Health of the population

WP4 – Studies on the causes of disease

The CISAS-WP4, coordinated by the Institute of Clinical Physiology of the CNR, is aimed at developing knowledge on the association between specific pollutants for each study area and selected diseases, identified on the basis of environmental and epidemiological knowledge.

Specifically, the research is structured as follows:

  • in the area of Augusta/Priolo Contaminated Site (CS), the association between exposure to organic contaminants and increased risk factors for liver diseases is investigated. In particular, blood and urine specimens of 500 healthy people living in contaminated areas and in reference neighboring areas (considered as of “no exposure”) are examined. The presence of biomarkers of hepatic damage and hepatocellular carcinoma (i.e., index of hepatic steatosis, index of liver fibrosis, de novo lipogenesis, inflammation biomarkers) is verified;
  • in the area of Milazzo CS the association between exposure to heavy metals and increased risk of thyroid diseases is studied. Specifically, blood and urine specimens of 480 healthy people who live in contaminated areas and in neighboring areas considered as of “no exposure” are analyzed. The presence of hypothyroidism, early indicators of thyroid cancer (i.e., measurements of thyroid hormones, cadmium -Cd and lead – Pb in blood and urine) or early signs of thyroid diseases (with ultrasonografic examination for thyroid nodules) is investigated;
  • in the area of Crotone CS, the association between exposure to heavy metals, and cardiovascular risk and kidney disease is investigated. In particular, blood and urine specimens of 300 healthy people living in contaminated areas and in neighboring areas considered of “no exposure” are examined. The presence of early indicators of cardiovascular risk (i.e., measurement of inflammation, oxidative stress and bone metabolism biomarkers, Cd and Pb in the blood, carotid Doppler ultrasonography, assessment of endothelial function, and electrocardiogram) and of renal damage (i.e., urinary Cd and Pb, measure of renal damage, estimation of glomerular filtration rate) is evaluated.

These are etiological* studies with cross-sectional design** aimed to the recruitment of voluntary subjects of predefined age groups resident in the CS (exposed to environmental pollution) and to an equal number of subjects residing in a reference area, considered of “no exposure”.

The study of healthy population groups at different levels of exposure will allow as follows:

  • to select reliable indicators of exposure to priority environmental pollutants;
  • to evaluate early indicators of health impairment (risk of disease and early damage);
  • to construct health surveillance systems to monitor the population at risk over time;
  • to work out evidence-based indications for primary prevention action (principally remediation interventions in risk areas).

* etiological epidemiology: research into the causes of higher-risk recognized diseases in contaminated sites.

** cross-sectional or transversal study: epidemiological study based on the observation of a phenomenon or a clinical event in a given period of time. Transversal studies can be considered as a snapshot of a disease in a population at a particular time.

WP5 – Birth cohort and exposure to environmental pollutants

As part of the analyzes planned by this Work Package, in order to evaluate the relationship between environmental pollutants and human health, an epidemiological study is planned aimed at the creation and evaluation of a birth cohort in the activity areas of the project and the realization of experiments in vivo, ex vivo and in vitro on animal models and on cell lines.

Task 5.1 – Epidemiological studies, aimed at investigating the effects of environmental contaminants on human health, tend to focus on the most vulnerable segments of the population: this demonstrates the analysis carried out in recent years on the evaluation of the effects of pollutants on the health of children.

More recently, the theory called “Intrauterine origins of health and disease susceptibility” has shown that many multifactorial pathologies can have an in utero origin, when because of suboptimal intrauterine conditions – including exposure to environmental contaminants – irreversible changes on the fetus are generated, that can predispose to the development of pathologies in adult life. In this context, the influence of environmental pollution on pregnant women is of particular interest and will be assessed within the Project (Figure 1).

A birth cohort will be created consisting of pregnant women residing in the SIN areas of Augusta-Priolo, Milazzo-Valle del Mela and Crotone and their children together with a population of pregnant women living in neighboring areas. The role of the placenta in mediating fetal exposure through biomolecular investigations will be evaluated as part of the research project. Maternal blood, cord blood and placenta will be used as biomatrix for the assessment of exposure to environmental toxicants. The activity will therefore be focused particularly on understanding i) the mechanisms by which toxic substances pass from maternal blood to the developing fetus; ii) changes induced by environmental pollutants on placental physiology; iii) the influence of these substances on pregnancy outcomes and on the predisposition to late-onset pathologies. For further details and supplemental information, visit the website www.neho.it.

