Induced Lung Cancer Research Articles

Effect of a 12-week submaximal swimming training in rats exposed to tobacco- derived nitrosamine ketone.

Background:Tobacco contains carcinogens such as NNK (nicotine-derived nitrosamine ketone) that makes induction of lung cancer by changing the stimulation of IL-10 expression. The aim of this study was to investigate changes in resting levels of IL-10 in lung tissues of rats exposed to NNK after a 12-week aerobic submaximal swimming training.Methods:For this purpose, 46 Wistar rats were randomly divided into five groups consisting training, training + NNK, NNK, saline and control. NNK-induced groups received NNK subcutaneously one day per week at a rate of 12/5 mg per kg body weight and the training groups performed submaximal swimming training for 12 weeks. The levels of IL-10 in homogenized lung tissue were measured by ELISA.Results:Findings indicated that a period of swimming training increased the IL-10 levels significantly in lung tissue of training group when compared to control (P=0.00) and NNK groups (P=0.00). Also, a significant increase of IL-10 level was observed in exercise + NNK group when compared to NNK group (p≤0/02). Furthermore, it was observed that IL-10 levels of NNK group had a significant decrease when compared to training group (P=0/00), training + NNK group (p≤0/02), but had insignificant increase when compared to saline group (p≤0/85).Conclusions:Generally, it could be confirmed that regular submaximal aerobic training plays an important role in the inhibition of the effects of lung inflammation induced by NNK via increasing IL-10 activity.

USP24 Induces IL-6 in Tumor-associated Microenvironment through Stabilizing p300 and beta-TrCP to Promote the Malignancy of Lung Cancer

Lung cancer is the leading cause of cancer mortality worldwide. In particular,tumor metastasis is the most life-threatening event of lung cancer patients. More and more effective drugs for clinical use in cancer therapy have been developed up to now. However,the drug resistance is always occurred after drug treatment for a long time. Therefore,how to conquer the drug resistance during drug treatment is an important issue. Our preliminary result show that over expression of ubiquitin-specific peptidase 24 (USP24),a deubiquitinase,causes the cell apoptosis and abnormal mitosis progression,implying that USP24 might be involved in genomic instability. Therefore,we hypothesize that USP24 might induce genomic instability and drug resistance through mediating DNA damage repair activity. Our previous study indicated that increase in USP24 level in lung cancer facilitates lung cancer metastasis. Our recent study found that higher USP24 level was found in tumor-associated microenvironment (TAM) than surrounding lung cancer cells in lung cancer patient specimens. USP24 level was significantly increased in M2-macrophages. Knockdown of USP24 in M2-macrophages decreased the migratory and chemotactic activity of lung cancer cells and angiogenetic ability of human umbilical vein endothelial cells (HUVEC) induced by M2-derived conditioned medium. In vivo experiments also showed USP24 knockdown decreased M2-macrophages induced lung cancer cells metastasis and angiogenesis. IL-6 mRNA and protein levels were significantly decreased in USP24 knockdown M2-macrophages and lung cancer cells. IL-6 replenishment can rescue migratory and chemotactic ability of conditioned medium derived from USP24 knockdown M2-macrophages,suggesting that increase in USP24 in M2-macrophages induce the metastasis activity of lung through promoting IL-6 expression. To address the molecular mechanism of how USP24 regulated IL-6 in M2-macrophage and lung cancer cells, we found that USP24 stabilized p300, thereby increased the transcriptional activity of IL-6. USP24 could also decreased DNMT1 and Ikapa-B levels through stabilizing beta-TrCP and resulted in upregulated IL-6. Metastatic cytokines in tumor-associated microenvironment are important for cancer progression,in this study,we provide the direct evidence to support that USP24 increased in M2-macrophages and lung cancer cells positive regulates the metastasis of lung cancer through enhancing the IL-6 expression, which might be beneficial for the drug development for cancer therapy in the future.

LncRNA DANCR Promotes Lung Cancer by Sequestering miR-216a.

Background:Long noncoding RNAs (lncRNAs) are a new class of cancer regulators. Here, we aimed to investigate the diagnostic and therapeutic values of an lncRNA, differentiation antagonizing noncoding RNA (DANCR), in lung cancer.Methods:Real-time polymerase chain reaction was used to compare DANCR levels in normal and cancerous lung tissues as well as lung cancer cells. Lentiviral transduction was used to induce DANCR overexpression or silencing in vitro, followed by monitoring cell proliferation, colony formation, and changes in microRNA-216a (miR-216a) expression. DANCR-specific small hairpin RNA transduction was used to establish cells with stable DANCR knockdown, and silenced cells were used to initiate lung tumor xenografts, followed by monitoring tumor growth.Results:DANCR upregulation was seen in lung cancer, particularly in high-grade lung cancer tissues and aggressive cancer cells. Ectopic DANCR expression induced lung cancer cell proliferation and colony formation, whereas DANCR silencing induced opposing effects. The miR-216a level in cancer cells was negatively correlated with DANCR expression. The DANCR knockdown reduced the growth of tumor xenografts in vivo.Conclusion:DANCR upregulation is a potential indicator of aggressive lung cancer. Silencing of DANCR has great potential as a potent therapeutic strategy in lung cancer.

Association between lung cancer risk and inorganic arsenic concentration in drinking water: a dose-response meta-analysis.

High dose arsenic in drinking water (≥100 μg L-1) is known to induce lung cancer, but lung cancer risks at low to moderate arsenic levels and its dose-response relationship remains inconclusive. We conducted a systematic review of cohort and case-control studies that quantitatively reported the association between arsenic concentrations in drinking water and lung cancer risks by searching the PubMed database till June 14, 2018. Pooled relative risks (RRs) of lung cancer associated with full range (10 μg L-1-1000 μg L-1) and low to moderate range (<100 μg L-1) of water arsenic concentrations were calculated using random-effects models. A dose-response meta-analysis was performed to estimate the pooled associations between restricted cubic splines of log-transformed water arsenic and the lung cancer risks. Fifteen studies (9 case-control and 6 cohort studies) involving a total of 218 481 participants met the inclusion criteria. Meta-analysis identified significantly increased risks of lung cancer on exposure to both full range (RR = 1.21; 95% confidence interval [CI] = 1.05-1.37; heterogeneity I 2 = 54.3%) and low to moderate range (RR = 1.18; 95%CI = 1.00-1.35; I 2 = 56.3%) of arsenic-containing water. In the dose-response meta-analysis of eight case-control studies, we found no evidence of non-linearity, although statistical power was limited. The corresponding pooled RRs and their 95%CIs for exposure to 10 μg L-1, 50 μg L-1, and 100 μg L-1 water arsenic were 1.02 (1.00-1.03), 1.10 (1.04-1.15), and 1.20 (1.08-1.32), respectively. We provide evidence on the association between increased lung cancer risks and inorganic arsenic in drinking water across low, moderate and high levels. Minimizing arsenic levels in drinking water may be of public health importance.

Volatile oil from alpinia officinarum promotes lung cancer regression in vitro and in vivo.

The anti-lung cancer activity of volatile oil from Alpinia officinarum (VOAO) and the underlying mechanism has not been studied. Herein, VOAO was extracted by steam distillation and its components were identified by GC-MS analysis. Cells viability was measured by an MTT assay and the cell survival capacity was tested via a colony formation assay. Apoptosis cells were detected using the Annexin V-FITC/PI method. Hoechst 33342 cell unclear staining was employed to evaluate the nuclear morphology change. The mitochondrial membrane potential was detected by a JC-1 staining assay. Bcl-2, Mcl-1 and cleaved caspase-3 proteins were quantified by immune blotting. Furthermore, VOAO anti-cancer activity was evaluated on an A549 cell xenograft nude mice model. Our results have revealed that VOAO could inhibit the cell viability of lung carcinoma cells and the colony formation capacity. VOAO downregulates Bcl-2 and Mcl-1 and triggers dysfunction of the mitochondrial membrane potential. VOAO further activates caspase-3 cleavage and induces lung cancer cells apoptosis. In addition, VOAO administration significantly suppresses lung cancer growth in xenograft mice without obvious hepatotoxicity. We conclude that VOAO could be an effective, low cytotoxicity natural component for treatment of lung carcinoma.

