METHODOLOGICAL APPROACHES TO THE STUDY OF THE DETERMINANTS OF SOMATIC MITOCHONDRIAL HETEROPLASMY IN THE ELDERLY

A.G. Goncharov¹, K.Yu. Popad’in¹, I.I. Kozenkov¹, A.G. Mikhailova¹,
V.V. Lobanova¹, M.A. Tatarkina¹, D.V. Ilyushchenko¹, I.V. Reverchuk¹,
A.M. Tynterova¹, A.A. Dvirsky², A.K. Dzhigkaev³,
E.V. Timofeeva¹, K.V. Gunbin¹

DOI: 10.22138/2500-0918-2022-19-3-322-332

¹ Immanuel Kant Baltic Federal University, Kaliningrad, Russian Federation;
² Crimean Federal University named after V.I. Vernadsky, Simferopol, Russian Federation;
³ Center for High Medical Technologies, Kaliningrad, Russian Federation

Abstract. The article substantiates the algorithm and methodology of the study aimed at search and analysis of the determinants of somatic mitochondrial heteroplasmy in the elderly people. Aging is considered as a process that has developed in the process of evolution, characterized by a decrease in all basic biological functions and accompanied by a gradual degradation of organs, tissues and systems of the body, a classification of the main hypotheses and theories of aging is given. The need to develop diagnostic criteria and markers to differentiate the concept of «healthy aging» from diseases of the elderly is noted. Separately, the authors analyze the role of mitochondria in the development of age-related changes associated with the development of neurodegeneration and muscular dystrophy. It has been noted that the development of senile changes is to some extent associated with the main function of mitochondria, namely the oxidation of nutrients and the production of ATP molecules, a universal energy source for most biological processes in cells. For the mitochondrial genome, a driving mechanism has been proposed to explain how mitochondrial somatic mutations can lead to certain aging phenotypes — neurodegeneration and muscular dystrophy. To establish the relationship between mitochondrial somatic deletions and age-related changes, an algorithm and methodology have been developed based on a comprehensive study of the determinants of somatic mitochondrial heteroplasmy in a cohort of elderly people. It is planned to analyze the dynamics of mitochondrial somatic mutations in muscle tissues (biopsies) for 450 elderly volunteers. The authors propose an original approach to the determination of mtDNA heteroplasmy and the detection of de-novo mutations, including the enrichment of mitochondrial fractions and the purification of mtDNA from muscle tissues, as well as a method for quantifying mtDNA copy number to normalize the amount of genomic material in the samples. The resulting material will be used to prepare libraries for sequencing on NGS platforms. Subsequently, the data after bioinformatic analysis will be compared with the results of neurological and psychological studies, including an assessment of the level of cognitive processes, the presence of asthenic disorders, and an assessment of the patient’s rehabilitation potential. It is assumed that this psychoneurological block will allow us to further distinguish at least three groups among volunteers, namely: persons with normal aging (healthy aging), patients with a weakening of mental and physical abilities associated with neurodegenerative or with vascular pathologies. In order to search for informative markers in the immune system that allow differentiating «healthy aging» from diseases of the elderly, it is proposed to perform a correlation analysis between the assessment of immune and cytokine status and the data of clinical and bioinformatic studies.

Keywords: methodology, healthy aging, mitochondrial heteroplasmy, neurological status, psychological testing, immune status

There is no conflict of interest.
Contact details of the corresponding author:
Andrey G. Goncharov
agoncharov59@mail.ru
Received 13.07.2022
For citation:
Goncharov A.G., Popad’in K.Yu., Kozenkov I.I., Mikhailova A.G., Lobanova V.V., Tatarkina M.A., Ilyushchenko D.V., Reverchuk I.V., Tynterova A.M., Dvirsky A.A., Dzhigkaev A.K., Timofeeva E.V., Gunbin K.V. Methodological approaches to the study of the determinants of somatic mitochondrial heteroplasmy in the elderly. Vestn. Ural. Med. Akad. Nauki. = Journal of Ural Medical Academic Science. 2022, Vol. 19, no. 3, pp. 322–332. DOI: 10.22138/2500-0918-2022-19-3-322-332 (In Russ)

