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Doctoral Thesis
DOI
https://doi.org/10.11606/T.5.2020.tde-31012020-105058
Document
Author
Full name
Nathalie Oliveira de Santana
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2019
Supervisor
Committee
Danilovic, Debora Lucia Seguro (President)
Marui, Suemi
Hoff, Ana Amélia Fialho de Oliveira
Kimura, Edna Teruko
Title in Portuguese
Perfil clínico e molecular dos carcinomas de células de Hürthle da tireoide
Keywords in Portuguese
Células oxífilas
Citologia
Neoplasias da glândula tireoide
Nódulo da glândula tireoide
Sequenciamento de nucleotídeos em larga escala
Transdução de sinais
Ultrassonografia
Via de sinalização Wnt
Abstract in Portuguese
INTRODUÇÃO: A caracterização genômica dos carcinomas de células de Hürthle da tireoide (CCHs) está em progressão. Estudos recentes identificam alterações cromossômicas e mutações de DNA mitocondrial, além de distinto padrão de expressão gênica das vias PI3K-AKT-mTOR e Wnt/Beta-catenina, particularmente em CCHs extensamente invasivos. Entretanto, as potenciais mutações condutoras ou as responsáveis pelo fenótipo rico em mitocôndrias não estão completamente definidas. OBJETIVOS: caracterizar clinicamente os pacientes e avaliar o perfil molecular dos CCHs por meio de sequenciamento paralelo em larga escala e correlacioná-lo com as características clínicas, histológicas e desfechos. MÉTODOS: Avaliação retrospectiva de 52 CCHs e 49 adenomas de células de Hürthle (ACHs). Analisaram-se dados clínicos, histológicos, exames de ultrassonografia de tireoide e desfechos. Realizou-se o sequenciamento do DNA extraído de 40 CCHs e 10 ACHs por painel de 97 genes das vias MAPK, PI3K-AKT-mTOR, Wnt/Beta-catenina e Notch e outros, como promotor do TERT e TP53. Os programas Lofreq, Mutect e Lancet identificaram variantes somáticas, e os achados foram revisados no "Integrative Genomics Viewer". As alterações genéticas identificadas foram avaliadas no banco de dados COSMIC70, e potenciais efeitos deletérios por ferramentas de predição "in silico". Os dados clínicos, histológicos e desfechos dos CCHs foram correlacionados com o perfil molecular. RESULTADOS: A idade média dos pacientes com CCHs foi de 57,3 ± 14,5 anos, com predomínio do sexo feminino (82,7%). Os nódulos malignos foram significativamente maiores que os benignos (47,4 ± 25,7 mm vs. 28,6 ± 19,5 mm, p < 0,001). Achados ultrassonográficos suspeitos foram infrequentes nos CCHs (sólido 54,2%, hipoecoico 37,5%, macrocalcificações 12,5%, microcalcificações 8,3%, contornos irregulares 4,2%, altura > largura 16,7% e vascularização predominante central 52,6%). Os carcinomas eram extensamente invasivos em 61,5%. Metástases linfonodais ocorreram em 9,6% e à distância em 13,5%. Em seguimento médio de 92,7 meses, recorrência e/ou persistência ocorreram em 17,3% e óbito pela doença em 3,8%. Houve variantes somáticas em 47,5% dos CCHs (190 SNPs e 5 inserções/deleções). A via Wnt/Beta-catenina foi a mais frequentemente alterada (30%), seguida das vias MAPK (27,5%) e PI3K-AKT-mTOR (25%). Os genes com maior número de alterações foram FAT1 e APC (12 e 18 mutações, respectivamente) em 17,5% dos casos. Ocorreram mutações de NRAS em 7,5%, HRAS e KRAS em um CCH cada. Não foram identificadas mutações de BRAF e TP53, e as duas mutações do promotor do TERT foram distintas das C228T e C250T. Mutações foram mais numerosas nos carcinomas extensamente invasivos. Não houve associação significativa entre perfil mutacional e dados clínicos e histopatológicos. CONCLUSÃO: O perfil clínico dos pacientes com CCHs demonstrou idade média avançada e predomínio do sexo feminino. Na ultrassonografia, predominaram nódulos sólidos, não-hipoecoicos, sem calcificações, com contornos regulares, halo periférico e vascularização predominantemente central. Apesar de a maior parte dos casos ter sido extensamente invasiva, metástases linfonodais e à distância foram infrequentes, assim como recorrência ou persistência de doença. No sequenciamento paralelo em larga escala, observaram-se diversas mutações destacando-se a via Wnt/Beta-catenina e seus genes FAT1 e APC. Não houve associação do perfil mutacional com características clínicas, histológicas e desfechos
