Science Advances

Supplementary Materials

This PDF file includes:

  • Fig. S1. Construction of a three-dimensional fluorescent cell barcoding matrix for multiplexing of 80 cellular treatments.
  • Fig. S2. Gating strategies for the functional analysis of 80 barcoded T cell populations.
  • Fig. S3. MFIs and CVs across 80 barcoded T cell populations for each functional fluorescence channel.
  • Fig. S4. Z factor analyses across 80 barcoded T cell populations for each functional fluorescence channel.
  • Fig. S5. Reproducibility across time and independent PBMC cohorts.
  • Fig. S6. Stain indices of antibody clones against T cell signaling epitopes used in the KP experiments.
  • Fig. S7. Kinetic induction of cell signaling responses across the ligand and epitope array (n = 5460 nodes; i.e., ligand-epitope combinations).
  • Fig. S8. Distribution of FCs for T cell signaling responses across time points.
  • Fig. S9. Dynamic regulation of JAK/STAT T cell signaling across time course.
  • Fig. S10. Gating strategies for the functional analysis of 64 barcoded T cell populations.
  • Fig. S11. Gating strategy for cell viability and immunophenotyping.
  • Fig. S12. Clinical response to antipsychotic treatment with olanzapine in patients with SCZ at 6 weeks.
  • Fig. S13. Altered T cell signaling nodes (ligand-epitope combinations) in pretreatment SCZ versus control and pretreatment versus posttreatment SCZ comparisons.
  • Fig. S14. Association between the drug target response to thapsigargin at PLC-γ1 in SCZ and the genome-wide significant SCZ risk SNP rs4766428 in the ATP2A2 gene.
  • Fig. S15. Normal regulatory response at PLC-γ1 to calcium release from the endoplasmic reticulum and hypothetical mechanism of action in SCZ, based on the altered response to thapsigargin at PLC-γ1 in T cells from patients with SCZ.
  • Fig. S16. Gating strategies for the functional analysis of PLC-γ1 expression in four barcoded T cell populations.
  • Fig. S17. Thapsigargin dose response at PLC-γ1.
  • Fig. S18. Selective potentiation of PLC-γ1 response in the presence of thapsigargin.
  • Fig. S19. Tanimoto structural similarity clustering of calcium channel blocker, antipsychotic, corticosteroid, and antibiotic compounds used in PLC-γ1 dose-response validation and selectivity testing.
  • Fig. S20. Validation and selectivity testing of calcium channel blocker, antipsychotic, corticosteroid, antibiotic, and other drug classes at PLC-γ1.
  • Fig. S21. Validation of top drug candidates in the SH-SY5Y neuronal cell line.
  • Fig. S22. Correlation of ex vivo drug-target activity with in vivo efficacy in the CV study.
  • Fig. S23. Potentiation of thapsigargin/PLC-γ1 dose response at 30 min by top drug candidates from the screening phase at 10 μM concentration in PBMCs from drug-naïve patients with SCZ.
  • Table S1. Antibodies used to detect intracellular cell signaling epitopes and PBMC subtypes.
  • Table S2. Ligands used to stimulate/alter cell signaling dynamics in PBMCs.
  • Table S3. Activity of ligands across the time course.
  • Table S4. Activity of epitopes across the time course.
  • Table S5. Demographic characteristics and matching of PBMC donors used in the TI study.
  • Table S6. Altered ligand responses at T cell signaling epitopes in healthy control versus pretreatment SCZ and pretreatment versus posttreatment SCZ comparisons.
  • Table S7. Altered basal expression of T cell signaling epitopes in pretreatment versus posttreatment SCZ comparison.
  • Table S8. Extended FDA-approved compound library.
  • Table S9. Extended FDA-approved library screening of compounds which selectively potentiate the PLC-γ1 response in the presence of 0.5 μM thapsigargin.
  • Table S10. Demographic characteristics and matching of PBMC donors used in the CV study.
  • Table S11. Prediction of in vivo response to treatment from ex vivo treatment activity.

Download PDF

Files in this Data Supplement: