Journal of Evolutionary Biochemistry and PhysiologyJournal of Evolutionary Biochemistry and PhysiologyЖурнал эволюционной биохимии и физиологии0044-45293034-5529The Russian Academy of Sciences64811310.31857/S0044452924030107YWSURREXPERIMENTAL ARTICLESЭКСПЕРИМЕНТАЛЬНЫЕ СТАТЬИResearch ArticleMitochondrial dynamics and metabolic remodeling in xenograft of IPSC-derived human neural precursorsМитохондриальная динамика и метаболическая перестройка в ксенотрансплантате индуцированных плюрипотентных стволовых клеток продуцированных нейрональных предшественников человекаVoronkovD. N.ВоронковД. Н.
Russian Federation
voronkov@neurology.ruEgorovaA. V.ЕгороваА. В.
Russian Federation
voronkov@neurology.ruFedorovaE. N.ФедороваЕ. Н.
Russian Federation
voronkov@neurology.ruStavrovskayaA. V.СтавровскаяА. В.
Russian Federation
voronkov@neurology.ruLebedevaO. S.ЛебедеваО. С.
Russian Federation
voronkov@neurology.ruOlshanskiyA. S.ОльшанскийА. С.
Russian Federation
voronkov@neurology.ruPodoprigoraV. V.ПодопригораВ. В.
Russian Federation
voronkov@neurology.ruSukhorukovV. S.СухоруковВ. С.
Russian Federation
voronkov@neurology.ruResearch Center of NeurologyФГБНУ НЦ неврологииN.I. Pirogov Russian National Research Mediвcal UniversityФГАОУ ВО РНИМУ им. Н.И. Пирогова Минздрава РоссииLopukhin Federal Research and Clinical Center of Physical-Chemical MedicineФНКЦ физико-химической медицины ФМБА1505202460332032828012025Copyright ©; 2024, Russian Academy of SciencesCopyright ©; 2024, Российская академия наук2024Russian Academy of SciencesРоссийская академия наукhttps://transsyst.ru/0044-4529/article/view/648113

It is well recognized that the regulation of mitochondrial functions affects the differentiation and maturation of neurons. The study of these processes is of both fundamental and practical importance for regenerative neurobiology. Aim of the study: to characterize the mitochondrial fission changes and their relation to the activation of oxidative phosphorylation (metabolic shift) during maturation of human IPSC-derived neural precursors grafted into rat striatum. Wistar rats (n = 15) were unilaterally injected into the caudate nucleus with neural precursors derived from human IPSCs. Changes in localization and expression of neuronal differentiation markers: nestin, NeuN, neuronal enolase, as well as mitochondrial outer membrane protein, ATP synthase and mitochondrial fission protein Drp1 were assessed by immunostaining. Measurements were performed on graft cells 2 weeks, 3 and 6 months after surgery. Maturation of grafted neurons was associated with fluctuations morphometric parameters of the mitochondrial fraction and Drp1 levels. Increased mitochondrial fission was detected 3 months after transplantation, before an increase in ATP synthase staining by 6th month and a switch of transplanted cells to oxidative phosphorylation. The conducted experiment demonstrated a link between mitochondrial dynamics and changes in the metabolic profile and maturation of transplanted neurons. The regulation of mitochondrial dynamics may have future implications for developing methods to improve the integration of transplanted neurons into recepient brain structures.

Известно, что регуляция митохондриальных функций влияет на дифференцировку и созревание нейронов. Изучение этих процессов имеет как фундаментальное, так и практическое значение для регенеративной нейробиологии. Цель исследования: охарактеризовать изменения деления митохондрий и их связь с активацией окислительного фосфорилирования (метаболического переключения) при созревании иПСК-продуцированных нейрональных предшественников при введении в стриатум крыс. Крысам Вистар (n = 15) в хвостатое ядро односторонне вводили нейрональные предшественники, полученные из иПСК человека. С помощью иммуноокрашивания оценивали изменения локализации и экспрессии маркеров дифференцировки нейронов: нестина, NeuN, нейрональной енолазы, а также белка наружной мембраны митохондрий, АТФ-синтазы и белка деления митохондрий Drp1. Измерения проводились на клетках трансплантата через 2 недели, 3 и 6 месяцев после операции. Созревание трансплантата было связано с колебаниями морфометрических параметров митохондриальной фракции и уровня Drp1. Усиление деления митохондрий выявляли через 3 месяца после трансплантации, что предшествовало увеличению содержания АТФ-синтазы к 6 месяцу и переключению трансплантированных клеток на окислительное фосфорилирование. Проведенный эксперимент показал связь митохондриальной динамики с изменениями метаболического профиля и созреванием трансплантированных нейронов. Регуляция митохондриальной динамики может иметь значение для разработки методов улучшения интеграции трансплантированных нейронов в структуры мозга.

