Gene Therapy For Lung Cancer Research Articles

Oncolytic adenovirus encoding LHPP exerts potent antitumor effect in lung cancer

LHPP has been shown to be a new tumor suppressor, and has a tendency to be under-expressed in a variety of cancers. Oncolytic virotheray is a promising therapeutics for lung cancer in recent decade years. Here we successfully constructed a new recombinant oncolytic adenovirus GD55-LHPP and investigated the effect of GD55-LHPP on the growth of lung cancer cells in vitro and in vivo. The results showed that LHPP had lower expression in either lung cancer cells or clinical lung cancer tissues compared with normal cells or tissues, and GD55-LHPP effectively mediated LHPP expression in lung cancer cells. GD55-LHPP could effectively inhibit the proliferation of lung cancer cell lines and rarely affected normal cell growth. Mechanically, the oncolytic adenovirus GD55-LHPP was able to induce stronger apoptosis of lung cancer cells compared with GD55 through the activation of caspase signal pathway. Notably, GD55-LHPP also activated autophagy-related signal pathway. Further, GD55-LHPP efficiently inhibited tumor growth in lung cancer xenograft in mice and prolonged animal survival rate compared with the control GD55 or PBS. In conclusion, the novel construct GD55-LHPP provides a valuable strategy for lung cancer-targeted therapy and develop the role of tumor suppress gene LHPP in lung cancer gene therapy.

Research on the gene therapy and prevention of lung cancer

In recent years, there have been significant advancements in molecular biology, gene therapy technology, and high-tech testing techniques. As our understanding of the pathogenesis of lung cancer continues to deepen, it has become evident that gene therapy plays a crucial role in its treatment. This article provides an overview of tumor cell development, various gene therapy techniques including suicide gene therapy, silent gene therapy, cancer suppressor genetic therapy, and antibody genetic therapy. Additionally, it analyzes the application value, treatment technology, and safety concerns associated with gene therapy. The discussion also covers the safety issues and current application status of viral and nonviral vectors in gene therapy, comparing the advantages and disadvantages of different vector types. Furthermore, the article summarizes the future directions for designing and improving gene delivery vectors, along with their clinical application prospects. Ultimately, this article offers insights into the significance and treatment approaches for early prevention of lung cancer.

Dual peptides-modified cationic liposomes for enhanced Lung cancer gene therapy by a gap junction regulating strategy

BackgroundGene therapy for lung cancer has emerged as a novel tumor-combating strategy for its superior tumor specificity, low systematical toxicity and huge clinical translation potential. Especially, the applications of microRNA shed led on effective tumor ablation by directly interfering with the crucial gene expression, making it one of the most promising gene therapy agents. However, for lung cancer therapy, the microRNA treatment confronted three bottlenecks, the poor tumor tissue penetration effect, the insufficient lung drug accumulation and unsatisfied gene transfection efficiency. To address these issues, an inhalable RGD-TAT dual peptides-modified cationic liposomes loaded with microRNA miR-34a and gap junction (GJ) regulation agent all-trans retinoic acid (ATRA) was proposed, which was further engineered into dry powder inhalers (DPIs).ResultsEquipped with a rough particle surface and appropriate aerodynamic size, the proposed RGD-TAT-CLPs/ARTA@miR-34a DPIs were expected to deposit into the deep lung and reach lung tumor lesions guided by targeting peptide RGD. Assisted by cellular transmembrane peptides TAT, the RGD-TAT-CLPs/ARTA@miR-34a was proven to be effectively internalized by cancer cells, enhancing gene transfection efficiency. Then, the GJ between tumor cells was upregulated by ARTA, facilitating the intercellular transport of miR-34a and boosting the gene expression in the deep tumor.ConclusionOverall, the proposed RGD-TAT-CLPs/ARTA@miR-34a DPIs could enhance tumor tissue penetration, elevate lung drug accumulation and boost gene transfection efficiency, breaking the three bottlenecks to enhancing tumor elimination in vitro and in vivo. We believe that the proposed RGD-TAT-CLPs/ARTA@miR-34a DPIs could serve as a promising pulmonary gene delivery platform for multiple lung local disease treatments.

Bioinformatics Analysis and Screening of Potential Target Genes Related to the Lung Cancer Prognosis.

