Advertisement

An innovative approach for a rare disease

Charcot-Marie-Tooth disease type 2A (CMT2A) is a rare, inherited neurodegenerative condition. Affected individuals develop severe progressive muscle weakness, motor deficits, and peripheral neuropathy. Although defects in the gene encoding mitofusin 2 (MFN2) are known to cause CMT2A, the disease remains incurable. Rocha et al. identified specific MFN2 residues contributing to the disease and developed a class of MFN2-agonist drugs. The small molecules restored mitochondrial fusion and activity in the sciatic nerves of mice; they may also help in other diseases linked to mitochondrial trafficking.
Science, this issue p. 336

Abstract

Mitofusins (MFNs) promote fusion-mediated mitochondrial content exchange and subcellular trafficking. Mutations in Mfn2 cause neurodegenerative Charcot-Marie-Tooth disease type 2A (CMT2A). We showed that MFN2 activity can be determined by Met376 and His380 interactions with Asp725 and Leu727 and controlled by PINK1 kinase–mediated phosphorylation of adjacent MFN2 Ser378. Small-molecule mimics of the peptide-peptide interface of MFN2 disrupted this interaction, allosterically activating MFN2 and promoting mitochondrial fusion. These first-in-class mitofusin agonists overcame dominant mitochondrial defects provoked in cultured neurons by CMT2A mutants MFN2 Arg94→Gln94 and MFN2 Thr105→Met105, as demonstrated by amelioration of mitochondrial dysmotility, fragmentation, depolarization, and clumping. A mitofusin agonist normalized axonal mitochondrial trafficking within sciatic nerves of MFN2 Thr105→Met105 mice, promising a therapeutic approach for CMT2A and other untreatable diseases of impaired neuronal mitochondrial dynamism and/or trafficking.
Get full access to this article

View all available purchase options and get full access to this article.

Already a Subscriber?

Supplementary Material

Summary

Materials and Methods
Figs. S1 to S31
Table S1
References (1730)
Movies S1 to S4
Data S1 to S4

Resources

File (aao1785-rocha-sm.pdf)
File (aao1785_datas1.xls)
File (aao1785_datas2.xlsx)
File (aao1785_datas3.xlsx)
File (aao1785_datas4.xlsx)
File (aao1785s1.mp4)
File (aao1785s2.mp4)
File (aao1785s3.mov)
File (aao1785s4.mov)

