Browsing by Author "Avery, Paul, committee member"
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Item Open Access Clinical and molecular characterization of canine small cell B-cell lymphocytosis disorders(Colorado State University. Libraries, 2020) Rout, Emily, author; Avery, Anne, advisor; Avery, Paul, committee member; Zabel, Mark, committee member; Weil, Michael, committee memberTo view the abstract, please see the full text of the document.Item Open Access Cytokines, antibodies and plasma viremia of cats infected with feline immunodeficiency virus(Colorado State University. Libraries, 2013) Wood, Britta Ann, author; VandeWoude, Sue, advisor; Avery, Paul, committee member; Zabel, Mark, committee member; Hussey, Gisela, committee memberFeline immunodeficiency viruses (FIVs) are naturally occurring lentiviruses (family Retroviridae) of felid species, including domestic and wild cats. Studies on FIVs are beneficial for understanding the host immune response associated with disease progression (e.g., domestic cat FIV) or the viral kinetics and molecular ecology associated with naturally occurring infections in wildlife (e.g., bobcat and mountain lion FIVs). Here we describe the development and validation of the following microsphere immunoassays (MIAs) for evaluating the cytokine and antibody response of domestic cats: i) the quantification of cytokines (interferon gamma (IFNγ), interleukin (IL)-10, and IL-12/IL-23) in cell culture supernatant, and ii) the quantification of these cytokines in plasma; iii) the quantification of total IgG and IgA in plasma, and iv) the detection of IgG and IgA antibodies to feline CD134 (the primary cell receptor for FIV), and FIV capsid (CA) and surface (SU) proteins in plasma. These assays were used to evaluate temporal cytokine and antibody responses of domestic cats experimentally infected with various FIV strains. To analyze viral RNA loads associated with naturally occurring FIV infections in bobcats or mountain lions, we are adapting existing quantitative PCR assays for use with plasma samples. The eight assays described here are/will be beneficial for addressing questions related to lentiviral immune response and viral kinetics.Item Open Access Molecular characterization of canine peripheral T-cell lymphoma(Colorado State University. Libraries, 2020) Harris, Lauren, author; Avery, Anne, advisor; Avery, Paul, committee member; Basaraba, Randall, committee member; Bailey, Susan, committee memberTo view the abstract, please see the full text of the document.Item Open Access Overactive NF-KB signaling as a druggable target and evaluation of parthenolide an NF-KB inhibitor in canine cancer(Colorado State University. Libraries, 2022) Schlein, Lisa Janelle, author; Thamm, Douglas H., advisor; Avery, Paul, committee member; Duval, Dawn, committee member; MacNeill, Amy, committee memberThis study provides a unique translational research opportunity to help both humans and dogs diagnosed with diseases that carry dismal prognoses in both species: histiocytic sarcoma (HS), hemangiosarcoma (HSA), and disseminated mastocytosis (MCT). Lymphoma is one of the most common cancer types affecting dogs and humans, and therefore, novel therapeutic approaches are always needed. For all of these cancer types, dogs and human cancers share common molecular abnormalities, consistent with a conserved pathogenesis between species. Relative to traditional murine models for human cancers, dogs are genetically diverse, large mammals with heterogeneous, spontaneous tumors. Dogs generally receive good medical care and share the environmental factors with humans, and accordingly, dogs with spontaneous tumors are an excellent model for human oncology generally. Additionally, although disseminated HS, MCT and visceral HSA are exceedingly rare diseases in humans, they are more common in some dog breeds, giving us the opportunity to study this disease in a larger population than would otherwise be available. Therapeutics evaluated in dogs with these diseases stand to benefit both canine and human patients. NF-kB proteins are a family of structurally related, eukaryotic transcription factors that have 400+ genetic targets, and are involved in many vital cellular processes, including innate immunity, inflammatory responses, development, cellular growth, and survival. Not surprisingly, overactivation of NF-kB is a feature of many chronic disease processes, including cardiac disease, neurodegenerative disease, immune-mediated disease, and cancer. While NF-kB overactivation has been documented extensively in human oncology, there is a relative paucity of data documenting the same phenomenon in veterinary medicine. As part of this study, large scale validation of NF-kB overactivation was