Prevention of Chronic Lung Disease: New High Risk Profile for Early Detection and Management Starting From Perinatal Life (CLD - Prevention Study)

Background Chronic Lung Disease (CLD) is one of the most common complications of perinatal lung injury which represents a systemic condition with long term sequelae, such as persistent pulmonary dysfunction, asthma-like symptoms, and BPCO. CLD recognizes many prenatal risk factors, including maternal smoking, chorioamnionitis, and intrauterine growth restriction (IUGR), besides postnatal risk factors such as hyperoxia, parenteral nutrition, inflammation and mechanical ventilation. Exposure to inflammation in utero alters neonatal immune development and predisposes the fetal lung to a dysregulated prolonged response to invasive mechanical ventilation and supraphysiological oxygen. Responses to injurious (and protective) influences early in life are modulated by genetic and epigenetic mechanisms, that could result in permanent structural changes with significant consequences in adulthood. IUGR pregnancies are characterized by increased oxidative stress and IUGR infants are at high risk of cardiovascular disease, metabolic syndrome, lung dysfunction and chronic kidney and respiratory diseases in adulthood Despite the constant increase in knowledge of the mechanisms leading to the progression of lung damage, so far, no effective management has been developed that would lead to CLD prevention.

Aims

1. prospectively identify potential biomarkers able to predict the severe course of pulmonary function in the first 12 months of life and realize a new profile to early identify high risk newborns.
2. detect genetic variance causation model (by MiSeq Illumina platform) correlating with severe pulmonary dysfunction and asthma development.
3. detect MicroRNAs as well as anti- and pro-inflammatory cytokine variations (MIP-1a, MCP-1, IL-8, TNF-a, IFN-g, IL-10) correlating with the severity of pulmonary dysfunction in the first 12 months of life and the risk of asthma development.

Experimental design This is a multicenter longitudinal study, whose primary aim is to identify a new high-risk neonatal profile for Chronic Lung Disease (CLD) development, starting from perinatal life. In order to reach this objective all participant Units will perform vaginal swab to the enrolled mothers, will collect placenta and cord blood at birth and peripheral blood, bronchoalveolar lavage fluid (BALF), meconium and feces samples of all enrolled newborn infants. The study will begin with the evaluation of prenatal risk factors.

After delivery the placentas will be fixed in 10% buffered formalin. Subsequently macroscopic and microscopic analyses will be performed according to the Amsterdam placental workshop group consensus statement. A vaginal swab will be collected at the time of delivery for all the enrolled women. Three ml of cord blood (artery and/or vein) will be performed for analysis of miRNA, genetic variance and cytokines. In intubated infants only, BALF sample will be obtained by instilling 1 ml/kg of 0.9% sodium chloride in the endotracheal tube and suctioning the fluid into a sterile mucus trap. The first BALF sample will be collected within the first 24 h of life, whereas further BALF samples will be collected at 7 and 28 days of life in newborns who will be still intubated. First meconium sample after birth and additional feces samples will be collected at 7, 28 days of life, 6 and 12 months of life. Vaginal swabs, placenta, BALF, meconium and feces samples will be stored at -80°C until further processing. For each sample, bacterial DNA extraction will be performed in a strictly controlled level-2 biological safety workplace using DANAGENE MICROBIOME DNA kits (Danagen-Bioted) according to manufacturers instructions.