Targeting adhesion signaling in KRAS, LKB1 mutant lung adenocarcinoma
Melissa Gilbert-Ross, … , Wei Zhou, Adam I. Marcus
Melissa Gilbert-Ross, … , Wei Zhou, Adam I. Marcus
Published March 9, 2017
Citation Information: JCI Insight. 2017;2(5):e90487. https://doi.org/10.1172/jci.insight.90487.
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Research Article Cell biology Oncology

Targeting adhesion signaling in KRAS, LKB1 mutant lung adenocarcinoma

Abstract

Loss of LKB1 activity is prevalent in KRAS mutant lung adenocarcinoma and promotes aggressive and treatment-resistant tumors. Previous studies have shown that LKB1 is a negative regulator of the focal adhesion kinase (FAK), but in vivo studies testing the efficacy of FAK inhibition in LKB1 mutant cancers are lacking. Here, we took a pharmacologic approach to show that FAK inhibition is an effective early-treatment strategy for this high-risk molecular subtype. We established a lenti-Cre–induced Kras and Lkb1 mutant genetically engineered mouse model (KLLenti) that develops 100% lung adenocarcinoma and showed that high spatiotemporal FAK activation occurs in collective invasive cells that are surrounded by high levels of collagen. Modeling invasion in 3D, loss of Lkb1, but not p53, was sufficient to drive collective invasion and collagen alignment that was highly sensitive to FAK inhibition. Treatment of early, stage-matched KLLenti tumors with FAK inhibitor monotherapy resulted in a striking effect on tumor progression, invasion, and tumor-associated collagen. Chronic treatment extended survival and impeded local lymph node spread. Lastly, we identified focally upregulated FAK and collagen-associated collective invasion in KRAS and LKB1 comutated human lung adenocarcinoma patients. Our results suggest that patients with LKB1 mutant tumors should be stratified for early treatment with FAK inhibitors.

Authors

Melissa Gilbert-Ross, Jessica Konen, Junghui Koo, John Shupe, Brian S. Robinson, Walter Guy Wiles IV, Chunzi Huang, W. David Martin, Madhusmita Behera, Geoffrey H. Smith, Charles E. Hill, Michael R. Rossi, Gabriel L. Sica, Manali Rupji, Zhengjia Chen, Jeanne Kowalski, Andrea L. Kasinski, Suresh S. Ramalingam, Haian Fu, Fadlo R. Khuri, Wei Zhou, Adam I. Marcus

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Figure 3

Treatment with a FAK inhibitor results in multiple therapeutic responses in Kras and Lkb1 mutant lung adenocarcinoma in vivo.

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Treatment with a FAK inhibitor results in multiple therapeutic responses...
(A) (Top panel) Dorsal bioluminescent images (BLI) of 2 KrasG12D/+ Lkb1fl/fl Rosa-luc mice induced with lenti-Cre (KLLLenti) on day 0 and imaged 10 weeks post-infection (wpi). Regions of interest (ROIs) indicate total flux in photons/second. (Bottom panel) H&E-stained primary lung tumors from mice pictured above (50×). (B) Graphic of preclinical trial design. (C) BLI images of vehicle- and GSK6098-treated mice after 5-week treatment protocol (Nec, necropsy). (D) Total flux (photons/second) at enrollment (Enr) vs. necropsy (Nec) for vehicle- (n = 10) and GSK6098-treated mice (n = 9) in 5-week treatment trial. Each box represents an individual mouse pictured in C; one-way ANOVA with Tukey’s multiple comparisons test, ****P < 0.0001, *P < 0.05). (E) Mean tumor burden (μm2) in vehicle- (n = 10) vs. GSK6098-treated mice (n = 9); 2-tailed t test, ****P < 0.0001. (Bottom panel) Representative images of lung tumors in vehicle- vs. GSK6098-treated mice. Scale bar: 3 mm. (F) Mean invasive burden in vehicle- (n = 10) vs. GSK6098-treated mice (n = 9); 2-tailed t test, P = 0.1089. (Bottom panel) Representative images of invasive tumors (outlined in yellow) in vehicle- vs. GSK6098-treated mice (40×; Nanozoomer; 8× digital zoom). (G) SHG imaging of collagen (white) in vehicle- vs. GSK6098-treated KLLLenti lung tumors. Images are 2 independent stage-matched (Inv Adc) fields per treatment group. (H) Kaplan Meier analysis of days from enrollment to the onset of clinical symptoms of chronically treated vehicle (n = 12) vs. GSK6098 (n = 11) KLLLenti mice. Wilcoxon test, P = 0.0647. (I) Representative lymph nodes from chronically treated vehicle (3 independent lymph nodes from a single mouse pictured [numbered 1–3] vs. GSK6098 [1 positive lymph node from a single mouse pictured]). Yellow outlines mark metastatic tumor cells. Scale bar: 2 mm. (J) Western blot analysis of the FAK/Src pathway in vehicle- vs. GSK6098-treated lung lobes. Numbers in parentheses indicate fold change in the vehicle/GSK6098 mean signal intensity ratio. SE, short exposure; LE, long exposure.