Therapy of pelizaeus-merzbacher disease in mice by feeding a cholesterol-enriched diet

ABSTRACT Duplication of _PLP1_ (proteolipid protein gene 1) and the subsequent overexpression of the myelin protein PLP (also known as DM20) in oligodendrocytes is the most frequent cause of

Pelizaeus-Merzbacher disease (PMD), a fatal leukodystrophy1 without therapeutic options2,3. PLP binds cholesterol and is contained within membrane lipid raft microdomains4. Cholesterol

availability is the rate-limiting factor of central nervous system myelin synthesis5. Transgenic mice with extra copies of the _Plp1_ gene6 are accurate models of PMD. Dysmyelination6,7,8

followed by demyelination9,10, secondary inflammation and axon damage contribute to the severe motor impairment in these mice9,10. The finding that in _Plp1_-transgenic oligodendrocytes, PLP

and cholesterol accumulate in late endosomes and lysosomes (endo/lysosomes)9,11,12,13, prompted us to further investigate the role of cholesterol in PMD. Here we show that cholesterol

itself promotes normal PLP trafficking and that dietary cholesterol influences PMD pathology. In a preclinical trial, PMD mice were fed a cholesterol-enriched diet. This restored

oligodendrocyte numbers and ameliorated intracellular PLP accumulation. Moreover, myelin content increased, inflammation and gliosis were reduced and motor defects improved. Even after onset

of clinical symptoms, cholesterol treatment prevented disease progression. Dietary cholesterol did not reduce _Plp1_ overexpression but facilitated incorporation of PLP into myelin

membranes. These findings may have implications for therapeutic interventions in patients with PMD. Access through your institution Buy or subscribe This is a preview of subscription

content, access via your institution ACCESS OPTIONS Access through your institution Subscribe to this journal Receive 12 print issues and online access $209.00 per year only $17.42 per issue

Learn more Buy this article * Purchase on SpringerLink * Instant access to full article PDF Buy now Prices may be subject to local taxes which are calculated during checkout ADDITIONAL

ACCESS OPTIONS: * Log in * Learn about institutional subscriptions * Read our FAQs * Contact customer support SIMILAR CONTENT BEING VIEWED BY OTHERS CORRECTION OF DYSREGULATED LIPID

METABOLISM NORMALIZES GENE EXPRESSION IN OLIGODENDROCYTES AND PROLONGS LIFESPAN IN FEMALE POLY-GA C9ORF72 MICE Article Open access 11 April 2025 DYSREGULATION OF METABOLIC PATHWAYS BY

CARNITINE PALMITOYL-TRANSFERASE 1 PLAYS A KEY ROLE IN CENTRAL NERVOUS SYSTEM DISORDERS: EXPERIMENTAL EVIDENCE BASED ON ANIMAL MODELS Article Open access 24 September 2020 THE TRANSLATIONAL

POTENTIAL OF CHOLESTEROL-BASED THERAPIES FOR NEUROLOGICAL DISEASE Article 29 August 2023 REFERENCES * Seitelberger, F. Neuropathology and genetics of Pelizaeus-Merzbacher disease. _Brain

Pathol._ 5, 267–273 (1995). Article  CAS  Google Scholar  * Garbern, J.Y. Pelizaeus-Merzbacher disease: Genetic and cellular pathogenesis. _Cell Mol. Life Sci._ 64, 50–65 (2007). Article 

CAS  Google Scholar  * Woodward, K.J. The molecular and cellular defects underlying Pelizaeus-Merzbacher disease. _Expert Rev. Mol. Med._ 10, e14 (2008). Article  Google Scholar  * Simons,

M., Kramer, E.M., Thiele, C., Stoffel, W. & Trotter, J. Assembly of myelin by association of proteolipid protein with cholesterol- and galactosylceramide-rich membrane domains. _J. Cell

Biol._ 151, 143–154 (2000). Article  CAS  Google Scholar  * Saher, G. et al. High cholesterol level is essential for myelin membrane growth. _Nat. Neurosci._ 8, 468–475 (2005). Article  CAS

  Google Scholar  * Readhead, C., Schneider, A., Griffiths, I. & Nave, K.A. Premature arrest of myelin formation in transgenic mice with increased proteolipid protein gene dosage.

