The canine transmissible venereal tumour (CTVT) is a contagious cancer that is naturally transmitted between dogs by the allogeneic transfer of living cancer cells during coitus. CTVT first arose several thousand years ago and has been reported in dog populations worldwide. The goals of this Thesis were (1) to gain further understanding of CTVT distribution patterns and prevalence around the world, (2) to use genetics to trace the historical spread of CTVT and (3) to map the genetic as well as phenotypic diversity of CTVT tumours around the world. To understand the distribution patterns of CTVT, I obtained information from 645 veterinarians and animal health workers in 109 countries, and generated a snapshot of the locations in which this disease is found. Additionally, as preparation for further genetic analysis, I collected samples from over one thousand CTVT cases from more than 50 countries, optimised methods for high-throughput DNA extraction and quantification and optimised a qPCR-based assay for CTVT diagnosis and host contamination detection. With the goal of tracing the historical spread of CTVT and learning about the genetic diversity of this disease, I sequenced complete mitochondrial genomes of 449 CTVT tumours and their matched hosts. The analysis of the CTVT mitochondrial diversity revealed that CTVT has captured mitochondrial DNA (mtDNA) through horizontal transfer events at least five times during the history of the lineage, delineating five tumour clades. CTVT appears to have spread rapidly around the world within the last 2,000 years, perhaps transported by dogs travelling along historic maritime trade routes. This work indicated that negative selection has operated to prevent accumulation of deleterious mutations in captured mtDNA, and that recombination has caused occasional mtDNA re-assortment. A histology-based screen of CTVT clades did not show any significant phenotypic differences between groups. In order to determine how the five mtDNA clades relate to each other, I analysed data from 539 CTVT exomes. This revealed that a single canine mtDNA haplogroup has recurrently and recently undergone multiple horizontal transfer events. Analysis of this haplotype highlighted a number of candidate genetic variants which may be conferring a selective advantage to this haplotype in CTVT, possibly by influencing mtDNA transcription or replication. Overall, genetic and phenotypic analysis of CTVT tumours from across the globe has broadened our understanding of CTVT diversity, and provided important insights into the biology of a unique transmissible cancer.

Orientador: Noeme Sousa Rocha / Resumo: O tumor venéreo transmissível canino (TVTC) ocorre naturalmente em cães, sem predileção por raça ou sexo sendo transmitido durante o coito ou hábitos sociais. É também um tumor transplantável experimentalmente e tem sido utilizado como modelo para o estudo da relação entre tumor e hospedeiro. Apesar da maior infiltração inflamatória intratumoral e da expressão de moléculas do complexo principal de histocompatibilidade (MHC) estar associada à regressão do tumor, o papel central das células imunes do hospedeiro na evolução clínica do TVTC ainda não está claro. Neste estudo nós buscamos analisar a interação entre TVTC natural e hospedeiro, especialmente sob o ponto de vista da imunidade celular tumoral. Aqui nós identificamos e quantificamos por citometria de fluxo células T (CD3+, CD4+ e CD8+), células NK, células B, macrófagos, em amostras de sangue e de tumor, além da expressão imunoistoquímica de moléculas do MHC de classe I e II, sobretudo nas diferentes fases clínicas do tumor, assim como classificamos os subtipos citológicos do tumor e avaliamos o comportamento tumoral frente ao tratamento quimioterápico com sulfato de vincristina em uma amostra de 22 cães com TVTC natural. A quimioterapia foi efetiva no tratamento da maioria dos casos. Encontramos predomínio de TVTC linfocitóide e que metástases e resistência quimioterápica ocorreram apenas nos tumores de fenótipo linfocitóide e misto. Identificamos aumento significativo na expressão de moléculas de MHC classe I e II na ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The transmissible venereal canine tumor (CTVT) occurs naturally in dogs, without predilection for race or gender being transmitted during intercourse or social habits. It is also an experimentally transplantable tumor and it has been used as a pattern for the study about the relationship between the tumor and the host. Despite the greater intratumoral inflammatory infiltration and the expression of major histocompatibility complex molecules (MHC) is associated with tumor regression, the central role of host immune cells in the CTVT clinical evolution is not clear yet. In this study we sought to analyze the interaction between natural and host CTVT, especially from the point of view of tumor cell immunity. Here we identify and quantify by flow cytometry cells T (CD3+, CD4+ e CD8+), cells NK, cells B, macrophages, in blood and tumor samples, besides the immune histochemical expression of MHC class I and II molecules, specially in the different clinical phases of the tumor as well as classifying the cytological subtypes of the tumor and evaluating the tumor behavior against the chemotherapy treatment with vincristine sulfate in a sample of 22 dogs with natural CTVT. Chemotherapy was effective in the treatment of most cases. We found a predominance of lymphocytoid CTVT and that metastases and chemotherapeutic resistance occurred only in tumors of lymphocytoid and mixed phenotype. We identified a significant increase in the expression of MHC class I and II molecules in the regress... (Complete abstract click electronic access below) / Mestre