Identification of inhibitors of the pathological Notch3 aggregation

• Applicant: Privatdozent Dr. Christof Haffner
• Subject Area: Molecular and Cellular Neurology and Neuropathology ; Molecular Biology and Physiology of Neurons and Glial Cells
• Term: from 2017 to 2023
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 387165796

CADASIL (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy) represents the most frequent monogenic cerebral small vessel disease and is characterized by the premature appearance of ischemic strokes and vascular dementia. The underlying mutations in the Notch3 receptor lead to the misfolding, aggregation and accumulation of the Notch3 extracellular domain (Notch3-ECD) in arterial vessel walls resulting in a fibrotic degeneration of small cerebral vessels through as yet incompletely understood mechanisms. Using the Notch3 C183R mutant and the single molecule detection technique SIFT (scanning for intensely fluorescent targets) we have recently succeeded in recapitulating the aggregation process in vitro and could demonstrate that its inhibition by synthetic drug-like substances is principally possible. Thus, antiaggregation represents a promising therapeutic approach for CADASIL which we intend to follow up and further develop until its applicability in a preclinical mouse model in the present project. For that purpose we will 1) validate the efficacy of identified inhibitors on additional mutations and in further in vitro aggregation assays (dynamic light scattering, filter-trap assay), 2) use the SIFT assay to conduct an extended primary screening of substance libraries for additional inhibitors, 3) investigate the effect of inhibitors on more complex aggregates (in the extracellular matrix of cultured cells and in isolated human vessels), and 4) test inhibitors suitable for in vivo application in an established CADASIL mouse model.

• DFG Programme: Research Grants