References and Notes

1
D. C. Chan, Fusion and fission: Interlinked processes critical for mitochondrial health. Annu. Rev. Genet. 46, 265–287 (2012).
2
M. Liesa, M. Palacín, A. Zorzano, Mitochondrial dynamics in mammalian health and disease. Physiol. Rev. 89, 799–845 (2009).
3
A. Franco, R. N. Kitsis, J. A. Fleischer, E. Gavathiotis, O. S. Kornfeld, G. Gong, N. Biris, A. Benz, N. Qvit, S. K. Donnelly, Y. Chen, S. Mennerick, L. Hodgson, D. Mochly-Rosen, G. W. I. I. Dorn, Correcting mitochondrial fusion by manipulating mitofusin conformations. Nature 540, 74–79 (2016).
4
Y. Chen, G. W. Dorn II, PINK1-phosphorylated mitofusin 2 is a Parkin receptor for culling damaged mitochondria. Science 340, 471–475 (2013).
5
G. Gong, M. Song, G. Csordas, D. P. Kelly, S. J. Matkovich, G. W. Dorn II, Parkin-mediated mitophagy directs perinatal cardiac metabolic maturation in mice. Science 350, aad2459 (2015).
6
V. H. Lawson, B. V. Graham, K. M. Flanigan, Clinical and electrophysiologic features of CMT2A with mutations in the mitofusin 2 gene. Neurology 65, 197–204 (2005).
7
K. Verhoeven, K. G. Claeys, S. Züchner, J. M. Schröder, J. Weis, C. Ceuterick, A. Jordanova, E. Nelis, E. De Vriendt, M. Van Hul, P. Seeman, R. Mazanec, G. M. Saifi, K. Szigeti, P. Mancias, I. J. Butler, A. Kochanski, B. Ryniewicz, J. De Bleecker, P. Van den Bergh, C. Verellen, R. Van Coster, N. Goemans, M. Auer-Grumbach, W. Robberecht, V. Milic Rasic, Y. Nevo, I. Tournev, V. Guergueltcheva, F. Roelens, P. Vieregge, P. Vinci, M. T. Moreno, H. J. Christen, M. E. Shy, J. R. Lupski, J. M. Vance, P. De Jonghe, V. Timmerman, MFN2 mutation distribution and genotype/phenotype correlation in Charcot-Marie-Tooth type 2. Brain 129, 2093–2102 (2006).
8
R. H. Baloh, R. E. Schmidt, A. Pestronk, J. Milbrandt, Altered axonal mitochondrial transport in the pathogenesis of Charcot-Marie-Tooth disease from mitofusin 2 mutations. J. Neurosci. 27, 422–430 (2007).
9
A. Misko, S. Jiang, I. Wegorzewska, J. Milbrandt, R. H. Baloh, Mitofusin 2 is necessary for transport of axonal mitochondria and interacts with the Miro/Milton complex. J. Neurosci. 30, 4232–4240 (2010).
10
A. L. Misko, Y. Sasaki, E. Tuck, J. Milbrandt, R. H. Baloh, Mitofusin2 mutations disrupt axonal mitochondrial positioning and promote axon degeneration. J. Neurosci. 32, 4145–4155 (2012).
11
K. W. Chung, S. B. Kim, K. D. Park, K. G. Choi, J. H. Lee, H. W. Eun, J. S. Suh, J. H. Hwang, W. K. Kim, B. C. Seo, S. H. Kim, I. H. Son, S. M. Kim, I. N. Sunwoo, B. O. Choi, Early onset severe and late-onset mild Charcot-Marie-Tooth disease with mitofusin 2 (MFN2) mutations. Brain 129, 2103–2118 (2006).
12
C. Casasnovas, I. Banchs, J. Cassereau, N. Gueguen, A. Chevrollier, J. A. Martínez-Matos, D. Bonneau, V. Volpini, Phenotypic spectrum of MFN2 mutations in the Spanish population. J. Med. Genet. 47, 249–256 (2010).
13
F. Bombelli, T. Stojkovic, O. Dubourg, A. Echaniz-Laguna, S. Tardieu, K. Larcher, P. Amati-Bonneau, P. Latour, O. Vignal, C. Cazeneuve, A. Brice, E. Leguern, Charcot-Marie-Tooth disease type 2A: From typical to rare phenotypic and genotypic features. JAMA Neurol. 71, 1036–1042 (2014).
14
K. Hirai, G. Aliev, A. Nunomura, H. Fujioka, R. L. Russell, C. S. Atwood, A. B. Johnson, Y. Kress, H. V. Vinters, M. Tabaton, S. Shimohama, A. D. Cash, S. L. Siedlak, P. L. Harris, P. K. Jones, R. B. Petersen, G. Perry, M. A. Smith, Mitochondrial abnormalities in Alzheimer’s disease. J. Neurosci. 21, 3017–3023 (2001).
15
K. F. Winklhofer, C. Haass, Mitochondrial dysfunction in Parkinson’s disease. Biochim. Biophys. Acta 1802, 29–44 (2010).
16
J. Kim, J. P. Moody, C. K. Edgerly, O. L. Bordiuk, K. Cormier, K. Smith, M. F. Beal, R. J. Ferrante, Mitochondrial loss, dysfunction and altered dynamics in Huntington’s disease. Hum. Mol. Genet. 19, 3919–3935 (2010).
17
H. Chen, S. A. Detmer, A. J. Ewald, E. E. Griffin, S. E. Fraser, D. C. Chan, Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion and are essential for embryonic development. J. Cell Biol. 160, 189–200 (2003).
18
Y. Zhang, I-TASSER server for protein 3D structure prediction. BMC Bioinformatics 9, 40 (2008).
19
H. H. Low, J. Löwe, A bacterial dynamin-like protein. Nature 444, 766–769 (2006).
20
Y. Qi, L. Yan, C. Yu, X. Guo, X. Zhou, X. Hu, X. Huang, Z. Rao, Z. Lou, J. Hu, Structures of human mitofusin 1 provide insight into mitochondrial tethering. J. Cell Biol. 215, 621–629 (2016).
21
L. Yan, Y. Ma, Y. Sun, J. Gao, X. Chen, J. Liu, C. Wang, Z. Rao, Z. Lou, Structural basis for mechanochemical role of Arabidopsis thaliana dynamin-related protein in membrane fission. J. Mol. Cell Biol. 3, 378–381 (2011).
22
M. G. Ford, S. Jenni, J. Nunnari, The crystal structure of dynamin. Nature 477, 561–566 (2011).
23
C. Fröhlich, S. Grabiger, D. Schwefel, K. Faelber, E. Rosenbaum, J. Mears, O. Rocks, O. Daumke, Structural insights into oligomerization and mitochondrial remodelling of dynamin 1-like protein. EMBO J. 32, 1280–1292 (2013).
24
C. Goujon, R. A. Greenbury, S. Papaioannou, T. Doyle, M. H. Malim, A triple-arginine motif in the amino-terminal domain and oligomerization are required for HIV-1 inhibition by human MX2. J. Virol. 89, 4676–4680 (2015).
25
E. F. Pettersen, T. D. Goddard, C. C. Huang, G. S. Couch, D. M. Greenblatt, E. C. Meng, T. E. Ferrin, UCSF Chimera—a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605–1612 (2004).
26
F. Sievers, A. Wilm, D. Dineen, T. J. Gibson, K. Karplus, W. Li, R. Lopez, H. McWilliam, M. Remmert, J. Söding, J. D. Thompson, D. G. Higgins, Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega. Mol. Syst. Biol. 7, 539 (2011).
27
F. C. Fiesel, R. Hudec, W. Springer, Non-radioactive in vitro PINK1 kinase assays using ubiquitin or Parkin as substrate. Bio Protoc. 6, e1946 (2016).
28
C. Sobieski, X. Jiang, D. C. Crawford, S. Mennerick, Loss of local astrocyte support disrupts action potential propagation and glutamate release synchrony from unmyelinated hippocampal axon terminals in vitro. J. Neurosci. 35, 11105–11117 (2015).
29
P. Bannerman, T. Burns, J. Xu, L. Miers, D. Pleasure, Mice hemizygous for a pathogenic mitofusin-2 allele exhibit hind limb/foot gait deficits and phenotypic perturbations in nerve and muscle. PLOS ONE 11, e0167573 (2016).
30
X. Yang, S. Arber, C. William, L. Li, Y. Tanabe, T. M. Jessell, C. Birchmeier, S. J. Burden, Patterning of muscle acetylcholine receptor gene expression in the absence of motor innervation. Neuron 30, 399–410 (2001).

