Fig. 4: Altered neuron signaling following safusidenib treatment.

a, Spatial metabolomics detecting adenosine intensity (top) in participant O-01 pre- and post-safusidenib. Bottom: kernel density plot of smoothed intensity. b, Barcode plot of the ‘Adora2b-mediated anti-inflammatory cytokine production’ pathway in LC–MS metabolomics data (GSEA permutation-based testing, n = 9 participants) and normalized metabolite ratio of cyclic AMP pre- and post-safusidenib (n = 9 participants, paired t-test). Each box indicates the IQR, the center line is the median and the whiskers extend to the furthest points within 1.5 × IQR. c, Barcode plot of the ‘Neuroinflammation and glutamatergic signaling’ pathway in LC–MS metabolomics data (GSEA permutation-based testing, n = 9 participants). d, Spatial map of transcript niches of three participants pre- and post-safusidenib computed using GraphSAGE. e, Cell assignments to transcript-based niches are represented as normalized proportions. Bar plot of the proportion of cells in each niche. f, Spatial maps of neurons (red) in pre- and post-safusidenib samples of participant O-01 with associated density contour plot (left) and key neuronal layer markers (right). g, Average log expression of the ‘synaptic signaling’ pathway for neurons in the T7 niche pre- and post-safusidenib from participant O-01, inferred through CytoSPACE integration. Insets: cell-type annotations, with donut plots of neuron subtype proportions within the region. h, Dot plots of the proportion of neurons per transcript niche expressing synaptic signaling genes in matched pre- and post-safusidenib samples. The dot colors represent scaled average log expression. i, Example layer 2/3 pyramidal neuron filled with 5-(and-6)-tetramethylrhodamine biocytin. Inset: whole-cell patch-clamp voltage response to current step injection (130 pA, representative of n = 25 for n = 7 participants). Scale, 20 mV, 200 ms. j, Example whole-cell patch-clamp recordings (120 pA steps) from pyramidal neurons obtained from a single patient pre- and post-safusidenib. k, Firing rate (20 pA steps, 1,200 ms) from (left) the neurons and patient shown in j and (right) average of all neurons (pre-safusidenib (n = 11 neurons; 4 participants) and post-safusidenib (n = 14 neurons; 3 participants), ANOVA). Mean ± s.e.m. l–n, Rheobase (t-test) (l), membrane resistance (t-test) (m) and resting membrane potential (RMP) (t-test) in neurons (n) from tissue analyzed pre- and post-safusidenib. Panels l and m show mean ± s.e.m.