Several genes have been validated as molecular targets for gene therapy in lung cancer. We screened target genes that affect survival of patients with lung cancer. Data on gene expression in normal lung tissues/lung adenocarcinoma (LUAD) samples were acquired from Genotype-Tissue Expression (GTEx)/The Cancer Genome Atlas (TCGA) databases and merged to expand the sample size, followed by differential analysis of the merged expression data and acquisition of differentially expressed genes. Survival and simple Cox analyses were used to screen for genes affecting LUAD survival. Protein-protein interaction/multivariable Cox analyses were utilized, and a risk model was established. Candidate genes expression levels in cancer/paracancerous tissues of lung cancer patients, and BEAS-2B/A549/HCC95 cells were measured by RT-qPCR/Western blot. Survival analysis of candidate genes was conducted in LUAD samples collected from TCGA. Among 947 genes differentially expressed in LUAD, 151 were correlated with patient survival, and 116 might act as risk factors for LUAD. The 7 identified candidate genes (TOP2A, TK1, KIF4A, ANLN, KIF2C, ASF1B, CCNB1) were high-risk genes playing possible roles in LUAD. These genes were differentially expressed in lung cancer and were associated with TNM stages (III - IV)/differentiation grade/lymph node metastasis/distant metastasis, which affected lung cancer patient survival. P2A, TK1, KIF4A, ANLN, KIF2C, ASF1B and CCNB1 were highly-expressed in LUAD/lung squamous cell carcinoma (LUSC) and correlated with LUAD patient survival. This study contributes to better understanding of the prognostic regulation mechanism in LUAD and the screening of target genes for clinical treatment.

Apoptosis-targeted gene therapy for non-small cell lung cancer using chitosan-poly-lactic-co-glycolic acid -based nano-delivery system and CASP8 and miRs 29A-B1 and 34A.

Non-small cell lung cancer (NSCLC) is a leading cause of cancer-related deaths worldwide, with resistance to apoptosis being a major driver of therapeutic resistance and aggressive phenotype. This study aimed to develop a novel gene therapy approach for NSCLC by targeting resistance to apoptosis. Loss of function mutations of caspase 8 (CASP8) and downregulation of microRNAs (miRs) 29A-B1 and 34A were identified as key contributors to resistance to apoptosis in NSCLC. A biodegradable polymeric nano-gene delivery system composed of chitosan-poly-lactic-co-glycolic acid was formulated to deliver initiator CASP8 and miRs 29A-B1 and 34A. The nano-formulation efficiently encapsulated the therapeutic genes effectively internalized into NSCLC cells and induced significant apoptosis. Evaluation of the nano-formulation in A549 tumor spheroids showed a significant increase in apoptosis within the core of the spheroids, suggesting effective penetration into the spheroid structures. We provide a novel nano-formulation that demonstrate therapeutic potential for suicidal gene therapy in NSCLC.

CRISPR-Cas9-mediated gene therapy in lung cancer.

As the largest cause of cancer-related deaths worldwide, pulmonary cancer is the most common form of the disease. Severalgenetic, epigenetic, and environmental factorscome into play during the multi-step mechanism of tumorigenesis. The heterogeneity that makes discovering successful therapeutics for pulmonary cancer problematic is significantly influenced by the epigenetic landscape, includingDNA methylation, chromatin architecture, histone modifications, and noncoding RNA control. Clinical activity of epigenetic-targeted medicines has been reported in hematological tumors, and these compounds may also have therapeutic effects in solid tumors. Over the course of the past few years, some researchers have successfully modified the expression of genes in cells using the clustered regularly interspaced short palindromic repeats (CRISPR)-Cas (CRISPR-associated proteins) technique. The utilization of this technology allows for the induction of site-specific mutagenesis, epigenetic alterations, and the regulation of gene expression. This study will present an overview of the primary epigenetic alterations seen in pulmonary cancer, as well as a summary of therapeutic implications for targeting epigenetics in the management of pulmonary cancer, with a particular emphasis on the technique known as CRISPR/Cas9.

The Potential Application of Branch-PCR Assembled PTEN Gene Nanovector in Lung Cancer Gene Therapy.

