Extended Data Fig. 1: PR006 transduces FTD-GRN mutation carrier neuronal cultures in a dose-dependent manner.

(a) PR006 consists of a non-replicating recombinant AAV of 4184 nucleotides which includes an expression cassette containing the codon optimized coding sequence of human Progranulin GRN. The viral particle (AAV capsid) is comprised of three capsid proteins, VP1, VP2, and VP3, with the theoretical molecular weights of approximately 87, 73, and 62 kDa. PR006 encapsidates the modified viral vector, including the GRN expression cassette. The vector contains flanking ITRs on each side of the expression cassette, the Cytomegalovirus enhancer (CMVe) and Chicken Beta Actin promoter (CBAp) to constitutively express the codon optimized coding sequence of human GRN. The 3′ region also contains a Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) element followed by a bovine growth hormone polyadenylation (bGH polyA) tail. Further information is provided in US Patent 10689625, which covers composition claims for PR006 vector and rAAV. (b) Neural stem cells (NSCs) were seeded at an equal density and differentiated into neurons. On day 7, neurons were transduced with excipient or the indicated amounts of PR006 for 72 hours. Secreted progranulin expression was measured from the cell media by ELISA and normalized to volume (n=3–4; mean ± SEM). Black dashed line represents endogenous levels of secreted progranulin from Control neurons (excipient-treated). Secreted progranulin was not detectable in excipient-treated FTD-GRN neurons. Statistics were determined using ANOVA followed by Tukey HSD and statistical comparison of each condition to excipient-treated Control neurons is indicated on the graph. MOI, multiplicity of infection.