ITRAQ-Based Proteomics Analysis of Triptolide On Human A549 Lung Adenocarcinoma Cells

Background/Aims: Triptolide (TP) is a diterpenoid triepoxide extracted from the traditional Chinese medical herb Tripterygium wilfordii that exerts prominent broad-spectrum anticancer activity to repress proliferation and induce cancer cell apoptosis through various molecular pathways. We previously observed that TP inhibits the progression of A549 cells and pancreatic cancer cells (PNCA-1) in vitro. However, the complex molecular mechanism underlying the anticancer activity of TP is not well understood. Methods: To explore the molecular mechanisms by which TP induces lung cancer cell apoptosis, we investigated changes in the protein profile of A549 cells treated with TP using a proteomics approach (iTRAQ [isobaric tags for relative and absolute quantitation] combined with NanoLC-MS/MS [nano liquid chromatography-mass spectrometry]). Changes in the profiles of the expressed proteins were analyzed using the bioinformatics tools OmicsBean and the Kyoto Encyclopedia of Genes and Genomes (KEGG) and were verified using western blotting. Apoptosis and cell cycle effects were analyzed using flow cytometry. Results: TP induced apoptosis in A549 cells and blocked A549 cells at the G2/M phase. Using iTRAQ technology, we observed 312 differentially expressed proteins associated in networks and implicated in different KEGG pathways. Gene Ontology (GO) analysis showed the overviews of dysregulated proteins in the biological process (BP), cell component (CC), and molecular function (MF) categories. Moreover, some candidate proteins involved in PARP1/AIF and nuclear Akt signaling pathways or metastasis processes were validated by western blotting. Conclusion: TP exerted anti-tumor activity on non-small cell lung cancer (NSCLC) A549 lung adenocarcinoma cells by dysregulating tumor-related protein expression. Herein, we provide a preliminary study of TP-related cytotoxicity on A549 cells using proteomics tools. These findings may improve the current understanding of the anti-tumor effects of TP on lung cancer cells and may reveal candidate proteins as potential targets for the treatment of lung cancer.

Snail1-expressing cancer-associated fibroblasts induce lung cancer cell epithelial-mesenchymal transition through miR-33b.

Lung cancer has a high propensity for metastasis. Cancer-associated fibroblasts (CAFs) are the main type of stromal cells in cancer tissue, are activated by tumor cells, and play a significant role in tumor development. However, whether CAFs induce lung cancer cell metastasis, as well as pathway involved in CAF-induced lung cancer cell metastasis, is uncertain. Snail1 is a transcriptional factor whose expression in the stroma is associated with lower survival rates in patients with cancer. However, how Snail1 regulates the crosstalk between stromal cells and tumor cells when it is expressed in the stroma has not been determined. Altered microRNA (miRNA) expression is correlated with lung cancer metastasis. Our previous study of microRNAs showed that miR-33b levels were clearly reduced in lung cancer cell lines and lung cancer tissues, and miR-33b suppressed tumor cell epithelial-mesenchymal transition (EMT) when its expression was elevated. In this study, we found that co-culturing CAFs with lung cancer cells induced miR-33b downregulation and promoted epithelial cells EMT. Moreover, we found that miR-33b overexpression in lung cancer cells counteracted CAF-induced EMT. Interestingly, Snail1 expression in fibroblasts activate the inductive effects of CAFs on lung cancer cell EMT. Hence, understanding the molecular mechanism underlying the communication between stromal cells and tumor cells mediated by miR-33b may lead to the identification of novel targets for the treatment of lung cancer. Additionally, understanding the role of Snail1 driving CAFs to induce lung cancer cell EMT may provide with a new perspective on the treatment of lung cancer.

The effects of NRF2 modulation on the initiation and progression of chemically and genetically induced lung cancer

Targeting the transcription factor NRF2 has been recognized as a feasible strategy for cancer prevention and treatment, but many of the mechanistic details underlying its role in cancer development and progression are lacking. Therefore, careful mechanistic studies of the NRF2 pathway in cancer initiation and progression are needed to identify which therapeutic avenue-activation or inhibition-is appropriate in a given context. Moreover, while numerous reports confirm the protective effect of NRF2 activation against chemical carcinogenesis little is known of its role in cancer arising from spontaneous mutations. Here, we tested the effects of NRF2 modulation (activation by sulforaphane or inhibition by brusatol) in lung carcinogenesis using a chemical (vinyl carbamate) model in A/J mice and a genetic (conditional KrasG12D oncogene expression, to simulate spontaneous oncogene mutation) model in C57BL/6J mice. Mice were treated with NRF2 modulators before carcinogen exposure or KrasG12D expression to test the role of NRF2 in cancer initiation, or treated after tumor development to test the role of NRF2 in cancer progression. Lung tissues were analyzed to determine tumor burden, as well as status of NRF2 and KRAS pathways. Additionally, proliferation, apoptosis, and oxidative DNA damage were assessed. Overall, NRF2 activation prevents initiation of chemically induced cancer, but promotes progression of pre-existing tumors regardless of chemical or genetic etiology. Once tumors are initiated, NRF2 inhibition is effective against the progression of chemically and spontaneously induced tumors. These results have important implications for NRF2-targeted cancer prevention and intervention strategies.

