Potential Lung Cancer Therapy Research Articles

Cell-Surface GRP78-Targeted Chimeric Antigen Receptor T Cells Eliminate Lung Cancer Tumor Xenografts.

Lung cancer is one of the most common and intractable malignancies. It is associated with low survival rates despite existing treatments, indicating that new and more effective therapies are urgently needed such as the chimeric antigen receptor-T (CAR-T) cell immunotherapy. The cell-surface glucose-regulated protein 78 (csGRP78) is expressed in various hematological malignancies and solid tumor cells including lung cancer in response to cancer-related endoplasmic reticulum stress, while GRP78 is restricted to inside the normal cells. Here, we detected the prominent expression of csGRP78 in both lung cancer cell lines, A549 and H1299, as well as cancer stemlike cells derived from A549 by immunofluorescence. Next, a csGRP78-targeted CAR was constructed, and the transduced CAR-T cells were tested for their potency to kill the two lung cancer cell lines and derived stemlike cells, which was correlated with specific interferon γ release in vitro. Finally, we found that csGRP78 CAR-T cells also efficiently killed both lung cancer cells and cancer stemlike cells, resulting into the elimination of tumor xenografts in vivo, neither with any evidence of relapse after 63 days of tumor clearance nor any detrimental impact on other body organs we examined. Our study reveals the capacity of csGRP78 as a therapeutic target and offers valuable insight into the development of csGRP78 CAR-T cells as potential therapy for lung cancer.

Tumor-derived GABA promotes lung cancer progression by influencing TAMs polarization and neovascularization

BackgroundGamma-aminobutyric acid (GABA), a common neurotransmitter, has been found in various cancers but its origin and its role in the tumor immune microenvironment remains unclear. MethodsHere, we reported the expression of glutamate decarboxylase 1 (GAD1, converting glutamate into GABA) in lung cancer tissues based on the publicly available database, and explored the effects and underlying mechanism of GABA on lung cancer progression. ResultsCompared with normal tissues, GAD1 was aberrantly overexpressed in lung adenocarcinoma (LUAD) based on TCGA database. Furthermore, the LUAD patients’ overall survival was negatively correlated with the GAD1 expression levels. Our work found that a GABAa receptor inhibitor had a therapeutic effect on mouse tumors and significantly reduced tumor size and weight. Further experiments showed that GABA derived from tumor cells promoted tumor progression not by directly affecting cancer cells but by affecting macrophages polarization in the tumor microenvironment. We found that GABA inhibited the NF-κB pathway and STAT3 pathway to prevent macrophages from polarizing towards M1 type, while promoting macrophage M2 polarization by activating the STAT6 pathway. GABA was also found to promote tumor neovascularization by increasing the expression of FGF2 in macrophages. ConclusionsThese results suggest that GABA affects tumor progression by regulating macrophage polarization, and targeting GABA and its signaling pathway may represent a potential therapy for lung cancer.

Bioinformatics Study: Syzygium samarangense Leaves Chalcone Extract as Lung Cancer Proliferation Inhibitor

Wax apple (Syzygium samarangense) is a plant commonly found in Indonesia with various health benefits, especially leaves. Anticancer effect in chalcone of the extract of Syzygium samarangense's leaves has been proven. This study identifies chalcone extract potential, a flavonoid active compound group in Syzygium samarangense, as a proliferation inhibitor in lung cancer based on bioinformatics analysis. The extract of Syzygium samarangense's leaves identification analysis shows that our extract contains flavonoids with a 3.63% w/w yield. We identified protein target from the main candidate of active compound from chalcone in Syzygium samarangense, Dimethyl Cardamonin (DMC). A total of 263 potential targets has been collected, and 21 of 263 genes were further analyzed, yielding five gene protein roles in cancer pathways in humans based on DAVID V6.8. Furthermore, expression level and survivabilities of 5 genes in lung cancer were analyzed, resulting in only HDAC1, a significant result. This result was further strengthened by molecular docking DMC with HDAC1 protein compared to the native ligand and inhibitor, showing no significant differences in binding energy between DMC and all comparator compounds. The compound interaction profile shows that DMC interacts similarly with zolinza. Thus, DMC was predicted to be potential lung cancer therapy by inhibiting HDAC1 hence cancer proliferation. However, additional research in the form of in vitro and in vivo proves our prediction.