Gene therapy offers an alternative and promising avenue to lung cancer treatment. Here, we used dibenzocyclooctyne (DBCO)-branched primers to construct a PTEN gene nanovector (NP-PTEN) through branch-PCR. NP-PTEN showed the nanoscale structure with biocompatible size (84.7±11.2 nm) and retained the improved serum stability compared to linear DNA. When transfected into NCI-H1299 cancer cells, NP-PTEN could overexpress PTEN protein to restore PTEN functions through the deactivation of PI3K-AKT-mTOR signaling pathway to inhibit cell proliferation and induce cell apoptosis. The apoptosis rate of NCI-H1299 cancer cells could reach up to 54.5 %±4.6 % when the transfection concentration of NP-PTEN was 4.0 μg/mL. In mice bearing NCI-H1299 tumor xenograft intratumorally administrated with NP-PTEN, the average tumor volume and tumor weight was separately reduced by 61.7 % and 63.9 %, respectively, compared with the PBS group on the 18th day of administration. The anticancer efficacy of NP-PTEN in NCI-H1299 tumor xenograft suggests the promising therapeutic potential of branch-PCR assembled PTEN gene nanovectors in lung cancer gene therapy and also provided more opportunities to introduce two or more tumor suppressor genes as an all-in-one gene nanovector for multiple gene-based cancer gene therapy.

Effects of inhalable gene transfection as a novel gene therapy for non-small cell lung cancer and malignant pleural mesothelioma

Gene therapy using vectors has attracted attention in recent years for the treatment of cancers caused by gene mutations. Besides, new treatments are imperative for lung cancer, including non-small cell lung cancer (NSCLC) and malignant pleural mesothelioma (MPM), due to its high mortality. We developed a minimally invasive and orally inhalable tumor suppressor gene drug (SFD-p16 and SFD-p53) with non-viral vectors for lung cancer treatment by combining tumor suppressor genes with an inhalant powder that can deliver active ingredients directly to the lung. We used NSCLC (A549 and H1299) and MPM (H2052) cell lines in an air–liquid interface culture. Transfection of A549 and H2052 cells with SFD-p16 significantly increased p16 mRNA expression levels and decreased cell proliferation in both cell lines. Similar results were obtained with transfection of H1299 with the inhalable gene drug SFD-p53. In an in vivo experiment, a mouse model of lung cancer with orthotopically transplanted luciferase-expressing A549 cells was subjected to intratracheal insufflation of SFD-p16. Consequently, SFD-p16 effectively and directly affected lung cancer. This study suggests that inhalable gene drugs are effective treatments for NSCLC and MPM. We expect inhalable gene drugs to present a novel gene therapy agent for lung cancer that patients can self-administer.

Polyethylenimine-Modified Bombyx mori Silk Fibroin as a Delivery Carrier of the ING4-IL-24 Coexpression Plasmid.

One of the major challenges for lung cancer gene therapy is to find a gene delivery vector with high efficiency and low toxicity. In this study, low-molecular-weight polyethyleneimine (PEI, 1.8 kDa) was grafted onto the side chains of Bombyx mori silk fibroin (BSF) to prepare cationized BSF (CBSF), which was used to package the plasmid DNA (pDNA) encoded by the inhibitor of growth 4 (ING4) and interleukin-24 (IL-24). FTIR and 1H-NMR spectra demonstrated that PEI was effectively coupled to the side chains of BSF by amino bonds. The results of the trinitrobenzene sulfonic acid method and zeta potential showed that the free amino group content on BSF increased from 125.1 ± 1.2 µmol/mL to 153.5 ± 2.2 µmol/mL, the isoelectric point increased from 3.68 to 8.82, and the zeta potential reversed from − 11.8 ± 0.1 mV to + 12.4 ± 0.3 mV after PEI grafting. Positively charged CBSF could package pDNA to form spherical CBSF/pDNA complexes. In vitro, human lung adenocarcinoma A549 cells and human embryonic lung fibroblast WI-38 cells were transfected with CBSF/pDNA complexes. Confocal laser scanning microscopy analysis and flow cytometry tests showed that CBSF/pDNA complexes can effectively transfect A549 cells, and the transfection efficiency was higher than that of 25 kDa PEI/pDNA complexes. CCK-8 assay results showed that CBSF/pDNA complexes significantly inhibited the proliferation of A549 cells but had no significant effect on WI-38 cells and exhibited lower cytotoxicity to WI-38 cells than 25 kDa PEI. Therefore, a gene delivery system, constructed with the low-molecular-weight PEI-modified silk fibroin protein and the ING4-IL-24 double gene coexpression plasmid has potential applications in gene therapy for lung cancer.

