Figure 2

CRIPSR-modified TIL expand to clinically relevant numbers while maintaining resistance to TGF-β signaling and TCR clonal diversity. (A) Control and TGFBR2-KO OvCa TIL expand with equal efficiency during the 14-day REP. Fold expansions ranging from 1000× to 3000× were observed across four independent patient samples. (B) Resistance of TGFBR2-KO TIL to TGF-β–mediated SMAD phosphorylation was maintained after the 14-day REP. (C) The majority of TCR clones are identified with equal abundance in control and CRISPR-modified TIL, with differentially abundant clones randomly distributed on either side of the line of frequency equity. These data indicate that CRISPR-modification does not skew the distribution of unique TCR clones in TIL samples. Data from patient #2 are presented and are representative of the results obtained from an additional three patient samples (shown in online supplemental figure S6). Left: Cas9 mock transfected TIL versus non-transfected TIL. Middle: TGFBR2-KO TIL with gRNA #3 versus non-transfected TIL. Right: TGFBR2-KO TIL with gRNA #4 versus non-transfected TIL. (D) Productive Simpson clonality is equivalent in TGFBR2-KO TIL compared with control TIL samples. ns, no significant; OvCa, ovarian cancer; REP, rapid expansion protocol; TIL, tumor-infiltrating lymphocytes.