In recent years, obesity-induced metabolic syndrome and chronic inflammation have been singled out as major risk factors driving non-communicable diseases, which display rising incidence world-wide (Saklayen 2018; Christ et al. 2018; Mangan et al. 2018). Although the mechanism of chronic inflammation caused by over-supply of carbohydrates and fat is not fully understood, mouse models of numerous non-communicable diseases point to a common mechanistic involvement of a single inflammatory key pathway,the NLRP3 inflammasome. While inflammasome-dependent cytokine release and pyroptosis have been a major focus of immunology research for the past 15 years, most genetic models were established in mice, and the mechanism of NLRP3 activation in human myeloid cells is still highly opaque. As an example, we and others have identified the kinase NEK7 to be critically involved in NLRP3 activation in mouse macrophages in 2016 (Schmid-Burgk et al. 2016; He et al. 2016), where it prevents NLRP3activation during mitosis (Shi et al. 2016), but this important function is clearly not mirrored inhuman myeloid cells (Schmacke et al. 2019) and therefore calls for systematic human genetic studies.The project we propose to pursue with excellent collaboration partners within the existing CRC 1454 “Metaflammation and Cellular Programming” will systematically clarify the molecular events of human NLRP3 activation in response to metabolic conditions, cellular danger signals, and pathogen infections. First, we will delineate the molecular function of three putative novel pathway members that we recently identified using an unpublished forward-genetic screen. Second, we aim to adapt our unique genetic screening platform to identify redundant or context-dependent genetic factors of NLRP3 activation. We will achieve this by performing genome-scale CRISPR screening under differential metabolic conditions and homozygous knock-out backgrounds.Third, we aim to establish genome-scale screening in human primary myeloid cells, which will unlock access to systematic genetic studies in a physiological background.