Techniques of female reproductive tissue morphology and viability assessment as tools for enhancing cancer research success

• Grant number: 16/22947-8
• Support Opportunities: Scholarships abroad - Research Internship - Doctorate
• Start date: September 01, 2017
• End date: August 31, 2018
• Field of knowledge: Biological Sciences - Morphology - Histology
• Principal Investigator: Sebastião Roberto Taboga
• Grantee: Ellen Cristina Rivas Leonel
• Supervisor: Christiani Andrade Amorim
• Host Institution: Instituto de Biociências, Letras e Ciências Exatas (IBILCE). Universidade Estadual Paulista (UNESP). Campus de São José do Rio Preto. São José do Rio Preto , SP, Brazil
• Institution abroad: Universitè Catolique de Louvain (UCL), Belgium
• Associated to the scholarship: 15/01548-5 - Evaluation of the Bisphenol-A carcinogenic effect in the mammary gland of Mongolian Gerbil (Merionis unguiculatus), BP.DR

Abstract

The incidence of cancer has been higher in the last few decades. The challenge of developing new studies amongst prevention and treatment for these diseases has motivated numerous research groups through the world. Even so, the knowledge about etiology is still scarce and treatments already established are known to cause several side effects in patients, especially in the gonads. Women submitted to chemotherapy may develop gonad dysfunction and become infertile and exhibit iatrogenic or induced menopause. For these patients, reproductive biotechnologies, like ovarian tissue cryopreservation and transplantation are an interesting source to help resume the gonad function and recover fertility. Thus, the objectives of this internship project are: 1) perform immunohistochemistry and western blotting in gerbil mammary and ovary tissue obtained from the experiments developed during the doctorate at UNESP and; 2) participate in an extra project designed by GYNE/IREC/UCL and gain expertise in techniques like tissue cryopreservation, ultrastructure evaluation with transmission electron microscopy (TEM), X-ray computed tomography, in vitro and in vivo tissue culture with human and bovine ovarian tissue after cryopreservation. For the gerbil mammary gland and ovarian tissue already obtained at UNESP, double immunohistochemistry for some markers not available in our institution and already developed at IREC, such as Cx43 and KI67 will be developed. We also aim to perform the western blotting technique in the gerbil mammary gland tissue for detection of the proteins EZH2 and Esr-1. In addition to that doctorate project complementation, it will be developed an experiment to establish an ovarian tissue vitrification protocol. Firstly, a test will be performed at bovine ovaries, which will be warmed, in vitro cultured and transplanted to nude mice. As second part of the experiment, human ovarian biopsies will be obtained from patients to compare the new vitrification protocol to a slow freezing method. Afterwards, the warmed or thawed tissue strips will be xenotransplanted to nude mice for 6 months. Evaluation of the xenografted ovarian tissue will include X-Ray computed tomography in order to certify the cryoprotectant perfusion before vitrification; immunohistochemistry for proliferation (KI67), apoptosis (TUNEL and active caspase-3), vascularization (CD-34) and vessel maturation (a-ASMA) antigen detection; and TEM for membrane integrity and lysosome evaluation. The development of all techniques proposed here must contribute to the improvement of our laboratory abilities and consequently the quality and visibility of publications. It will also enable the establishment of a new research field in our group, such as female reproductive tract morphology.

Scientific publications

(6)

(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)

ASIABI, P.; LEONEL, E. C. R.; MARBAIX, E.; DOLMANS, M. M.; AMORIM, C. A.. Immunodetection and quantification of enzymatic markers in theca cells: the early process of ovarian steroidogenesis. BIOLOGY OF REPRODUCTION, v. 102, n. 1, p. 145-155, JAN 2020. (16/22947-8)

RIVAS LEONEL, ELLEN CRISTINA; PEGORIN CAMPOS, SILVANA GISELE; BARALDI BEDOLO, CAROLINA MARQUES; ALVES GUERRA, LUIZ HENRIQUE; LEITE VILAMAIOR, PATRICIA SIMONE; CALMON, MARILIA FREITAS; RAHAL, PAULA; AMORIM, CHRISTIANI ANDRADE; TABOGA, SEBASTIAO ROBERTO. Perinatal exposure to bisphenol A impacts in the mammary gland morphology of adult Mongolian gerbils. Experimental and Molecular Pathology, v. 113, APR 2020. (15/01548-5, 16/22947-8)

RIVAS LEONEL, ELLEN CRISTINA; CORRAL, ARIADNA; RISCO, RAMON; CAMBONI, ALESSANDRA; TABOGA, SEBASTIAO ROBERTO; KILBRIDE, PETER; VAZQUEZ, MARINA; MORRIS, JOHN; DOLMANS, MARIE-MADELEINE; AMORIM, CHRISTIANI A.. Stepped vitrification technique for human ovarian tissue cryopreservation. SCIENTIFIC REPORTS, v. 9, DEC 27 2019. (16/22947-8)

RIVAS LEONEL, ELLEN CRISTINA; LUCCI, CAROLINA M.; AMORIM, CHRISTIANI A.. Cryopreservation of Human Ovarian Tissue: A Review. TRANSFUSION MEDICINE AND HEMOTHERAPY, v. 46, n. 3, p. 173-181, 2019. (16/22947-8)

AMORIM, CHRISTIANI A.; RIVAS LEONEL, ELLEN CRISTINA; AFIFI, YOUSRI; COOMARASAMY, ARRI; FISHEL, SIMON. Cryostorage and retransplantation of ovarian tissue as an infertility treatment. BEST PRACTICE & RESEARCH CLINICAL ENDOCRINOLOGY & METABOLISM, v. 33, n. 1, p. 89-102, FEB 2019. (16/22947-8)

RIVAS LEONEL, ELLEN CRISTINA; PEGORIN CAMPOS, SILVANA GISELE; ALVES GUERRA, LUIZ HENRIQUE; BEDOLO, CAROLINA MARQUES; LEITE VILAMAIOR, PATRICIA SIMONE; CALMON, MARILIA FREITAS; RAHAL, PAULA; AMORIM, CHRISTIANI ANDRADE; TABOGA, SEBASTIDO ROBERTO. Impact of perinatal bisphenol A and 17 beta estradiol exposure: Comparing hormone receptor response. ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY, v. 188, JAN 30 2020. (15/01548-5, 16/22947-8)