neurogenesisgraftinghuman IPSCmitochondrial dynamicsDrp1Oxidative phosphorylationнейрогенезтрансплантацияиндуцированные плюрипотентные клетки человекамитохондриальная динамикаDrp1окислительное фосфорилированиеРНФRussian Science Foundation24-45-00052Doss MX, Sachinidis A (2019) Current Challenges of iPSC-Based Disease Modeling and Therapeutic Implications. Cells 8: 403. https://doi.org/10.3390/cells8050403Bigarreau J, Rouach N, Perrier AL, Mouthon F, Charvériat M (2022) Modeling and Targeting Neuroglial Interactions with Human Pluripotent Stem Cell Models. Int J Mol Sci 23: 1684. https://doi.org/10.3390/ijms23031684Crane AT, Voth JP, Shen FX, Low WC (2019) Concise Review: Human-Animal Neurological Chimeras: Humanized Animals or Human Cells in an Animal? Stem Cells 37: 444–452. https://doi.org/10.1002/stem.2971Aleksandrova MA, Marey MV (2015) [Stem Cells in the Brain of Mammals and Human: Fundamental and Applied Aspects]. Zh Vyssh Nerv Deiat Im I P Pavlova 65: 271–305Iwata R, Casimir P, Vanderhaeghen P (2020) Mitochondrial dynamics in postmitotic cells regulate neurogenesis. Science (80) 369:858–862. https://doi.org/10.1126/science.aba9760Folmes CDL, Terzic A (2016) Energy metabolism in the acquisition and maintenance of stemness. Semin Cell Dev Biol 52: 68–75. https://doi.org/10.1016/j.semcdb.2016.02.010Sun X, St John JC (2016) The role of the mtDNA set point in differentiation, development and tumorigenesis. Biochem J 473: 2955–2971. https://doi.org/10.1042/BCJ20160008Maffezzini C, Calvo-Garrido J, Wredenberg A, Freyer C (2020) Metabolic regulation of neurodifferentiation in the adult brain. Cell Mol Life Sci 77: 2483–2496. https://doi.org/10.1007/s00018-019-03430-9Zheng X, Boyer L, Jin M, Mertens J, Kim Y, Ma L, Ma L, Hamm M, Gage FH, Hunter T (2016) Metabolic reprogramming during neuronal differentiation from aerobic glycolysis to neuronal oxidative phosphorylation. Elife 5. https://doi.org/10.7554/eLife.13374Khacho M, Slack RS (2018) Mitochondrial dynamics in the regulation of neurogenesis: From development to the adult brain. Dev Dyn 247: 47–53. https://doi.org/10.1002/dvdy.24538Khacho M, Harris R, Slack RS (2019) Mitochondria as central regulators of neural stem cell fate and cognitive function. Nat Rev Neurosci 20:34–48. https://doi.org/10.1038/s41583-018-0091-3Vantaggiato C, Castelli M, Giovarelli M, Orso G, Bassi MT, Clementi E, De Palma C (2019) The Fine Tuning of Drp1-Dependent Mitochondrial Remodeling and Autophagy Controls Neuronal Differentiation. Front Cell Neurosci 13. https://doi.org/10.3389/fncel.2019.00120Singh M, Denny H, Smith C, Granados J, Renden R (2018) Presynaptic loss of dynamin‐related protein 1 impairs synaptic vesicle release and recycling at the mouse calyx of Held. J Physiol 596: 6263–6287. https://doi.org/10.1113/JP276424Voronkov DN, Stavrovskaya A V., Lebedeva OS, Li W, Olshansky AS, Gushchina AS, Kapkaeva MR, Bogomazova AN, Lagarkova MA, Illarioshkin SN (2023) Morphological Changes in Neural Progenitors Derived from Human Induced Pluripotent Stem Cells and Transplanted into the Striatum of a Parkinson’s Disease Rat Model. Ann Clin Exp Neurol 17: 43–50. https://doi.org/10.54101/ACEN.2023.2.6Holmqvist S, Lehtonen S, Chumarina M, Puttonen KA, Azevedo C, Lebedeva O, Ruponen M, Oksanen M, Djelloul M, Collin A, Goldwurm S, Meyer M, Lagarkova M, Kiselev S, Koistinaho J, Roybon L (2016) Creation of a library of induced pluripotent stem cells from Parkinsonian patients. npj Park Dis 2: 16009. https://doi.org/10.1038/npjparkd.2016.9Voronkov DN, Stavrovskaya A V., Guschina AS, Olshansky AS, Lebedeva OS, Eremeev A V., Lagarkova MA (2022) Morphological Characterization of Astrocytes in a Xenograft of Human iPSC-Derived Neural Precursor Cells. Acta Naturae 14: 100–108. https://doi.org/10.32607/actanaturae.11710Lebedeva OS, Sharova EI, Grekhnev DA, Skorodumova LO, Kopylova I V., Vassina EM, Oshkolova A, Novikova IV, Krisanova AV, Olekhnovich EI, Vigont VA, Kaznacheyeva E V, Bogomazova AN, Lagarkova MA (2023) An Efficient 2D Protocol for Differentiation of iPSCs into Mature Postmitotic Dopaminergic Neurons: Application for Modeling Parkinson’s Disease. Int J