Monogenic forms of Inflammatory Bowel Disease (IBD) offer valuable insights into key regulators of intestinal inflammation. Building on our previous advancements in characterizing very early onset IBD (VEO-IBD) patients with monogenic CASP8 and RIPK1 deficiencies, we aim to elucidate the key factors and molecular mechanisms of disrupted cell death responses (necroptosis) in these life-threatening conditions.
Our hypothesis proposes that unbiased and simultaneous exploration of thousands of genes by employing discovery-oriented pooled CRISPR screens will illuminate the signaling network governing perturbed inflammatory and cell death responses, shedding light on novel therapeutic targets for RIPK1 and CASP8 deficiencies. In the proposed CRISPR screen, we will assess the enrichment of guide RNAs from genome-wide libraries that are capable of rescuing uncontrolled LPS-induced necroptosis and inflammatory responses in RIPK1- and CASP8-deficient macrophages using next-generation sequencing and bioinformatics analysis. A set of prioritized targets will be characterized in advanced genetically engineered induced pluripotent stem cell-derived human disease models.
Given the potency of monogenic IBD as a powerful model for defining key regulators of intestinal homeostasis, we anticipate uncovering critical insights into the molecular pathomechanisms and druggable targets of inflammation, contributing valuable knowledge for rare and common forms of IBD according to the translational genomics principle bedside-to-bench and back.
