850 JAK-STAT platform for immune-related drug discovery

<h3>Background</h3> The Janus kinase (JAK) and signal transducer and activator of transcription (STAT) pathways are evolutionarily conserved transmembrane signaling mechanisms that ensure the ability of normal cellular communication with the external environment. Various cytokines, interferons, growth factors, and other specific molecules can activate JAK-STAT signaling to drive a range of physiological and pathological processes, including proliferation, metabolism, immune response, inflammation, and malignancy. However, the JAK-STAT signaling pathway and related gene mutations are closely associated with aberrant immune activation and cancer progression. Through extensive research into the structure and function of the JAK-STAT pathway, a variety of clinical drugs have emerged and been approved for the treatment of disease, primarily targeting cytokines and their receptors, JAK inhibitors and STAT inhibitors. <h3>Methods</h3> Both JAK and STAT, once activated, cause STAT phosphorylation and the amount of downstream cytokines secreted. Therefore by using different cytokines such as IL-6, IL-23, IFNα, TPO or GM-CSF, etc., stimulating PBMC, T cells or whole blood, and detecting STAT phosphorylation as well as secreted cytokines, the activity of various drugs on the JAK-STAT pathway can be assessed. <h3>Results</h3> Whether it is THP-1 cells, PBMC or monocyte-induced macrophages, once NLRP3 is inhibited after LPS stimulation, it can effectively inhibit the secretion of IL-1β or IL-18, as well as caspase-1 activity, and by analyzing the amount of cytokine secretion or caspase1 activity, we can assess the inhibitory activity of different types of compounds on NLRP3. <h3>Conclusions</h3> Both in PBMC and in sorted T cells as well as whole blood, various JAK-STAT-related inhibitors can be validated for their activity by assaying STAT phosphorylation as well as cytokines. Currently, ICE has established various in vitro assays for the JAK-STAT signaling pathway to facilitate drug discovery.