Laboratory of Cytogenetics

The study of structural genome variability caused by pathogenetically significant variations in the number of DNA copies (CNVs) at the microscopic and submicroscopic level using high-tech molecular cytogenetic research methods such as fluorescent in situ hybridization (FISH), multi-color FISH technologies and comparative genomic hybridization (CGH) is one of the fields of scientific research of the cytogenetics laboratory. Unbalanced structural variants or variations in the number of copies DNA units as one of the mutational forms of structural genome variability is the basis of the genomic imbalance. This imbalance is often accompanied by impairment of a large number of dose-sensitive genes that regulate and control the normal development of the body, which leads to the development of chromosomal diseases. The main task of the laboratory in this field is development of approaches and strategies for identification of chromosomal micro and macro rearrangements, as well as the study of mechanisms of genomic imbalance, which is an etiological factor of chromosomal diseases.

Although variations in the number of DNA copies are detected in the genome everywhere, there are areas most prone to this kind of rearrangement. This is due to the peculiarities of the human genome architecture, namely, the presence of a special class of highly homologous duplicated DNA sequences or segmental duplications, which tend to cluster in the pericentromeric and subtelomeric regions of chromosomes. Such blocks of segmental duplications are a substrate for formation of CNVs, which, in turn, can be both a source of structural polymorphism and a cause of genetically determined comditions, for example, congenital heart disease (CHD). Therefore, special attention is paid to the study of associations of CHD in combination with extracardiac pathology and pathogenetically significant CNVs in subtelomeric and pericentromeric regions of chromosomes, as well as development of a strategy for selective screening of DNA copy number variations in these patients.

Information about the topography and structure of the genomic imbalance allows not only to clarify the mechanism of its development, but is also important for genetic family counseling and for identifying the nature of the rearrangement (hereditary or de novo). Determining the degree of genetic risk and the severity of medical and social consequences of the suspected pathology permits to develop correct tactics of preventive measures to prevent the birth of a sick child. Therefore, the study of meiotic segregation in men with genomic structural variants in the form of small supernumerary marker chromosomes and in carriers of balanced rare chromosomal abnormalities, as well as factors affecting the formation of gametes due to pathological meiotic segregation is another area of scientific activity of the cytogenetics laboratory.

In continuation of the scientific traditions of the laboratory of prenatal diagnosis, attention is paid to such an important problem as non-invasive prenatal diagnosis in the study of fetal cells circulating in the blood of women during pregnancy. At present, development, optimization and implementation in clinical practice of non-invasive methods of prenatal diagnosis allowing the analysis of the fetal genetic status without any damage to the developing fetus and the pregnant woman are especially important. During pregnancy, there is an amazing phenomenon of transplacental migration of cells in mother's body, so that they can be detected in the peripheral blood of a pregnant woman. This unique availability of genetic material of the fetus has led to the intensive development of its research methods.

At present, the scientific activities of the laboratory are focused on the development of a protocol for the study of the genome of cytotrophoblast cells to identify chromosomal abnormalities in the fetus. Much work is being done to optimize the protocols of isolation and detection of cytotrophoblast cells from the blood of pregnant women using laser microdissection of single cells and their subsequent analysis by the CGH technique.

The practical work includes cytogenetic and molecular genetic diagnosis of chromosomal rearrangements in patients with mental retardation, developmental abnormalities or reproductive problems. In addition, the laboratory has developed an algorithm for the study of abortive material using the metaphase CGH method in non-developing pregnancy, which also includes interphase FISH for the detection of polyploidy in the fetus. This whole-genome approach makes it possible to identify all chromosomal abnormalities that are an etiological factor in non-developing pregnancy.

