ABSTRACT
Lung cancer is the leading reason of cancer-related death from cancer globally for both men and women. Recently, tumor immune heterogeneity has been implicated in cancer clinical outcome. However, this prognostic significance of immune cell types in lung squamous cell carcinoma (LUSC) is unclear and should be systematically investigated. Two microarray datasets (GSE67061 and GSE2088) from the Gene Expression Omnibus (GEO) database were downloaded and then integrated to estimate the fraction of 22 immune cell types by CIBERSORT algorithm. To validate the estimation for LUSC, the data of LUSC TCGA were also assessed in order to determine specific infiltrating immune cell type closely correlated with LUSC patients' survival determined by Cox regression analyses. Immunotherapeutic and chemotherapeutic response between the LUSC patients were also evaluated. T follicular helper cells were obtained by Cox regression analysis to develop the prognostic signature. According to this immune prognostic risk score, immune signature of T follicular helper cells is an independent and specific prognostic signature for predictions of LUSC patient overall survival. Moreover, high-risk group exhibited less expression of N6-methyladenosine (m6A) RNA methylation regulator including ALKBH5, METTL3, HNRNPC and KIAA1429 and was much more sensitive to immunotherapy and chemotherapy. This study suggests that this immune signature is important determinants of prognosis in LUSC and may provide potential prognostic biomarker or therapeutic target for immunotherapeutic and chemotherapeutic development.
Subject(s)
Biomarkers, Tumor/genetics , Carcinoma, Non-Small-Cell Lung/immunology , Carcinoma, Squamous Cell/immunology , Lung Neoplasms/immunology , T Follicular Helper Cells/immunology , Adenosine/analogs & derivatives , Adenosine/metabolism , Aged , AlkB Homolog 5, RNA Demethylase/genetics , AlkB Homolog 5, RNA Demethylase/immunology , Antineoplastic Agents, Immunological/pharmacology , Antineoplastic Agents, Immunological/therapeutic use , Biomarkers, Tumor/immunology , Carcinoma, Non-Small-Cell Lung/drug therapy , Carcinoma, Non-Small-Cell Lung/genetics , Carcinoma, Non-Small-Cell Lung/mortality , Carcinoma, Squamous Cell/drug therapy , Carcinoma, Squamous Cell/genetics , Carcinoma, Squamous Cell/mortality , Datasets as Topic , Female , Gene Expression Regulation, Neoplastic/drug effects , Gene Expression Regulation, Neoplastic/immunology , Heterogeneous-Nuclear Ribonucleoprotein Group C/genetics , Heterogeneous-Nuclear Ribonucleoprotein Group C/immunology , Humans , Kaplan-Meier Estimate , Lung Neoplasms/drug therapy , Lung Neoplasms/genetics , Lung Neoplasms/mortality , Lymphocytes, Tumor-Infiltrating/drug effects , Lymphocytes, Tumor-Infiltrating/immunology , Lymphocytes, Tumor-Infiltrating/metabolism , Male , Methylation/drug effects , Methyltransferases/genetics , Methyltransferases/immunology , Middle Aged , Prognosis , RNA-Binding Proteins/genetics , RNA-Binding Proteins/immunology , RNA-Seq , T Follicular Helper Cells/drug effects , T Follicular Helper Cells/metabolism , Transcriptome/drug effects , Transcriptome/immunologyABSTRACT
BACKGROUND: Ribonucleoproteins particles that form the spliceosomes are among the most frequently targeted molecules of the autoimmune response. In the last few years, autoantibodies against all A/B hnRNP proteins have been found in the sera of patients with rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), mixed connective tissue disease (MCTD), and serve as diagnostic markers for several rheumatic diseases. However, the functional role of hnRNP C1/C2 in autoimmune diseases is still not clearly understood. OBJECTIVE: To identify hnRNP C1/C2 as an autoantigen in patients with Behcet's Disease (BD). METHODS: First, HaCaT and EA.hy926 cells were cultured and RNA was extracted. Second, amplification of the corresponding gene by RT-PCR, cloning, and purification techniques was applied to acquire the recombinant protein hnRNP C1/C2. Third, the target protein band was excised from gel electrophoresis, digested with trypsin, and analyzed by (MALDI-TOF/). Finally, Western blotting and ELISA were performed to verify the immunoreactivity of BD serum with recombinant hnRNPC1/C2. RESULTS: Results demonstrated that the reactivity of BD serum against recombinant hnRNP C1/C2 protein was significantly higher as compared to healthy control (P<0.001). CONCLUSION: hnRNP C1/C2 can be considered as a self antigen which might be involved in BD pathology in Hans Chinese population.