ROP and ATRP fabricated redox sensitive micelles based on PCL-SS-PMAA diblock copolymers to co-deliver PTX and CDDP for lung cancer therapy

Combining dual drugs in one vehicle to cancer cells offers spatiotemporal localization of drug at the site of action, leading to synergistic therapeutic effects and reduced side effects. To improve pH/redox responsiveness to the tumor microenvironments for cancer therapy, a pH/redox-responsive micelle based on poly(ε-caprolactone)-SS-poly(methacrylic acid) (PCL-SS-PMAA) diblock copolymer was fabricated for dual drug delivery. The PCL-SS-PMAA was formulated into a core-shell micelle (PSPm) in an aqueous solution. The critical micelle concentration (CMC) values of PSPm were 7.94 × 10−3 mg mL-1 at pH 5.0 and 1.00 × 10-2 mg mL-1 at pH 7.4. The hydrodynamic diameters of PSPm were within 210–270 nm, depending on pH values. Changes in morphology and size of PSPm were clearly observed before and after exposure to a reducing agent. Paclitaxel (PTX) was encapsulated into the core and cisplatin (CDDP) was chelated on the shell of PSPm, with both PTX and CDDP being efficiently released from PSPm in the presence of a reducing agent in an acid condition. MTT and annexin V/propidium iodide dual staining results demonstrated that co-loading of CDDP and PTX into PSPm had a synergistic effect in killing lung cancer cells and exerted superior antitumor activity over the combination of single drug-loaded PSPm or the combination of free-CDDP and free-PTX at equivalent drug amounts. Hence, encapsulating the dual drugs into PSPm exhibits a synergistic effect for potential lung cancer therapy.

154 Poster - Potential lung cancer therapy using plant derived cholesterol structural analogs

154 Poster - Potential lung cancer therapy using plant derived cholesterol structural analogs

FCHO1560−571 peptide, a PKB kinase motif, inhibits tumor progression

BackgroundCell division is regulated by protein kinase B (PKB)-mediated FCH domain only 1 (FCHO1) phosphorylation. MethodsFCHO1560−571, a synthetic water-soluble peptide, was generated from the PKB substrate motif 560PPRRLRSRKVSC571 found in the human FCHO1 protein. ResultsIn this study, we found that in vitro FCHO1560−571 inhibits cell proliferation via PKB/ERK/SMAD4 pathways in KRAS-mutated A549 lung cancer cells. In addition, FCHO1560−571, at effective doses of 15 and 30 mg/kg, significantly suppressed tumor growth and decreased the size and weight of tumors in A549-xenograft mice. ConclusionThese results suggest that the FCHO1560−571 peptide could be a potential therapy for lung cancer.

Potential lung cancer therapy using plant derived cholesterol structural analogs

The acquired multidrug resistance syndrome (MDRS) is one of the leading causes for the failure of chemotherapy in lung cancer. Recently plant‐derived triterpenoids (eg, cholesterol‐structural analogs) have showed a wide spectrum of pharmacological activities. In addition, in different studies, anticancer agents have been combined to cholesterol moieties to improve their cellular uptake and target specificity. Herein, we investigated the cytotoxic activity of various triterpenoids cholesterol analogs: oleanolic acid (OleA), ursolic acid (UrA), betulinic acid (BeA), asiatic acid (AsA), lupeol (Lupe), stigmasterol (Stg) and β‐sitosterol (β‐Sito) on non‐small lung adenocarcinoma cells (A549) and normal lung cells (MRC5). From our studies, the IC 50 for most of these cholesterol‐analogs were in the low μM range after 24h of incubation. β‐Sito and Stg did not show any significant cytotoxic pattern in this μM range. In addition BeA and OleA showed the highest therapeutic indexes comparing the cytotoxic effect in A549 vs MRC5 cells. Metabolic activity assays were performed to determine the mechanism of action of these natural compounds. Caspases activity and cell membrane integrity assays were performed and all the cytotoxic triterpenoids induced no activation of caspase‐3, and high membrane permeabilization. Gene expression assays of MDRS‐related genes (EGF, VEGF, Pgp, Bcl‐2, P 53 , NDRG1, GLIPR1) demonstrated the downregulation of some of these genes after the incubation with these triterpenoids. These results demonstrated that even slight structural changes in these cholesterol analogs can influence the cytotoxic response and cellular mechanistic pathways. Complete results will be presented. This study opens promising perspectives for further research on the role of phytochemical triterpenoids, which ultimately will contribute to a more rational application in cancer therapy.Support or Funding InformationSJB‐Research Center Seed

Corosolic acid impairs human lung adenocarcinoma A549 cells proliferation by inhibiting cell migration.