143 - Benzo(a)Pyrene Induced Lung Carcinogenesis in Mice: Plausible role of CEA

Background Lung cancer is identified as the most widespread type of cancer with high mortality rate. Despite the increase in early diagnosis and treatments, most patients have a poor prognosis, with an overall very low 5-year survival rate. Acacia catechu is an indigenous plant found in India and shows various properties such as antiproliferative, anti-oxidative and anti-inflammatory properties. The present study showed the chemopreventive effects of methanolic bark extract of Acacia catechu (MEBA) against B(a)P induced lung cancer in Swiss albino mice. Methods B(a)P was given orally (50 mg/kg body weight) twice a week for four successive weeks to induce lung cancer in mice. The mice were treated with MEBA at doses of 200 and 400mg/kg b.wt. The lung weight, body weight, lipid peroxidation, lactate dehydrogenase, xanthine oxidase, carcinoembryonic antigen, antioxidants armory activities were estimated. Furthermore, histopathological analysis of lung tissue and immunohistochemistry analysis of VEGF, PCNA, NF-kB andCOX-2 were also carried out Results B(a)P exposure resulted in an increase in LDH, LPO and XO with subsequent decrease in anti-oxidant armory. It caused an increase in the expression of PCNA, VEGF, Cox-2 and NF-kB. The CEA level was also increased in the B(a)P-induced lung cancer group. Treatment with MEBA at a dose of 200 and 400 mg/kg b.wt. significantly decreased in LPO, LDH, XO and increased in anti-oxidant armory activities. This decrease in expression of hyperproliferation and inflammation markers reveals that MEBA treatment seems effective against lung cancer. Elisa kit analysis also revealed that MEBA treatment showed significantly decrease in the CEA level. Furthermore, the antiproliferative effect of MEBA was confirmed by histopathological analysis. Conclusion The present study suggests the chemo-preventive effects of methanolic bark extract of Acacia catechuagainst B(a)P-induced lung tumor by attenuating theoxidative stress, hyperproliferation and inflammation.

P1.03-025 Combination Therapy with Carboplatin and Hyperoxia Synergistically Enhances Suppression of Benzo[a]Pyrene Induced Lung Cancer

P1.03-025 Combination Therapy with Carboplatin and Hyperoxia Synergistically Enhances Suppression of Benzo[a]Pyrene Induced Lung Cancer

119 - Penta-azamacrocyclic SOD Mimetics Enhance Pharmacological Ascorbate Oxidation and Anticancer effects in an H2O2-dependent Manner

Lung cancer accounts for approximately one-fourth of cancer deaths worldwide; new therapeutic approaches are desperately needed. An emerging adjuvant therapy currently in clinical trials is pharmacological ascorbate (P-AscH‒). The anti-cancer effects of P-AscH‒ are a result of its oxidation, resulting in increased flux and steady-state levels of H2O2 in tumors. P-AscH‒ oxidation can be catalyzed by transition metal ions. P-AscH‒ has been shown to reduce Mn(III)-porphyrin superoxide dismutase (SOD) mimetics, increasing the rate of P-AscH‒ oxidation and enhancing the anti-tumor effect of P-AscH‒. However, the ability of Mn(II)-containing SOD mimetics to enhance the anticancer effects of P-AscH‒ is unknown. The current study shows that Mn(II)-containing penta-azamacrocylic SOD mimetics GC4419 and GC4401 can enhance P-AscH‒ oxidation, as determined by increased oxygen consumption and steady-state concentrations of ascorbate radical in complete cell culture media. These mimetics also enhance the toxicity of P-AscH‒ in H292 and H1299 lung cancer cell lines. This enhanced P-AscH‒-induced lung cancer cell killing was shown to be dependent on the catalytic activity of the SOD mimics and the subsequent generation of H2O2, as determined using conditional overexpression of catalase in H1299 cells. GC4419 combined with P-AscH‒ was also capable of enhancing radiation-induced cancer cell killing in a manner that appears to be greater-than-additive. Since P-AscH‒ and GC4419 are both being tested in individual phase II cancer clinical trials in combination with radiation therapy, these results support the proposal that the combination of GC4419 and P-AscH‒ may provide an effective means by which to significantly enhance radiation responses.