Lentiviral shRNA Construction in Gene Therapy for Knock Down CYPIA1, and YWHAZ gene in Lung Cancer

Lung cancer is a disease that is in the top 10 causes of death globally. It is caused by CYP1A1 and YWHAZ genes expressed in the lungs. One of the solutions offered for lung cancer sufferers is gene therapy. Gene therapy for lung cancer is focused on the knock-down of expressed CYP1A1 and YWHAZ genes. It can be using shRNA transfected into lung cells by lentivirus. The lentivirus envelope is modified using glycoprotein filovirus, one of which is ebolavirus. Lentivirus pseudotyping can recognize certain cells. Especially, Ebolavirus can recognize lung cells. The lentiviral gene is designed to carry the gene of interest (shRNA) but no the infectious gene. The shRNA gene transfused into lung cancer cells will recognize and complement the specific target mRNA sites of the CYP1A1 and YWHAZ genes. After that, shRNA will cut the mRNA which should be expressed into a cancer protein. Unexpressed mRNA into protein causes cancer cell growth inhibited. Thus, this gene therapy solution is expected to be able to reduce the rate of death caused by lung cancer. Keywords: Lung cancer, CYP1A1 and YWHAZ genes, gene therapy, lentiviral, shRNA

Targeting stanniocalcin-1-expressing tumor cells elicits efficient antitumor effects in a mouse model of human lung cancer.

Lung cancer is the most common cause of cancer‐related death in developed countries; therefore, the generation of effective targeted therapeutic regimens is essential. Recently, gene therapy approaches toward malignant cells have emerged as attractive molecular therapeutics. Previous studies have indicated that stanniocalcin‐1 (STC‐1), a hormone involved in calcium and phosphate homeostasis, positively regulates proliferation, apoptosis resistance, and glucose metabolism in lung cancer cell lines. In this study, we investigated if targeting STC‐1 in tumor cells could be a promising strategy for lung cancer gene therapy. We confirmed that STC‐1 levels in peripheral blood were higher in lung cancer patients than in healthy donors and that STC‐1 expression was observed in five out of eight lung cancer cell lines. A vector expressing a suicide gene, uracil phosphoribosyltransferase (UPRT), under the control of the STC‐1 promoter, was constructed (pP STC‐1‐UPRT) and transfected into three STC‐1‐positive cell lines, PC‐9, A549, and H1299. When stably transfected, we observed significant cell growth inhibition using 5‐fluorouracil (5‐FU) treatment. Furthermore, growth of the STC‐1‐negative lung cancer cell line, LK‐2 was significantly arrested when combined with STC‐1‐positive cells transfected with pP STC‐1‐UPRT.We believe that conferring cytotoxicity in STC‐1‐positive lung cancer cells using a suicide gene may be a useful therapeutic strategy for lung cancer.

Recombinant Adeno-associated Virus 9-mediated Expression of Kallistatin Suppresses Lung Tumor Growth in Mice.

Lung cancer remains the most common cause of cancer-related deaths in China and worldwide. Traditional surgery and chemotherapy do not offer an effective cure, although gene therapy may be a promising future alternative. Kallistatin (Kal) is an endogenous inhibitor of angiogenesis and tumorigenesis. Recombinant adeno-associated virus (rAAV) is considered the most promising vector for gene therapy of many diseases due to persistent and long-term transgenic expression. The aim of this study was to investigate whether rAAV9-Kal inhibited NCI-H446 subcutaneous xenograft tumor growth in mice. The subcutaneous xenograft mode was induced by subcutaneous injection of 2×107 H446 cells into the dorsal skin of BALB/c nude mice. The mice were administered with ssrAAV9-Kal (single- stranded rAAV9) or dsrAAV9-Kal (double-stranded rAAV9) by intraperitoneal injection (I.P.). Tumor microvessel density (MVD) was examined by anti-CD34 staining to evaluate tumor angiogenesis. Compared with the PBS (blank control) group, tumor growth in the high-dose ssrAAV9-Kal group was inhibited by 40% by day 49, and the MVD of tumor tissues was significantly decreased. The results indicate that this therapeutic strategy is a promising approach for clinical cancer therapy and implicate rAAV9-Kal as a candidate for gene therapy of lung cancer.

