Morphological, functional and molecular study of the cellular differentiation of the adrenal cortex: the role of the SOCS3 gene

• Grant number: 18/19035-2
• Support Opportunities: Regular Research Grants
• Start date: June 01, 2019
• End date: August 31, 2021
• Field of knowledge: Biological Sciences - Physiology - Physiology of Organs and Systems
• Principal Investigator: Claudimara Ferini Pacicco Lotfi
• Grantee: Claudimara Ferini Pacicco Lotfi
• Host Institution: Instituto de Ciências Biomédicas (ICB). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated scholarship(s): 19/14925-2 - Technical training to support the research project, BP.TT

Abstract

The development of the adrenal starts with the bilateral formation of the adrenogonadal primordium (AGP) marked by the expression of steroidogenic factor 1 (SF-1). SF-1 positive cells will form the adrenal (AP) primordium that will give rise to the fetal adrenal. The fetal adrenal expands rapidly due to fetal zone growth (ZFe) followed by the definitive zone (ZDe), which will give rise to adult adrenal. In humans, ZFe undergoes apoptosis and progressive differentiation to form glomerulosa and fasciculate zones between 2-4 years of age, while the reticular zone will differentiate only in the adrenarche, between 6-9 years of age. In mice the ZFe only disappears in male puberty and in females after the first gestation, and during this period this remaining zone is denominated zone X (ZX). Recent work by our group described that SF-1/SOCS3 knockout (KO) animal present retention of ZX in male and female mice, becoming a model to study the process of adrenal cortex differentiation, and the importance of SOCS3 gene and accordingly the JAK/STAT/ SOCS3 on this process. Therefore, we hypothesized that the JAK/STAT/SOCS3 pathway plays a role in the adrenal gland differentiation, and to test this hypothesis we have the following goals: 1) to characterize morphologically and functionally the adrenals of male and female SF-1/SOCS3KO mice at different ages, through histochemical, immunohistochemical analyzes of the entire gland; 2) to analyze ZX and ZDe marker genes from animals at different ages by microdissection and qPCR; 3) to analyze parameters related to cell differentiation, such as SF-1, DAX-1 and FAdE, by qPCR and immunoblotting in primary adrenal cortex cells from SF-1/SOCS3KO and control mice; 4) to analyze the activation of the JAK/STAT pathway by analyzing the phosphorylation of the STATs in primary adrenal cortex cells from SF-1/SOCS3KO and control mice; 5) to modulate the expression of SOCS3 by overexpressing it using lentiviral vector or by silencing it through siRNA in primary adrenal cortex cell from, respectively, SF-1/SOCS3 KO and control mice and 6) to analyze differentiation markers such as SF-1, DAX-1 and FAdE, and also StAR protein and steroidogenic enzymes CYP11B1 and CYP11B2 expression in cells overexpressed or silenced SOCS3 by qPCR. At the end of this project we will have a better understanding of the still unexplored participation of the JAK/STAT/SOCS3 pathway in the differentiation and function of the adrenal cortex of mice, as well as insights into the role of the human fetal zone in the formation of pediatric tumors, since maintenance of embryonic cells from the fetal zone in humans and their characteristics were found in pediatric adrenocortical tumors. (AU)

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)

PASSAIA, BARBARA DOS SANTOS; KREMER, JEAN LUCAS; VILLARES FRAGOSO, MARIA CANDIDA; PACICCO LOTFI, CLAUDIMARA FERINI. N-type calcium channel v2.2 is a target of TCF21 in adrenocortical carcinomas. NEOPLASMA, v. 69, n. 4, p. 13-pg., 2022-01-01. (18/19035-2, 16/17285-6, 16/12381-7)

KREMER, JEAN LUCAS; AURICINO, THAIS BARABBA; DOS SANTOS PASSAIA, BARBARA; LOTFI, CLAUDIMARA FERINI PACICCO. pregulation of TCF21 inhibits migration of adrenocortical carcinoma cell. HORMONES & CANCER, v. 12, n. 1, JUL 23 2021. (16/17285-6, 18/19035-2)