Canine DH-28 cells persistant infection with CDV, starving and hypoxic treatments
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ABSTRACT: 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: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: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:The recently developed COXEN method (PMID: 17666531) has been used to successfully extrapolate gene signatures of drug sensitivity across different tumor histotypes. We wanted to explore the utility of COXEN to predict chemosensitivity in canine cancer, specifically if we could extrapolate gene signatures identified in human datasets over to canine osteosarcoma tumors. This dataset of canine osteosarcoma tumor samples has available clinical outcome data after patients had infected limbs amputated and were treated with doxorubicin and/or carboplatin. We performed microarray analysis on this panel of tumor samples for validating our COXEN prediction models for doxorubicin or carboplatin sensitivity.
Project description:The recently developed COXEN method (PMID: 17666531) has been used to successfully extrapolate gene signatures of drug sensitivity across different tumor histotypes. We wanted to explore the utility of COXEN to predict chemosensitivity in canine cancer, specifically if we could extrapolate gene signatures identified in human datasets over to canine osteosarcoma tumors. This dataset of canine osteosarcoma tumor samples has available clinical outcome data after patients had infected limbs amputated and were treated with doxorubicin and/or carboplatin. We performed microarray analysis on this panel of tumor samples for validating our COXEN prediction models for doxorubicin or carboplatin sensitivity. Chemotherapy naive primary tumors were collected at the time of amputation and archived at the Flint Animal Cancer Center. RNA was extracted from 33 frozen archived tumor samples, followed by microarray analysis. The gene expression data was RMA preprocessed, scaled and were used as a independent test set to evaluate developed prediction models of sensitivity to doxorubicin or carboplatin. Drug predictions were than compared to clinical outcome in these patients that received doxorubicin and/or carboplatin.