Extended Data Fig. 2: Suppression of NK immunity specific to pancreas TME following T/P-induced senescence.
a-b, KPC2 PDAC or KP2 LUAD tumor cells expressing GFP were injected i.v. or orthotopically into the pancreas of 8-12 week old C57BL/6 female mice. Following tumor formation, mice were treated with vehicle (V) or combined trametinib (1 mg/kg) and palbociclib (100 mg/kg) (T/P) for 2 weeks. Flow cytometry analysis of NK cell numbers and degranulation in PDAC (PIP, PIL) (a) and LUAD-derived tumors (LIL, LIP) (b) are shown (PIP V, n = 3; PIP T/P, n = 4; PIL V and PIL T/P, n = 8; LIL V, n = 6; LIL T/P, n = 7; LIP V and T/P, n = 9 mice). c, Flow cytometry analysis of NK cell numbers and degranulation in spleens of mice with KPC1-derived PIP tumors treated as in (a) (n = 5 mice per group). d, Kaplan-Meier survival curve of mice with KPC2-derived PIP tumors treated with vehicle, combined trametinib (1 mg/kg) and palbociclib (100 mg/kg), and/or depleting antibodies against NK1.1 (PK136; 250 μg) or CD8 (2.43; 200 μg) (V, n = 5; T/P, T/P + αNK1.1 and T/P + αCD8, n = 8 mice). e, IVIS images showing luciferase signaling in KPC1-derived PIL tumors following treatment as in (a). Right, quantification of total luminescence in the thoracic region (V, n = 5; T/P and T/P + αNK1.1, n = 8 mice). f, Waterfall plot of the response of KPC1-derived PIP tumors following 2 week treatment with vehicle, combined trametinib (1 mg/kg) and palbociclib (100 mg/kg), and/or an NK1.1 depleting antibody (PK136; 250 μg) (V and T/P, n = 5; T/P + αNK1.1, n = 6 mice). g, Waterfall plot of the response of KPC2-derived PIP tumors following 2 week treatment with vehicle, combined trametinib (1 mg/kg) and palbociclib (100 mg/kg), and/or an NK1.1 (PK136; 250 μg) or CD8 (2.43; 200 μg) depleting antibody (V, n = 5; T/P, n = 7; T/P + αNK1.1 and T/P + αCD8, n = 8 mice). h, Kaplan-Meier survival curve of mice with KPC1-derived PIL tumors treated with vehicle, combined trametinib (1 mg/kg) and palbociclib (100 mg/kg), and/or depleting antibodies against CD8 (2.43; 200 μg) or CD4 (GK1.5; 200 μg) (V, n = 5; T/P, T/P + αCD4 and T/P + αCD8, n = 7 mice). i, Flow cytometry analysis of CD4+ and CD8+ T cell numbers and degranulation in KPC1 PDAC (PIP, PIL, PILiver) and KP1 LUAD-derived tumors (LIL, LIP, LILiver) grown in different organs and treated as in (a) (PIP V and T/P, n = 5 or 10; PIL V, n = 5 or 9; PIL T/P, n = 5 or 10; LIL V, n = 3; LIL T/P, n = 5; LIP V, n = 5 or 13; LIP T/P, n = 6 or 15; PILiver V, n = 9; PILiver T/P, n = 10; LILiver V, n = 8; LILiver T/P, n = 10 mice). Data represents pool of 3 independent experiments. P values in a-c, e-g, and i were calculated using two-tailed, unpaired Student’s t-test, and those in d and h were calculated using log-rank test. Error bars, mean ± SEM.