A novel tumour suppressor lncRNA F630028O10Rik inhibits lung cancer angiogenesis by regulating miR-223-3p.

Lung cancer is the world's leading cause of cancer‐related morbidity and mortality despite advances in surgery, chemotherapy and immunotherapy; thus, there is an urgent need to find new molecules to develop novel treatment strategies. Although ncRNAs were found to account for 98% transcripts, the number of lncRNAs with distinct function in lung cancer is extremely limited. We previously demonstrated that Plasmodium infection inhibits tumour growth and metastasis, but the exact mechanisms involved have not been fully understood. In this study, we carried out RNA sequencing (RNA‐Seq) of tumour tissues isolated from LLC tumour‐bearing mice treated with either Plasmodium yoelli (Py)‐infected red blood cells or uninfected red blood cells. We found that F630028O10Rik (abbreviated as F63) is a novel lncRNA that was significantly up‐regulated in tumours isolated from mice treated with Py‐infected red blood cells compared to the control. By using gene silencing technique, F63 was found to inhibit both tumour Vascular Endothelial Growth Factor A (VEGFA) secretion and endothelial cells clone formation, migration, invasion and tube formation. Injection of cholesterol‐modified siRNA‐F63 into mice tumour tissues produced a significant increase in tumour volume, blood vessel formation and angiogenesis 17 days after injection. We further showed that inhibiting miR‐223‐3p results in the down‐regulation of VEGFA and VEGFR2 which are vital molecules for angiogenesis. These results reveal that F63 inhibit tumour growth and progression by modulating tumour angiogenesis suggesting F63 can be a novel lncRNA with great potential as a candidate molecule for gene therapy in lung cancer.

Research progress of driving gene and molecular targeting therapy for non-small cell lung cancer

The morbidity and mortality of lung cancer are the highest in China, and the non-small cell lung cancer(NSCLC) is the first one, accounting for about 80% of lung cancer.NSCLC includes squamous cell carcinoma, adenocarcinoma, etc.Because of the slow growth of lung cancer cells and the late diffusion and metastasis, most of the patients were in late stage and lost the best period of surgical treatment.In recent years, molecular targeted therapy for specific genes has become an important strategy for the treatment of NSCLC.Multiple target genes, such as EGFR, KRAS, BRAF and EML4-ALK have been identified.This article reviews the related driving genes and molecular targeted therapy of NSCLC, expecting to be helpful to the individualized treatment of advanced NSCLC patients. Key words: Lung neoplasms; Carcinoma, non-small-cell lung; Driving genes; Molecular targeted therapy; Driving gene related inhibitor; Molecular detection technique; Genic mutation; Clinical application; Individualized treatment; Drug resistance

Non-small-cell lung cancer homing peptide-labeled dendrimers selectively transfect lung cancer cells.

Lung cancer gene therapies require reagents to selectively transfect lung tumors after systemic administration. We created a reagent called NSCLC-NP by attaching a peptide with binding affinity for lung cancer to polyamidoamine dendrimers. The positively charged dendrimers electrostatically bind negatively charged nucleic acids, inhibit endogenous nucleases and transfect cells targeted by the attached peptide. In vitro, NSCLC-NP complexed to DNA plasmids bound and transfected three human lung cancer cell lines producing protein expression of the plasmid's gene. In vivo, systemically administered NSCLC-NP selectively transfected lung cancer cells growing in RAG1KO mice. The capability of NSCLC-NP to selectively transfect lung cancer allows its future use as a vehicle to implement human lung cancer gene therapy strategies.