Ferulenol, a Sesquiterpene Coumarin, Induce Apoptosis via Mitochondrial Dysregulation in Lung Cancer Induced by Benzo[a]pyrene: Involvement of Bcl2 Protein.

Ferulenol, a sesquiterpene coumarin, was extracted from "Ferula vesceritensis" and possesses pro-oxidative and anticancer effects in different types of cancer. The objective of this study is to determine whether ferulenol has an anticancer effect by regulation the Bcl2 protein expression in lung cancer induced by benzo[a]pyrene in Wistar rats. Rats in group 1 have received, intraperitoneally, olive oil and considered as controls, animals of group 2 were treated with 100 mg/kg of benzo[a]pyrene intraperitoneally in order to induce lung cancer for 24 weeks, the 3rd groups of rats received the ferulenol 50 mg/kg intraperitoneally after 24 weeks of administration of benzo[a]pyrene and the last group, the rats were treated with ferulenol alone 50 mg/kg. Treatment with ferulenol significantly increased the rate of lipid peroxidation and decrease enzymatic (CAT and GST) and non-enzymatic (GSH) anti-oxidants in benzo[a]pyrene induced lung cancer. Anticancer efficacy of ferulenol was evaluated by down-regulation of Bcl2 protein and up-regulation of Bax protein, Therefore, ferulenol can alter the functionality of lung mitochondria by increasing the production of superoxide anion and mitochondrial swelling. Together, our results depict that ferulenol can be used as pro-oxidant and chemotherapeutic agent against lung cancer.

HAT1 induces lung cancer cell apoptosis via up regulating Fas.

The dysfunction of apoptosis is one of the factors contributing to lung cancer (LC) growth. Histone acetyltransferase HAT1 can up regulate cell apoptosis. This study aims to investigate the mechanism by which HAT1 induces LC cell (LCC) apoptosis via up regulating the expression of Fas. In this study, the surgically removed human LC tissues were collected. LCCs were isolated from the LC tissues and analyzed for the expression of HAT1 and Fas by RT-qPCR and Western blotting. We observed that the expression of Fas was negatively correlated with PAR2 in LCCs. Activation of PAR2 suppressed the expression of Fas in normal lung epithelial cells. The expression of HAT1 was lower and positively correlated with Fas expression and negatively correlated with PAR2 expression in LCCs. Activation of PAR2 suppressed Fas expression in lung epithelial cells via inhibiting HAT1. Restoration of HAT1 expression restored Fas expression in LCCs and induced LCC apoptosis. In conclusion, less expression of HAT1 in LCCs was associated with the pathogenesis of LC. Up regulation of HAT1 expression in LCCs can induce LCCs apoptosis, which may be a potential novel therapy for the treatment of LC.

CANADIAN POPULATION RISK OF RADON INDUCED LUNG CANCER-VARIATION RANGE ASSESSMENT BASED ON VARIOUS RADON RISK MODELS.

To address public concerns regarding radon risk and variations in risk estimates based on various risk models available in the literature, lifetime lung cancer risks were calculated with five well-known risk models using more recent Canadian vital statistics (5-year averages from 2008 to 2012). Variations in population risk estimation among various models were assessed. The results showed that the Canadian population risk of radon induced lung cancer can vary from 5.0 to 17% for men and 5.1 to 18% for women based on different radon risk models. Averaged over the estimates from various risk models with better radon dosimetry, 13% of lung cancer deaths among Canadian males and 14% of lung cancer deaths among Canadian females were attributable to long-term indoor radon exposure.

Gemcitabine and Chk1 Inhibitor AZD7762 Synergistically Suppress the Growth of Lkb1-Deficient Lung Adenocarcinoma.