Information & Authors

Information

Published In

Science
Volume 360 | Issue 6386
20 April 2018

Submission history

Received: 22 June 2017
Accepted: 27 February 2018
Published in print: 20 April 2018

Permissions

Request permissions for this article.

Acknowledgments

We gratefully acknowledge discussions with P. Needleman and the assistance of L. Zhang, P. Erdmann-Gilmore, Y. Mi, and R. Connors. Funding: This work was supported by NIH grants R35HL135736 (G.W.D.), R01HL128071 (R.N.K. and G.W.D.), and R01CA178394 and P30CA013330 (E.G.); a McDonnell Center for Cellular and Molecular Neurobiology postdoctoral fellowship (A.F.); and the Washington University Proteomics Shared Resource, supported by National Center for Advancing Translational Sciences grants UL1TR000448, NIGMSP41, GM103422, and NCIP30 CA091842. G.W.D. is the Philip and Sima K. Needleman–endowed professor. Author contributions: G.W.D., A.M.K., D.M.-R, and R.R.T. conceived of or designed the research, except the initial in silico screen. E.G., R.N.K., N.B., and E.Z. conceived of the small-molecule screen, designed the original pharmacophore model, and performed the initial in silico screen. G.W.D. wrote the manuscript. J.M.R. and R.R.T. performed phosphoprotein mass spectroscopy analyses. A.M.K. performed peptide nuclear magnetic resonance studies. A.G.R. screened compounds for activity and characterized agonists. A.G.R., A.F., J.M.A., and W.C.K. performed mitochondrial studies. A.F. performed cultured neuron and ex vivo sciatic nerve studies. J.W.J. analyzed and purified compounds. R.N.K., E.G., and R.H.B. provided essential reagents. Competing interests: D.M.-R. and G.W.D. are inventors on patent application 15/710,696, submitted by Stanford University, which covers the use of peptide regulators of mitochondrial fusion and small-molecule peptidomimetics derived from them. G.W.D. is an inventor on provisional patent applications 62/488,787 and 62/584,515, submitted by Washington University, which cover the use of novel small-molecule mitofusin agonists to treat chronic neurodegenerative diseases. E.G., R.N.K., N.B., and E.Z. are inventors on patent application 62/573,217, submitted by Albert Einstein College of Medicine, which covers compositions of mitofusin agonists and their uses for the treatment of diseases and disorders. D.M.-R. is the founder of Mitoconix Bio, a company focused on improving mitochondrial health as a therapeutic approach for neurodegenerative diseases. None of the research conducted in D.M.-R.’s laboratory is supported by or performed in collaboration with Mitoconix. The other authors declare no competing interests. Data and materials availability: All data are available in the manuscript or the supplementary materials. There are no material transfer agreements associated with this study.