_Neuron_ 12, 583–595 (1994). Article  CAS  Google Scholar  * Anderson, T.J. et al. Distinct phenotypes associated with increasing dosage of the PLP gene: implications for CMT1A due to PMP22

gene duplication. _Ann. NY Acad. Sci._ 883, 234–246 (1999). Article  CAS  Google Scholar  * Anderson, T.J. et al. Late-onset neurodegeneration in mice with increased dosage of the

proteolipid protein gene. _J. Comp. Neurol._ 394, 506–519 (1998). Article  CAS  Google Scholar  * Edgar, J.M. et al. Demyelination and axonal preservation in a transgenic mouse model of

Pelizaeus-Merzbacher disease. _EMBO Mol. Med._ 2, 42–50 (2010). Article  CAS  Google Scholar  * Karim, S.A. et al. PLP/DM20 expression and turnover in a transgenic mouse model of

Pelizaeus-Merzbacher disease. _Glia_ 58, 1727–1738 (2010). Article  Google Scholar  * Krämer, E.M., Schardt, A. & Nave, K.A. Membrane traffic in myelinating oligodendrocytes. _Microsc.

Res. Tech._ 52, 656–671 (2001). Article  Google Scholar  * Karim, S.A. et al. PLP overexpression perturbs myelin protein composition and myelination in a mouse model of Pelizaeus-Merzbacher

disease. _Glia_ 55, 341–351 (2007). Article  Google Scholar  * Simons, M. et al. Overexpression of the myelin proteolipid protein leads to accumulation of cholesterol and proteolipid protein

in endosomes/lysosomes: implications for Pelizaeus-Merzbacher disease. _J. Cell Biol._ 157, 327–336 (2002). Article  CAS  Google Scholar  * Björkhem, I. & Meaney, S. Brain cholesterol:

long secret life behind a barrier. _Arterioscler. Thromb. Vasc. Biol._ 24, 806–815 (2004). Article  Google Scholar  * Alberti, S. et al. Hepatic cholesterol metabolism and resistance to

dietary cholesterol in LXRbeta-deficient mice. _J. Clin. Invest._ 107, 565–573 (2001). Article  CAS  Google Scholar  * Ip, C.W. et al. Origin of CD11b+ macrophage-like cells in the CNS of

PLP-overexpressing mice: low influx of haematogenous macrophages and unchanged blood-brain-barrier in the optic nerve. _Mol. Cell Neurosci._ 38, 489–494 (2008). Article  CAS  Google Scholar

  * Stolp, H.B. & Dziegielewska, K.M. Review: Role of developmental inflammation and blood-brain barrier dysfunction in neurodevelopmental and neurodegenerative diseases. _Neuropathol.

Appl. Neurobiol._ 35, 132–146 (2009). Article  CAS  Google Scholar  * Xie, C., Lund, E.G., Turley, S.D., Russell, D.W. & Dietschy, J.M. Quantitation of two pathways for cholesterol

excretion from the brain in normal mice and mice with neurodegeneration. _J. Lipid Res._ 44, 1780–1789 (2003). Article  CAS  Google Scholar  * Liu, B. et al. Reversal of defective lysosomal

transport in NPC disease ameliorates liver dysfunction and neurodegeneration in the _Npc1_−/− mouse. _Proc. Natl. Acad. Sci. USA_ 106, 2377–2382 (2009). Article  CAS  Google Scholar  *

Brown, M.S. & Goldstein, J.L. The SREBP pathway: regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor. _Cell_ 89, 331–340 (1997). Article  CAS 

Google Scholar  * Saher, G. et al. Cholesterol regulates the endoplasmic reticulum exit of the major membrane protein P0 required for peripheral myelin compaction. _J. Neurosci._ 29,

6094–6104 (2009). Article  CAS  Google Scholar  * Qin, C., Nagao, T., Grosheva, I., Maxfield, F.R. & Pierini, L.M. Elevated plasma membrane cholesterol content alters macrophage

signaling and function. _Arterioscler. Thromb. Vasc. Biol._ 26, 372–378 (2006). Article  CAS  Google Scholar  * Shepard, L.A. et al. Identification of a membrane-spanning domain of the

thiol-activated pore-forming toxin Clostridium perfringens perfringolysin O: an α-helical to β-sheet transition identified by fluorescence spectroscopy. _Biochemistry_ 37, 14563–14574

(1998). Article  CAS  Google Scholar  * Karten, B., Peake, K.B. & Vance, J.E. Mechanisms and consequences of impaired lipid trafficking in Niemann-Pick type C1–deficient mammalian cells.

