Abstract Drosophila larval hematopoiesis takes place at the lymph gland, where blood cell progenitors differentiate into two possible cell types: plasmatocytes, analogous to mammalian macrophages, or crystal cells that share features with mammalian megakaryocytes; a third cell type, the lamellocytes, can develop only upon specific immune challenges. In this work, we investigate the role of autophagy in Drosophila hematopoiesis. We found that autophagy inhibition in blood cell progenitors results in augmented crystal cell differentiation due to accumulation of high levels of Notch protein. Notch activation during hematopoiesis depends on the endocytic pathway, which cross-talks with autophagy: While endocytosis and endosomal maturation are essential for Notch activation, autophagosomes are required for Notch lysosomal degradation. TOR signaling inhibits autophagosome biogenesis, which in turn prevents the formation of Notch-containing amphisomes, being the latter necessary for Notch lysosomal destruction. Reduction of Notch lysosomal degradation shifts the balance towards Notch activation at late endosomal membranes, thereby enhancing differentiation of crystal cells. Our work defines a novel mechanism of regulation of immune cell differentiation in response to the nutritional status of the organism: High nutrient availability induces TOR activation, thereby inhibiting autophagy, hindering lysosomal degradation of Notch, and promoting crystal cell differentiation.