REFERENCES
1. Lobachev A.N. Mitochondrial biogenesis during cell differentiation and aging. M., 1985. VINITI 19.09.85, No. 6756-B85 (28 p.) (The article is cited after Berezina T.N. — A probabilistic approach to life expectancy in modern theories of aging // Psychologist. — 2016. — No. 1. — pp. 93 — 106. doi: 10.7256/2409-8701.2016.1.18716 (in Russ)
2. Amorim JA, Coppotelli G, Rolo AP, Palmeira CM, Ross JM, Sinclair DA. Mitochondrial and metabolic dysfunction in ageing and age-related diseases. Nat Rev Endocrinol. 2022 Apr;18(4):243-258 doi: 10.1038/s41574-021-00626-7.
3. Arbeithuber B, Hester J, Cremona MA, Stoler N, Zaidi A, Higgins B, Anthony K, Chiaromonte F, Diaz FJ, Makova KD. Age-related accumulation of de novo mitochondrial mutations in mammalian oocytes and somatic tissues. PLoS Biol. 2020 Jul 15;18(7):e3000745 doi: 10.1371/journal.pbio.3000745
4. Brunk UT, Terman A. Lipofuscin: mechanisms of age-related accumulation and influence on cell function. Free Radic Biol Med. 2002 Sep 1;33(5):611-9 doi: 10.1016/s0891-5849(02)00959-0
5. Guo X, Popadin KY, Markuzon N, Orlov YL, Kraytsberg Y, Krishnan KJ, Zsurka G, Turnbull DM, Kunz WS, Khrapko K. Repeats, longevity and the sources of mtDNA deletions: evidence from ‘deletional spectra’. Trends Genet. 2010 Aug;26(8):340-3. doi: 10.1016/j.tig.2010.05.006
6. Herbst A, Wanagat J, Cheema N, Widjaja K, McKenzie D, Aiken JM. Latent mitochondrial DNA deletion mutations drive muscle fiber loss at old age. Aging Cell. 2016 Dec;15(6):1132-1139 doi: 10.1016/j.arr.2019.100947
7. Johnson AA, Shokhirev MN, Shoshitaishvili B. Revamping the evolutionary theories of aging. Ageing Res Rev. 2019 Nov;55:100947 doi: 10.1016/j.arr.2019.100947
8. Kraytsberg Y, Kudryavtseva E, McKee AC, Geula C, Kowall NW, Khrapko K. Mitochondrial DNA deletions are abundant and cause functional impairment in aged human substantia nigra neurons. Nat Genet. 2006 May;38(5):518-20 doi: 10.1038/ng1778.
9. Levy G, Levin B. An Evolution-Based Model of Causation for Aging-Related Diseases and Intrinsic Mortality: Explanatory Properties and Implications for Healthy Aging. Front Public Health. 2022 Feb 18;10:774668 doi: 10.3389/fpubh.2022.774668.
10. Liu C, Zhang X, Chai H, Xu S, Liu Q, Luo Y, Li S. Identification of Immune Cells and Key Genes associated with Alzheimer’s Disease. Int J Med Sci. 2022 Jan 1;19(1):112-125. doi: 10.7150/ijms.66422
11. Maresova P, Javanmardi E, Barakovic S, Barakovic Husic J, Tomsone S, Krejcar O, Kuca K. Consequences of chronic diseases and other limitations associated with old age — a scoping review. BMC Public Health. 2019 Nov 1;19(1):1431 doi: 10.1186/s12889-019-7762-5
12. Popadin K, Safdar A, Kraytsberg Y, Khrapko K. When man got his mtDNA deletions? Aging Cell. 2014 Aug;13(4):579-82 doi: 10.1111/acel.12231
13. Regan T, Conway R, Bharath LP. Regulation of immune cell function by nicotinamide nucleotide transhydrogenase. Am J Physiol Cell Physiol. 2022 Apr 1;322(4):C666-C673 doi: 10.1152/ajpcell.00607.2020.
14. Schiller M, Ben-Shaanan TL, Rolls A. Neuronal regulation of immunity: why, how and where? Nat Rev Immunol. 2021 Jan;21(1):20-36 doi: 10.1038/s41577-020-0387-1
15. Tidball JG, Flores I, Welc SS, Wehling-Henricks M, Ochi E. Aging of the immune system and impaired muscle regeneration: A failure of immunomodulation of adult myogenesis. Exp Gerontol. 2021 Mar;145:111200 doi: 10.1016/j.exger.2020.111200

Authors
Andrey G. Goncharov
Immanuel Kant Baltic Federal University
Candidate of Medical Sciences, Senior Researcher, Center for Immunology and Cellular Biotechnologies
189-15 Pobedy av. Kaliningrad Russian Federation 236010
agoncharov59@mail.ru

Konstantin Yu. Popad’in
Immanuel Kant Baltic Federal University
Candidate of Biological Sciences, Senior Researcher, Center for Genomic Research
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Ivan I. Kozenkov
Immanuel Kant Baltic Federal University
Junior Researcher, Center for Genomic Research
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Alina G. Mikhailova
Immanuel Kant Baltic Federal University
Junior Researcher, Center for Genomic Research
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Valeria V. Lobanova
Immanuel Kant Baltic Federal University
Research Laboratory Assistant, Center for Genomic Research
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Maria A. Tatarkina
Immanuel Kant Baltic Federal University
Research Laboratory Assistant, Center for Genomic Research
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Dmitry V. Ilyushchenko
Immanuel Kant Baltic Federal University
Laboratory Researcher, Center for Genomic Research
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Igor V. Reverchuk
Immanuel Kant Baltic Federal University
Doctor of Medical Sciences, Professor of the Medical Institute
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Anastasia M. Tynterova
Immanuel Kant Baltic Federal University
Candidate of Medical Sciences, Associate Professor of the Medical Institute
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Alexander A. Dvirsky
Federal State Autonomous Educational Institution of Higher Education «Crimean Federal University named after V.I. Vernadsky, Russian Ministry of Education and Science
Doctor of Medical Sciences, Professor, Professor of the Medical Academy. S.I. Georgievsky
4 Academician Vernadsky av. Simferopol Russian Federation

Akhsarbek Kh. Dzhigkaev
Center for High Medical Technologies of the Ministry of Health of the Russian Federation
Candidate of Medical Sciences, Head of the Department of Traumatology and Orthopedics
4 Kaliningradskoye shosse pos. Rodniki Gurʹyevskiy rayon Kaliningradskaya obl. 238312

Elena V. Timofeeva
Immanuel Kant Baltic Federal University
PhD student of the Medical Institute
189-15 Pobedy av. Kaliningrad Russian Federation 236010

Konstantin V. Gunbin
Immanuel Kant Baltic Federal University
Candidate of Biological Sciences, Senior Researcher, Center for Genomic Research
189-15 Pobedy av. Kaliningrad Russian Federation 236010