Cells lacking the tumor suppressor gene LKB1/STK11 alter their metabolism to match the demands of accelerated growth, leaving them highly vulnerable to stress. However, targeted therapy for LKB1-deficient cancers has yet to be reported. In both Kras/p53/Lkb1 cell lines and a genetically engineered mouse model of Kras/p53/Lkb1-induced lung cancer, much higher rates of DNA damage occur, resulting in increased dependence on Chk1 checkpoint function. Here we demonstrate that short-term treatment with the Chk1 inhibitor AZD7762 reduces metabolism in pembrolizumab tumors, synergizing with the DNA-damaging drug gemcitabine to reduce tumor size in these models. Our results offer preclinical proof of concept for use of a Chk1 inhibitor to safely enhance the efficacy of gemcitabine, particularly in aggressive KRAS-driven LKB1-deficient lung adenocarcinomas. Cancer Res; 77(18); 5068-76. ©2017 AACR.

The Role of Radiation Induced Injury on Lung Cancer.

This manuscript evaluates the role of cell killing, tissue disorganization, and tissue damage on the induction of lung cancer following low dose rate radiation exposures from internally deposited radioactive materials. Beagle dogs were exposed by inhalation to 90Y, 91Y, 144Ce, or 90Sr in fused clay particles. Dogs lived out their life span with complete pathology conducted at the time of death. The radiation dose per cell turnover was characterized and related to the cause of death for each animal. Large doses per cell turnover resulted in acute death from lung damage with extensive cell killing, tissue disorganization, chronic inflammatory disease, fibrosis, and pneumonitis. Dogs with lower doses per cell turnover developed a very high frequency of lung cancer. As the dose per cell turnover was further decreased, no marked tissue damage and no significant change in either life span or lung cancer frequency was observed. Radiation induced tissue damage and chronic inflammatory disease results in high cancer frequencies in the lung. At doses where a high frequency of chromosome damage and mutations would be predicted to occur there was no decrease in life span or increase in lung cancer. Such research suggests that cell killing and tissue damage and the physiological responses to that damage are important mechanisms in radiation induced lung cancer.

HPV16 E6/E7 upregulates HIF-2α and VEGF by inhibiting LKB1 in lung cancer cells.

Long-term persistent infection of HPV16 E6/E7 is frequently associated with lung cancers, especially in non-smokers and in Asians. However, molecular mechanisms of HPV16 E6/E7 induction of lung cancer are not fully understood. Using bi-directional genetic manipulation and four well-established lung cancer cell lines, we showed HPV16 E6/E7 downregulated expression of liver kinase B1 at both protein and messenger RNA levels; liver kinase B1 downregulated hypoxia-inducible factor 2α at protein level but not at messenger RNA level, and hypoxia-inducible factor 2α upregulated vascular endothelial growth factor at both protein and messenger RNA levels. This is the first study to show hypoxia-inducible factor 2α as a downstream effector of liver kinase B1 in lung cancer cells. Our results indicate that HPV16 E6/E7 indirectly upregulated the expression of vascular endothelial growth factor by inhibition of liver kinase B1 expression and upregulation of hypoxia-inducible factor 2α expression, thus propose a human papillomavirus-liver kinase B1-hypoxia-inducible factor 2α-vascular endothelial growth factor axis for the tumorigenesis of lung cancer. Our study also provides new evidence to support the critical role of liver kinase B1 in the pathogenesis of human papillomavirus-related lung cancer and suggests novel therapeutic targets.

Abstract 5619: Development of a TK1 specific chimeric antigen receptor T-cell for the treatment of non-small-cell lung cancer

