Γενετικοί πολυμορφισμοί και έκβαση της κύησης

Εισαγωγή: Πρόσφατα ευρήματα δείχνουν ότι ένας αριθμός μονών νουκλεοτιδικών πολυμορφισμών (SNPs) εντός της περιοχής του υποκινητή του γονιδίου αννεξίνης-Α5 (ΑΝΧΑ5) μειώνει την έκφραση του γονιδίου αναφοράς και έτσι αυτά παρουσιάζουν σημαντική συσχέτιση με την ύπαρξη επαναλαμβανόμενων απωλειών εγκυμοσύνης (RPL). Σκοπός: Στόχος της παρούσας μελέτης ήταν να μελετηθεί η συσχέτιση του απλοτύπου M2/ ANXA5 που αποτελείται από τέσσερα υπολειπόμενα αλληλόμορφα: [(SNP1: (-) 467G> A, SNP2: (-) 448A> C, SNP3: (-) 422T> C και SNP4: (-) 373G> A)] με την εμφάνιση επαναλαμβανόμενων απωλειών εγκυμοσύνης στον ελληνικό πληθυσμό, καθώς και των υπολειπόμενων αλληλόμορφων δύο περαιτέρω πολυμορφισμών (SNPs): του SNP5: (-) 302T> G και του SNP6: (-) 1C> T, ως ανεξάρτητων παραγόντων κινδύνου για RPL. Μέθοδοι: Μια γονιδιωματική περιοχή 752bp του υποκινητή του ANXA5 γονιδίου ενισχύθηκε με PCR χρησιμοποιώντας ειδικούς εκκινητές. Για τα έξι αυτά SNPs (υπολειπόμενα αλλήλια) στην περιοχή του υποκινητή γονιδίου της ANXA5, έγινε γονοτυπική ανάλυση με Sanger ακολουθία σε εκατό (100) Ελληνίδες γυναίκες με επαναλαμβανόμενες αποβολές (διάμεση τιμή = 3) και εβδομήντα (70) γόνιμες γυναίκες. Η στατιστική ανάλυση έγινε χρησιμοποιώντας το SAS 9.3 για τα Windows (SAS Institute Inc. NC, ΗΠΑ) και τα πακέτα SPSS για Windows (C.DiMaggio 2013, SAS Institute 2014). Αποτελέσματα: Αυτή η μελέτη ελέγχου αποκάλυψε ότι δεν υπάρχει στατιστικά σημαντικά αυξημένος κίνδυνος εμφάνισης καθ’έξιν αποβολών (RPL) μεταξύ των φορέων του M2 / ANXA5 (υπολειπόμενα αλληλόμορφα των SNP1-4) στον ελληνικό πληθυσμό, καθώς δεν υπήρχαν στατιστικές διαφορές μεταξύ των ασθενών με επαναλαμβανόμενες απώλειες εγκυμοσύνης και των γόνιμων μαρτύρων (11,5% στο RPL περιπτώσεις έναντι 9,29% στους μάρτυρες, ρ-τιμή: 0,6364). Δεν υπήρχε στατιστικά σημαντική διαφορά όσο αφορά τον ανεξαρτητο προγνωστικό παράγοντα κινδύνου για τα SNP5 και SNP6 μεταξύ των δύο ομάδων. Συγκεκριμένα, οι φορείς των SNP5 και SNP6 είχαν αυξημένο κίνδυνο για εμφάνισηκαθ’έξιν αποβολών (RPL) με αναλογία πιθανοτήτων: 1,2472 και 1,3846 αντίστοιχα, αλλά χωρίς στατιστικά σημαντική σημασία. Συμπεράσματα: Ο απλότυπος M2 / ANXA5 δεν διαφέρει όσο αφορά το μέγεθος κινδύνου μεταξύ των ασθενών με RPL και των γόνιμων γυναικών στον ελληνικό πληθυσμό. Επίσης, είναι η πρώτη φορά που τα υπολειπόμενα αλλήλια των SNP5 και SNP6 αξιολογήθηκαν εκτενώς σε γυναίκες ευρωπαϊκής προέλευσης με επαναλαμβανόμενες απώλειες εγκυμοσύνης (RPL) και δεν φαίνεται να αποτελούν ανεξάρτητους παράγοντες κινδύνου στην εμφάνιση RPL στον ελληνικό πληθυσμό. Ωστόσο, αυτό πρέπει να επιβεβαιωθεί σε περαιτέρω και μεγαλύτερες κλινικές δοκιμές σε γυναίκες ευρωπαϊκής προέλευσης.Background: Recent findings show that a number of single nucleotide polymorphisms (SNPs) within the promoter region of the annexin A5-gene (ANXA5) reduce the expression of the reporter gene and so they display a significant association with recurrent pregnancy loss (RPL). Objective: The objective of the present study aimed to address the contribution of ANXA5 M2 haplotype consisting of four minor alleles: (SNP1: (-)467G>A, SNP2: (-)448A>C, SNP3: (-)422T>C and SNP4:(-)373G>A) in the occurrence of recurrent pregnancy losses in the Greek population, and the role of further two minor alleles: SNP5:(-)302 T>G and SNP6: (-)1C>T as independent risk factors for RPL. Methods: A 752bp genomic region of ANXA5 promoter was amplified by PCR using specific primers. Genotypic analysis by Sanger sequencing was performed for these six SNPs (minor alleles) in the promoter region of ANXA5 gene,in one hundred (100) Greek women with recurrent miscarriages (median=3) and seventy (70) fertile controls.Statistical analysis was done using the SAS 9.3 for Windows (SAS Institute Inc. NC, USA) and SPSS packages for Windows (C.DiMaggio 2013, SAS Institute 2014). Results: This case-control study revealed that there is not any significantly increased risk of RPL among the M2/ANXA5 haplotype carriers in the Greek population, as there were no statistical differences between the patients with recurrent pregnancy losses and the fertile controls (11.5% in RPL cases vs 9.29% in controls, p-value: 0.6364). There was no difference in SNP5 and SNP6 minor carriership between the two groups. In particular, carriers of SNP5 and SNP6 had an increased risk for RPL state with odds ratio: 1.2472 and 1.3846 respectively, however without statistically significant importance. Conclusion: The M2/ANXA5 haplotype does not differ between RPL patients and controls in the Greek population. Also, it is the first time that SNP5 and SNP6 minor alleles were evaluated extensively in women of European origin with recurrent pregnancy losses (RPL), and they do not seem to be independent risk factors in the occurrence of RPL in the Greek population. Though, this has to be confirmed in further and larger clinical trials with women of European origin

