Role of epstein-barr virus in the severity of recurrent respiratory papillomatosis.

OBJECTIVE: The objective was to investigate the prevalence of the Epstein-Barr virus (EBV) and its association with human papilloma virus (HPV) detection, clinicopathological features, and the severity of recurrent respiratory papillomatosis (RRP). METHODS: Cases of juvenile recurrent respiratory papillomatosis (JRRP) (n = 36) and adult recurrent respiratory papillomatosis (ARRP) (n = 44) were collected retrospectively and subdivided into low- and high-risk severity groups based on the Derkay score. We performed HPV detection and genotyping using a reverse hybridization protocol and investigated the presence of EBV by polymerase chain reaction (PCR) and in situ hybridization. CD21 levels were accessed by immunohistochemistry. RESULTS: All samples were HPV-positive, including 49 cases of HPV 6, 26 cases of HPV 11, four cases of HPV 6 and 11 coinfections, and one case of HPV 16. EBV-DNA was detected in nine samples by PCR, although none of the cases were positive by means of in situ hybridization. CD21 immunoexpression was not statistically associated with any of the variables analyzed. HPV 6 detection was significantly higher in ARRP cases (P = 0.03), whereas HPV 11 was more prevalent in JRRP cases (P = 0.02) and was even more prevalent in JRRP cases of greater severity (Derkay laryngoscopic scale ≥20) (P = 0.04). CONCLUSION: The presence of EBV does not seem to play an important role in the progression/severity of RRP. LEVEL OF EVIDENCE: 4 Laryngoscope, 130:E611-E618, 2020.

Busca de marcadores moleculares tecido-associados em regiões transcritas não caracterizadas do genoma humano / Search for tissue-associated molecular markers in non-characterized transcribed regions of the human genome

Com foco na atividade transcricional do genoma humano, foi desenvolvido um trabalho de mestrado, em que construímos um microarranjo de cDNAs composto por sequências ORESTES resultantes do Projeto Genoma do Câncer Humano (FAPESP/LICR - HCGP) que não se alinharam com sequências de cDNA geradas por outros projetos. Este arranjo foi hibridizado contra cDNAs derivados de diferentes tecidos humanos, normais ou tumorais, resultando na identificação de 3.421 regiões transcritas do genoma humano (83,3% da plataforma) não descritas por outros projetos de sequênciamento como transcritos. Acreditando que parte dessas sequências pudessem representar RNAs não codificadores, variantes de splicing ou transcritos antisenso naturais fizemos uma reanálise computacional das sequências avaliadas no trabalho de mestrado e também uma análise de expressão diferencial das mesmas, buscando variações de expressão de transcritos tumor- e/ou tecido-associadas. Identificamos como possíveis ncRNAs 28% das sequências analisadas. Mil e sete sequências foram identificadas como diferencialmente expressas, sendo que 291 representam potenciais ncRNAs. Além disso, três potenciais marcadores tumorais de próstata foram validados por PCR em tempo real. Estudos adicionais de um desses marcadores, PCA3, revelaram um possível papel desse ncRNA na regulação do gene PRUNE2 e a existência de uma retenção intrônica não descrita na sequência de PCA3, aparentemente mais frequente em amostras normais de próstata. Também pudemos contribuir com análises iniciais de um potencial novo marcador de câncer de próstata a ser explorado para a complementação de marcadores já existentes, mas falhos em alguns aspectos. Um artigo referente a parte desse trabalho foi publicado no periódico Nucleic Acids Research (MELLO et al. 2009).

With focus on transcriptional activity of the human genome, we developed a master's work, in which we built a cDNA microarray composed of ORESTES sequences resulting from the Human Cancer Genome Project (FAPESP / LICR - HCGP) that did not align with cDNA sequences generated by other projects. This array was hybridized against cDNAs derived from different normal or tumor tissues, resulting in the identification of 3,421 transcribed regions of the human genome (83.3% of the slide) not described by other sequencing projects as transcripts. Believing that some of these sequences may represent non-coding RNAs, and splicing variants of natural antisense transcripts we did a new computational analysis of the sequences found in the master's work and also an analysis of differential expression of these sequences, seeking changes in expression of tumor- and/or tissue-associated transcripts. We identified as possible ncRNAs 28% of the sequences analyzed. One thousand and seven sequences were identified as differentially expressed, and from these 291 represent potential ncRNAs. In addition, three potential tumor markers for prostate cancer were validated by real-time PCR. Further studies of one of these markers, PCA3, revealed a possible role of this ncRNA in the regulation of PRUNE2 gene and the existence of an intron retention not described within PCA3 sequence, apparently more common in normal samples from prostate. We could also contribute with initial analysis of a potential new marker for prostate cancer to be explored in complementing currently markers not very acurated. An article containing part of this work was published in the journal Nucleic Acids Research (MELLO et al. 2009).