<i>c9orf72</i> and <i>smcr8</i> mutant mice reveal MTORC1 activation due to impaired lysosomal degradation and exocytosis

How lysosome and MTORC1 signaling interact remains elusive in terminally differentiated cells. A G4C2 repeat expansion in <i>C9orf72</i> is the most common cause of familial amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD) (C9ALS-FTD). We previously identified a C9orf72-SMCR8-containing complex. Here we found that <i>c9orf72</i> and <i>smcr8</i> double-knockout (dKO) mice exhibit similar but more severe immune defects than the individual <i>knockouts</i>. In <i>c9orf72</i> or <i>smcr8</i> mutant macrophages, lysosomal degradation and exocytosis were impaired due to the disruption of autolysosome acidification. As a result of impaired lysosomal degradation, MTOR protein was aberrantly increased, resulting in MTORC1 signaling overactivation. Inhibition of hyperactive MTORC1 partially rescued macrophage dysfunction, splenomegaly and lymphadenopathy in <i>c9orf72</i> or <i>smcr8</i> mutant mice. Pharmacological inhibition of lysosomal degradation upregulated MTOR protein and MTORC1 signaling in differentiated wild-type macrophages, which resemble phenotypes in KO mice. In contrast, <i>C9orf72</i> or <i>Smcr8</i> depletion in proliferating macrophages decreased MTORC1 signaling. Our studies causatively link C9orf72-SMCR8’s cellular functions in lysosomal degradation, exocytosis, and MTORC1 signaling with their organism-level immune regulation, suggesting cell state (proliferation vs. differentiation)-dependent regulation of MTOR signaling via lysosomes.