Phd in MOLECULAR MEDICINE

Thesis title: Loss of ATP2C1 function promotes endocytosis and degradation of NOTCH1: implications for Hailey-Hailey- disease.

Hailey-Hailey Disease (HHD) is a rare autosomal dominantly inherited disorder caused by mutations in the ATP2C1 gene that encodes an adenosine triphosphate (ATP)-powered calcium channel pump. HHD is characterized by impaired epidermal cell-to-cell adhesion and defective keratinocyte growth/differentiation. The mechanism by which mutant ATP2C1 causes HHD is unknown and current treatments for affected individuals do not address the underlying defects and are ineffective. NOTCH signaling is a direct determinant of keratinocyte growth and differentiation. We found that loss of ATP2C1 leads to impaired NOTCH1 signaling, thus deregulation of the NOTCH signaling response is therefore likely to contribute to HHD manifestation. NOTCH1 is a transmembrane receptor and upon ligand binding, NOTCH intracellular domain (NICD) translocates to the nucleus activating its target genes. In the context of HHD we found that loss of ATP2C1 function promotes upregulation of the active NOTCH1 protein, (NICD-Val1744). Here, deeply exploring this aspect, we observed that NOTCH1 activation is not associated with the transcriptional enhancement of its targets. Moreover, in agreement with these results, we found a cytoplasmic localization of NICD-Val1744. We have also observed that ATP2C1-loss is associated with the degradation of NICD-Val1744 through the lysosomal/proteasome pathway. These results show that ATP2C1-loss could promote a mechanism by which NOTCH1 is endocytosed and degraded by the cell membrane. The deregulation of this phenomenon, finely regulated in physiological conditions, could in HHD lead to the deregulation of NOTCH1 with alteration of skin homeostasis and disease manifestation.