Spontaneous chromosomal instability in cells with a ring chromosome as the basis for chromosomal therapy
Abstract
Introduction: Extensive chromosomal aberrations are often associated with psychomotor development delay, intellectual disability, and congenital malformations. Despite the high frequency of mutations and due to the involvement, as a rule, of several genes, as well as the wide variety of chromosomal abnormalities themselves, there are currently no ways to effectively treat such patients. Aim: To study spontaneous chromosomal instability in patients with ring chromosomes in differentiated and induced pluripotent stem cells (iPSC). Materials and methods: In patients with intellectual disability and developmental abnormalities, ring chromosomes 13 and 22 were identified during standard karyotyping. A number of additional chromosomal mutations in their karyotype were identified using Agilent 860K microarrays. FISH analysis confirmed ring chromosomes and found mosaicism for them in lymphocytes, fibroblasts, and iPSCs of the patients. iPSCs were obtained from skin fibroblasts by exogenous expression of transcription factors (KLF4, OCT4, SOX2, and human c-MYC). Results: Terminal 13q34 and 22q13.32-q13.33 deletions were identified in patients, resulting in the formation of ring chromosomes 13 and 22, respectively. FISH analysis confirmed the presence of ring chromosomes and found that 47% and 8% of lymphocytes of the patients had monosomies 13 and 22, respectively. At the first passage 50% and 24% of the fibroblasts were monosomic for chromosomes 13 and 22, respectively. On the 9th passage 56% of fibroblasts demonstrated monosomy 13. Among the lymphocytes and fibroblasts at the 9th passage, 1.8% and 1% of the cells, respectively, had a normal karyotype. By the 33rd passage, the number of fibroblasts with monosomy 22 reached 44% and statistically significantly exceeded the initial level ( P < 0.05). The proportion of iPSC monosomic for chromosome 22 varied between 6.3-17% for different clones. Conclusion: The presence of monosomic cells in patients with a ring chromosome indicates chromosomal instability already in vivo. In vitro a statistically significant increase in the number of cells with monosomy 22 is observed. Cells with normal karyotype can be evidence of chromosomal defect correction processes, which are the basis of the chromosomal therapy of genetic diseases that is beginning to be developed.
About the Authors
A. A. Kashevarova
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
E. O. Belyaeva
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
A. M. Nikonov
Diagnostic Center of the Altai Region
Russian Federation
Russian Federation
O. V. Plotnikova
Diagnostic Center of the Altai Region
Russian Federation
Russian Federation
I. G. Gergert
Altai Regional Clinical Perinatal Center "Dar"
Russian Federation
Russian Federation
T. V. Nikitina
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
N. A. Skryabin
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
A. G. Menzorov
Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences
Russian Federation
Russian Federation
M. M. Gridina
Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences
Russian Federation
Russian Federation
S. A. Vasilyev
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
M. E. Lopatkina
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
R. R. Savchenko
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
A. V. Churilova
Tomsk State University
Russian Federation
Russian Federation
E. N. Tolmacheva
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
O. L. Serov
Institute of Cytology and Genetics, Siberian Branch of Russian Academy of Sciences
Russian Federation
Russian Federation
L. P. Nazarenko
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
I. N. Lebedev
Research Institute of Medical Genetics, Tomsk National Research Medical Center, Russian Academy of Sciences
Russian Federation
Russian Federation
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Review
For citations:
Kashevarova A.A., Belyaeva E.O., Nikonov A.M., Plotnikova O.V., Gergert I.G., Nikitina T.V., Skryabin N.A., Menzorov A.G., Gridina M.M., Vasilyev S.A., Lopatkina M.E., Savchenko R.R., Churilova A.V., Tolmacheva E.N., Serov O.L., Nazarenko L.P., Lebedev I.N. Spontaneous chromosomal instability in cells with a ring chromosome as the basis for chromosomal therapy. Medical Genetics. 2017;16(12):18-26. (In Russ.)
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