Orientador: Konradin Metze / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-10T11:27:03Z (GMT). No. of bitstreams: 1 Valladao_MariaLuizadeCastroRamos_M.pdf: 884922 bytes, checksum: 2e058daa788641932fbfccd1408d3d69 (MD5) Previous issue date: 2007 / Resumo: O Tumor Venéreo Transmissível Canino (TVTC) é uma neoplasia muito comum em caninos com livre acesso as ruas. O TVTC pode provocar metástases que levam o animal à morte. A presente tese realizou estudo exploratório e prospectivo em 100 caninos portadores de TVTC de ocorrência natural. Foram anotados dados clínicos como raça, sexo, idade, peso corporal e escore na Escala de Desempenho Karnofsky Adaptada (EDKA), bem como amostras citológicas. O objetivo do estudo foi elaborar fatores prognósticos relacionados à sobrevida e fatores preditivos da resposta da monoterapia com vincristina. Os resultados demonstraram que o a idade, o peso corporal do cão e o escore na EDKA são fatores independentes prognósticos da sobrevida durante o tratamento. Cães com escore na EDKA abaixo de 50 tiveram um péssimo prognóstico de sobrevida. O tempo de interrupção do tratamento, época do ano em que se iniciou o tratamento, área do núcleo e a "rugosidade" da cromatina em preparações citológicas mostraram ser fatores independentes preditivos do sucesso da terapia. A sobrevida durante a terapia depende do estado clínico do animal, enquanto a resposta terapêutica depende tanto de características da neoplasia como de fatores ambientais externos que modificam o estado clínico do animal / Abstract: The veneral transmissible tumor of dogs (CTVT) is a common neoplasia in free roaming dogs. Since this tumor may metastasize it is a possible tread for the animals. In a prospective exploratory study based on 100 dogs with naturally occuring CTVT we collected data on breed, sex, age, weight, tumor size and the score on a modified Karnofsky Performance Scale, as well cytologic smears. The objective of the investigation was study to elaborate prognostic factors related to survival and predicitive factors related to the response to vincristine monochemotherapy. Age, sex and the Karnofsky score showed to be independent prognostic factors for survival during chemotherapy. A score below 50 indicated poor survival. Time of interruption of the treatment, season of the year, nuclear area and "roughness" of the cromatin in cytologic preparations showed to be independent prognostic factors for the success of therapy. Thus survival during therapy depends on the performance status of the dog, whereas therapy success depends both on tumor characteristics and external factors modificating the animals performance / Mestrado / Biologia Estrutural, Celular, Molecular e do Desenvolvimento / Mestre em Fisiopatologia

Made available in DSpace on 2014-07-29T15:07:28Z (GMT). No. of bitstreams: 1 NIVIA faria.pdf: 1723183 bytes, checksum: 8c7fa557f1d9e528a75b069ae5bd4fbf (MD5) Previous issue date: 2009-12-18 / The transmissible venereal tumor (TVT) is among the most common cancers in dogs in Brazil. This is a sexually transmitted cancer through a mechanism of transplantation of viable tumor cells, it has high incidence among dogs of both sexes in reproductive age, with high occurrence in areas of high concentration of dogs with poor control of reproduction. The diagnosis made in the routine of clinical medicine is usually based only on clinical symptoms and the chemotherapy of choice has been the vincristine sulfate. This drug has been shown in most cases a good response to treatment with complete regression of the tumor, but there are also cases where the tumor appears resistant to treatment. This study aimed to demonstrate the practicality of using fine needle aspiration cytology as a method of diagnosis and cytomorphologic classification, being effective and safe as well as measuring the response of different cytomorphologic groups forward to treatment with vincristine sulfate. Also evaluating the clinical response, hematologic and biochemical tests using serum liver function (ALT) and renal (creatinine), in the therapy instituted. Were evaluated and treated 18 dogs of both sexes, carrying TVT, which received six consecutive weeks during the administration of vincristine sulfate in the dosage of 0.03 mg / kg administered intravenously. The animals underwent weekly prior to medication, clinical assessment, haematological and biochemical. It was observed in this study that cytological examination is a simple technique, minimally invasive and painless, and a safe method that provides no risk to the patient's life and does not require the use of sophisticated equipment, sedation or anesthesia for their achievement. Animals carrying the TVT plasmacytoid type, had higher rate of tumor resistance to treatment in relation to established groups and linfocytic mixed, showing a greater resistance to treatment with this group of vincristine sulfate. The efficiency of response to treatment was presented by the linfocytic group. The vincristine sulfate showed few side effects, low hematological, hepatic and renal toxicity, it is therefore an effective option in the control of transmissible venereal tumor in not resistant cases. / O tumor venéreo transmissível (TVT) é uma das neoplasias mais freqüentes em cães no Brasil. Trata-se de uma neoplasia sexualmente transmissível, através de um mecanismo de transplantação de células tumorais viáveis, apresenta