4:10 PM Abstract No. 50 Intratumoral radiofrequency hyperthermia-enhanced herpes simplex virus thymidine kinase (HSV-TK) gene therapy of orthotopic lung cancer through the augmentation of apoptosis and heat shock protein (HSP70) expression pathway

To investigate the associated biomolecular mechanisms of intratumoral RFH- enhanced local HSV-TK/ ganciclovir (GCV)-mediated suicide gene therapy of rat orthotopic lung cancers. Human lung cancer cells (A549) transduced with Luciferase/mCherry/lentivirus and 24 nude rats with orthotopic lung cancers were divided into four study groups with treatments of (i) intratumoral HSV-TK/GCV gene therapy followed by RFH at 41-42 °C for 30 minutes; (ii) HSV-TK/GCV therapy alone; (iii) RFH alone; (iv) PBS as a control. Flow cytometry, bioluminescence assay and confocal microscopy were used to determine the apoptosis and viability of cells. Bioluminescence optical imaging was used to evaluate the changes of bioluminescent signals among the groups over 2 weeks. The expression of Bcl-2/Bax, Caspase-3 and HSP70 in both cells and tumor tissue were quantified by western blotting (WB) and immunohistochemistical (IHC) staining respectively. Compared with gene therapy alone, RFH alone or PBS, combination therapy induced the lowest cell viability (30.56 ± 9.12% VS 65.47 ± 8.02% VS 89.95 ± 6.33% VS 100% P<0.05), the highest cell apoptosis (20.05 ± 2.12% VS 9.87 ± 2.02% VS 4.31 ± 4.31% VS 3.12 ± 1.03%, P<0.05), and a remarkable decrease of relative bioluminescence signal (16.54 ± 0.12% VS 54.8 ± 0.10% VS 85.01 ± 0.08% VS 100%, P<0.01). Optical imaging showed a markedly decreased bioluminescence signal of the tumor in combination therapy (1.29 ± 0.18 VS 1.76 ± 0.13 VS 2.26 ± 0.17 VS 2.94 ± 0.15, P<0.05). Both WB analysis and IHC staining displayed the significantly decreased expression of Bcl-2 (1.03 ± 0.28 VS 2.15 ± 0.33 VS 3.48 ± 0.12 VS 3.87 ± 0.17×107 IOD, P<0.01), as well as increased expression of Bax (3.30 ± 0.0.36 VS 2.02 ± 0.20 VS 1.48 ± 0.31 VS 1.26 ± 0.07, 107 IOD, P<0.01), Caspase-3 (5.58 ± 0.52 VS 3.36 ± 0.29 VS 1.42 ± 0.34VS 1.21 ± 0.52, 107 IOD, P<0.05) and HSP70 (8.79 ± 0.62 VS 3.97 ± 0.31 VS 4.48 ± 0.42 VS 1.60 ± 0.23, 107 IOD, P<0.01) in cancer tissues of combination therapy group. Intratumoral RFH-enhanced suicide gene therapy on rat orthotopic lung cancers through the mechanisms of augmenting apoptosis and the expression of HSP-70 pathway, which may provide additional insights to further improving treatment of lung cancers.

Down-regulated REIC expression in lung carcinogenesis: a molecular target for gene therapy.

REIC (Reduced Expression in Immortalized Cells) gene is down-regulated in immortalized cells, compared with the normal parental counterparts. Its encoding protein could inhibit colony formation, tumor growth, and induce apoptosis. To investigate the roles of REIC expression in lung cancer, we examined REIC expression in lung cancer cells and tissues by RT-PCR or Western blot, and observed the effects of both recombinant REIC exposure and REIC overexpression on the aggressive phenotypes of lung cancer cells. It was found that the demethylation of REIC promoter by 5-Aza-dC could reserve its mRNA expression in lung cancer cells (P<0.05). There was a lower REIC mRNA expression in lung cancer than that in matched normal tissue (P<0.05). Recombinant REIC treatment enhanced the proliferation of lung cancer cells (P<0.05), but versa for REIC overexpression (P<0.05). Both recombinant REIC treatment and REIC overexpression induced apoptosis, and inhibited the migration and invasion of SQ-5 and KJ cells (P<0.05). Immunohistochemically, there was a positive correlation between REIC and Caspase-3 expression in lung cancer (P<0.05). According to Kaplan-Meier plotter, REIC mRNA overexpression was found to positively correlate with overall, progression-free and post- progression survival rates of lung cancer patients (P<0.05), even stratified by sex, histological subtyping, grading, TNM staging, chemotherapy, radiotherapy, or smoking. These findings suggested that the down-regulated REIC expression might be involved in lung carcinogenesis due to its promoter methylation. Both recombinant REIC exposure and REIC overexpression might reverse the aggressive phenotypes of lung cancer cells. REIC may be employed as a potential target of gene therapy for lung cancer.