performed in canine cancer via immunohistochemistry of 215 tumor samples (lymphoma, HS, HSA, and MCT). Antibodies were validated for use via western blot, immortalized cell pellets, and evaluation of normal canine tissues. In addition to validation of NF-kB overactivation, assays were performed to assess the therapeutic potential of parthenolide (PTL), a known, canonical NF-kB signaling inhibitor with additional mechanisms of antineoplastic activity, including alteration of cellular redox balance. Growth inhibition assays were performed with canine cell lines and primary lymphoma cells isolated from canine patients, using PTL alone or in combination with redox-perturbing standard-of-care therapeutics. Cell death was assessed using flow cytometry. Immunofluorescence was used to assess NF-kB localization, western blot was used to assess NF-kB activity with and without PTL, and canine cells were transfected with a reporter gene cassette containing the NFkB consensus sequence followed by firefly luciferase gene to study the effect of PTL on NF-kB-related luminescence. PTL's effects on glutathione and reactive oxygen species generation were assessed with a colorimetric assay and a fluorescent H2DCFDA assay, respectively. Genetic expression changes were assessed with RNA sequencing of HS cells, with and without PTL treatment. A mouse model of disseminated HS was created with NF-kB luminescent cells to study the effect of PTL on this disease in vivo. Many spontaneous canine tumor samples have nuclear p65 and p100/p52 IHC staining that is of greater magnitude than observed in comparable, normal cell populations, indicating the promise of PTL and other therapeutics that target aberrant NF-kB signaling. Canine cell lines and primary cells are sensitive to PTL and undergo dose-dependent apoptosis following exposure to drug. PTL exposure also leads to glutathione depletion, reactive oxygen species generation, and NF-kB inhibition in canine cells. Standard-of-care therapeutics broadly synergize with PTL. In two canine HS cell lines, genetic expression of NF-kB pathway signaling partners is downregulated with PTL therapy. Preliminary data suggest that PTL inhibits NF-kB activity of cells in a mouse model of disseminated HS. Overall, these data support further investigation of compounds that can antagonize canonical and alternative NF-kB pathway signaling, which are overactivated in canine lymphoma, HS, HSA, and MCT disease. PTL is one promising therapeutic that acts, in part, via canonical NF-kB antagonism in canine neoplasms. Further investigation of this compound in vivo is underway in a mouse model of disseminated HS, and if this study is successful, it will provide strong justification for clinical trials with this compound in dogs.Item Embargo Using gene expression and mutational profiling to characterize canine acute myeloid leukemia and assess their comparative features with human acute myeloid leukemia(Colorado State University. Libraries, 2023) Harris, Adam, author; Avery, Anne, advisor; Avery, Paul, committee member; Dow, Steven, committee member; Duval, Dawn, committee memberAcute myeloid leukemia (AML) is an aggressive heterogenous hematopoietic neoplasm that afflicts both dogs and people. Over 10,000 individuals (about the seating capacity of Cameron basketball stadium at Duke University) in the United States succumb to AML-related deaths every year. Treatment options for AML have made little progress in the past few decades and prognosis for both human and canine AML (cAML) remains dismal. However, there are large ongoing multi-institutional studies devoted to advancing medical management for human AML (hAML) by providing targeted therapeutics to patients based on their molecular characteristics. A preclinical model for testing novel therapies could accelerate the development of better treatments in people. We hypothesize that cAML will have similar underlying molecular features as human AML and dogs could be a translational model for developing therapeutics focused on treating AML. The goals of this thesis were to assess the gene expression programs and mutational profiles of cAML and compare our findings with available human AML data. First, we established diagnostic criteria for defining cAML using flow cytometry. Next, we globally assessed normal hematopoiesis in dogs using single cell transcriptomics to generate a hematopoietic tree for defining the cellular composition of cAML. Additionally, we investigated the mRNA expression and genetic variants in cAML to ultimately compare the molecular features with pediatric and adult AML subtypes. We hope this work advances our knowledge of cAML molecular characteristics and adds further credentials to the dog as spontaneous model for human AML.