New syngeneic inflammatory‐related lung cancer metastatic model harboring double KRAS/WWOX alterations

Abstract

New mouse models with specific drivers of genetic alterations are needed for preclinical studies. Herein, we created and characterized at the genetic level a new syngeneic model for lung cancer and metastasis in Balb-c mice. Tumor cell lines were obtained from a silica-mediated airway chronic inflammation that promotes tumorigenesis when combined with low doses of N-nitrosodimethylamine, a tobacco smoke carcinogen. Orthotopic transplantation of these cells induced lung adenocarcinomas, and their intracardiac injection led to prominent colonization of various organs (bone, lung, liver and brain). Driver gene alterations included a mutation in the codon 12 of KRAS (G-A transition), accompanied by a homozygous deletion of the WW domain-containing oxidoreductase (WWOX) gene. The mutant form of WWOX lacked exons 5-8 and displayed reduced protein expression level and activity. WWOX gene restoration decreased the in vitro and in vivo tumorigenicity, confirming the tumor suppressor function of this gene in this particular model. Interestingly, we found that cells displayed remarkable sphere formation ability with expression of specific lung cancer stem cell markers. Study of non-small-cell lung cancer patient cohorts demonstrated a deletion of WWOX in 30% of cases, with significant reduction in protein levels as compared to normal tissues. Overall, our new syngeneic mouse model provides a most valuable tool to study lung cancer metastasis in balb-c mice background and highlights the importance of WWOX deletion in lung carcinogenesis.Lung cancer remains the leading cause of cancer mortality, being responsible of about 1.3 million annual deaths worldwide. 1 Non-small-cell lung cancer (NSCLC), the most frequent lung cancer type, is characterized by defined genetic burden. The most commonly mutated genes for adenocarcinomas include the tumor suppressor genes p53, STK11/LKB1 as well as EGFR, KRAS and PIK3CA oncogenes. In addition to BRAF and NRAS mutations, ALK, RET and ROS1 translocations also stand for a considerable part of the mutational spectrum. 2 Hallmarks for squamous cell carcinoma consist in PTEN, PIK3CA and DDR2 mutations, along with amplifications of the PDGFRA, FGFR1 and SOX2 genes. Currently, EGFR-TK inhibitors are used routinely for lung cancer patients showing well characterized somatic mutations. Due to the manifest complexity of the disease and the development of new targeted therapies, future treatment strategies

A genome-wide Drosophila epithelial tumorigenesis screen identifies Tetraspanin 29Fb as an evolutionarily conserved suppressor of Ras-driven cancer

“…Interestingly, several genes and pathways identified in our primary screen have an effect on oxidative stress levels or proper mitochondrial function: detoxifying enzymes such as Gutathione S Transferases or Wwox [ 64 ], and many mitochondrial genes, the loss of which can potentially impede proper mitochondrial function and generate oxidative stress. Interestingly, we have recently shown that increased oxidative stress levels induced by autophagy impairment can indeed cooperate with oncogenic Ras by triggering the JNK stress response pathway [ 28 ].…”

A Combined PD-1/C5a Blockade Synergistically Protects against Lung Cancer Growth and Metastasis

“…LLC cells were obtained from the American Type Culture Collection. The Lacun3 cell line was previously established by our group from a chemically induced lung adenocarcinoma and stably transfected with the luciferase gene (24). Nontransduced Lacun3 cells were used for the survival experiment.…”

“…The metastasis model based on LLC cells was performed by intravenous injection of 1 × 10 6 cells into the tail vein of C57BL/6J mice. The multiorgan metastatic potential of luciferase-transfected Lacun3 cells was evaluated after inoculation of 2 × 10 5 cells into the left cardiac ventricle of BALB/c mice as previously described (24,25). In these two metastasis models, treatments were performed as described above, except for the injections of anti-PD-1 blocking antibody, which were performed at days 3, 7, and 10 after cell inoculation.…”

Sphere-derived tumor cells exhibit impaired metastasis by a host-mediated quiescent phenotype

“…To determine the prometastatic activity of TSC, we used two different cell models. First, murine Lacun.3 cells were obtained from a chemically-induced lung adenocarcinoma developed in mice; it is an aggressive cell line that forms spontaneous metastases in different organs [ 39 ]. Second, we used the human lung cancer cell line H460 that develops spontaneous bone metastases in athymic nude mice [ 40 ].…”

“…The mouse lung adenocarcinoma cell line, Lacun.3, was isolated and characterized at the University of Navarra (Pamplona, Spain) from a combined silica and N-Nitrosodimethylamine (NDMA)-induced lung cancer model [ 39 ]. The human cell line Mai9 was also isolated and characterized by our group from a malignant pleural effusion (MPE) of a patient diagnosed with NSCLC [ 43 ].…”