Fig. 4: AAV-mini-PCDH15s rescue hearing in the Myo15-Cre mouse model of USH1F.
From: Mini-PCDH15 gene therapy rescues hearing in a mouse model of Usher syndrome type 1F
a Schematic of the three mini-PCDH15 constructs that are short enough to be packaged into AAV capsids. The lengths of the DNA sequences are shown. Vectors used a CMV promoter and BGH polyadenylation sequence but not a WPRE. b AAV9-PHP.B was used to deliver the different mini PCDH15 versions at P1 via round-window membrane injection. IF immunofluorescence, EM electron microscopy. c Auditory brainstem response (ABR) testing showed differential rescue with different AAV-mini-PCDH15 versions. Representative click ABR traces from P35 Pcdh15fl/fl hearing control mice, from uninjected Pcdh15fl/fl,Myo15-Cre+/− deaf knockout mice, and from knockout mice injected with AAV-mini-PCDH15-V4, -V7, or -V8. SP, Summating potential. d Average click ABR thresholds for P35 uninjected Pcdh15fl/fl control mice (n = 4), uninjected Pcdh15fl/fl,Myo15-Cre+/− conditional knockout mice (n = 6), and conditional knockout mice injected with AAV-mini-PCDH15-V4 (n = 7), AAV- HA-mini-PCDH15-V4 (n = 14), AAV-mini-PCDH15-V7 (n = 12), or AAV-mini-PCDH15-V8 (n = 5). Significance was determined by a two-tailed unpaired t-test. P values: uninjected Pcdh15fl/fl,Myo15-Cre+/− vs. AAV-mini-PCDH15-V7, p = 0.0008 (***); Pcdh15fl/fl,Myo15-Cre+/− vs. AAV-mini-PCDH15-V4, p < 0.0001 (****); Pcdh15fl/fl vs. AAV-mini-PCDH15-V4, p = 0.08 (ns); AAV-HA-mini-PCDH15-V4 vs. AAV-mini-PCDH15-V4, p = 0.84 (ns). e Average tone ABR thresholds as a function of frequency for P35 uninjected Pcdh15fl/fl control mice (n = 9), uninjected Pcdh15fl/fl,Myo15-Cre+/− conditional knockout mice (n = 10), and conditional knockout mice injected with AAV-mini-PCDH15-V4 (n = 7), HA-tagged mini-PCDH15-V4 (n = 14), mini-PCDH15-V7 (n = 12), or mini-PCDH15-V8 (n = 5). f, g Average distortion-product otoacoustic emission (DPOAE) thresholds and average DPOAE emission amplitudes at 11.3 kHz, for P35 hearing control mice (n = 9) and uninjected Pcdh15fl/fl,Myo15-Cre+/− mice (n = 6), and for those injected with AAV-mini-PCDH15-V4 (n = 7), -V7 (n = 12), or -V8 (n = 5). All data are presented as mean values ± SEM. Source data are provided as a Source Data file.