Subject(s)
Autoantigens/immunology , Autoimmunity , Behcet Syndrome/immunology , Heterogeneous-Nuclear Ribonucleoprotein Group C/immunology , Adolescent , Adult , Aged , Aged, 80 and over , Arthritis, Rheumatoid/immunology , Autoantibodies/blood , Autoantibodies/immunology , Autoantigens/genetics , Autoantigens/isolation & purification , Behcet Syndrome/genetics , Case-Control Studies , Cell Line , Cloning, Molecular , Enzyme-Linked Immunosorbent Assay , Female , Gene Expression , Heterogeneous-Nuclear Ribonucleoprotein Group C/genetics , Heterogeneous-Nuclear Ribonucleoprotein Group C/isolation & purification , Humans , Lupus Erythematosus, Systemic/immunology , Male , Mass Spectrometry , Middle Aged , ROC Curve , Young AdultABSTRACT
Spinal muscular atrophy (SMA) is caused by loss of SMN1. A nearly identical gene, SMN2, fails to compensate for the loss of SMN1 because SMN2 produces mainly an exon 7-skipped product. The +6C in SMN1 exon 7 proceeds to include exon 7 into mRNA, while the +6U in SMN2 causes skipping of exon 7. Here, approximately 45kD proteins bound to the SMN exon 7 RNA probe was found, and identified as hnRNP C1/C2. In gel-shift assay, hnRNP C1/C2 had a greater affinity for the RNA probe with +6C than for the RNA probe with +6U. In vitro splicing assay showed that anti-hnRNP C1/C2 antibody hampered splicing of SMN1 exon 7, but did not affect splicing of SMN2 exon 7. In conclusion, we showed the possibility that hnRNP C1/C2 enhanced SMN1 exon 7 splicing specifically.
Subject(s)
Heterogeneous-Nuclear Ribonucleoprotein Group C/metabolism , Muscular Atrophy, Spinal/genetics , RNA Splicing , Survival of Motor Neuron 1 Protein/genetics , Amino Acid Sequence , Antibodies , Exons , Gene Silencing , HeLa Cells , Heterogeneous-Nuclear Ribonucleoprotein Group C/immunology , Humans , Mass Spectrometry , Molecular Sequence Data , RNA Probes , Survival of Motor Neuron 2 Protein/geneticsABSTRACT
Heterogeneous nuclear ribonucleoproteins (hnRNPs) are among the most abundant proteins in the eukaryotic cell nucleus and play a direct role in several aspects of the RNA life including splicing, export of the mature RNAs and translation. To date, approximately 30 proteins have been identified. A growing body of evidence points to hnRNPs as an important target of the autoimmune response in rheumatic diseases. Autoantibodies to A and B proteins of the hnRNP complex have been detected in late 1980s in rheumatoid arthritis (RA), systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD). Beyond their role as diagnostic test in clinical practice, these autoantibodies are starting to be regarded as important tools to obtain deeper insight into the pathogenesis of autoimmune rheumatic diseases. Furthermore, new anti-hnRNP antibodies have been recognized in the last ten years extending the spectrum of anti-hnRNP reactivity in different autoimmune disorders.