List of theses defended in the Cytogenetics Laboratory (prenatal diagnosis):

1. Kostyuk Edmund Vladimirovich
   "Study of serum markers of pregnant women to form high genetic risk groups for congenital malformations." Thesis for a Candidate of medical science, 1993. (supervisor - Zolotukhina T.V.)
2. Zolotukhina Tatyana Vladimirovna
   "Prenatal diagnosis of chromosomal diseases (scientific and practical aspect)." Thesis for a Doctor of Biological Science, 1994.
3. Chivilev Igor Viktorovich
   "Hereditary factors impact on variability of fetal pathologies marker levels." Thesis for a Candidate of medical science, 1997 (supervisor - T. Zolotukhina)
4. Shilova Nadezhda Vladimirovna
   "Research on fetal cells in the blood of mother: a new non-invasive approach in prenatal diagnosis." Thesis for Candidate of medical science, 1999 (Supervisor - T. Zolotukhina)
5. Klimova Marina Ivanovna
   “Comprehensive prenatal prevention of malformations and chromosomal diseases (according to Voronezh Interregional Counseling for Medical Genetics).” Thesis for a Candidate of biological science, 2002 (supervisor - Zolotukhina T.V.)
6. Markova Zhanna Gennadyevna
   "Development of an automated software package for calculating the risk of Down Syndrome in fetus." Thesis for a Candidate of biological science, 2005 (supervisor - Zolotukhina T.V.)
7. Miroshnikova Irina Varlenovna
   “Optimization of the algorithm for prenatal diagnosis of Down Syndrome using screening markers of the first trimester of pregnancy.” Thesis for a Candidate of medical science, 2007 (supervisor - Zolotukhina T.V.)
8. Kozlova Yulia Olegovna
   "Development of technology for pre- and postnatal diagnosis of a group of syndromes caused by 22q11.2. microdeletion." Thesis for a Candidate of medical science, 2014 (supervisor - T. Zolotukhina)
9. Min’zhenkova Marina Evgenyevna
   "Metaphase comparative genomic hybridization in the diagnosis of chromosomal imbalance." Thesis for a Candidate of medical science, 2014 (supervisor - Shilova N.V.)
10. Shilova Nadezhda Vladimirovna
    "Improving approaches to the diagnosis of chromosomal abnormalities in personalized medicine." Thesis for a Doctor of Medical Science, 2016

