Supplemental Methods, Tables 1 - 3, Figures 1 - 16 from Identification of Chimeric Antigen Receptors That Mediate Constitutive or Inducible Proliferation of T Cells

Table S1: Expression of c-Met and Mesothelin mRNA in activated CD4 T Cells. Table S2: Genes upregulated in CD4+ T cells of healthy human adults expressing c-Met IgG4 continuous CAR or CD19 CD8α non-continuous CAR on day 11. Table S3: Genes downregulated in CD4+ T cells of healthy human adults expressing c-Met IgG4 continuous CAR or CD19 CD8α non-continuous CAR on day 11. Figure S1. Chimeric antigen receptor constructs and relative expression levels. Figure S2. CAR T cells with constitutive proliferation retain specific cytotoxicity. Figure S3. c-Met and mesothelin expression are not detected on human CD4+ T cells. Figure S4. Supernatant from CARs displaying the growth phenotype induces activation of naïve unstimulated T cells. Figure S5.The role of fetal bovine serum in the constitutive growth of c-Met IgG4 28ζ CAR T cells. Figure S6. A) CARs with a constitutive growth phenotype display a unique gene signature. B) CAR T cells with a constitutive growth phenotype display distinct transcription factors. Figure S7. Genome-wide microarray analysis of CAR T cells with constitutive proliferation. Figure S8. Distinct gene expression signature of CAR T cells with constitutive proliferation. Figure S9. CAR T cells with constitutive proliferation have ligand-independent NFAT activation. Figure S10. Transgene expression levels are sufficient to convey the constitutive CAR growth phenotype. Figure S11. Constitutive CAR T cell proliferation results in differentiation and evolution of a distinct cell surface phenotype. Figure S12. Effects of stimulation and cell culture on differentiation of non-transduced T cells. Figure S13. Temporal patterns of telomere restriction fragment length (TRF) in continuous CAR T cells and mock transduced T cells. Figure S14. CAR T cells with a constitutive growth phenotype retain a diverse TCR Vβ repertoire. Figure S15. Engraftment and proliferation of continuous CAR T cells in NSG mice. Figure S16. PGK100 promoter results in antigen driven accumulation of CAR T cells in tissue but not blood.