Study of expression of the transcription factor POD-1/TCF21 in adrenocortical tumors, identifying new regulatory targets of Pod-1 and its role in cell migration of tumors cells

• Grant number: 15/14199-9
• Support Opportunities: Regular Research Grants
• Duration: June 01, 2016 - November 30, 2018
• 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): 18/00090-3 - Technical training to support the research project, BP.TT

Abstract

POD-1/TCF21 can inhibit the transcription factor SF-1/NR5A1 adrenal during development. Such inhibition may lead to alterations in the development of adrenal, however its increase is associated with increased proliferation and adrenal tumorigenesis. A previous study from our group showed that overexpression of POD-1 inhibits the expression of endogenous SF-1 by binding in the E-box site of the Sf-1 promoter, which resulted in decreased steroidogenic StAR protein. Although it was not affected cell viability, POD-1 analysis adrenocortical carcinoma (ACC) patients showed an inverse relationship between the expression of this gene and various cell cycle control genes, including BUB1B, one of the molecular markers for prognosis ACC in adults. Therefore, in view of the importance of SF-1 in adrenocortical (ACT), and given the evidence that POD-1 can regulate the expression of SF-1 gene and other cell cycle genes, we hypothesized that the expression of POD-1 may have value in the differential diagnosis and prognosis of ACC adrenal masses. Therefore we aim to 1) analyze the POD-1 and SF-1 expression using cDNA samples of ACT adult and pediatric by reaction TaqMan and quantitative qPCR, and correlate this data with clinical outcomes of patients. We also will analyze the combined expression of BUB1B and PINK1 and POD-1 and BUB1B in ACC of adult and pediatric, to test its prognostic value. Other results from our group showed that Pod1 promotes increased gene expression of LRH-1 and SHP decreased expression, a negative regulator of LRH-1, in adrenocortical cells and hepatocellular carcinomas. Together, these and other findings suggest that Pod1 may have a broad role as a transcription regulator in tumor cells and is a tumor suppressor gene candidate. Therefore, 2) identification of other targets controlled by Pod1 can contribute to the understanding of its importance and its role in tumor cells. To this end, we sequencing the immunoprecipitated DNA adrenal tumor cells and hepatocarcinoma, respectively, the NCI-H295R and HepG2 lines permanently transduced with lentiviral particles expressing POD-1, the ChIP-Seq method followed by validation of targets found by ChIP-PCR, as qPCR and immunoblotting. As the cell viability was not affected by increasing the POD-1 expression in ACT, and in melanomas downregulation POD-1 was related to invasion and metastasis, 3) we will examine whether increased POD-1 expression in adrenocortical cultures diminishes the ability to migrate and invade these cells. Permentemente cells infected with lentivirus expressing POD-1 will be used, followed by migration tests in a chemotaxis chamber containing porous micro-membrane, and invasion tests on plates with insert containing Matrigel. In summary, in the end we will have: 1) established the importance of POD-1 transcription factor in adrenocortical adults and pediatric patients, if this factor can be used in the diagnosis and prognosis of tumors and if cell cycle control genes are involved; 2) we identified new targets, i.e. novel DNA sequences, that may be being controlled by POD-1, increasing our knowledge of their significance in tumor cells and 3) we will know if POD-1 has ability to control cell migration and thus the progression of adrenocortical tumors and other 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)

CLAUDIMARA FERINI PACICCO LOTFI; JEAN LUCAS KREMER; BARBARA DOS SANTOS PASSAIA; ISADORA PONTES CAVALCANTE. The human adrenal cortex: growth control and disorders. Clinics, v. 73, 2018-09-06. (16/17285-6, 15/14199-9)

PACICCO LOTFI, CLAUDIMARA FERINI; KREMER, JEAN LUCAS; PASSAIA, BARBARA DOS SANTOS; CAVALCANTE, ISADORA PONTES. The human adrenal cortex: growth control and disorders. Clinics, v. 73, n. 1, 2018. (16/17285-6, 15/14199-9)

BARBARA BRITO DA CONCEIÇÃO; ISADORA PONTES CAVALCANTE; JEAN LUCAS KREMER; THAIS BARABBA AURICINO; EDUARDA CORRÊA BENTO; MARIA CLAUDIA NOGUEIRA ZERBINI; MARIA CANDIDA BARISSON VILLARES FRAGOSO; CLAUDIMARA FERINI PACICCO LOTFI. ARMC5 mutations are associated with high levels of proliferating cell nuclear antigen and the presence of the serotonin receptor 5HT4R in PMAH nodules. ARCHIVES OF ENDOCRINOLOGY METABOLISM, v. 64, n. 4, p. 390-401, 2020-04-06. (15/50192-9, 15/14199-9)