Abstract Our current research explores the development and tumoricidal activity of chimeric antigen receptor T-cells targeting a new immunotherapeutic target, thymidine kinase 1 (TK1), against non-small lung cancer (NSCLC), both in vitro and in vivo. There has been recent success utilizing CAR T-cell therapy in clinical trials, but it has been mainly focused on the treatment of haematological malignancies targeting CD19. Lung cancer is the most common cause of cancer mortality globally and is responsible for more than one million of deaths every year. Moreover, NSCLC comprises approximately 85 % of all lung cancers. Clinical trials against NSCLC using engineered T-cells targeting NY-ESO-1, VEGFR2, MAGE-A3, and mesothelin are currently ongoing. In spite of efforts to find new molecular targets, CAR T-cell therapy still faces several challenges in the treatment of solid malignancies due to the lack of specific molecular targets. We have previously reported the up-regulation of TK1 in multiple malignant tissues including lung cancer tissues and the presence of TK1 on the cell surface of different NSCLC cell lines, such as H460 and A549. Flow cytometry, scanning electron microscopy and confocal microscopy showed evidence of TK1 on the surface of these cancer cells lines. We have built third generation TK1-CARs with lentiviral and retroviral vectors. The constructions include a single chain variable fragment for TK1, a CD28 and 4-1BB moieties connected with a CD3ζ signaling domain. Confirmation of the CAR expression on transduced human T cells was performed by flow cytometry and confocal microscopy, and approximately 50 % of the transduced T-cells expressed TK1-CARs. Upon co-culturing TK1-CAR T-cells there was a significant increase in T-cell activity and cancer cell lysis elevated as high as 48% in comparison to negative controls. Cytokine profiles revealed a significant increase of the levels of IL-2 and IFN-γ after 24 hrs of co-culturing, indicating T-cell activation. TK1-CAR T-cells, untransduced T-cells, and transduced T-cells with empty vectors were co-cultured with H460 cells and time-lapse videos were recorded, every 5 minutes between 12 and 24 hrs post transduction. Clustering of TK1-CAR T-cells around lung cancer cells and induction of cell death after T-cell synapsis with target cells was observed. Preliminary in vitro data has shown that TK1-CAR T-cells induce specific cell lysis in NSCLC cells. In vivo experiments using xenografts models in SNG mice will be performed. Statistical differences between survival curves of mice treated with TK1 CAR T-cells, untransduced T-cells and transduced T-cells with empty vectors are expected. Note: This abstract was not presented at the meeting. Citation Format: Edwin J. Velazquez, Kiara Vaden, Michelle H. Townsend, Evita G. Weagel, Scott Weber, Richard A. Robison, Kim L. O'Neill. Development of a TK1 specific chimeric antigen receptor T-cell for the treatment of non-small-cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 5619. doi:10.1158/1538-7445.AM2017-5619

Abstract 1890: PLEK2 contributes to lung cancer progression through upregulation of NDRG1 expression

Abstract PLEK2 is one of the pleckstrin homology domain (PH domain) proteins. Recently studies suggested that PLEK2 may play a critical role in T lymphocyte migration through by PI3K/AKT dependent and independent pathways. However, the biological functions of PLEK2 in lung cancer is not clear. We detected the expression of PLEK2 in various lung adenocarcinoma cell lines using quantitative RT-PCR methods. The results indicated that PLEK2 expression significantly positive correction with cancer cell migration and invasion ability. Forced expression of PLEK2 in lung cancer cells exhibited markedly promoting in the proliferation rate, anchorage independent growth, cell motility and invasion, whereas knockdown of PLEK2 was found to suppress lung cancer cell progression. In addition, we also reported that PLEK2 expression induce lung cancer cell epithelial-mesenchymal transition (EMT) phenomenon, and involve in chemotherapeutic drug resistance through upregulation NDRG1 gene expression. Overexpression of NDRG1 in PLEK2-knockdown cell line restored cancer cell proliferation, migration, and invasion. These results will provide information and help us to understand the roles of NDRG1 in lung cancer cells. Citation Format: Yi-Pin Hsieh, Hui-Chia Liu, Hao-Yu Huang, Wan-Jiun Tang, Tai-Lin Lee, Meng-Feng Tsai. PLEK2 contributes to lung cancer progression through upregulation of NDRG1 expression [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1890. doi:10.1158/1538-7445.AM2017-1890

A Positive Feedback Loop between Sestrin2 and mTORC2 Is Required for the Survival of Glutamine-Depleted Lung Cancer Cells

Proper regulation of mTORC1 and mTORC2 upon nutrient starvation is critical for cancer cell survival.Upregulation of Sestrin2 in response to glutamine deprivation rescues cell death by suppressing mTORC1. However, the contribution of mTORC2 to Sestrin2-mediated mTORC1 suppression remains unclear. Here, we report that both Sestrin2 and mTORC2 are upregulated in glutamine-depleted lung cancer cells. Moreover, glutamine depletion caused Sestrin2 to associate with mTORC2, which was required for the increase in Sestrin2 protein stability and the reduction in mTORC1 activity. Ultimately, differential regulation of mTORC1 and 2 by Sestrin2 reprogramed lipid metabolism and enabled glutamine-depleted lung cancer cells to survive by maintaining energy and redox balance. Importantly, combined inhibition of glutamine utilization and Sestrin2 induced lung cancer cell death both invitro and invivo. This study shows that differential Sestrin2-mediated regulation of mTORC1 and mTORC2 is necessary for the survival of glutamine-depleted lung cancer cells.