alta incidência entre cães de ambos os sexos em idade reprodutiva, tendo grande ocorrência em ambientes de alta concentração de cães errantes e com deficiente controle de reprodução. O diagnóstico realizado na rotina da clínica médica geralmente se baseia apenas na sintomatologia clínica e o quimioterápico de eleição tem sido o sulfato de vincristina. Este quimioterápico tem demonstrado na maioria dos casos boa resposta ao tratamento com regressão completa da neoplasia, mas também há casos onde o tumor se mostra resistente ao tratamento. O presente trabalho visou demonstrar a praticidade do uso da citologia aspirativa por agulha fina como método de diagnóstico e de classificação citomorfológica eficaz e seguro, assim como mensurar a resposta dos diferentes grupos citomorfológicos frente ao tratamento com sulfato de vincristina. Avaliando também a resposta clínica, hematológica e por meio de provas bioquímicas séricas a função hepática (ALT) e renal (creatinina), frente à terapia instituída. Foram avaliados e tratados 18 cães de ambos os sexos, portadores de TVT, os quais receberam durante seis semanas consecutivas a administração de sulfato de vincristina na dosagem de 0,03 mg/kg administrado via intravenosa. Os animais foram submetidos semanalmente previamente a medicação, avaliação clínica, hematológica e bioquímica. Observou-se com este estudo que o exame citológico constitui-se em uma técnica simples, minimamente invasiva e indolor, sendo um método seguro, que não proporciona riscos à vida do paciente e não requer o uso de equipamento sofisticado, sedação ou anestesia para sua realização. Os animais portadores do TVT do tipo plasmocitóide, apresentaram maior índice de resistência tumoral ao tratamento estabelecido em relação aos grupos linfocitóide e misto, demonstrando uma maior resistência deste grupo ao tratamento com sulfato de vincristina. A maior eficácia de resposta ao tratamento foi apresentada pelo grupo linfocitóide. O sulfato de vincristina apresentou poucos efeitos colaterais, baixa toxicidade hematológica, hepática e renal constituindo, portanto em opção eficaz no controle do tumor venéreo transmissível em casos não resistentes.

Title: Electromyographic and clinical evaluation of vincristine-induced peripheral neuropathy in pediatric patients after treatment of acute lymphoblastic leukemia and correlation with the Bruinkins-Oseretsky Test of Motor Proficiency Second Edition Objectives: The aim of this work was to characterize the neurological consequences of vincristine-induced peripheral neuropathy (VIPN) clinically and electromyographically and to evaluate motor skills of pediatric patients after the end of treatment of acute lymphoblastic leukemia. We also determined the relationship between the results of the clinical and electromyographic evaluation of VIPN and the correlation with the results of motor skills tests. Methods: The study involved 35 probands (19 girls and 16 boys) with a mean age of 10.7 years (SD ± 4.3) and a mean time since the last dose of vincristine of 2.3 years (SD ± 1.2). VIPN was assessed using a clinical pediatric-modified Total Neuropathy Score (ped-mTNS) and nerve conduction studies (NCS). Motor skills were assessed using the Bruinkins-Oseretsky Test of Motor Proficiency, Second Edition (BOT-2). Results: The clinical presence of VIPN, according to ped-mTNS, was found in 20 % of probands. Abnormalities in nerve conduction studies were reported by 60.9 % of probands. Of these, 92.9 % had motor...

Le traitement de la leucémie lymphoblastique aiguë (LLA) de l’enfant, une affection d'origine maligne des cellules progénitrices lymphoïdes, s’est considérablement amélioré au cours des dernières décennies. En effet, le taux de succès du traitement a dépassé 90% dans des conditions favorables. Cependant, des toxicités liées au traitement peuvent être fatales et entrainer l’interruption ou la cessation du traitement. L'allergie, la pancréatite et la thrombose sont des complications fréquentes du traitement de la LLA et sont associées à l'utilisation de l'asparaginase (ASNase), tandis qu’une toxicité fréquente due à la vincristine (VCR) induit la neuropathie périphérique (VIPN). Étant donné que l’ajustement du schéma posologique afin d’augmenter l'efficacité et diminuer la toxicité est un processus sensible, ceci demeure un défi majeur dans plusieurs protocoles de traitement. La pharmacogénétique étudie comment des altérations de la composante génétique peuvent influer sur la variabilité interindividuelle observée dans la réponse au traitement. Une meilleure compréhension de la base moléculaire de cette variabilité pourrait améliorer considérablement les résultats du traitement, en permettant la personnalisation de ce dernier en fonction du profil génétique du patient. Des études récentes suggèrent l’avantage d’appliquer l’analyse de l’exome à la découverte de variants associés à des traits humains complexes ainsi qu’à des phénotypes de réactions médicamenteuses. L'objectif de notre travail était d'utiliser les données de séquençage pour réaliser des études d'association à l'échelle de l'exome, y compris des étapes de filtrage et de validation, afin d'identifier de nouveaux variants génétiques susceptibles de moduler le risque de développer des complications associées à ASNase et à VIPN. Douze SNP étaient associés à des complications due à l’ASNase dans la cohorte initiale, dont 3 