Confirmatory proteomic analysis of biomarkers of sperm functional alteration and seminal oxidative stress

• Grant number: 14/17185-6
• Support Opportunities: Regular Research Grants
• Duration: February 01, 2015 - January 31, 2018
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Ricardo Pimenta Bertolla
• Grantee: Ricardo Pimenta Bertolla
• Host Institution: Escola Paulista de Medicina (EPM). Universidade Federal de São Paulo (UNIFESP). Campus São Paulo. São Paulo , SP, Brazil
• Associated researchers: Agnaldo Pereira Cedenho

Abstract

Conventional semen analysis, although routinely utilized during infertility investigation, presents low sensitivity and predictive power for pregnancy. Therefore, new diagnostic methods for male infertility are necessary, among which tests to evaluate sperm functional alterations (DNA fragmentation, mitochondrial activity, acrosome integrity, among others) and semen oxidative stress are of particular interest. Despite recent advances in the study of sperm functional integrity, underlying molecular mechanisms and post-genomic pathways remain unexplored. In that sense, a previous study from our laboratory performed proteomics analysis of seminal plasma from normozoospermic men and identified a panel of seminal biomarkers of sperm functional alterations and semen oxidative stress. The next step in a proteomics discovery pipeline would be confirmatory analysis of these biomarkers. Thus, the objective of this study is to validate these biomarkers as potential diagnostic methods of alterations in sperm mitochondrial activity, acrosome defects, sperm DNA fragmentation, and of high levels of seminal lipid peroxidation. Validation will be performed in three cohorts: (i) validation cohort, with semen samples from our previous screening study, which were grouped according to sperm functional integrity and seminal levels of lipid peroxidation (n=114); (ii) confirmatory prospective cohort, with samples from normozoospermic men without clinical factors of infertility; and (iii) confirmatory prospective cohort in different biological conditions (varicocele, obesity, and smoking). For seminal biomarkers evaluation, ELISA or western blotting methods will be performed. For the prospective cohorts, sperm functional integrity and seminal levels of lipid peroxidation will also be analyzed, as in the previous study. For comparison between the groups, unpaired Student's T-test will be used, and an alpha of 5% will be adopted. With these findings, we expect to validate seminal biomarkers of alterations in sperm function and seminal oxidative stress, and to better understand the underlying molecular mechanisms of male infertility. (AU)

Scientific publications

(10)

(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)

BELARDIN, LARISSA BERLOFFA; ANTONIASSI, MARIANA PEREIRA; CAMARGO, MARIANA; INTASQUI, PAULA; FRAIETTA, RENATO; BERTOLLA, RICARDO PIMENTA. Semen levels of matrix metalloproteinase (MMP) and tissue inhibitor of metallorproteinases (TIMP) protein families members in men with high and low sperm DNA fragmentation. SCIENTIFIC REPORTS, v. 9, JAN 29 2019. (14/17185-6, 16/05487-3)

CAMARGO, M.; INTASQUI, P.; BELARDIN, L. B.; ANTONIASSI, M. P.; CARDOZO, K. H. M.; CARVALHO, V. M.; FRAIETTA, R.; BERTOLLA, R. P.. Molecular pathways of varicocele and its repair - A paired labelled shotgun proteomics approach. JOURNAL OF PROTEOMICS, v. 196, p. 22-32, MAR 30 2019. (12/15039-7, 14/17185-6)

BELARDIN, L.; CAMARGO, M.; INTASQUI, P.; ANTONIASSI, M.; FRAIETTA, R.; BERTOLLA, R.. Cysteine-rich secretory protein 3: inflammation role in adult varicocoele. ANDROLOGY, v. 7, n. 1, p. 53-61, JAN 2019. (16/05487-3, 14/17185-6)

CAMARGO, MARIANA; INTASQUI, PAULA; BERTOLLA, RICARDO PIMENTA. Understanding the seminal plasma proteome and its role in male fertility. BASIC AND CLINICAL ANDROLOGY, v. 28, JUN 4 2018. (14/17185-6, 14/11493-0)

CAMARGO, MARIANA; IBRAHIM, EMAD; ABALLA, TEODORO C.; CARDOZO, KARINA H. M.; CARVALHO, VALDEMIR M.; LYNNE, CHARLES M.; BRACKETT, NANCY L.; BERTOLLA, RICARDO P.. Proteomic insight of seminal plasma in spinal cord injured men submitted to oral probenecid treatment for improved motility. JOURNAL OF SPINAL CORD MEDICINE, v. 44, n. 6, p. 966-971, NOV 2 2021. (12/15039-7, 14/17185-6, 17/14346-7)

DEL GIUDICE, P. T.; BELARDIN, L. B.; CAMARGO, M.; ZYLBERSZTEJN, D. S.; CARVALHO, V. M.; CARDOZO, K. H. M.; BERTOLLA, R. P.; CEDENHO, A. P.. Determination of testicular function in adolescents with varicocoele - a proteomics approach. ANDROLOGY, v. 4, n. 3, p. 447-455, MAY 2016. (14/17185-6, 14/11636-6, 12/15039-7)

CAMARGO, MARIANA; INTASQUI, PAULA; BERTOLLA, RICARDO PIMENTA. Proteomic profile of seminal plasma in adolescents and adults with treated and untreated varicocele. ASIAN JOURNAL OF ANDROLOGY, v. 18, n. 2, p. 194-201, MAR-APR 2016. (14/17185-6, 14/11493-0, 12/15039-7)

BELARDIN, LARISSA BERLOFFA; DEL GIUDICE, PAULA TONI; CAMARGO, MARIANA; INTASQUI, PAULA; ANTONIASSI, MARIANA PEREIRA; BERTOLLA, RICARDO PIMENTA; CEDENHO, AGNALDO PEREIRA. Alterations in the proliferative/apoptotic equilibrium in semen of adolescents with varicocele. JOURNAL OF ASSISTED REPRODUCTION AND GENETICS, v. 33, n. 12, p. 1657-1664, DEC 2016. (14/17185-6, 14/11636-6)

JEREMIAS, JESSICA TIMOTEO; BELARDIN, LARISSA BERLOFFA; OKADA, FATIMA KAZUE; ANTONIASSI, MARIANA P.; FRAIETTA, RENATO; BERTOLLA, RICARDO PIMENTA; INTASQUI, PAULA. Oxidative origin of sperm DNA fragmentation in the adult varicocele. INTERNATIONAL BRAZ J UROL, v. 47, n. 2, p. 275-283, MAR-APR 2021. (19/05879-7, 14/17185-6, 14/11493-0)

CAMARGO, MARIANA; IBRAHIM, EMAD; ABALLA, TEODORO C.; CARDOZO, KARINA H. M.; CARVALHO, VALDEMIR M.; LYNNE, CHARLES M.; BRACKETT, NANCY L.; BERTOLLA, RICARDO P.. Proteomic insight of seminal plasma in spinal cord injured men submitted to oral probenecid treatment for improved motility. JOURNAL OF SPINAL CORD MEDICINE, FEB 2020. (12/15039-7, 14/17185-6, 17/14346-7)