The present study aimed to investigate the anticancer effects of corosolic acid (CA) in the human lung adenocarcinoma A549 cell line. A549 cells were treated with increasing concentrations of CA, prior to assessing cell viability, migration rate, vascular endothelial growth factor receptor 2 (VEGFR2) kinase activity and cytoskeleton structure. In addition, in vivo imaging system was used to analyze the anticancer effects of CA in vivo. Results demonstrated that CA exhibited a low cytotoxicity with a half maximal inhibitory concentration of 65 µM. In addition, 4 µM CA efficiently inhibited A549 cell migration. Furthermore, CA inhibited VEGFR2 kinase activity and disrupted tubulin structure. Data also revealed that CA inhibited A549 cell proliferation in a xenograft mouse model. In conclusion, results from the present study suggested that CA may be used as a novel potential therapy for lung cancer.

Specific inhibition of Notch1 signaling suppresses properties of lung cancer stem cells.

Lung cancer is the leading cause of cancer-related death worldwide with a relatively low 5-year relative survival rate of 16%. Novel and efficient therapeutic approach for lung cancer is desperately needed. Targeting cancer stem cells (CSCs) provides a promising strategy to eradicate malignancies. The Notch signaling pathway plays an important role in the control of cell fates and developmental processes including CSCs. The function of Notch1 in the regulation of CSCs and whether targeting Notch1 could be a potential therapy for lung cancer were explored in this study. Lung CSCs (LCSCs) were isolated from A549 cells and identified as CD44+/CD24- cells by magnetic-assisted cell sorting, then the putative LCSCs were treated with Notch1 inhibitor and Notch1 small interfering RNAs (siRNAs); the growth and proliferation of LCSCs were investigated to test the effect of Notch1 blocking on the growth and viability of LCSCs. CD44+/CD24- cells isolated from A549 cells possessed stem cell-like properties with high expression of Notch1. Blocking Notch1 by inhibitor DAPT or siRNA both inhibited the growth capacity of LCSCs. Our discovery demonstrated a depression of growth in CD44+/CD24- and A549 cells caused by blockade of Notch signaling pathway. Further studies are needed to demonstrate the detailed effects of Notch1 blocking on the LCSCs. Nevertheless, targeting the Notch pathway has exhibited great potential to be an improved lung cancer treatment.

Synthesis and evaluation of osimertinib derivatives as potent EGFR inhibitors

Osimertinib has been identified as a promising therapeutic drug targeting for EGFR T790M mutant non-small cell lung cancer (NSCLC). A new series of N-oxidized and fluorinated osimertinib derivatives were designed and synthesized. The cellular anti-proliferative activity, kinase inhibitory activity and the activation of EGFR signaling pathways of 1–6 in vitro were determined against L858R/T790M and wild-type EGFR, the antitumor efficacy in NCI-H1975 xenografts in vivo were further studied. Compound 2, the newly synthesized N-oxide metabolite in N,N,N′-trimethylethylenediamine side chain of osimertinib, showed a comparable kinase selectivity in vitro and a slightly better antitumor efficacy in vivo to osimertinib, making it valuable and suitable for the potential lung cancer therapy.

Silencing Receptor EphA2 Enhanced Sensitivity to Lipoplatin™ in Lung Tumor and MPM Cells

ABSTRACTReceptor EphA2 is overexpressed in lung cancer and malignant pleural mesothelioma (MPM) which promote tumorogenesis. Lipoplatin™, a new liposomal cisplatin formulation, is used against resistant tumors. Use of cisplatin-based drugs leads to unacceptable toxicities. To improve the effectiveness of Lipoplatin, enhancing the cellular sensitivity of lung tumor and MPM cells is critical. Therefore, we targeted receptor EphA2 by silencing interference RNA (siRNA) and treated tumor cells with Lipoplatin. The combined effects of siRNA-EphA2 and Lipoplatin were determined. We report that silencing EphA2 significantly enhanced the cellular sensitivity of lung tumor and MPM cells to Lipoplatin and maybe a potential therapy for lung cancer.

Hyaluronic acid-modified zirconium phosphate nanoparticles for potential lung cancer therapy.