The areas selected for the study are as follows:

Area SIN Comuni SIN Comuni limitrofi
Milazzo/Valle del Mela Milazzo
Pace del Mela
San Filippo del Mela
Barcellona P.d.G.
Terme Vigliatore
Villafranca Tirrena
Spadafora
Augusta/Priolo Augusta
Priolo Gargallo
Melilli
Solarino
Floridia
Avola
Lentini
Carlentini
Canicattini Bagni
Crotone Crotone Torretta
Isola di C. Rizzuto
Torre Melissa
Le Castella
Strongoli
Botricello
Steccato di Cutro

 

Fig. 1: Schematic representation of the activities planned for the birth cohort.

Task 5.2 – In vitro/ex vivo studies have shown that cell cultures (cell lines and primary cells derived from organ) differentiated in mono-layer or two-dimensional (2D) or three-dimensional (3D) models are a very useful model for the understanding of physiological properties of pulmonary tissue and its alterations that determine the origin of pathogenetic mechanisms of chronic inflammatory diseases and cancer of the upper and lower airways (Figure 2A). In order to develop and deepen the knowledge on the alterations of biomolecular mechanisms that can generate on the fetus irreversible molecular changes, which then predispose to the development of airway pathologies in adults, the role of environmental pollutants on cellular activities concerning oxidative stress (ROS, NOX4, INOS), DNA repair mechanisms (DNA repair markers: p53, PARP, BRCA1/2, H2AX, CHK1/2), cell proliferation regulation (OCT3/4, SOX-2, Nano, CD144, CD105), the genotoxic effects (γ-H2AX, AKT, EZH2, comet assay), and the cellular mechanisms involved in the activation of many genes, biomarkers, proteins and other factors (intracellular signals) will be studied (Figure 2B). The application of appropriate techniques of molecular biology and biochemistry is expected (Real-time PCR, immunoassay test, immunoprecipitation, western blot analysis, elisa methods, immunocytochemistry, immunohistochemistry, immunofluorescence, microscopy, fluorimetry, etc).

Fig. 2: Schematic representation of the activities included in the study on the alteration of biomolecular mechanisms underlying the airway diseases.

Task 5.3 – L’esposizione ad inquinanti ambientali è in grado di causare effetti avversi anche sui sistemi biologici, quali il sistema nervoso centrale, il fegato, il sistema riproduttivo e quello endocrino.

Task 5.3 – Exposure to environmental pollutants can cause adverse effects even on biological systems, such as the central nervous system, the liver, the reproductive system and the endocrine system.

Furthermore, some environmental pollutants have an immunotoxic potential. Studies on isolated cell lines of both human and murine origin can provide important information but it is necessary to make use of an animal model from which isolate the relevant immune populations, coming from different organs and tissues. Moreover, the use of in vitro and ex vivo systems cannot provide functional information, such as the humoral and cellular response as a whole, since for its evaluation it is necessary to resort to in vivo immunization with standard antigens and parallel treatment with pollutants agents. (Figure 3).

Fig. 3: Activities planned in the study of in vitro and in vivo mechanisms of immunomodulation and immunotoxicity.

The objectives are i) to evaluate the immunotoxic and immunomodulatory potential of environmental pollutants in murine and human cell lines and in ex vivo isolated primary immune cells, to study their impact on the innate and acquired immune system; ii) to evaluate the immunotoxicity of environmental pollutants, selected on the basis of the results obtained in the previous point, on the immune cellular and humoral response induced by immunization with standard antigens; iii) to evaluate the correlation between the results obtained in the ex vivo, in vitro and in vivo studies in the mouse model, and with those generated by the studies on human cells, in vivo functional studies, with particular attention to the potential and criticality of each system.

Task 5.4 – Many diseases related to exposure to environmental contaminants, including chronic diseases, diseases linked to oxidative stress and cancer, have a multifactorial origin. Carcinogenesis is a complex process characterized by multiple phases; thanks to in vitro studies and alternative methods to animal experimentation it is possible to isolate and analyze progressively the effect of some experimental variables and therefore understand the modalities of action and the effects of contaminants in a dose and time dependent manner and to identify early warning markers for cancer. Therefore, it is important to evaluate, in in vitro cellular systems, the single or combined action of the contaminants of interest of the project, selected in a coordinated way, in the promotion of biochemical processes that induce the cells towards pathways due to the cellular transformation, evaluating signal transduction pathways,  biochemical markers related to toxicity, inflammation, oxidative stress, control of the cell cycle, angiogenesis and apoptosis, on various human, primary and immortalized cell lines (hepatocytes, melanocytes, preadipocytes, vascular endothelial cells, etc. – Figure 4).

Fig. 4: Schematic representation of the activities planned in the study on exposure and tumors in vitro biomolecular markers.

 

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