutrophils. Thus, we found novel potential therapeutic targets that were validated both in vitro in functional neutrophil assays and in vivo in murine models of EBA Here, we demonstrate that both systemic and topical treatment with several individual STIs is effective in a prophylactic approach in these models. Furthermore, therapeutic treatment with the BTK inhibitor ibrutinib in the immunization-induced EBA model reduced disease severity by approximately 85 % and showed efficacy in additional experimental models of EBA, arthritis, and ITP. Together, the present study contributes to the elucidation of Fc{gamma}R-dependent signaling in neutrophils and identifies multiple novel promising treatment options including inhibition of PLC, PDK-1, PKC, p38, DNA-PK, KSP, c-Met, TBK-1 and BTK for IC-mediated autoimmune diseases.

immunology↗

bioRxiv2025

CDK7, CDK9, or CDK11 Inhibition Reduces Neutrophil-Driven Inflammation and Tissue Damage in Experimental Autoimmune Models

BackgroundCyclin-dependent kinases (CDKs) are involved in basic cellular processes like regulation of cell-cycle progression and transcription. However, recent data also indicate a specific role in terminally differentiated neutrophils by promoting reactive oxygen species (ROS) release, degranulation, neutrophil extracellular trap formation, or apoptosis. Since these mechanisms are implicated in multiple autoimmune diseases, we aimed to delineate the role of CDKs in IC-mediated autoimmune diseases both in vitro and in vivo. MethodsWe analyzed CDK gene expression in unstimulated and immune complex (IC)-stimulated neutrophils. Subsequently, we investigated the effect of pharmacological CDK inhibition on IC-activated neutrophil functions. To analyze the inhibitors in a more translational approach, we proceeded with the systemic and topical application of the effective inhibitors in a murine antibody transfer-induced local epidermolysis bullosa acquisita (EBA) model. The most efficient inhibitor, MC180295, was validated in two other IC-mediated models of autoimmune disease: Serum-transfer arthritis (STA), which also, but not exclusively, depends on neutrophils and immune thrombocytopenia (ITP), which is considered less neutrophil-dependent. ResultsWe found 14 CDKs expressed in unstimulated cells, while the IC-stimulation showed an upregulation of CDK2 and CDK4 expression. Inhibitors selectively targeting CDK1, CDK2, CDK4/6, CDK7, CDK9, CDK11, and CDK12 showed effects on different neutrophil functions (surface activation marker expression, ROS release, adhesion, apoptosis) in vitro. In the predominantly neutrophil-driven EBA model, we observed a reduction of disease severity upon treatment with CDK7, CDK9, or CDK11 inhibitors. Inhibiting these CDKs with topical THZ2, MC180295, or OTS964, respectively, also improved the clinical phenotype. In line with our hypothesis, MC180295 impaired the development of STA, but not ITP. ConclusionsCDK7, CDK11, and especially CDK9 inhibition show therapeutic potential in IC-driven neutrophil-mediated diseases such as rheumatoid arthritis and EBA.

immunology↗

bioRxiv2023

Cholangiocyte primary cilia transduce a fluid shear signal to increase KLF2 via the actin cytoskeleton

Cholangiocarcinoma is an aggressive solid tumor formed in the bile duct epithelium. Often this tumor obstructs bile flow, known as cholestasis. Normal cholangiocytes detect bile flow in the ductal lumen with an extension of the apical membrane called the primary cilium. However, these sensory organelles are often lost in malignant cells. Kruppel-like factor 2 (KLF2) is an important flow-sensitive transcription factor involved in shear stress response in endothelial cells, and has anti-proliferative, anti-inflammatory, and anti-angiogenic effects. The potential role of KLF2 in cholangiocyte flow detection and in cholangiocarcinoma is unknown. We hypothesized that reduced bile flow contributes to malignant features in cholangiocarcinoma through regulation of KLF2 signaling. We observed that primary cilia were expressed in normal cholangiocytes but were absent in malignant cells. KLF2 expression was higher in normal cells compared to malignant. Depletion of cilia in normal cells led to a decrease in KLF2 expression and increased cilia number was associated with increased KLF2. Enforced KLF2 expression inhibited cell proliferation, migration, and also decreased cell death induction in malignant cells. Applied media flow over cholangiocytes increased KLF2 and cilia depletion completely blocked flow-induced KLF2 expression. Disruption of filamentous actin decreased KLF2 expression, suggesting the cilium may communicate through a cytoskeletal mechanotransduction pathway. Our studies demonstrate that cilia positively regulated KLF2 protein levels and increased fluid flow induced KLF2 expression for the first time in cholangiocytes, emphasizing the importance of reestablishing bile flow in cholestatic cholangiocarcinoma.

cancer biology↗