The Combination of MEK Inhibitor With Immunomodulatory Antibodies Targeting Programmed Death 1 and Programmed Death Ligand 1 Results in Prolonged Survival in Kras/p53-Driven Lung Cancer.
Lee Jong Woo,Zhang Yu,Eoh Kyung Jin,Sharma Roshan,Sanmamed Miguel F,Wu Jenny,Choi Justin,Park Hee Sun,Iwasaki Akiko,Kaftan Edward,Chen Lieping,Papadimitrakopoulou Vali,Herbst Roy S,Koo Ja Seok
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
INTRODUCTION:This study aimed to characterize the tumor-infiltrating immune cells population in Kras/tumor protein 53 (Trp53)-driven lung tumors and to evaluate the combinatorial antitumor effect with MEK inhibitor (MEKi), trametinib, and immunomodulatory monoclonal antibodies (mAbs) targeting either programmed death -1 (PD-1) or programmed cell death ligand 1 (PD-L1) in vivo. METHODS:Trp53;Kras;Rosa26 (PKL) genetically engineered mice were used to develop autochthonous lung tumors with intratracheal delivery of adenoviral Cre recombinase. Using these tumor-bearing lungs, tumor-infiltrating immune cells were characterized by both mass cytometry and flow cytometry. PKL-mediated immunocompetent syngeneic and transgenic lung cancer mouse models were treated with MEKi alone as well as in combination with either anti-PD-1 or anti-PD-L1 mAbs. Tumor growth and survival outcome were assessed. Finally, immune cell populations within spleens and tumors were evaluated by flow cytometry and immunohistochemistry. RESULTS:Myeloid-derived suppressor cells (MDSCs) were significantly augmented in PKL-driven lung tumors compared to normal lungs of tumor-free mice. PD-L1 expression appeared to be highly positive in both lung tumor cells and, particularly MDSCs. The combinatory administration of MEKi with either anti-PD-1 or anti-PD-L1 mAbs synergistically increased antitumor response and survival outcome compared with single-agent therapy in both the PKL-mediated syngeneic and transgenic lung cancer models. Theses combinational treatments resulted in significant increases of tumor-infiltrating CD8 and CD4 T cells, whereas attenuation of CD11b/Gr-1 MDSCs, in particular, Ly6G polymorphonuclear-MDSCs in the syngeneic model. CONCLUSIONS:These findings suggest a potential therapeutic approach for untargetable Kras/p53-driven lung cancers with synergy between targeted therapy using MEKi and immunotherapies.
OCT4&SOX2-specific cytotoxic T lymphocytes plus programmed cell death protein 1 inhibitor presented with synergistic effect on killing lung cancer stem-like cells in vitro and treating drug-resistant lung cancer mice in vivo.
Zhang Xueyan,Hu Fang,Li Changhui,Zheng Xiaoxuan,Zhang Bo,Wang Huimin,Tao Guangyu,Xu Jianlin,Zhang Yanwei,Han Baohui
Journal of cellular physiology
This study aimed to investigate the synergistic effect of octamer-binding transcription factor 4 and sex determining region Y-box 2 (OCT4&SOX2)-specific cytotoxic T lymphocytes (CTLs) and programmed cell death protein 1 (PD-1) inhibitor on killing lung cancer stem-like cells (LCSCs) and their efficacy in treating drug-resistant lung cancer (DRLC) mice. OCT4&SOX2-specific CTLs and PD-1 inhibitor with differed doses were applied to treat PC9 cells and PC9 LCSCs. Cell counting kit-8 (CCK8) assay and flow cytometry (FCM) assay with carboxyfluorescein diacetate/succinimidyl ester staining target cells before treatment and propidium iodide (PI) staining dead cells after treatment were conducted to detect the cytotoxic activity. DRLC mice were constructed by injection of PC9 LCSCs suspension and Matrigel into left lung of SD mice. DRLC mice were randomly divided into five groups: control group, cytomegalovirus (CMV) pp65 CTLs group, OCT4&SOX2 CTLs group, PD-1 inhibitor group, and OCT4&SOX2 CTLs + PD-1 inhibitor group. In vitro, both CCK8 assay and FCM assay disclosed that OCT4&SOX2-specific CTLs plus PD-1 inhibitor presented with elevated cytotoxic activity on PC9 cells and PC9 LCSCs. In vivo, tumor volume and tumor weight were decreased, while tumor necrosis and tumor apoptosis were increased in OCT4&SOX2 CTLs group than CMV pp65 CTLs group and control group, and in OCT4&SOX2 CTLs + PD-1 inhibitor group than OCT4&SOX2 CTLs group and PD-1 inhibitor group. In addition, CD8 expression was increased while OCT4&SOX2 expressions were decreased in OCT4&SOX2 CTLs + PD-1 inhibitor group than OCT4&SOX2 CTLs group and PD-1 inhibitor group. In conclusion, OCT4&SOX2-specific CTLs and PD-1 inhibitor presented with the synergistic effect on killing LCSCs in vitro and treating DRLC mice in vivo.
[Establishment of a dual-fluorescence-traced lung cancer subcutaneous transplantation model in nude mice].
Fan H L,Liu M Z,Min J T,Li H J,Yang X H,Li Y H,Li Z Z
Zhonghua zhong liu za zhi [Chinese journal of oncology]
To establish a nude mouse model of subcutaneous lung cancer using dual fluorescence reporting genes of luciferase (Luc) and near-infrared fluorescent protein (iRFP). The Luc and iRFP expressed lentiviral vector was constructed by Gateway method. After verified by sequencing, the lentivirus particle was prepared and infected into lung cancer A549 cells. Successfully infected A549 (mA549) cells were selected by puromycin and amplified. The expression of Luc and iRFP were observed under fluorescence microscope, and the expression of c-Met protein on the cell surface was detected by immunofluorescence. Twelve female nude mice were randomly divided into 2 groups, 6 in each group. A549 and mA549 cells were inoculated subcutaneously into the right forelimb of nude mice. The growth and fluorescence expression of the tumor were observed by in vivo imaging. The tumor formation was evaluated by hematoxylin-eosin (HE) staining and immunohistochemistry. The Luc and iRFP stably expressed mA549 cell line was successfully constructed. The expressions of iRFP and Luc in mA549 cells were observed under fluorescence microscope. The results of immunofluorescence showed that c-Met protein expressed in both A549 cells and mA549 cells. The growth period of mA549 xenograft in nude mice was moderate and the tumorigenesis rate was 100%. The growth trend of mA549 cells in vivo was not significantly different from that of A549 cells (>0.05). HE staining and immunohistochemistry results showed that the tumor issues displayed typical histopathological features of tumor. Immunohistochemistry results showed that both A549 and mA549 tumors expressed c-Met protein. A stable, real-time monitoring model of iRFP-Luc-A549 lung cancer cell xenograft in nude mice was successfully constructed.