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Herpesvirus oral shedding ín oral mucosa of patients under radiotherapy tò oral squamous cell carcinoma of “The head and neck treatment

Grant number: 15/03906-6
Support type:Scholarships in Brazil - Scientific Initiation
Effective date (Start): June 01, 2015
Effective date (End): December 31, 2015
Field of knowledge:Health Sciences - Medicine - Medical Clinics
Principal Investigator:Paulo Henrique Braz da Silva
Grantee:Victor Adriano de Oliveira Martins
Home Institution: Faculdade de Odontologia (FO). Universidade de São Paulo (USP). São Paulo , SP, Brazil

Abstract

The malignant tumor of head and neck represents the 6th place in the world rank, responding for 15000 new cases in Brazil every year. The squamous cell carcinoma (CEC) correspond to 90% of these head and neck malignant neoplasias. Radiotherapy (RT) is a modality of antineoplastic treatment capable of controlling 80% of tumor growth, for tumors level 1, and 70% for level 2 tumors (being able to preserve important anatomic structures). The oral mucositis is an acute complication, frequently observed in patients undergoing head and neck radiation therapy, which can even lead to an interruption of the antineoplastic treatment. The correlation between salivary excretion of herpes simplex virus 1 (HSV-1), and the developing of ulcerative mucositis in Chemotherapy patients, have been well documented in the literature. However, no study was published, that identify the virus from the Herpesviridae family, in the development of oral radiation mucositis. The identification of Herpesviridae family virus excretion, in the mouth of patients undergoing head and neck radiation therapy, and the eventual association with the replication of these virus in the development of mucositis, will make it possible to elaborate strategies that are more efficient, to control, prevent or delay the severe oral radiation mucositis, improving the quality of life of these patients. The purpose of the present study is to evaluate the oral excretion of human herpes virus (HSV- 1, HSV - 2, EBV, CMV, VZV, HH6, HHV7 e HHV8) and its possible association with the development of oral mucositis, in patients diagnosed with squamous cell carcinoma of oral cavity and oropharynx, undergoing head and neck radiation therapy. Twenty adults, male and female patients, will be included in this study. All patients must be confirmed with head and neck squamous cell carcinoma, be candidates for radiotherapy, with dose of 60Gy or higher and agreed to sign the written consent form. Salivary samples, will be collected from all patients, along with clinical evaluation of oral mucosa. Blood samples, will also be collected every 15 days. The patients involved in this study will be oriented to mouthwash with 10 ml of Chlorhexidine 0.12% for 30 sec. and deposit the liquid in a Falcon ® tube collector (15mL). The first sample, will be collected, prior the first session of RT and after that, two samples will be collected per week, until the end of treatment. Sixty days after the end of RT, an extra sample will be collected. The samples will be kept in the refrigerator for 3 hours and then stored at -80 ° C until further processing of the material at the Instituto de Medicina Tropical de São Paulo (IMTSP). Sample extraction of viral DNA, will be performed using the techniques specified in QIAamp DNA Mini Kit. The viral DNA amplification reaction, will be developed, using two sets of primers that align to a well-conserved region of DNA polymerase gene from the virus of the Herpesviridae family. The first set of primers, HSVP1 and HSVP2, is going to be used, for DNA amplification of HSV-1, HSV-2, EBV, CMV and HHV8, and the second sets of primers VZVP1 and VZVP2, will be used for DNA amplification of VZV, HHV-6 (A and B) and HHV-7.The positive samples, for the presence of viral strains of the Herpesviridae family, will be submitted to digestion, with restriction enzymes BamHI and BstUI. Oral mucosa evaluation will be observed in all samples, by the same dentist. The dentist will make a clinical examination of the mucosa, through direct visual observation and an assessment of patient's mucositis degree, following the criteria of classification described by the World Health Organization. To evaluate the association between the presence of the Herpesviridae family virus and the development of oral mucositis, it will be applied the Mann-Whitney Test, that belongs to the category of non-parametric statistical procedures or distribution free procedures, using the Biostat 5.3 program.

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
PALMIERI, MICHELLE; ORNAGHI, MARIANA; DE OLIVEIRA MARTINS, VICTOR ADRIANO; CORREA, LUCIANA; BRANDAO, THAIS BIANCA; DO PRADO RIBEIRO, ANA CAROLINA; SUMITA, LAURA MASAMI; TOZETTO-MENDOZA, TANIA REGINA; PANNUTI, CLAUDIO SERGIO; BRAZ-SILVA, PAULO HENRIQUE. Oral shedding at human herpesviruses in patients undergoing radiotherapy/chemotherapy for head and neck squamous cell carcinoma is not affected by xerostomia. JOURNAL OF ORAL MICROBIOLOGY, v. 10, MAY 28 2018. Web of Science Citations: 1.
DE SANTANA SARMENTO, DMITRY JOSE; TOZETTO-MENDOZA, TANIA REGINA; SUMITA, LAURA MASAMI; PIERROTI, LIGIA CAMARA; PALLOS, DEBORA; CALIENTO, RUBENS; PALMIERI, MICHELLE; DE OLIVEIRA MARTINS, VICTOR ADRIANO; GALLOTTINI, MARINA; PANNUTI, CLAUDIO SERGIO; BRAZ-SILVA, PAULO HENRIQUE. Oral shedding of human herpesviruses in renal transplant recipients. CLINICAL ORAL INVESTIGATIONS, v. 22, n. 2, p. 885-891, MAR 2018. Web of Science Citations: 4.
PALMIERI, MICHELLE; DE OLIVEIRA MARTINS, VICTOR ADRIANO; SUMITA, LAURA MASAMI; TOZETTO-MENDOZA, TANIA REGINA; ROMANO, BRUNA BARALDI; MACHADO, CLARISSE MARTINS; PANNUTI, CLAUDIO SERGIO; BRANDAO, THAIS BIANCA; PRADO RIBEIRO, ANA CAROLINA; CORREA, LUCIANA; BRAZ-SILVA, PAULO HENRIQUE. Oral shedding of human herpesviruses in patients undergoing radiotherapy/chemotherapy treatment for head and neck squamous cell carcinoma. CLINICAL ORAL INVESTIGATIONS, v. 21, n. 7, p. 2291-2301, SEP 2017. Web of Science Citations: 2.

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