TTC7A organizes glandular lumen formation in the intestine through a Class II phosphatidylinositol 3-kinase

ABSTRACT The formation of a single central lumen is a critical step for glandular morphogenesis. Patients with loss of function variants in the chaperone protein TTC7A have multiple lumen formation in colonic crypt glands. We show that trafficking and localization of TTC7A to the plasma membrane is required for directionally specifying the apical membrane with TTC7A patient loss-of-function variants leading to mislocalized membrane and lumen formation. Our experiments show that TTC7A, in early stages of apical membrane development, functions as a molecular chaperone for the Class II phosphatidylinositol 3-kinase, PIK3C2A and is trafficked in Rab11a positive vesicles to generate phosphatidylinositol 3,4-bisphosphate (PI(3,4)P2). We show that the apical specification process is dependent on PIK3C2A dependent generation of PI(3,4)P2 in intestinal epithelia and that defective lumen formation can be rescued by exogenous PI(3,4)P2 or small molecules that modulate phosphoinositide homeostasis. Competing Interest Statement SBS declares the following interests: Scientific advisory board participation for Pfizer, Merck, Sonoma Biotherapeutics, Spyre Therapeutics, Biolojic Design, Trex Bio.