In Vitro Seeding Activity of Glycoform-Deficient Prions from Variably Protease-Sensitive Prionopathy and Familial CJD Associated with PrP^V180I Mutation

Both sporadic variably protease-sensitive prionopathy (VPSPr) and familial Creutzfeldt-Jakob disease linked to the prion protein (PrP) V180I mutation (fCJD^V180I) have been found to share a unique pathological prion protein (PrP^Sc) that lacks the protease-resistant PrP^Sc glycosylated at residue 181 because two of four PrP glycoforms are apparently not converted into the PrP^Sc from their cellular PrP (PrP^C). To investigate the seeding activity of these unique PrP^Sc molecules, we conducted in vitro prion conversion experiments using serial protein misfolding cyclic amplification (sPMCA) and real-time quaking-induced conversion (RT-QuIC) assays with different PrP^C substrates. We observed that the seeding of PrP^Sc from VPSPr or fCJD^V180I in the sPMCA reaction containing normal human or humanized transgenic (Tg) mouse brain homogenates generated PrP^Sc molecules that unexpectedly exhibited a dominant diglycosylated PrP isoform along with PrP monoglycosylated at residue 181. The efficiency of PrP^Sc amplification was significantly higher in non-CJDMM than in non-CJDVV human brain homogenate, whereas it was higher in normal TgVV than in TgMM mouse brain homogenate. PrP^C from the mixture of normal TgMM and Tg mouse brain expressing PrP^V180I mutation (Tg180) but not TgV180I alone was converted into PrP^Sc by seeding with the VPSPr or fCJD^V180I. The RT-QuIC seeding activity of PrP^Sc from VPSPr and fCJD^V180I was significantly lower than that of sCJD. Our results suggest that the formation of glycoform-selective prions may be associated with an unidentified factor in the affected brain and the glycoform-deficiency of PrP^Sc does not affect the glycoforms of in vitro newly amplified PrP^Sc.