Genome-wide epigenetic changes in compartments of genetically unbalanced human blastocysts

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Abstract

BACKGROUND: Epigenetic genome reprogramming is an important determinant of human embryo development. However, its mechanisms remain poorly elucidated, especially in genetically unbalanced embryos.

AIM: The aim of this study is the analysis of DNA methylation and hydroxymethylation levels in trophectoderm and inner cell mass of genetically balanced and unbalanced human blastocysts.

MATERIALS AND METHODS: Twenty-two IVF-derived human blastocysts were enrolled in the study; of these blastocysts, 15 were genetically unbalanced and 7 — genetically balanced. Detection of 5-methylcytosine and 5-hydroxymethylcytosine was performed on trophectoderm and inner cell mass nuclei by indirect immunofluorescence.

RESULTS: In genetically unbalanced blastocysts, the DNA methylation level was elevated in both compartments. The DNA hydroxymethylation level, in contrast, was elevated only in inner cell mass, whereas trophectoderm cells retained the same level as in genetically balanced embryos. These changes equalized the inner cell mass and trophectoderm DNA hydroxymethylation levels in genetically unbalanced blastocysts, while in genetically balanced ones the 5-hydroxymethylcytosine content in inner cell mass lagged behind that in trophectoderm.

CONCLUSIONS: Genetic imbalance is associated with differential epigenetic changes in trophectoderm and inner cell mass cells of human blastocysts: DNA methylation level increases in both compartments while DNA hydroxymethylation level increases only in inner cell mass. The trophectoderm cells in genetically unbalanced blastocysts retain the same hydroxymethylation level as in genetically balanced ones, suggesting a possible explanation of the ability of karyotypically abnormal embryos to implant.

About the authors

Andrei V. Tikhonov

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology

Author for correspondence.
Email: tixonov5790@gmail.com
ORCID iD: 0000-0002-2557-6642
SPIN-code: 3170-2629
Scopus Author ID: 57191821068
ResearcherId: Q-1380-2016

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Mikhail I. Krapivin

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology

Email: krapivin-mihail@mail.ru
ORCID iD: 0000-0002-1693-5973
SPIN-code: 4989-1932
Scopus Author ID: 56507166200
ResearcherId: F-4166-2017
Russian Federation, Saint Petersburg

Olga V. Malysheva

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology

Email: omal99@mail.ru
ORCID iD: 0000-0002-8626-5071
SPIN-code: 1740-2691
Scopus Author ID: 6603763549
ResearcherId: O-9897-2014

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Alla S. Koltsova

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology

Email: rosenrot15@yandex.ru
ORCID iD: 0000-0002-6587-9429
SPIN-code: 3038-4096
Scopus Author ID: 57189621865
ResearcherId: O-1814-2017
Russian Federation, Saint Petersburg

Evgeniia M. Komarova

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology

Email: evgmkomarova@gmail.com
ORCID iD: 0000-0002-9988-9879
SPIN-code: 1056-7821
Scopus Author ID: 57191625749

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Arina V. Golubeva

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology

Email: AlikovaAV1504@yandex.ru
ORCID iD: 0000-0003-1613-222X
SPIN-code: 4610-3686
Russian Federation, Saint Petersburg

Olga A. Efimova

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology

Email: efimova_o82@mail.ru
ORCID iD: 0000-0003-4495-0983
SPIN-code: 6959-5014
Scopus Author ID: 14013324600
ResearcherId: F-5764-2014

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg

Anna A. Pendina

D.O. Ott Research Institute of Obstetrics, Gynecology and Reproductology; Saint Petersburg State University

Email: pendina@mail.ru
ORCID iD: 0000-0001-9182-9188
SPIN-code: 3123-2133
Scopus Author ID: 6506976983

Cand. Sci. (Biology)

Russian Federation, Saint Petersburg; Saint Petersburg

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2. Fig. 1. Interphase nuclei of the inner cell mass (ВКМ) and trophectoderm (ТЭ) of genetically balanced and unbalanced human blastocysts after immunocytochemical detection of 5-methylcytosine (AT-5mC) and 5-hydroxymethylcytosine (AT-5hmC)

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3. Fig. 2. The comparison of immunofluorescently assessed DNA methylation and hydroxymethylation level in trophectoderm (ТЭ) and inner cell mass (ВКМ) cells between genetically balanced and unbalanced human blastocysts. The comparisons were performed with the Mann–Whitney U-test

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4. Fig. 3. The comparison of immunofluorescently assessed DNA methylation and hydroxymethylation levels between trophectoderm (ТЭ) and inner cell mass (ВКМ) in genetically balanced and unbalanced human blastocysts. The comparisons were performed with the Mann–Whitney U-test

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