_Biochim. Biophys. Acta_ 1791, 659–670 (2009). Article  CAS  Google Scholar  * Otomo, T., Higaki, K., Nanba, E., Ozono, K. & Sakai, N. Lysosomal storage causes cellular dysfunction in

mucolipidosis II skin fibroblasts. _J. Biol. Chem._ 286, 35283–35290 (2011). Article  CAS  Google Scholar  * Tang, Y., Leao, I.C., Coleman, E.M., Broughton, R.S. & Hildreth, J.E.

Deficiency of niemann-pick type C-1 protein impairs release of human immunodeficiency virus type 1 and results in Gag accumulation in late endosomal/lysosomal compartments. _J. Virol._ 83,

7982–7995 (2009). Article  CAS  Google Scholar  * Sobo, K. et al. Late endosomal cholesterol accumulation leads to impaired intra-endosomal trafficking. _PLoS ONE_ 2, e851 (2007). Article 

Google Scholar  * Hawes, C.M., Wiemer, H., Krueger, S.R. & Karten, B. Pre-synaptic defects of NPC1-deficient hippocampal neurons are not directly related to plasma membrane cholesterol.

_J. Neurochem._ 114, 311–322 (2010). CAS  Google Scholar  * Bauer, N.G., Richter-Landsberg, C. & Ffrench-Constant, C. Role of the oligodendroglial cytoskeleton in differentiation and

myelination. _Glia_ 57, 1691–1705 (2009). Article  Google Scholar  * Prusky, G.T., Alam, N.M., Beekman, S. & Douglas, R.M. Rapid quantification of adult and developing mouse spatial

vision using a virtual optomotor system. _Invest. Ophthalmol. Vis. Sci._ 45, 4611–4616 (2004). Article  Google Scholar  * Goetze, B. et al. Vision and visual cortical maps in mice with a

photoreceptor synaptopathy: reduced but robust visual capabilities in the absence of synaptic ribbons. _Neuroimage_ 49, 1622–1631 (2010). Article  Google Scholar  * Stromnes, I.M. &

Goverman, J.M. Active induction of experimental allergic encephalomyelitis. _Nat. Protoc._ 1, 1810–1819 (2006). Article  CAS  Google Scholar  * Jung, M., Sommer, I., Schachner, M. &

Nave, K.A. Monoclonal antibody O10 defines a conformationally sensitive cell-surface epitope of proteolipid protein (PLP): evidence that PLP misfolding underlies dysmyelination in mutant

mice. _J. Neurosci._ 16, 7920–7929 (1996). Article  CAS  Google Scholar  * Hölttä-Vuori, M. et al. BODIPY-cholesterol: a new tool to visualize sterol trafficking in living cells and

organisms. _Traffic_ 9, 1839–1849 (2008). Article  Google Scholar  * Möbius, W. et al. Immunoelectron microscopic localization of cholesterol using biotinylated and non-cytolytic

perfringolysin O. _J. Histochem. Cytochem._ 50, 43–55 (2002). Article  Google Scholar  * Waheed, A.A. et al. Selective binding of perfringolysin O derivative to cholesterol-rich membrane

microdomains (rafts). _Proc. Natl. Acad. Sci. USA_ 98, 4926–4931 (2001). Article  CAS  Google Scholar  Download references ACKNOWLEDGEMENTS We dedicate this work to the memory of Jim Garbern

for his support of PMD research. We are grateful to A. Fahrenholz, T. Freerck, M. Schildmann, M. Matthes and A. Kanbach for technical support and thank J. Edgar for helpful comments on the

manuscript. We thank M. Grebe (Umea University) for providing the BCθ expression construct and Pfizer for the generous gift of the squalestatin CP-340868. This work was funded by the PMD

foundation and the Deutsche Forschungsgemeinschaft (SA2114/1-1) to G.S. K.-A.N. is supported by the EU-FP7 (Leukotreat, Ngidd), the Bundesministerium für Bildung und Forschung (BMBF;

