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Energy Metabolism and Mitochondrial Superoxide Anion Production in Pre-symptomatic Striatal Neurons Derived from Human-Induced Pluripotent Stem Cells Expressing Mutant Huntingtin

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Abstract

In the present study, we investigated whether mutant huntingtin (mHTT) impairs mitochondrial functions in human striatal neurons derived from induced pluripotent stem cells (iPSCs). Striatal neurons and astrocytes derived from iPSCs from unaffected individuals (Ctrl) and Huntington’s disease (HD) patients with HTT gene containing increased number of CAG repeats were used to assess the effect of mHTT on bioenergetics and mitochondrial superoxide anion production. The human neurons were thoroughly characterized and shown to express MAP2, DARPP32, GABA, synapsin, and PSD95. In human neurons and astrocytes expressing mHTT, the ratio of mHTT to wild-type huntingtin (HTT) was 1:1. The human neurons were excitable and could generate action potentials, confirming successful conversion of iPSCs into functional neurons. The neurons and astrocytes from Ctrl individuals and HD patients had similar levels of ADP and ATP and comparable respiratory and glycolytic activities. The mitochondrial mass, mitochondrial membrane potential, and superoxide anion production in human neurons appeared to be similar regardless of mHTT presence. The present results are in line with the results obtained in our previous studies with isolated brain mitochondria and cultured striatal neurons from YAC128 and R6/2 mice, in which we demonstrated that mutant huntingtin at early stages of HD pathology does not deteriorate mitochondrial functions. Overall, our results argue against bioenergetic deficits as a factor in HD pathogenesis and suggest that other detrimental processes might be more relevant to the development of HD pathology.

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Abbreviations

mHTT:

Mutant huntingtin

HTT:

Wild-type huntingtin

iPSCs:

Induced pluripotent stem cells

neurons:

Human medium spiny neurons

HD:

Huntington’s disease

ROS:

Reactive oxygen species

BDNF:

Brain-derived neurotrophic factor

GDNF:

Glia-derived neurotrophic factor

DAPI:

2-(4-Amidinophenyl)-1H-indole-6-carboxamidine

OCRs:

Oxygen consumption rates

ECARs:

Extracellular acidification rates

TMRM:

Tetramethylrhodamine, methyl ester

TH:

Tyrosine hydroxylase

AP:

Action potentials

2,4-DNP:

2,4-Dinitrophenol

FCCP:

Carbonyl cyanide p-trifluoromethoxyphenylhydrazone

(hESCs):

Human embryonic stem cells

polyQ:

Poly-glutamine

Ant:

Antimycin A

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Acknowledgments

We are very thankful to Dr. George Daley (Harvard University, Cambridge, MA) and Dr. David Gamm (University of Wisconsin, Madison, WI) for providing human undifferentiated induced pluripotent stem cells.

Funding

This study was supported by National Institutes of Health grant R01 NS098772 and in part by a grant from Indiana Traumatic Spinal Cord & Brain Injury Research Fund to N.B.

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Authors and Affiliations

  1. Department of Pharmacology and Toxicology, School of Medicine, Indiana University, Indianapolis, IN, 46202, USA

    James Hamilton, Tatiana Brustovetsky & Nickolay Brustovetsky

  2. Program in Medical Neuroscience, School of Medicine, Indiana University, Indianapolis, IN, 46202, USA

    Akshayalakshmi Sridhar, Yanling Pan, Theodore R. Cummins, Jason S. Meyer & Nickolay Brustovetsky

  3. Paul and Carole Stark Neurosciences Research Institute, School of Medicine, Indiana University, Indianapolis, IN, 46202, USA

    Theodore R. Cummins, Jason S. Meyer & Nickolay Brustovetsky

  4. Department of Biology, School of Science, IUPUI, Indianapolis, IN, 46202, USA

    Theodore R. Cummins & Jason S. Meyer

  5. Indiana University School of Medicine, 635 Barnhill Dr., Medical Science Building, Room 362, Indianapolis, IN, 46202, USA

    Nickolay Brustovetsky

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  1. James Hamilton
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Correspondence to Nickolay Brustovetsky.

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Hamilton, J., Brustovetsky, T., Sridhar, A. et al. Energy Metabolism and Mitochondrial Superoxide Anion Production in Pre-symptomatic Striatal Neurons Derived from Human-Induced Pluripotent Stem Cells Expressing Mutant Huntingtin. Mol Neurobiol 57, 668–684 (2020). https://doi.org/10.1007/s12035-019-01734-2

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