étaient associés à une allergie, 3 à une pancréatite et 6 à une thrombose. Parmi ceux-ci, les variants rs3809849, rs11556218 et rs34708521 des gènes MYBBP1A, IL16 et SPEF2 respectivement ont été associés à des complications multiples et leur association à une pancréatite a été répliquée dans une cohorte de validation indépendante. En ce qui concerne la VCR, trois variantes ont été associées à la modulation du risque de VIPN: rs2781377 dans SYNE2, rs10513762 dans MRPL47 et rs3803357 dans BAHD1. Nous démontrons également le puissant effet combiné de la présence de plusieurs variants de risque pour chacune des toxicités étudiées et fournissons des modèles de prédiction du risque pour la pancréatite et le VIPN basés sur la méthode d’évaluation du risque génétique pondérée et qui ont été validés à l’interne. De plus, étant donné une association du polymorphisme du gène MYBBP1A avec de multiples issus de traitement, nous avons cherché à comprendre comment cette altération génétique se traduit par des variabilités de réponse aux traitements à l’ASNase. En utilisant la technique CRISPR-CAS9 pour induire l'inactivation de gènes dans des lignées cellulaires cancéreuses PANC1 (pancréatiques) nous avons testé la différence de viabilité entre les cellules inactivées et les cellules du type sauvage à la suite de la suppression du gène et du traitement par ASNase. Nos résultats suggèrent un rôle fonctionnel de ce gène dans la modulation de la viabilité, de la capacité de prolifération et de la morphologie des cellules knock-out, ainsi que dans leur sensibilité à l'ASNase, et plaident en outre pour que le gène influence l’issus du traitement de la LLA par ASNase. Le présent travail démontre que l’utilisation de l’approche de séquençage de l’exome entier dans le contexte d’une étude d’association à l’échelle de l’exome est une stratégie valide « sans hypothèse » pour identifier de nouveaux marqueurs génétiques modulant l’effet du traitement de la LLA de l’enfant, et souligne l’importance de l'effet synergique de la combinaison des locus à risque. / Treatment of childhood acute lymphoblastic leukemia (ALL), a malignant disorder of lymphoid progenitor cells has improved significantly over the past decades and treatment success rates have surpassed 90% in favorable settings. However, treatment-related toxicities can be life-threatening and cause treatment interruption or cessation. Allergy, pancreatitis and thrombosis are common complications of ALL treatment associated with the use of asparaginase (ASNase), while vincristine-induced peripheral neuropathy (VIPN) is a frequent toxicity of vincristine (VCR). It is a sensitive process and a constant struggle to adjust the dosing regimen to ensure maximum efficacy and minimum toxicity. Pharmacogenetics studies show alterations in the genetic component between individuals can influence the observed variability in treatment response. A better understanding of the molecular basis of this variability in drug effect could significantly improve treatment outcome by allowing the personalization of ALL treatment based on the genetic profile of the patient. Emerging reports suggest the benefit of applying exome analysis to uncover variants associated with complex human traits as well as drug response phenotypes. Our objective in this work was to use available whole-exome sequencing data to perform exome-wide association studies followed by stepwise filtering and validation processes to identify novel variants with a potential to modulate the risk of developing ASNase complications and VIPN. Twelve SNPs were associated with ASNase complications in the discovery cohort including 3 associated with allergy, 3 with pancreatitis and 6 with thrombosis. Of those, rs3809849 in MYBBP1A, rs11556218 in IL16 and rs34708521 in SPEF2 genes were associated with multiple complications and their association with pancreatitis was replicated in an independent validation cohort. As for VCR, three variants were associated with modulating the risk of VIPN: rs2781377 in SYNE2, rs10513762 in MRPL47 and rs3803357 in BAHD1. We also demonstrate a strong combined effect of harbouring multiple risk variants for each of the studied toxicities, and provide internally-validated risk-prediction models based on the weighted genetic risk score method for pancreatitis and VIPN. Furthermore, given the association of the polymorphism in MYBBP1A gene with multiple treatment outcomes, we aimed at understanding how this genetic alteration translates into differences in ASNase treatment response through cell-based functional analysis. Using CRISPR-CAS9 technology we produced gene knockout of PANC1 (pancreatic) cancer cell-lines and tested the difference in viability between the knockouts and wild-type cells following gene deletion and ASNase treatment. Our results suggest a functional role of this gene in modulating the viability, proliferation capacity and the morphology of the knockout cells as well as their sensitivity to ASNase and further advocates the implication of the gene in influencing the outcome of ALL treatment with ASNase. The present work demonstrates that using whole-exome sequencing data in the context of exome-wide association study is a successful “hypothesis-free” strategy for identifying novel genetic markers modulating the effect of childhood ALL treatment and highlights the importance of the synergistic effect of combining risk loci.