Major publications of the Cytogenetics laboratory (prenatal diagnosis) 2005-2018

1. Zolotukhina T, Shilova N, Voskoboeva E. – Analysis of cell-free fetal DNA in plasma and serum of pregnant women // J Histochem a Cytochem. – 2005 – V.53 (2) - P.297-299.
2. Shilova N.V., Zolotukhina T.V. - Interphase fluorescence in situ hybridization (FISH) in the diagnosis of numerical chromosomal aberrations // Medical Genetics. - 2007 – V. 6, No. 10 (64) - P. 53-57.
3. Shilova N.V., Chernykh V.B., Tsvetkova T.G., Galkina V.A., Osipova E.V., Moshkova M.Yu., Polyakov A.V., Zolotukhina T.V. A comprehensive approach to the diagnosis of unique autosomal gonosomal rearrangement // Medical Genetics - 2009. - No. 2. - P. 44-47.
4. Chernykh V., Kurilo L., Shilova N., Zolotukhina T., Ryzhkova O., Bliznetz E., Polyakov A. Hidden X Chromosomal Mosaicism in a 46,XX Male // Sex Dev. – 2009. V. 3. – P.183-187.
5. Zolotukhina T.V., Shilova N.V., Min’zhenkova M.E., Kozlova Yu.O., Tsvetkova T.G., Markova Zh.G., Antonenko V.G. Using the FISH-method for diagnosis of constitutional chromosomal aberrations // Medical Genetics. - 2011. - No. 11. - P.3-8
6. Shilova N.V., Kozlova Yu.O., Demina N.A., Petukhova M.S., Min’zhenkova M.E., Miroshnikova I.V., Yudina E.V., Zolotukhina T.V. Pallister-Killian Syndrome: Features of Pre- and Postnatal Diagnosis // Medical Genetics. - 2012. - No. 3. - P. 20-26.
7. Zolotukhina T.V., Shilova N.V., Yudina E.V., Min’zhenkova M.E., Kozlova Yu.O. Prenatal diagnosis of rare chromosomal abnormalities // Medical Genetics. - 2012. - T.11. - No. 9 (123). - P.19-24
8. Zolotukhina T.V., Shilova N.V., Yudina E.V. Prenatal diagnosis // "Hereditary Diseases" National Guide edited by Acad. RAMS Bochkov N.P., Acad. RAMS Ginter E.K., Acad. RAMS Puzyrev.- M .: GEOTAR-Media, 2012 .-- p.936 pp. 798-832
9. Min’zhenkova M.E., Shilova N.V., Markova Zh.G., Antonenko V.G., Lebedev I.N., Kozlova Yu.O., Zemlyakova V.V., Zolotukhina T.V. Generation and application of dynamic standard reference intervals for analyzing the results of comparative genomic hybridization // Genetics. 2013. - Volume 49 No. 10. - P. 1229-1235.
10. M.E. Min’zhenkova, N.V. Shilova, Zh.G. Markova, V.G. Antonenko, I.N. Lebedev, Yu.O.Kozlova, V.V. Zemlyakova, T.V. Zolotukhina. Generation and Application of Dynamic Standard Reference Intervals for Analysing Results of Comparative Genomic Hybridization // ISSN 1022-7954 Russian Journal of Genetics. – 2013. - Vol. 49. - No. 10. P. 1072–1077.
11. Raygorodskaya N., Chernykh V., Morozov D., Olutoye O., Bolotova N., Averyanov A., Maslyakova G., Shilova N., Zolotukhina T. A 3-year-old boy with ovotestes: gender reassignment and surgical management //J Pediatr Endocr Met. - 2011. – V. 24(7-8). - P:587-589.
12. Min’zhenkova M.E., Shilova N.V., Markova Zh.G., Kozlova Yu.O., Zolotukhina T.V. Efficiency of various diagnostic methods for chromosomal abnormalities in reproductive losses // Medical Genetics. - 2014. - Volume 13 No. 2 (140). - P. 25-30
13. Yu. O. Kozlova, V. V. Zabnenkova, N. V. Shilova, M. E. Min’zhenkova, V. G. Antonenko, N. P. Kotlukova, L. V. Simonova, I. A. Kazantseva, E. G. Levchenko, T. D. Bombardirova, T. V. Zolotukhin, A. V. Polyakov. Clinical and Genetic Diagnosis of 22q11.2 Deletion Syndrome // Russian Journal of Genetics. – 2014. – V.50. – N.5. P. 602-610.
14. Yu. O. Kozlova, V. V. Zabnenkova, N. V. Shilova, M. E. Min’zhenkova, V. G. Antonenko, N. P. Kotlukova, L. V. Simonova, I. A. Kazantseva, E. G. Levchenko, T. D. Bombardirova, T. V. Zolotukhin, A. V. Polyakov. Clinical and Genetic Diagnosis of 22q11.2 Deletion Syndrome // Russian Journal of Genetics. – 2014. – V.50. – N.5. – P.528-535.
15. Zolotukhina T.V., Kanivets I.V., Korostelev S.A., Shilova N.V., Min’zhenkova M.E., Kozlova Yu.O., Demina N.A., Bessonova L.A., Galkina V .A., Markova Zh.G. The experience with a complex of modern research methods in constitutional cytogenetics // Medical Genetics, - 2014.- No. 12. - P.22-28
16. Musatova E.V., Shilova N.V. Fetal cells in a pregnant woman blood - a potential object of non-invasive prenatal diagnosis (historical and modern aspects) // Medical Genetics. - 2015.- V.14. - No. 7. - P. 9-16.
