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Investigation of pro-inflammatory cytokine genes polymorphisms as genetic risk factors for prematurity complications
Serbia (IFMSA-Serbia) - University of Belgrade, Belgrade
Prof. dr Tatjana Damnjanović
Prof. dr Tatjana Damnjanović, Prof. dr Nela Maksimović
Type of Research Project
- Clinical Project with Laboratory work
What is the background of the project?
Preterm birth is the leading cause of neonatal mortality. The risk of complications in premature babies increases with lower gestational age (GA) and lower birth weight (BW). The most frequent complications are respiratory distress syndrome (RDS), bronchopulmonary dysplasia (BPD), intraventricular hemorrhage (IVH), retinopathy of prematurity (ROP) and sepsis. Studies on genetic variations in cytokine genes such as tumor necrosis factor-alpha(TNFα), interleukin-1 beta(IL-1β) and interleukin-6(IL-6) genes and their association with premature birth have been reviewed. TNFα and IL-1β are pro-inflammatory cytokines capable of inducing preterm labor. Also, increased maternal levels of IL-1β and IL-6 have been associated with prematurity. Polymorphisms in genes for cytokines may modulate their expression and may have additional impact on immature innate immune response of premature babies. That may lead to severe form of complications in preterm neonates.
What is the aim of the project?
Our objective is to analyze TNFα, IL-1β and IL-6 genes polymorphisms in order to explore their association with complications in premature babies.
What techniques and methods are used?
Genomic DNA from buccal swabs taken from neonatal patients will be extracted using the conventional phenol – chloroform based method. We will analyze single nucleotide polymorphisms TNF-α - 308 G/A (rs1800629), IL-6 -174 G/C (rs1800795) and IL- 1β -511 G/A (rs16944). Genotyping will be performed with custom TaqMan® SNP Genotyping Assays. Genotype analyses will be carried out in Applied Biosystems' 7500 Real-Time PCR System. Differences in genotypes and alleles frequencies between group with expressed complications sepsis (RSD, BOD, IVH, ROP and sepsis) and group without complication will analyzed by Chi-square test and Fisher exact test. Association between analyzed polymorphisms and preterm infants with complications will be explored by logistic regression analysis after the adjustment for BW, and GA. Statistical analyses will performed using SPSS statistical package, version 16.0.
What is the role of the student?
- The student will mainly observe
- The student will observe the practical experiments but will be highly involved in the analysis of the results
- The tasks will be done under supervision
What are the tasks expected to be accomplished by the student?
Student will learn how to isolate DNA from buccal swabs by phenol-chloroform metho and measure the concentration of DNA with fluorimeter and spectrofotometer. Student will also learn how to prepare samples for real-time PCR using TaqMan reagents and analyze the results. After the lab work student will apply statistical tests including linear and logistic regression analysis in order to determine the association between genotypes and analyzed variables. Determination of genotypes and statistical analyses will be performed under supervision.
Will there be any theoretical teaching provided (preliminary readings, lectures, courses, seminars etc)
We will provide the material for preliminary reading and lecture about the project and the metodology applied in the project. What is expected from the student at the end of the exchange? And what
What is expected from the student at the end of the research exchange? What will be the general outcome of the student?
- No specific outcome is expected
What skills are required of the student? Is there any special knowledge or a certain level of studies needed?
Are there any legal limitations in the student’s involvement
Type of students accepted
This project accepts: - Medical students - Graduated students (less than 6 months)
- Esposito S; Zampiero A; Pugni L; Tabano S; Pelucchi C; Ghirardi B at all. Genetic Polymorphisms and Sepsis in Premature Neonates; PLOS ONE; 2014: 9; e101248
- Paananen R; Husa AK; Vuolteenaho R; Herva R; Kaukola T; Hallman M. Blood cytokines during the perinatal period in very preterm infants: relationship of inflammatory response and bronchopulmonary dysplasia. J Pediatr 2009;154:39–43. e33.
- Cooke WR; Drury JA; Mountford RC; Clark D. Genetic Polymorphisms and Retinopathy of Prematurity. Investigative Ophthalmology & Visual Science 45(6):1712-5
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