Extended Data Fig. 6: TDP-43 depleted iPSC-derived spinal motor neurons exhibit hyperexcitability that can be rescued by splice-modulating KCNQ2 ASOs.
From: TDP-43-dependent mis-splicing of KCNQ2 triggers intrinsic neuronal hyperexcitability in ALS/FTD
(a) Schematic illustrating splice modulating ASO (smASO) screen strategy in iPSC-MNs. (b) qRT-PCR of TARDBP expression in iPSC-MNs at day 32 of the screen; KD: knockdown. Data are presented as mean ± SEM; Statistical significance was determined by 2-tailed, unpaired t-test. Circles represent a biological replicate. (c) qRT-PCR of mis-spliced KCNQ2∆E5 after treatment with smASO-KCNQ2 candidate #2 at three doses (1μM, 3μM, 10μM). Data were presented as mean ± SEM; p value is the result of an unpaired, 2-tailed t-test. Circles represent a biological replicate. (d) Representative ICC images of iPSC-derived MNs stained for DAPI, TDP-43 and MAP2. Scale bar: 10 μm. (e) Cell capacitance recorded from all 3 groups; number of neurons: TDP-KD+smCTRL n = 55, TDP-KD+smCTRL n = 60, TDP-KD+smKCNQ2 n = 39 and (f) Input resistance; number of neurons: TDP-KD+smCTRL n = 49, TDP-KD+smCTRL n = 53, TDP- KD+smKCNQ2 n = 27. Statistical significance determined by a one-way ANOVA; no significant differences between groups. Data represented as mean ± SEM. (g) Representative traces for spontaneous APs recorded from neurons for all 3 groups. Scale bar: 10 mV/5 ms. (h-n) Spontaneous AP properties from neurons for all 3 groups; number of neurons: CTRL+smCTRL n = 51, TDP-KD+smCTRL n = 57, TDP-KD+smKCNQ2 n = 34. Each circle represents the average of all spontaneous APs recorded from one cell over the first 15 seconds of whole cell configuration. Cells recorded from n = 4 independent biological replicates. Statistical significance determined by a one-way ANOVA followed by a Tukey’s multiple comparisons test when applicable. Data presented as the mean ± SEM.