Genetic Polymorphisms Implicated in Major Pregnancy Complications: a Review

Pregnancy short- or long-term complications may involve the mother’s health, the fetus’s health, or both. A systematic literature review was performed, including studies up to October 2018 from Medline (PubMed), Science Direct, Web of Science and Google Scholar. The following inclusion criteria were applied: studies published until 2018 concerning the genetic background of pregnancy complications such as high blood pressure, gestational diabetes, preeclampsia, pregnancy loss, endometrial death, placental abruption, premature labor, and intrauterine growth retardation which may render pregnancy a high risk condition.We identified 164 articles that met the inclusion criteria and reviewed and analyzed them. The results so far are contradictory and the pathogenicity of these pregnancy complications remains unclear. For most of the polymorphisms studied so far, data refer to small studies size but research is on-going.The identification of genetic polymorphisms with strong correlations with certain pregnancy complications could provide us with useful tools which could be incorporated in diagnostic algorithms that could facilitate early detection and treatment of major pregnancy complications

Successful Pregnancy and Persistent Polyclonal B Cell Lymphocytosis (PPBL): A Case Study of a Rare Co-Existence

Objective: Unknown etiology Background: Persistent polyclonal B cell lymphocytosis (PPBL) is a benign clinical condition, which is characterized by persistent absolute polyclonal B lymphocytosis (>4.0 K/mu L), with the presence of circulating binucleated lymphocytes on the peripheral blood smear and an extra 3 chromosome long arm i(3q) in most cases. Immunophenotype reveals the polyclonal population of B cell lymphocytes with expression of CD19, CD20, and CD22 antigens, and k and l immunoglobulin light chains. Patients are mostly asymptomatic. Although PPBL has a benign clinical course and does not affect the survival expectancy of most patients, pregnancy seems to be extremely rare in these patients, as only 1 case reported so far. Although the real role of immunologic disorders, possibly PPBL, in recurrent pregnancy losses remains unclear, the rarity of successful pregnancy in PPBL patients could be attributed to the possible association of PPBL with infertility or recurrent miscarriages. Case Report: In the present study we present the second published case of a woman with a typical PPBL and recurrent pregnancy loss with a successful pregnancy outcome. Close clinical and laboratory monitoring in combination with the administration of thromboprophylaxis and the induction of mild immunosuppression with low-dose prednisolone may have contributed to the successful outcome of the pregnancy. Conclusions: In conclusion and taking all these findings into consideration, pregnancy in patients with PPBL seems to be extremely rare and the contribution of PPBL to the 2 previous miscarriages in our case could not be excluded

Bone marrow ribonucleotide reductase mRNA levels and methylation status as prognostic factors in patients with myelodysplastic syndrome treated with 5-Azacytidine

Ribonucleotide Reductase (RNR) is a two-subunit (RRM1, RRM2) enzyme, responsible for the conversion of ribonucleotides to deoxyribonucleotides required for DNA replication. To evaluate RNR as a biomarker of response to 5-azacytidine, we measured RNR mRNA levels by a quantitative real-time PCR in bone marrow samples of 98 patients with myelodysplastic syndrome (MDS) treated with 5-azacytidine with parallel quantification of the gene promoter's methylation. Patients with low RRM1 levels had a high RRM1 methylation status (p = 0.005) and a better response to treatment with 5-azacytidine (p = 0.019). A next-generation sequencing for genes of interest in MDS was also carried out in a subset of 61 samples. Splicing factor mutations were correlated with lower RRM1 mRNA levels (p = 0.044). Our results suggest that the expression of RNR is correlated with clinical outcomes, thus its expression could be used as a prognostic factor for response to 5-azacytidine and a possible therapeutic target in MDS