Impact of perinatal exposure to plastic waste on functional and oncogenic targets in the rat prostate and human prostate cells

• Grant number: 23/12506-8
• Support Opportunities: Scholarships in Brazil - Doctorate
• Start date: April 01, 2024
• End date: March 31, 2028
• Field of knowledge: Biological Sciences - Morphology - Cytology and Cell Biology
• Principal Investigator: Wellerson Rodrigo Scarano
• Grantee: Vanessa Aguiar Rocha
• Host Institution: Instituto de Biociências (IBB). Universidade Estadual Paulista (UNESP). Campus de Botucatu. Botucatu , SP, Brazil

Abstract

The intrauterine microenvironment has been the target of significant changes due to interference from environmental and/or socio-economic factors, which can have late consequences. The problem of plastic is one of the biggest public health challenges on the planet. Research has shown that global plastic production amounted to 400 million tons in 2015, and according to estimates, 12 billion tons of plastic waste will be produced by 2050. When plastics enter the environment, they degrade to form microplastics (MPs) and nanoplastics (NPs) that enter the body through breathing, ingesting contaminated water and food, and through the skin. Phthalates are plastic additives that are not covalently bound to the polymer and are therefore easily released into the environment. Phthalates represent a class of molecules detected in different proportions and concentrations in breast milk, the urine of pregnant women, amniotic fluid and the placenta of humans and rodents. Due to the extreme importance of the subject, the study seeks to analyze the histopathological aspects and the proteomic profile of the prostate of adult and old rats exposed in the perinatal period to a mixture of phthalate diesters (MFs) and NPs, in addition to investigating changes in immortalized normal human prostate PNT-2 cells subjected to similar treatment with a mixture of phthalate monoesters and NPs. For the in vivo study, pregnant rats of the Sprague Dawley strain will be divided into 5 experimental groups: G1: (control; vehicle); T1: 20µg/kg/day MF; T2: 200mg/kg/day MF; T3: NPs; T4: 20µg/kg/day MF + NPs; T5: 200mg/kg/day MF + NPs. The proportion of the mixture was based on a study that determined the amount of different phthalates in the urine of pregnant women, and the doses were based on the dose of environmental and occupational exposure. Polystyrene nanospheres of 100 nm will be used at a concentration of 1.0 mg/kg/day. Treatment will be carried out by gavage from gestational day 10 (GD10) to postnatal day 21 (PND21). On PND120 and PND540, ventral prostates will be collected from F1 males for histological analysis and proteomic analysis, where the samples will be analyzed by mass spectrometry (LC-Ms/Ms). For the in vitro study, PNT-2 human prostate cells will be exposed to MFs and NPs continuously for 15 and 30 passages and gene expression (RTq-PCR) and protein abundance will be assessed by WB. The genes and proteins to be tested will be selected based on the results of our in vivo experiments and in silico analysis.