Mol Sci 24: 7297. https://doi.org/10.3390/ijms24087297Song W, Bossy B, Martin OJ, Hicks A, Lubitz S, Knott AB, Bossy-Wetzel E (2008) Assessing mitochondrial morphology and dynamics using fluorescence wide-field microscopy and 3D image processing. Methods 46: 295–303. https://doi.org/10.1016/j.ymeth.2008.10.003Son G, Han J (2018) Roles of mitochondria in neuronal development. BMB Rep 51: 549–556. https://doi.org/10.5483/BMBRep.2018.51.11.226Iwata R, Vanderhaeghen P (2021) Regulatory roles of mitochondria and metabolism in neurogenesis. Curr Opin Neurobiol 69: 231–240. https://doi.org/10.1016/j.conb.2021.05.003Steib K, Schäffner I, Jagasia R, Ebert B, Lie DC (2014) Mitochondria Modify Exercise-Induced Development of Stem Cell-Derived Neurons in the Adult Brain. J Neurosci 34: 6624–6633. https://doi.org/10.1523/JNEUROSCI.4972-13.2014Kim HJ, Shaker MR, Cho B, Cho HM, Kim H, Kim JY, Sun W (2015) Dynamin-related protein 1 controls the migration and neuronal differentiation of subventricular zone-derived neural progenitor cells. Sci Rep 5: 15962. https://doi.org/10.1038/srep15962Rossi MJ, Pekkurnaz G (2019) Powerhouse of the mind: mitochondrial plasticity at the synapse. Curr Opin Neurobiol 57: 149–155. https://doi.org/10.1016/j.conb.2019.02.001Zhang S, Zhao J, Quan Z, Li H, Qing H (2022) Mitochondria and Other Organelles in Neural Development and Their Potential as Therapeutic Targets in Neurodegenerative Diseases. Front Neurosci 16. https://doi.org/10.3389/fnins.2022.853911Berthet A, Margolis EB, Zhang J, Hsieh I, Zhang J, Hnasko TS, Ahmad J, Edwards RH, Sesaki H, Huang EJ, Nakamura K (2014) Loss of Mitochondrial Fission Depletes Axonal Mitochondria in Midbrain Dopamine Neurons. J Neurosci 34: 14304–14317. https://doi.org/10.1523/JNEUROSCI.0930-14.2014Itoh K, Murata D, Kato T, Yamada T, Araki Y, Saito A, Adachi Y, Igarashi A, Li S, Pletnikov M, Huganir RL, Watanabe S, Kamiya A, Iijima M, Sesaki H (2019) Brain-specific Drp1 regulates postsynaptic endocytosis and dendrite formation independently of mitochondrial division. Elife 8. https://doi.org/10.7554/eLife.44739Beckervordersandforth R, Ebert B, Schäffner I, Moss J, Fiebig C, Shin J, Moore DL, Ghosh L, Trinchero MF, Stockburger C, Friedland K, Steib K, von Wittgenstein J, Keiner S, Redecker C, Hölter SM, Xiang W, Wurst W, Jagasia R, Schinder AF, Ming G, Toni N, Jessberger S, Song H, Lie DC (2017) Role of Mitochondrial Metabolism in the Control of Early Lineage Progression and Aging Phenotypes in Adult Hippocampal Neurogenesis. Neuron 93: 560–573.e6. https://doi.org/10.1016/j.neuron.2016.12.017Paumard P, Vaillier J, Coulary B, Schaeffer J, Soubannier V, Mueller DM, Brèthes D, di Rago J-P, Velours J (2002) The ATP synthase is involved in generating mitochondrial cristae morphology. EMBO J 21: 221–230. https://doi.org/10.1093/emboj/21.3.221Fame RM, Shannon ML, Chau KF, Head JP, Lehtinen MK (2019) Concerted metabolic shift in early forebrain alters the CSF proteome and depends on cMYC downregulation for mitochondrial maturation. Development 146(20):dev182857. https://doi.org/10.1242/dev.182857Haque A, Polcyn R, Matzelle D, Banik NL (2018) New Insights into the Role of Neuron-Specific Enolase in Neuro-Inflammation, Neurodegeneration, and Neuroprotection. Brain Sci 8: 33. https://doi.org/10.3390/brainsci8020033Tarazona OA, Pourquié O (2020) Exploring the Influence of Cell Metabolism on Cell Fate through Protein Post-translational Modifications. Dev Cell 54: 282–292. https://doi.org/10.1016/j.devcel.2020.06.035Dai Z, Ramesh V, Locasale JW (2020) The evolving metabolic landscape of chromatin biology and epigenetics. Nat Rev Genet 21: 737–753. https://doi.org/10.1038/s41576-020-0270-8Iwata R, Casimir P, Erkol E, Boubakar L, Planque M, Gallego López IM, Ditkowska M, Gaspariunaite V, Beckers S, Remans D, Vints K, Vandekeere A, Poovathingal S, Bird M, Vlaeminck I, Creemers E, Wierda K, Corthout N, Vermeersch P, Carpentier S, Davie K, Mazzone M, Gounko NV, Aerts S, Ghesquière B, Fendt S-M, Vanderhaeghen P (2023) Mitochondria metabolism sets the species-specific tempo of neuronal development. Science (80) 379. https://doi.org/10.1126/science.abn4705