Leukonet) and a European Research Council (ERC) Advanced Grant. Support of the BMBF to S.L. and K.-F.S. is gratefully acknowledged (01GQ0810). AUTHOR INFORMATION Author notes * Fabian

Rudolphi and Kristina Corthals: These authors contributed equally to this work. AUTHORS AND AFFILIATIONS * Department of Neurogenetics, Max Planck Institute of Experimental Medicine,

Göttingen, Germany Gesine Saher, Fabian Rudolphi, Kristina Corthals, Torben Ruhwedel, Payam Dibaj, Benoit Barrette, Wiebke Möbius & Klaus-Armin Nave * Bernstein Focus Neurotechnology and

Johann-Friedrich-Blumenbach Institute of Zoology and Anthropology, Georg-August University, Göttingen, Germany Karl-Friedrich Schmidt & Siegrid Löwel Authors * Gesine Saher View author

publications You can also search for this author inPubMed Google Scholar * Fabian Rudolphi View author publications You can also search for this author inPubMed Google Scholar * Kristina

Corthals View author publications You can also search for this author inPubMed Google Scholar * Torben Ruhwedel View author publications You can also search for this author inPubMed Google

Scholar * Karl-Friedrich Schmidt View author publications You can also search for this author inPubMed Google Scholar * Siegrid Löwel View author publications You can also search for this

author inPubMed Google Scholar * Payam Dibaj View author publications You can also search for this author inPubMed Google Scholar * Benoit Barrette View author publications You can also

search for this author inPubMed Google Scholar * Wiebke Möbius View author publications You can also search for this author inPubMed Google Scholar * Klaus-Armin Nave View author

publications You can also search for this author inPubMed Google Scholar CONTRIBUTIONS G.S. supervised the project, conducted experiments and wrote the manuscript; F.R., K.C. and T.R.

conducted experiments, K.-F.S. and S.L. performed visual acuity measurements, P.D. conducted the two-photon microscopic analysis, B.B. implemented the beam test, W.M. supervised and

contributed to electron microscopic studies, and K.-A.N. contributed to the discussion and the writing of the manuscript. CORRESPONDING AUTHORS Correspondence to Gesine Saher or Klaus-Armin

Nave. ETHICS DECLARATIONS COMPETING INTERESTS The authors declare no competing financial interests. SUPPLEMENTARY INFORMATION SUPPLEMENTARY TEXT AND FIGURES Supplementary Figures 1–8 and

Supplementary Table 1 (PDF 1330 kb) SUPPLEMENTARY VIDEO 1 Motor performance of treated and untreated PMD mice. This video shows an example of a beam test that was used to evaluate motor

abilities of PMD mice. (MPG 3093 kb) SUPPLEMENTARY VIDEO 2 Motor disabilities of cSQS KO and cSQS KO*PLP+/66 mice. (WMV 3826 kb) RIGHTS AND PERMISSIONS Reprints and permissions ABOUT THIS

ARTICLE CITE THIS ARTICLE Saher, G., Rudolphi, F., Corthals, K. _et al._ Therapy of Pelizaeus-Merzbacher disease in mice by feeding a cholesterol-enriched diet. _Nat Med_ 18, 1130–1135

(2012). https://doi.org/10.1038/nm.2833 Download citation * Received: 22 November 2011 * Accepted: 15 May 2012 * Published: 17 June 2012 * Issue Date: July 2012 * DOI:

https://doi.org/10.1038/nm.2833 SHARE THIS ARTICLE Anyone you share the following link with will be able to read this content: Get shareable link Sorry, a shareable link is not currently

available for this article. Copy to clipboard Provided by the Springer Nature SharedIt content-sharing initiative