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Aggregation of mutant cysteine string protein-α via Fe–S cluster binding is mitigated by iron chelators

Nature Structural & Molecular Biology volume 27, pages 192–201 (2020)Cite this article

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Abstract

Point mutations in cysteine string protein-α (CSPα) cause dominantly inherited adult-onset neuronal ceroid lipofuscinosis (ANCL), a rapidly progressing and lethal neurodegenerative disease with no treatment. ANCL mutations are proposed to trigger CSPα aggregation/oligomerization, but the mechanism of oligomer formation remains unclear. Here we use purified proteins, mouse primary neurons and patient-derived induced neurons to show that the normally palmitoylated cysteine string region of CSPα loses palmitoylation in ANCL mutants. This allows oligomerization of mutant CSPα via ectopic binding of iron–sulfur (Fe–S) clusters. The resulting oligomerization of mutant CSPα causes its mislocalization and consequent loss of its synaptic SNARE-chaperoning function. We then find that pharmacological iron chelation mitigates the oligomerization of mutant CSPα, accompanied by partial rescue of the downstream SNARE defects and the pathological hallmark of lipofuscin accumulation. Thus, the iron chelators deferiprone (L1) and deferoxamine (Dfx), which are already used to treat iron overload in humans, offer a new approach for treating ANCL.

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Fig. 1: Fe–S cluster binding and oligomerization of CSPα in vitro and in mammalian cells.
Fig. 2: Mechanism of Fe–S cluster binding to the ANCL mutants of CSPα in neurons.
Fig. 3: Mislocalization of ANCL mutants of CSPα leads to deficiencies in SNAP-25 levels and SNARE complex assembly in neurons.
Fig. 4: Iron chelation in neurons alleviates oligomerization of mutant CSPα and the downstream SNARE defects.
Fig. 5: In ANCL patient-derived iNs iron chelators alleviate CSPα oligomerization, the SNAP-25 chaperoning defect and lipofuscin accumulation.
Fig. 6: Model depicting the mechanism of CSPα oligomerization in ANCL and its alleviation by Fe chelators.

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Data availability

The data that support the findings of this study are available from the corresponding author upon reasonable request. Source data for Figs. 1–5 are provided with this paper.

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Acknowledgements

We thank T. C. Südhof for kindly sharing the CSPα knockout mouse line and antibodies against CSPα and neuronal SNARE proteins, and G. Petsko for his advice on experimental approaches and the manuscript. Cornell High Energy Synchrotron Source (CHESS) experiments were supported by the National Science Foundation under award DMR-1332208. This work was supported by grants from Alzheimer’s Association (NIRG-15-363678 to MS), American Federation for Aging Research (New Investigator in Alzheimer’s Research Grant, to M.S.), NIH National Institute for Aging (1R01AG052505, to M.S.) and National Institute for Neurological Disorders and Stroke (1R01NS095988, to M.S.) and (1R01NS102181, to J.B.), as well as F31 studentship (NS098623, to N.N.N.).

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Authors and Affiliations

  1. Appel Institute for Alzheimer’s Disease Research, and Brain & Mind Research Institute, Weill Cornell Medicine, New York, NY, USA

    Nima N. Naseri, Burçe Ergel, Parinati Kharel, Yoonmi Na, Jacqueline Burré & Manu Sharma

  2. Cornell High Energy Synchrotron Source (CHESS), Cornell University, Ithaca, NY, USA

    Qingqiu Huang & Rong Huang

  3. New York State Institute for Basic Research in Developmental Disabilities, Staten Island, NY, USA

    Natalia Dolzhanskaya & Milen T. Velinov

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  1. Nima N. Naseri
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Contributions

N.N.N., B.E., P.K., Y.N., J.B. and M.S. designed, performed and analyzed all experiments except the X-ray fluorescence experiment, which was performed and analyzed by R.H. and Q.H. N.D. and M.T.V. contributed ANCL patient fibroblasts to the study. N.N.N., J.B. and M.S. wrote the manuscript. M.S. conceived the project and directed the research.

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Correspondence to Manu Sharma.

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Naseri, N.N., Ergel, B., Kharel, P. et al. Aggregation of mutant cysteine string protein-α via Fe–S cluster binding is mitigated by iron chelators. Nat Struct Mol Biol 27, 192–201 (2020). https://doi.org/10.1038/s41594-020-0375-y

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