Science - USA (2018-12-21)

Ruscettiet al.,Science 362 , 1416–1422 (2018) 21 December 2018 2of7

G

E

D

A

B

Palbociclib (μM)

0.1 0.3 0.5 1

A549, trametinib (nM)

1 5 10 25 100

Ribociclib (μM)

A549, trametinib (nM)

0.1 0.3

0.5 1

1 5 10 25 100

Abemaciclib (μM)

A549, trametinib (nM)

0.1

0.3

0.5

1

1 5 10 25 100

F

H

0

50

100

0

0

0

P <0.0001

Vehicle

Trametinib

Palbociclib

Combo

Survival (%)

10 15 20 25 30

Time (d)

3.99 5.14 5.32

Vehicle Trametinib Palbociclib Combo

3.98 5.14 5.32

NK1.1-BV605

(^2) 10 10 0 10 10
CD3-BV650
0
10
10
10
10
2
9.44
3
3 4 5
5
4

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---

**

---

---

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**

• 
  


• 
  

  CD107a

  
  

• 
  


• cells) (%)
  5
  10
  15
  20
  25
  (per 10
  3 GFP


• tumor cells) (%)
  100
  200
  300
  400
  2
  4
  6
  8
  10
  NK1.1


• (of CD45


• cells) (%)
  0 0 0
  NK1.1


• 
  

  (of NK1.1
  Vehicle Tramet Palbo Combo Vehicle Tramet Palbo Combo Vehicle Tramet Palbo Combo
  Time (d)
  10 20 30
  0
  50
  100
  Survival (%)
  Isotype
  Combo + NK1.1
  Combo + Isotype
  NK1.1
  D
  Mouse KP C57BL/6
  (KrasG12D/+;p53-/-)
  Trametinib
  Palbociclib
  MSCV-Luc-GFP
  P = 0.0001
  9.44
  Time (d)
  Vehicle
  Trametinib (lo)
  Combo (lo)
  (^0 0 10 20 30 40 50)
  500
  1000
  1500
  Trametinib (hi)
  Combo (hi) n.s.

  
  

• 
  

  ---

  
  

  ---

  
  

  Palbociclib ****
  C
  Volume (mm
  3 )
  Time (d)
  Volume (mm
  3
  0
  500
  1000
  1500
  2000
  2500
  0 7 14 21 28 35 42 49
  )

  
  

  ---

  
  

  Vehicle
  Trametinib
  Combo
  Palbociclib

  
  


• 
  

  ---

  
  

  Fig. 1. NK cell immunity is required for the efficacy of combination
  MEK and CDK4/6 inhibitor therapy.(A) Clonogenic assay of
  A549 lung cancer cells treated with MEK (trametinib) and/or various
  CDK4/6 inhibitors (palbociclib, ribociclib, abemaciclib); representative
  of three biological replicates. (B) Tumor volumes of mice bearing
  KRAS-mutant MSK-LX27 PDX lung tumors treated with vehicle, trametinib
  (3 mg/kg body weight), palbociclib (150 mg/kg body weight), or both
  in combination (Combo) for indicated times (n=5micepergroup).
  (C) Tumor volumes of mice bearing KRAS-mutant MSK-LX68 PDX lung
  tumors treated with vehicle, trametinib [1 mg/kg (lo) or 3 mg/kg (hi)
  body weight], palbociclib (150 mg/kg body weight), or both in
  combination for indicated times (n= 8 mice per group). n.s., not
  significant. (D) Syngeneic KP transplant lung cancer model. (E)Kaplan-
  Meier survival curve of KP transplant mice treated with vehicle,
  trametinib (1 mg/kg body weight), palbociclib (100 mg/kg body
  weight), or both in combination (n≥8 per group) (log-rank test).
  (F) Representative flow cytometry plots of NK cell populations
  in lung tumors from KP transplant mice treated for 1 week as in (E).
  (G) Percentage of NK cells within the CD45+population (left), total NK
  cells relative to tumor cell number (middle), and percentage of CD107a+
  degranulating NK cells (right) (n≥4 mice per group). Palbo, palbociclib;
  Tramet, trametinib. (H) Kaplan-Meier survival curve of KP transplant
  mice treated with vehicle or combined trametinib (1 mg/kg body
  weight) and palbociclib (100 mg/kg body weight) and either an isotype
  control antibody (C1.18.4) or NK1.1-depleting antibody (PK136) (n≥ 8
  per group) (log-rank test). (B and C) Two-way ANOVA. (G) One-way
  ANOVA. Error bars, mean ± SEM. P<0.05,P<0.01,P<0.001,
  ****P< 0.0001.
  RESEARCH | REPORT
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