Title in English
Clinical and molecular profile of Hürthle cell thyroid carcinomas
Keywords in English
Cytology
High-throughput nucleotide sequencing
Oxyphil cells
Signal transduction
Thyroid neoplasms
Thyroid nodule
Ultrasonography
Wnt signaling pathway
Abstract in English
BACKGROUND: Genomic characterization of thyroid Hürthle cell carcinomas (HCCs) has evolved recently. Studies have identified recurrent driver mutations, chromosomal alterations and mitochondrial DNA mutations. PI3K-AKT-mTOR and Wnt/Beta-catenin pathways present distinct expression pattern, particularly in widely invasive HCCs. However, potential novel driver mutations or alterations responsible for mitochondrion-rich cells are not fully defined. OBJECTIVES: To clinically characterize patients with HCCs; to assess molecular profile of HCCs through next generation sequencing and to correlate it with clinical, pathologic characteristics and outcomes. METHODS: Retrospective assessment of 52 HCCs and 49 Hürthle cell adenomas (HCAs). Clinical, histological, ultrasound and outcome data were analyzed. Extracted DNA from 40 HCCs and 10 HCAs was sequenced through a 97-gene panel targeting MAPK, PI3K-AKT-mTOR, Wnt/Beta-catenin and Notch pathways and other genes, such as a TERT promoter and TP53. Variant callers Lofreq, Mutect and Lancet identified somatic variants, which were reviewed using Integrative Genomics Viewer. Genetic alterations were evaluated in COSMIC70 database and potential deleterious effects were assessed by in silico prediction tools. Clinical, histological and outcome data from HCCs were correlated with molecular profile. RESULTS: Mean age of patients with HCC was 57.3 ± 14.5 years, with a predominance of females (82.7%). Malignant nodules were significantly larger than the benign ones (47.4 ± 25.7 mm vs. 28.6 ± 19.5 mm, p < 0.001). Suspicious ultrasound features were infrequent in HCCs (solid 54.2%, hypoechoic 37.5%, coarse calcifications 12.5%, microcalcifications 8.3%, irregular contours 4.2%, taller-than-wide shape 16.7%, central vascularization 52.6%). HCCs were widely invasive in 61.5% of cases. Lymph node metastasis occurred in 9.6% and distant metastasis in 13.5%. At mean follow-up of 92.7 months, disease relapsed or persisted in 17.3%, and cancer-related death occurred in 3.8%. Somatic variants were identified in 47.5% of HCCs (190 SNPs and 5 insertions/deletions). Wnt/Beta-catenin was the most frequently altered pathway (30%), followed by MAPK (27.5%) and PI3K-AKT-mTOR (25%) pathways. FAT1 and APC were the most frequently mutated genes (12 and 18 mutations, respectively) in 17.5% of cases. NRAS mutations were present in 7.5% of HCCs, HRAS and KRAS in one HCC each. No BRAF or TP53 mutation was found, and there were two mutations in TERT promoter, different from C228T and C250T. Although mutations were more numerous in extensively invasive HCCs, no significant association was found between mutational profile and clinical, pathologic characteristics or outcomes. CONCLUSION: HCC clinical profile demonstrated advanced age at diagnosis and female predominance. Ultrasound showed mostly solid, regular, non-hypoechoic nodules with peripheral halo and central vascularization and without calcifications. Most HCCs were widely invasive, but persistent disease, recurrence, lymph node and distant metastasis were uncommon. HCC molecular profile depicted several mutations, mainly in Wnt/Beta-catenin pathway, particularly in FAT1 and APC genes. There was no association between mutational profile and clinical, pathologic characteristics and outcomes
 
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Publishing Date
2020-01-31
 
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