Authors

Affiliations

Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO, USA.
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO, USA.
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO, USA.
Jeanne M. Rumsey
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO, USA.
Justin M. Alberti
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO, USA.
William C. Knight
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO, USA.
Departments of Biochemistry and Medicine, Wilf Family Cardiovascular Research Institute, Albert Einstein Cancer Center, Albert Einstein College of Medicine, Bronx, NY, USA.
Emmanouil Zacharioudakis
Departments of Biochemistry and Medicine, Wilf Family Cardiovascular Research Institute, Albert Einstein Cancer Center, Albert Einstein College of Medicine, Bronx, NY, USA.
Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, St. Louis, MO, USA.
Department of Neurology, Cedars-Sinai Medical Center, Los Angeles, CA, USA.
Departments of Medicine and Cell Biology, Wilf Family Cardiovascular Research Institute, Albert Einstein Cancer Center, and Einstein-Mount Sinai Diabetes Research Center, Albert Einstein College of Medicine, Bronx, NY, USA.
Department of Chemical and Systems Biology, Stanford University School of Medicine, Stanford, CA, USA.
R. Reid Townsend
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO, USA.
Departments of Biochemistry and Medicine, Wilf Family Cardiovascular Research Institute, Albert Einstein Cancer Center, Albert Einstein College of Medicine, Bronx, NY, USA.
Gerald W. Dorn II [email protected]
Department of Internal Medicine, Washington University School of Medicine, St. Louis, MO, USA.

Funding Information

Notes

*
These authors contributed equally to this work.
†Corresponding author. Email: [email protected]

Metrics & Citations

Metrics

Article Usage
Altmetrics

Citations

Export citation

Select the format you want to export the citation of this publication.

Cited by
  1. Treatment and Management of Hereditary Neuropathies, Neuromuscular Disorders, (278-311), (2022).https://doi.org/10.1016/B978-0-323-71317-7.00014-7
    Crossref
  2. Updated review of therapeutic strategies for Charcot-Marie-Tooth disease and related neuropathies, Expert Review of Neurotherapeutics, 21, 6, (701-713), (2021).https://doi.org/10.1080/14737175.2021.1935242
    Crossref
  3. Pharmacological advances in mitochondrial therapy, EBioMedicine, 65, (103244), (2021).https://doi.org/10.1016/j.ebiom.2021.103244
    Crossref
  4. Mitochondria Dynamics: Definition, Players and Associated Disorders, Mitochondrial Diseases, (119-142), (2021).https://doi.org/10.1007/978-3-030-70147-5
    Crossref
  5. The Present and Future of Mitochondrial-Based Therapeutics for Eye Disease, Translational Vision Science & Technology, 10, 8, (4), (2021).https://doi.org/10.1167/tvst.10.8.4
    Crossref
  6. TDP-43 and PINK1 mediate CHCHD10S59L mutation–induced defects in Drosophila and in vitro, Nature Communications, 12, 1, (2021).https://doi.org/10.1038/s41467-021-22145-9
    Crossref
  7. Benefit of a single simulated hypobaric hypoxia in healthy mice performance and analysis of mitochondria-related gene changes, Scientific Reports, 11, 1, (2021).https://doi.org/10.1038/s41598-020-80425-8
    Crossref
  8. USP30 protects against oxygen-glucose deprivation/reperfusion induced mitochondrial fragmentation and ubiquitination and degradation of MFN2, Aging, 13, 4, (6194-6204), (2021).https://doi.org/10.18632/aging.202629
    Crossref
  9. Impaired mitochondrial dynamics in disease, Mitochondrial Dysfunction and Nanotherapeutics, (57-90), (2021).https://doi.org/10.1016/B978-0-323-85666-9.00011-5
    Crossref
  10. Mitofusin-2 modulates the epithelial to mesenchymal transition in thyroid cancer progression, Scientific Reports, 11, 1, (2021).https://doi.org/10.1038/s41598-021-81469-0
    Crossref
  11. See more
Loading...

View Options

Get Access

Log in to view the full text

AAAS Log in

AAAS login provides access to Science for AAAS members, and access to other journals in the Science family to users who have purchased individual subscriptions.

Log in via OpenAthens.
Log in via Shibboleth.
More options

Register for free to read this article

As a service to the community, this article is available for free. Login or register for free to read this article.

Purchase this issue in print

Buy a single issue of Science for just $15 USD.

View options

PDF format

Download this article as a PDF file

Download PDF

Media

Figures

Multimedia

Tables

Share

Share

Share article link

Share on social media