Current Gene Therapy - Volume 21, Issue 4, 2021

Background: Colorectal cancer (CRC) is a kind of tumor with high incidence and its treatment situation is still very difficult despite the constant renewal and development of treatment methods. Objective: To assist the prognosis, monitoring and survival of CRC patients with a model. Methods: In this study, we established a new prognostic model for CRC. Four groups of CRC data were accessed from the GEO database, and then differential analysis (logFoldChange>1, adjust- P<0.05) was carried out by using the limma package along with the RobustRankAggreg package used to identify the overlapping differentially expressed genes (DEGs). Univariate and multivariate Cox regression analyses were performed on the DEGs to screen the genes related to the patient’s prognosis, and a five-gene prognostic prediction model (including RPX, CXCL13, MMP10, FABP4 and CLDN23) was constructed. Then, we further plotted ROC curves to evaluate the predictive performance of the five-gene prognostic signature in the TCGA data sets (the AUC values of 1, 3, 5-year survival were 0.68, 0.632, 0.675, respectively) and an external independent data set GSE2962 (the AUC values of 1, 3, 5-year survival were 0.689, 0.702, 0.631, respectively). Results: The results showed that the model could effectively predict the prognosis of CRC patients, which provides a robust predictive model for the prognosis of CRC patients. Conclusion: The model could effectively predict the prognosis of CRC patients, which provides a robust predictive model for the prognosis of CRC patients.

Objective: The purpose of this study was to explore the mechanism of the miR-375/XPR1 axis in esophageal squamous cell carcinoma (ESCC) and provide a new idea for targeted therapy of ESCC. Methods: Differentially expressed genes in GEO and TCGA databases were analyzed by bioinformatics. The expression levels of miR-375 and XPR1 mRNA were detected by qRT-PCR. Protein expression of XPR1 was detected by western blot. Bioinformatics analysis and dual luciferase assay were conducted to confirm the target relationship between miR-375 and XPR1. The viability, proliferation, migration and invasion of cells in each treatment group were detected by CCK-8, colony formation, wound healing and Transwell assays. Results: Significantly down-regulated miR-375 and remarkably up-regulated XPR1 were observed in ESCC tissue and cells. Overexpression of miR-375 inhibited proliferation, invasion and migration of ESCC cells, and greatly reduced the promoting effect of XPR1 overexpression on cell proliferation, migration and invasion. Dual luciferase assay confirmed that miR-375 targeted and inhibited XPR1 expression in ESCC. Conclusion: These results demonstrate the regulatory role of the miR-375/XPR1 axis in ESCC cells and provide a new potential target for the precise treatment of patients with ESCC.

Aims: This study aims to identify the biomarkers for chronic hypersensitivity pneumonitis (CHP) and facilitate the precise gene therapy of CHP. Background: Chronic hypersensitivity pneumonitis (CHP) is an interstitial lung disease caused by hypersensitive reactions to inhaled antigens. Clinically, the task of differentiating CHP and other interstitial lung diseases, especially idiopathic pulmonary fibrosis (IPF), was challenging. Objective: In this study, we analyzed the publically available gene expression profile of 82 CHP patients, 103 IPF patients, and 103 control samples to identify the CHP biomarkers. Methods: The CHP biomarkers were selected with advanced feature selection methods: Monte Carlo Feature Selection (MCFS) and Incremental Feature Selection (IFS). A Support Vector Machine (SVM) classifier was built. Then, we analyzed these CHP biomarkers through functional enrichment analysis and differential co-expression analysis. Results: There were 674 identified CHP biomarkers. The co-expression network of these biomarkers in CHP included more negative regulations and the network structure of CHP was quite different from the network of IPF and control. Conclusion: The SVM classifier may serve as an important clinical tool to address the challenging task of differentiating between CHP and IPF. Many of the biomarker genes on the differential coexpression network showed great promise in revealing the underlying mechanisms of CHP.

Objective: This study presents a discussion regarding the mechanism affecting the malignant progression of LUAD and the potential therapeutic targets, so as to provide more effective therapeutic strategies for LUAD patients. Methods: Expression data from TCGA-LUAD were extracted to identify target miRNA, with its downstream target mRNA predicted using bioinformatics analysis. Gene expression in transcript level and protein level were separately examined by qRT-PCR and western blot. Cell malignant phenotypes were assessed via MTT and Transwell assays. Luciferase reporter plasmids carrying target gene sequences were constructed to verify the targeting association between the target miRNA and its downstream mRNA. Results: miR-1-3p showed decreased expression in LUAD. Over-expressing miR-1-3p suppressed cancer cells to proliferate, migrate and invade. CELSR3, directly regulated by miR-1-3p, presented significantly elevated expression in LUAD and could foster LUAD cells to proliferate, migrate and invade. The rescue experiment identified that miR-1-3p-induced inhibition on LUAD cell malignant phenotypes could be reversed by over-expressing CELSR3. Conclusion: This study uncovered that miR-1-3p could suppress the malignant phenotypes of LUAD cells by targeting CELSR3, which will help to provide novel therapeutic strategies for LUAD sufferers and new references for the targeted therapy of LUAD.