Novel tumor-targeting zirconium phosphate (ZP) nanoparticles modified with hyaluronic acid (HA) were developed (HA-ZP), with the aim of combining the drug-loading property of ZP and the tumor-targeting ability of HA to construct a tumor-targeting paclitaxel (PTX) delivery system for potential lung cancer therapy. The experimental results indicated that PTX loading into the HA-ZP nanoparticles was as high as 20.36%±4.37%, which is favorable for cancer therapy. PTX-loaded HA-ZP nanoparticles increased the accumulation of PTX in A549 lung cancer cells via HA-mediated endocytosis and exhibited superior anticancer activity in vitro. In vivo anticancer efficacy assay revealed that HA-ZP nanoparticles possessed preferable anticancer abilities, which exhibited minimized toxic side effects of PTX and strong tumor-suppression potential in clinical application.

Enhanced antitumor effect of combining TRAIL and MnSOD mediated by CEA-controlled oncolytic adenovirus in lung cancer.

Lung cancer, especially adenocarcinoma, is one of the leading causes of death in the world. Carcinoembryonic antigen (CEA), a superb non-small-cell lung cancer marker candidate, showed a beneficial effect in cancer therapy with oncolytic adenovirus in recent studies. Cancer-targeting dual gene-virotherapy delivers two therapeutic genes, linked by a connexon, in the replication-deficient vector instead of one gene so that they can work in common. In this study, we constructed a tumor-specific oncolytic adenovirus, CD55-TRAIL-IETD-MnSOD. The virus has the fusion protein complementary DNAs for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and for manganese superoxide dismutase (MnSOD) complementary DNA linked through a 4-amino acid caspase-8 cleavage site (IETD), and uses a CEA promoter to control virus E1A express. This is the first work to use a CEA promoter-regulated oncolytic adenovirus carrying two therapeutic genes for cancer research. Its targeting and anticancer capacity was evaluated by in vitro and in vivo experiments. The results indicated that CD55-TRAIL-IETD-MnSOD caused more cell apoptosis than CD55-TRAIL or CD55-MnSOD alone, or their combination in vitro, with low cytotoxicity of normal cells. In the A549 tumor xenograft model in nude mice, data showed that CD55-TRAIL-IETD-MnSOD could effectively suppress tumor growth than single gene groups, with no histological damage in liver, spleen or kidney tissues. Thus, the CEA-regulated dual-gene oncolytic virus CD55-TRAIL-IETD-MnSOD may be a novel potential therapy for lung cancer.

Candesartan-graft-polyethyleneimine cationic micelles for effective co-delivery of drug and gene in anti-angiogenic lung cancer therapy

A multifunctional polymer-drug conjugate candesartan-graft-polyethyleneimine (CD-PEI, CP) containing low molecular weight polyethyleneimine (PEI) and candesartan (CD) conjugated via an amide bond was fabricated as a co-delivery micelle of drug and siRNA for potential lung cancer therapy. Here, CD as an angiotensin II type 1 receptor blocker rich in imidazole and tetrazole rings was utilized to strengthen endosomal buffering capacity of CP and suppress tumor angiogenesis. The selfassembled CP/siRNA complexes exhibited desirable and homogenous particle size, moderate positive charges, and efficient release of drug and siRNA in vitro. In addition, CD and siRNA could readily detached from nanovectors in tumor cells via an amidase-responsive mechanism and they achieved synergistic anti-angiogenesis efficacy by effectively downregulating the expression of vascular endothelial growth factor (VEGF) mRNA and protein via different pathways in vitro. In vivo investigation on nude mice bearing A549 tumor xenografts revealed that CP/siRNA complexes possessed strong antitumor activity. These findings suggested that CP could be an ideal nanovector for simultaneous transfer of drug and siRNA, and a multifunctional CP/siRNA co-delivery system with enhanced endosomal buffering capacity, amidase-responsive drug release and synergistic anti-angiogenesis efficacy might be a new promising strategy for effective lung cancer therapy.

175 POSTER Chemoresistnce induction by tumor stroma interactions - molecular and epigenetic regulation of caspases

Osimertinib has been identified as a promising therapeutic drug targeting for EGFR T790M mutant non-small cell lung cancer (NSCLC). A new series of N-oxidized and fluorinated osimertinib derivatives were designed and synthesized. The cellular anti-proliferative activity, kinase inhibitory activity and the activation of EGFR signaling pathways of 1–6 in vitro were determined against L858R/T790M and wild-type EGFR, the antitumor efficacy in NCI-H1975 xenografts in vivo were further studied. Compound 2, the newly synthesized N-oxide metabolite in N,N,N′-trimethylethylenediamine side chain of osimertinib, showed a comparable kinase selectivity in vitro and a slightly better antitumor efficacy in vivo to osimertinib, making it valuable and suitable for the potential lung cancer therapy.