Project description:MicroRNAs (miRNAs), a class of noncoding RNAs measuring 18 to 23 nucleotides (nt) that play important regulatory roles in host-virus interactions. Avian-origin H3N2 canine influenza virus (CIV) has emerged as the most prevalent subtype among dogs in Asia since 2007. To evaluate the roles of host miRNAs in H3N2 CIV infection, here, miRNA profiles obtained from primary canine bronchiolar epithelial cells (CBECs) and canine alveolar macrophages (CAMCs) were compared between infected and mock-infected cells with the H3N2 CIV JS/10. It was found that cfa-miR-125b, cfa-miR-151 and cfa-miR-423a expressions were significantly decreased in CIV-infected canine primary cells. Bioinformatics prediction indicated that 5’ seed regions of three miRNAs are partially complementary to the mRNAs of nucleoprotein (NP) and non-structural protein 1 (NS1) of JS/10. As determined by virus titration, quantitative real-time PCR (qRT-PCR) and western blotting, overexpression of cfa-miR-125b and cfa-miR-151 inhibited CIV infection, whereas overexpression or inhibition of cfa-miR-423a inhibited this infection. These results indicated that CIV replication could be regulated by miRNAs from host cells infected with CIV. Our findings support the notion that cellular miRNAs can inhibit virus infection, help to elucidate the resistance of host cells to viral infection and to clarify the pathogenesis of H3N2 CIV. We used microarrays to detail the global programme of gene expression of primary canine alveolar macrophages (CAMCs) compared between infected and mock-infected cells with the H3N2 canine influenza virus (CIV) JS/10.
Project description:We demonstrated canine influenza virus (H3N2) pathogenicity to dogs using microarray analysis. Many genes related to innate immunity, such as toll-like receptors, immune cells of natural killer cells, macrophages, neutrophils, nitric oxide and reactive oxygen species, and interferon, were induced. RNA was extracted from canine influenza virus H3N2-infected dogs. The lung RNA of uninfected dogs was used as a negative control. We compared gene expression levels between infected and uninfected dogs using microarray analysis.
Project description:The objective of this study was to compare the transcriptomes of uninfected DH82 cells, a canine histiocytic sarcoma cell line, with DH82 cells, persistently infected with the Onderstepoort strain of canine distemper virus in order to identify effects of canine distemper virus infection upon the transcriptome of this tumor cell line and hereby draw conclusions on possible paramyxovirus-induced oncolytic mechanisms
Project description:Our previous study revealed that influenza virus matrix protein 1 (M1)and matrix 2 (M2) interacted with canine RIG-I. However, the influence of these interactions remains in the mist. To explore these cellular transcriptome modulation induced by the interaction between CIV M1/M2 and canine RIG-I, here we carried out the comparative transcriptional RNA analysis of the overexpression of CIV M1/M2 along with canine RIG-I in HEK 293T cells.
Project description:Analysis of the covalent attachment of GMP to the RNA dependent RNA polymerase proteins of equine arteritis virus and SARS-CoV-2 proteins using heavy-isotope assisted MS and MS/MS peptide sequencing.
Project description:RNAseq analysis of canine DH-28 tumor cells persistently infected with CDV canine distemper virus, starving and hypoxic conditions. Gene expression changes in infected and control cells were studied. Results: Many differences between infection and controls and different treatments
Project description:Spontaneously occurring canine mammary cancer represents an excellent model of human breast cancer, but is greatly understudied. To better use this valuable resource, we performed whole-genome sequencing, whole-exome sequencing, RNA-seq, and/or high-density arrays on twelve canine mammary cancer cases, including seven simple carcinomas and four complex carcinomas. Canine simple carcinomas, which histologically match human breast carcinomas, harbor extensive genomic aberrations, many of which faithfully recapitulate key features of human breast cancer. Canine complex carcinomas, which are characterized by proliferation of both luminal and myoepithelial cells and are rare in human breast cancer, seem to lack genomic abnormalities. Instead, these tumors have about 35 chromatin-modification genes downregulated and are abnormally enriched with active histone modification H4-acetylation, whereas aberrantly depleted with repressive histone modification H3K9me3. Our findings indicate the likelihood that canine simple carcinomas arise from genomic aberrations, whereas complex carcinomas originate from epigenomic alterations, reinforcing their unique value. Canine complex carcinomas offer an ideal system to study myoepithelial cells, the second major cell lineage of the mammary gland. Canine simple carcinomas, which faithfully represent human breast carcinomas at the molecular level, provide indispensable models for basic and translational breast cancer research.
Project description:Targeting PRMT5 in canine lymphoma: we characterized expression patterns of PRMT5 in canine lymphomas and correlated these with histological subtypes using tissue microarrays. We characterized expression of PRMT5 in three canine lymphoma-derived cell lines and primary lymph node biopsies. We have demonstrated that inhibition of PRMT5 leads to growth suppression and induction of apoptosis in canine lymphoma cell lines and primary canine lymphoma cells in a time and dose-dependent manner, while selectively decreasing global marks of symmetric dimethylarginine (SDMA) and histone H4 arginine 3 symmetric di-methylation. We performed ATAC-sequencing with pathway enrichment analysis to characterize genome-wide changes in chromatin accessibility after PRMT5 inhibition. We performed gene expression microarrays with pathway analysis to characterize whole transcriptome changes in canine lymphoma cell lines treated with PRMT5 inhibitors. This work validates PRMT5 as a promising therapeutic target for canine lymphoma and supports the continued used of the spontaneously occurring canine lymphoma model for the preclinical development of PRMT5 inhibitors for the treatment of human NHL.
