Targeting adhesion signaling in KRAS, LKB1 mutant lung adenocarcinoma
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
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
View: Text | PDF
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

×

Figure 2

Lkb1 mutant tumor cells exhibit collective, FAK-dependent 3D cell invasion and collagen remodeling.

Options: View larger image (or click on image) Download as PowerPoint
Lkb1 mutant tumor cells exhibit collective, FAK-dependent 3D cell invasi...
(A) (Top) Representative brightfield images of Lkb1 WT and Lkb1-null mouse tumor cell spheroids that were embedded in a collagen I matrix and allowed to invade for 24 hours. Area in white boxes is magnified to the right. Dashed line with asterisks marks single invasive cells, and sold line marks collectively invading cells. Scale bars: 50 μm. (Bottom) Representative immunofluorescence images of pYFAK397 at the invasive front in Lkb1 WT and Lkb1-null spheroids after 48 hours of invasion into a collagen matrix. Area in white boxes is magnified to the right. Scale bars: 50 μm. (B) Representative images of spheroids formed from Lkb1 WT and Lkb1-null cells analyzed for total invasion into a collagen matrix in the presence of FAK inhibitor PF-562271 at 250 nM. DMSO was used as a vehicle control. Scale bars: 100 μm. (C) Quantification of spheroid invasion after treatment with either PF-562271 at 250 nM or GSK6098 at 2 μM. n = 4 spheroids (FAK inhibitors) or n = 7 spheroids (DMSO). Data are represented as mean ± SD. P values were calculated using 1-way ANOVA with Tukey’s multiple comparisons test, *P < 0.05 ***, P < 0.0001. (D) Second harmonic generation imaging of collagen after invasion by 3D tumor cell spheroids of the indicated genotypes. Scale bar: 50 μm. (E) Lkb1-null 3D tumor spheroids treated with vehicle (DMSO, top panels) or PF-562271 (bottom panels) embedded in a collagen matrix and visualized 24 hours later by CellTracker (tumor cell spheroids, red in merge) and second harmonic generation (SHG) to visualize collagen fibers. Scale bar: 50 μm. (F) Collagen alignment coefficient of DMSO and PF-562271-treated Lkb1-null tumor cell spheroids shown in E.