Figure 7. Tpi1 is required for tumor growth in KP and KPL LUAD models and humanizing TPi1 regresses tumor growth and burden.

(A) Cartoon schematic depicting generated conditional genetic mouse models and subsequent derived tumor derived cell lines. (B) C57Bl/6J mice survival following tail vein injection of KPCas9 cell line (BS7341) infected with lentiviruses encoding control or targeting sgRNAs (n=5/group). (C) Subcutaneous tumor growth of KP or KPL derived tumor cell lines (BS7432) infected with lentiviruses encoding control or targeting sgRNAs 30d following engraftment in C57Bl/6J mice (n=5/group); paired t-test provided above. (D) Histogram quantifying tumor lesions per lobe in tail vein injected KPcas9 (BS7431) Tpi1 allelic variants in C57Bl/6J mice (N=5/group) and reported as the mean (−/+ s.e.m.). Statistical significance determined by two-tailed paired t-test. (E) Histogram quantifying tumor lesion diameter in tail vein injected KPcas9 (BS7431) Tpi1 allelic variants in C57Bl/6J mice (N=5/group) and reported as the mean (−/+ s.e.m.). Statistical significance determined by two-tailed paired t-test. (F) Subcutaneous tumor growth of KPCas9 derived tumor cell line (BS7432) first infected with control or Lkb1 targeting guide RNA to produce isogenic KP and KPL respectively. Derived lines were then infected with lentiviruses encoding Tpi1 sgRNA with subsequent lentiviral expression of transgenic guide RNA resistant mTpi1 WT, C21A or C21S and measured 30d following engraftment. Paired flank injections in C57Bl/6J mice were conducted (KP/KPL for each transgene) per mouse (n=5/group). (G) KLCas9v2 autochthonous tumor model survival following intratracheal administration of lentiviruses encoding cre-recombinase and control (sgNT) or targeting (sgTpi1) sgRNAs (n=12/group). Representative lung tumor burden in groups of mice at 8 weeks (sgNT and sgTpi1) and 14.5 weeks (sgTpi1) following intubation; histologic appearance of tumor lesions.