Identification and validation of key host cellular factors directing SARS-CoV-2 assembly and consequences for neuronal proteostasis

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We have previously demonstrated that replication of neurotropic viruses leads to cellular host protein recruitment, thereby to disturbed proteostasis that can unleash self-sustaining pathological signaling cascades known from the cellular pathology of neurodegenerative diseases. Dissecting the exact molecular pathway that SARS-CoV-2 runs through after entering the cells is key for understanding its pathogenesis. In this proposal, we focus on identifying cellular host proteins aiding in SARS-CoV-2 assembly through purifying the target proteins of a drug that potently inhibits SARS-CoV-2 assembly by binding to host cell cofactors. Candidate host proteins identified by drug affinity chromatography combined with a mass spectrometric will be validated by a variety of experiemtns including CRISPR/Cas9 knockout of corresponding genes and measuring virus replication in gene-deficient cells. Furthermore, specific effects on proteostasis in neuron-like cells will be investigated based on our observations that viruses can lead to protein aggregation observed also in neurodegeneration. Our insights from these proposed cellular studies will be backed by (co-) staining of human brain sections of COVID19 patients to confirm neurotropism and the possibility of long term effects. Our studies will be important for public health polocies regarding SARS-CoV-2.