Network analysis of DEGs and verification experiments reveal the notable roles of PTTG1 and MMP9 in lung cancer.

Lung cancer, a malignant tumor, is the most frequently fatal cancer, with poor survival rates in the advanced stages. In order to improve the understanding of this disease, and to improve the outcomes of patients, additional studies are required. In the present study, differentially expressed genes (DEGs) in patients with lung cancer compared with controls were identified. To understand how these DEGs act together to account for the initiation of lung cancer, a protein interaction network and a transcriptional regulatory network were constructed to explore the clusters and pathways in lung cancer, and the results indicated that PTTG1 and MMP9 served major roles in the development of lung cancer in the regulatory system. Consistent with this, mRNA and protein expression levels of PTTG1 and MMP9 were significantly upregulated in lung cancer tissues compared with normal lung tissues. The overexpression of PTTG1 or MMP9 was induced in the human bronchial epithelial BEAS-2B cell line, indicating that increased PTTG1 or MMP9 alone may not only facilitate cell migration, proliferation and induce colony formation, but also suppress cell apoptosis. In summary, PTTG1 and MMP9 were identified as potential targets for therapeutic intervention through gene therapy in lung cancer.

MA16.01 Targeted Gene Therapy for Tobacco Carcinogen-Induced Lung Cancer

Rab25, a member of Rab family of small GTPases, is associated with progression of various types of human cancer including lung cancer that is the leading cause of cancer-associated deaths around the globe. In this study, we report the gene therapeutic effect of short hairpin Rab25 (shRab25) on 4-(Methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK)-induced lung tumorgenesis in female A/J mice. Initially, Mice (six-week-old) were injected with single dose of NNK (2 mg/0.1 ml saline/mouse) by intraperitoneal injection to induce the tumor. 8 weeks later, shRab25 was delivered with GPT-SPE (glycerol propoxylate triacrylate (GPT) and spermine) complex into tobacco-induced lung cancer models through a nose-only inhalation system twice a week for 2 month. Remarkably, aerosol-delivered shRab25 significantly decreased the expression level of Rab25 and other prominent apoptosis related proteins in female A/J mice. The apoptosis in these mice were determined by detecting the expression level of Bcl-2, PCNA, Bax and further confirmed by TUNEL assay. Our results strongly confirm the tumorigenic role of Rab25 in tobacco carcinogen induced-lung cancer and hence demonstrate aerosol delivery of shRab25 as a therapeutic target for lung cancer treatment.

Lung cancer gene therapy: Transferrin and hyaluronic acid dual ligand-decorated novel lipid carriers for targeted gene delivery.

To achieve lung cancer gene therapy, nanocarriers decorated with different ligands were used. Surface decoration and nanoparticulate system will assist in targeting the gene to specific cells and tissues, such as cancers and diseased organs. The aim of this research was to develop a dual ligand-decorated nanocarriers, which could target the tumor cells through receptor-mediated pathways to increase the uptake of genetic materials. Transferrin (Tf) and hyaluronic (HA) containing polyethylene glycol-distearoylphosphatidylethanolamine (Tf-PEG-DSPE and HA-PEG-DSPE) ligands were synthesized. Novel Tf and HA ligand-decorated, plasmid-enhanced green fluorescent protein loaded nanostructured lipid carriers (Tf/HA-pDNA NLC) was constructed. Physicochemical properties such as morphology, size, and ζ-potential as well as release properties were evaluated. The invitro and invivo gene transfection efficiency of Tf/HA-pDNA NLC was evaluated in lung adenocarcinoma A549 cells and lung cancer bearing animal models. Tf/HA-pDNA NLC displayed significantly higher transfection efficiency than undecorated DNA-NLCs and single ligand-decorated NLCs invitro and invivo. The newly constructed NLCs could successfully load gene; and Tf and HA functioned as excellent targeting ligands to improve the cell targeting ability of the gene-loaded nanocarriers. The resulting dual ligands decorated vectors could be a promising targeted gene delivery system for the lung cancer treatment.