RESUMO
Comparative studies of naturally occurring canine cancers have provided new insight into many areas of cancer research. Development and validation of circulating tumor DNA (ctDNA) analysis in pet dogs can help address diagnostic needs in veterinary as well as human oncology. Dogs have high incidence of naturally occurring spontaneous cancers, demonstrate molecular heterogeneity and clonal evolution during therapy, allow serial sampling of blood from the same individuals during the course of disease progression, and have relatively compressed intervals for disease progression amenable to longitudinal studies. Here, we present a feasibility study of ctDNA analysis performed in 48 dogs including healthy dogs and dogs with either benign splenic lesions or malignant splenic tumors (hemangiosarcoma) using shallow whole genome sequencing (sWGS) of cell-free DNA. To enable detection and quantification of ctDNA using sWGS, we adapted two informatic approaches and compared their performance for the canine genome. At the time of initial clinical presentation, mean ctDNA fraction in dogs with malignant splenic tumors was 11.2%, significantly higher than dogs with benign lesions (3.2%; p = 0.001). ctDNA fraction was 14.3% and 9.0% in dogs with metastatic and localized disease, respectively (p = 0.227). In dogs treated with surgical resection of malignant tumors, mean ctDNA fraction decreased from 11.0% prior to resection to 7.9% post-resection (p = 0.047 for comparison of paired samples). Our results demonstrate that ctDNA analysis is feasible in dogs with hemangiosarcoma using a cost-effective approach such as sWGS. Additional studies are needed to validate these findings, and determine the role of ctDNA to assess burden of disease and treatment response in dogs with cancer.
Assuntos
DNA Tumoral Circulante , Hemangiossarcoma , Neoplasias Esplênicas , Animais , Biomarcadores Tumorais/genética , DNA Tumoral Circulante/genética , Progressão da Doença , Cães , Estudos de Viabilidade , Hemangiossarcoma/genética , Hemangiossarcoma/veterinária , Mutação , Neoplasias Esplênicas/genética , Neoplasias Esplênicas/veterináriaRESUMO
Abstract Equine influenza is one of the major respiratory infectious diseases in horses. An equine influenza virus outbreak was identified in vaccinated and unvaccinated horses in a veterinary school hospital in São Paulo, SP, Brazil, in September 2015. The twelve equine influenza viruses isolated belonged to Florida Clade 1. The hemagglutinin and neuraminidase amino acid sequences were compared with the recent isolates from North and South America and the World Organisation for Animal Health recommended Florida Clade 1 vaccine strain. The hemagglutinin amino acid sequences had nine substitutions, compared with the vaccine strain. Two of them were in antigenic site A (A138S and G142R), one in antigenic site E (R62K) and another not in antigenic site (K304E). The four substitutions changed the hydrophobicity of hemagglutinin. Three distinct genetic variants were identified during the outbreak. Eleven variants were found in four quasispecies, which suggests the equine influenza virus evolved during the outbreak. The use of an out of date vaccine strain or updated vaccines without the production of protective antibody titers might be the major contributing factors on virus dissemination during this outbreak.
Assuntos
Animais , Variação Genética , Surtos de Doenças , Infecções por Orthomyxoviridae/veterinária , Evolução Molecular , Vírus da Influenza A Subtipo H3N8/isolamento & purificação , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/virologia , Orthomyxoviridae , Proteínas Virais/genética , Brasil/epidemiologia , Análise de Sequência de DNA , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Substituição de Aminoácidos , Vírus da Influenza A Subtipo H3N8/classificação , Vírus da Influenza A Subtipo H3N8/genética , Genótipo , Cavalos , Hospitais Veterinários , Neuraminidase/genéticaRESUMO
Equine influenza is one of the major respiratory infectious diseases in horses. An equine influenza virus outbreak was identified in vaccinated and unvaccinated horses in a veterinary school hospital in São Paulo, SP, Brazil, in September 2015. The twelve equine influenza viruses isolated belonged to Florida Clade 1. The hemagglutinin and neuraminidase amino acid sequences were compared with the recent isolates from North and South America and the World Organisation for Animal Health recommended Florida Clade 1 vaccine strain. The hemagglutinin amino acid sequences had nine substitutions, compared with the vaccine strain. Two of them were in antigenic site A (A138S and G142R), one in antigenic site E (R62K) and another not in antigenic site (K304E). The four substitutions changed the hydrophobicity of hemagglutinin. Three distinct genetic variants were identified during the outbreak. Eleven variants were found in four quasispecies, which suggests the equine influenza virus evolved during the outbreak. The use of an out of date vaccine strain or updated vaccines without the production of protective antibody titers might be the major contributing factors on virus dissemination during this outbreak.
Assuntos
Surtos de Doenças , Evolução Molecular , Variação Genética , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/virologia , Vírus da Influenza A Subtipo H3N8/isolamento & purificação , Infecções por Orthomyxoviridae/veterinária , Substituição de Aminoácidos , Animais , Brasil/epidemiologia , Genótipo , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Cavalos , Hospitais Veterinários , Vírus da Influenza A Subtipo H3N8/classificação , Vírus da Influenza A Subtipo H3N8/genética , Neuraminidase/genética , Orthomyxoviridae , Infecções por Orthomyxoviridae/epidemiologia , Infecções por Orthomyxoviridae/virologia , Análise de Sequência de DNA , Proteínas Virais/genéticaRESUMO
Equine influenza virus (EIV) (H3N8 and H7N7) is the causative agent of equine influenza, or equine flu. The H7N7 subtype has been considered to be extinct worldwide since 1980. Affected animals have respiratory symptoms that can be worsened by secondary bacterial respiratory infection, thereby leading to great economic losses in the horse-breeding industry. In Brazil, equine influenza outbreaks were first reported in 1963 and studies on hemagglutination antibodies against viral subtypes in Brazilian horses have been conducted since then. The objective of the present review was to present the history of the emergence of EIV around the world and in Brazil and the studies that have thus far been developed on EIV in Brazilian equines...
O vírus da influenza equina (EIV) (H3N8 e H7N7) é o agente causador da influenza equina, ou gripe equina. O subtipo viral H7N7 é considerado como mundialmente extinto desde 1980. Os animais afetados têm sintomas respiratórios característicos que podem ser agravados por uma infecção respiratória bacteriana secundária causando grandes prejuízos no ramo equestre. No Brasil, os surtos da EI têm sido relatados desde 1963 e desde então vem sendo efetuados estudos sobre a presença de anticorpos hemaglutinantes contra os subtipos virais nos equídeos brasileiros. O presente artigo tem o objetivo de apresentar um histórico sobre o surgimento do EIV no mundo e no Brasil destacando os estudos conduzidos no Brasil até o momento acerca da infecção pelo EIV nos equídeos brasileiros...
