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MiRNAs and polymorphisms in the angiotensin 2 converting enzyme receptor as possible biomarkers of disease induced by SARS-CoV-2

Grant number: 21/04669-9
Support type:Regular Research Grants
Duration: March 01, 2022 - February 29, 2024
Field of knowledge:Health Sciences - Medicine - Pathological Anatomy and Clinical Pathology
Principal researcher:Eder de Carvalho Pincinato
Grantee:Eder de Carvalho Pincinato
Home Institution: Faculdade de Ciências Médicas (FCM). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
Assoc. researchers:José Luiz da Costa ; Luis Salazar Navarrete ; Marília Berlofa Visacri ; Maurício Wesley Perroud Júnior ; Patricia Moriel

Abstract

The new coronavirus (SARS-CoV-2), the causative agent of Coronavirus Disease 2019 (COVID-19), has become the latest worldwide emergency. It is known that SARS-CoV-2 enters the cells of the respiratory mucosa through the Angiotensin-Converting Enzyme 2 (ECA2) receptor, which is highly expressed in the epithelial cells of the lung where the virus is capable of causing a primary inflammatory process, characterizing the severe acute respiratory syndrome. At this time, we seek to use our expertise in genetics and epigenetics to contribute significantly in the context of the SARS-CoV-2 pandemic. Until January 2021, only six articles were conducted in humans to identify possible microRNAs as biomarkers for the diagnosis or severity of COVID-19 in human samples. To date, there are no studies to validate these or other microRNAs (miRNAs) as biomarkers of COVID-19. Thus, the objectives of this project are 1) To evaluate miRNAs as possible diagnostic biomarkers for COVID-19 and 2) To identify, through sequencing, the primary plasma miRNAs and polymorphisms in the ECA receptor related to gravity-induced by COVID-19. To achieve objective 1, an observational, analytical, cross-sectional study will be carried out, whose sampling is non-probabilistic of the consecutive type. Two groups will be formed to identify miRNAs that may be diagnostic biomarkers of COVID-19 (cross-sectional study). The Case group (50 patients): sample of subjects from the State Hospital of Sumaré (HES) who had their examination for SARS-CoV- 2 positives in the reference test [Real Time Polymerase Chain Reaction (RT-PCR)]; and Control group (50 patients): sample of volunteers who tested negative for SARS-CoV-2 in the reference test (RT-PCR). The miRNAs will be extracted from plasma, using a commercial kit, 6 samples from the Case group, and six samples from the Control group to perform the library's assembly, sequencing, and differential analysis of the expressed miRNAs. After analysis, silica analysis will be carried out, and the most promising miRNAs will be established for validation. The validation of the selected miRNAs will be performed on the other samples. 2) To achieve objective 2, an observational, analytical, retrospective cohort study will be carried out, whose sampling is non-probabilistic of the consecutive type. To identify miRNAs and polymorphisms in the ACE2 gene that may be biomarkers of COVID-19 severity (retrospective cohort), samples from the Case group (200 patients) will be stratified by the severity of the disease (mild-moderate disease and severe-critical disease) through clinical parameters and computed tomography of the lung. The miRNAs will be extracted from 6 plasma samples from patients who presented mild-moderate conditions and six samples from patients who presented severe-critical disease for later assembly of the library, sequencing, and differential analysis. After analysis, silica analysis will be performed, and the most promising miRNAs will be established for validation. The validation of the selected miRNAs will be performed on the other samples. RT-PCR will be used to analyze polymorphisms in the ECA2 gene. (AU)

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