The risk of heart failure is 2-5 times higher in diabetic patients as compared to non-diabetic patients with similar comorbidities. Recent reports suggest that nearly half of the diabetic population remains undiagnosed, making diabetic cardiomyopathy (DCM) a clinically relevant entity. In the myocardium, chronic hyperglycemia elicits structural and functional abnormalities characterized by ventricular dilation, diastolic dysfunction, fibrosis, and hypertrophy leading to heart failure. Since diabetes is a multifactorial heterogeneous metabolic disorder which cannot be diagnosed or controlled along with coronary artery disease or hypertension, there is an urgent need to understand the underlying molecular mechanisms that leads to DCM and identify potential therapeutic targets. Small non-coding RNAs, in particular, microRNAs (miRNAs), have emerged as key regulators for several life-threatening diseases, including DCM. Recent studies have reported that miRNAs not only regulate the fundamental mechanisms of DCM such as insulin resistance, MAPK pathway, PI3K-AkT pathway, oxidative stress, also inflammatory signaling, but also possess the potential to be a therapeutic or diagnostic target. This review examines the role of critical miRNAs in the onset and pathogenesis of DCM, which also depicts high potential as therapeutic and diagnostic molecule in preclinical studies. Further, it highlights the completed and on-going clinical trials around the globe for diabetes and miRNAs to provide a outlook about the upcoming miRNA therapeutics.

Recently, CRISPR-based techniques have significantly improved our ability to make desired changes and regulations in various genomes. Among them, targeted base editing is one of the most powerful techniques in making precise genomic editing. Base editing enabled the irreversible conversion of a specific single DNA base, from C to T or and from A to G, in desired genomic loci. This technique has important implications in the study of human genetic diseases, considering that many of them resulted from point mutations. More importantly, the high efficiency of these editing tools also provided great promise in clinical applications. In this review, we discuss the recent progress and challenges of base editing tools.

Background: Lung adenocarcinoma (LADC) is the most common type of lung cancer and is a subtype of non-small-cell lung cancer (NSCLC). Approximately 40% of LADC patients experience brain metastases (BMs) during the course of the disease. In this study, integrated bioinformatics methods were applied to identify key genes related to brain metastasis in lung adenocarcinoma. Methods: We derived and characterized genes differentially expressed between the primary tumour and brain metastases using tumour cells isolated from two lung cancer Patient-derived xenografts (PDX) cases (GSE 69405). Gene ontology (GO) and KEGG pathway enrichment analyses were applied, and protein-protein interaction (PPI) networks and Cytoscape software were utilized to identify key genes. Results: Four key genes, including CKAP4 (Cytoskeleton Associated Protein 4), SERPINA1 (Serpin Family A Member 1), SDC2 (Syndecan 2) and GNG11 (G Protein Subunit Gamma 11) were identified for BM-LADC by the Venn diagram. Conclusion: We believe these key genes may be potential biomarkers for improved prognosis and treatment of lung adenocarcinoma.

Background: Successful delivery of gene-editing tools using nano-carriers is dependent on the ability of nanoparticles to pass through the cellular membrane, move through the cytoplasm, and cross the nuclear envelope to enter the nucleus. It is critical that intracellular nanoparticles interact with the cytoskeletal network to move toward the nucleus, and must escape degradation pathways including lysosomal digestion. Without efficient intracellular transportation and nuclear entry, nanoparticles-based gene-editing cannot be effectively used for targeted genomic modification. Objective: We have developed nanoparticles with a low molecular weight branched polyethylenimine lipid shell and a PLGA core that can effectively deliver plasmid DNA to macrophages for gene editing while limiting toxicity. Methods: Core-shell nanoparticles were synthesized by a modified solvent evaporation method and were loaded with plasmid DNA. Confocal microscopy was used to visualize the internalization, intracellular distribution and cytoplasmic transportation of plasmid DNA loaded nanoparticles (pDNA-NPs) in bone marrow-derived macrophages. Results: Core-shell nanoparticles had a high surface charge of +56 mV and narrow size distribution. When loaded with plasmid DNA for transfection, the nanoparticles increased in size from 150 nm to 200 nm, and the zeta potential decreased to +36 mV, indicating successful encapsulation. Further, fluorescence microscopy revealed that pDNA-NPs crossed the cell membrane and interacted with actin filaments. Intracellular tracking of pDNA-NPs showed successful separation of pDNA- NPs from lysosomes, allowing entry into the nucleus at 2 hours, with further nuclear ingress up to 5 hours. Bone marrow-derived macrophages treated with pDNA/GFP-NPs exhibited high GFP expression with low cytotoxicity. Conclusion: Together, this data suggests pDNA-NPs are an effective delivery system for macrophage gene-editing.