17. Kozlova Yu.O., Zabnenkova V.V., Antonenko V.G., Shilova N.V. Atypical size deletions in SD22q11.2 patients: clinical characteristics and diagnosis // Medical Genetics. - 2015.- V.14. - No. 11.- P.17-22
18. Antoneneko V.A., Markova T.V., Markova Zh.G., Musatova E.V., Kozlova Yu.O., Shilova N.V. A case of interstitial deletion of a short arm of chromosome 4 in a 13-year-old boy with a weight deficit and learning difficulties // Medical Genetics. - 2016. - No. 6 (168). - Page 44-48.
19. Kozlova Yu.O., Kanivets I.V., Musatova E.V., Shilova N.V. Reciprocal translocation between chromosomes 1 and 2: modern diagnostic methods // Medical Genetics. - 2016. - - No. 7. - Page 33-35.
20. Markova Zh.G., Musatova E.V., Tarlycheva A.A., Shilova N.V. Effective diagnosis of chromosomal abnormalities during miscarriage // Modern Problems of Science and Education. - 2016. - No. 5; URL: http://www.science-education.ru/article/view?id=25286
21. Shilova N.V. Neocentrometers // Medical Genetics. - 2016. - No. 12 - V.15 - No. 11 (173). - Page 3-9
22. Shilova N.V. Prenatal diagnosis of chromosomal diseases. “Hereditary Diseases” National Guide (short edition) Moscow, GOETAR-Media, 2017. p. 92-117
23. N.A. Lyapunova, L.N. Porokhovnik, N.V. Kosyakova, I.A. Mandron, T.G. Tsvetkova.  Effects of the copy number of ribosomal genes (genes of rRNA) on a viability of subjects with chromosomal abnormalities // Gene. - 2017. - V.611. - P.47-53. 
24. Lyapunova N.A., Porokhovnik L.N., Kosyakova N.V., Mandron I.A., Tsvetkova T.G. Viability of carriers of chromosomal abnormalities depends on genomic dosage of active ribosomal genes (rRNA genes) // Russian Journal of Genetics. - 2017. - V.53. - No. 6. - P.722-731.
25. Lyapunova N.A., Porokhovnik L.N., Kosyakova N.V., Mandron I.A., Tsvetkova T.G. Viability of carriers of chromosomal abnormalities depends on genomic dosage of active ribosomal genes (rRNA genes) // Russian Journal of Genetics. - 2017. - V.53. - No. 6. - P. 703-711.
26. Antonenko V.G., Markova Zh.G., Shilova N.V., Glazkova A.V., Tsayuk Yu.V., Petrova O.V. A rare case of the X chromosome long arm rearrangement - an inverted duplication / deletion in women with gonadal dysgenesis // Medical Genetics. - 2017. - Volume 16. - No. 8 (182). - P.36-41
27. Tveleneva A.A., Musatova E.V., Shilova N.V. Methods of genome-wide amplification of the single cells genetic material// Medical Genetics. - 2017. - Volume 16. - No. 11 (185). - P.3-7.
28. Markova Zh.G., Min’zhenkova M.V., Tarlycheva A.A., Tsvetkova T.G., Galkina V.A., Shilova N.V. Phenotypic variability in patients with de novo X-autosomal unbalanced translocations. // Medical Genetics. - 2017. - Volume 16. - No. 12 (186). - P. 43-47
29. Shilova N.V. Autosomal reciprocal translocations: prenatal selection, segregation and assessment of the empirical risk of the birth of any viable child with chromosomal imbalance in family carriage // Medical Genetics. - 2018. - Volume 17. - No. 1 (187). - P. 41-49.
30. Min’zhenkova M.E., Markova Zh.G., Dadali E.L., Shilova N.V. Interchromosomal and intrachromosomal insertions involving chromosome 2 // Medical Genetics. - 2018. - Volume 17. - No. 2 (188). - P.12-17
31. Musatova E.V., Tveleneva A.A., Martynov A.V., Markov J.G., Shilova N.V. Comparison of the methods of single trophoblast cells amplification for the purpose of their subsequent analysis using metaphase comparative genomic hybridization // Genetics. - 2018. - Volume 54. - No. 9. - P.1092-1099.

1. Костюк Эдмунд Владимирович
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8. Козлова Юлия Олеговна
   «Разработка технологии пре- и постнатальной диагностики группы синдромов, обусловленных микроделецией 22q11.2». Диссертация на соискание ученой степени кандидата медицинских наук , 2014 г. (научный руководитель – Золотухина Т.В.)
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   «Метафазная сравнительная геномная гибридизация в диагностике хромосомного дисбаланса». Диссертация на соискание ученой степени кандидата медицинских наук, 2014 г. (научный руководитель – Шилова Н.В.)
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