Extended Data Figure 8. Ogdh inhibition reduces tumor cell competitive fitness in vivo.

a, αKG/succinate ratio of KPC^floxRIK and KPC^R172HRIK cells expressing dox-inducible shRNAs targeting Renilla, Ogdh, and Sdha grown 4 days with or without dox. b, Schematic of in vivo competition assay. Kate positive KP^floxRIK and KP^R172HRIK cells were infected with retroviruses encoding dox inducible, GFP linked shRNAs targeting Renilla, Ogdh, or Sdha. Cells were selected for viral integration, induced with dox for 2 days, mixed with uninfected parental cellsat a ratio of 8:2 and analyzed by flow cytometry to determine initial ratio of shRNA expressing cells to uninfected cells. This cell mixture was injected orthotopically into dox fed recipient mice. After 3 weeks of tumor growth, pancreatic tumors were removed, weighed, dissociated, and analyzed by flow cytometry to measure final ratio of shRNA expressing to uninfected cells. Data are presented in Fig. 3c. c,d, Tumor mass of orthotopically injected KP^floxRIK (c) or KP^R172HRIK (d) cells expressing a mixture of shRNAs targeting Renilla, Ogdh, or Sdha from Fig. 3c. KP^floxRIK: n= 5 mice shRenilla, n=4 mice shOgdh, shSdha. KP^R172KRIK : n= 5 mice shRenilla, n=4 mice shOgdh, shSdha-1,2, n=3 mice shSdha-3. e, Representative gross images of pancreatic tumors arising in dox fed mice 3 weeks days following orthotopic transplant of KPC^floxRIK (top) and KPC^R172HRIK (bottom) cells expressing dox-inducible shRNAs targeting Renilla, Ogdh, and Sdha mixed 8:2 from Fig. 3c. a was performed once. Data are presented as mean ± SD of n=3, independently treated wells of a representative experiment with individual data points shown. Significance assessed compared to shRenilla controls by 1-way ANOVA with Dunnet’s multiple comparison post-test (c). Scale bar 1 cm.