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Pharmacological chaperones stabilize retromer to limit APP processing

Abstract

Retromer is a multiprotein complex that trafficks cargo out of endosomes. The neuronal retromer traffics the amyloid-precursor protein (APP) away from endosomes, a site where APP is cleaved into pathogenic fragments in Alzheimer's disease. Here we determined whether pharmacological chaperones can enhance retromer stability and function. First, we relied on the crystal structures of retromer proteins to help identify the 'weak link' of the complex and to complete an in silico screen of small molecules predicted to enhance retromer stability. Among the hits, an in vitro assay identified one molecule that stabilized retromer against thermal denaturation. Second, we turned to cultured hippocampal neurons, showing that this small molecule increases the levels of retromer proteins, shifts APP away from the endosome, and decreases the pathogenic processing of APP. These findings show that pharmacological chaperones can enhance the function of a multiprotein complex and may have potential therapeutic implications for neurodegenerative diseases.

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Figure 1: Characterization of the weak link in retromer complex stability.
Figure 2: Identification of a retromer stabilizing pharmacological chaperone.
Figure 3: The pharmacological chaperone increases retromer levels in neurons.
Figure 4: The pharmacological chaperone decreases Aβ peptide accumulation and reduces the pathogenic pathway of APP.
Figure 5: The pharmacological chaperone shifts the endosomal localization of APP and SorL1.
Figure 6: The pharmacological chaperone specifically modulates Aβ production through the retromer pathway.

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Acknowledgements

We thank the US National Institute on Aging/US National Institutes of Health (NIH) grants AG025161 and AG08702, The Alzheimer's Association, Developmental Therapeutics Program of the National Cancer Institute, Medkoo Biosciences, The Fidelity Biosciences Research Initiative and give special thanks to S. Weninger for advice and encouragement. We also thank The McKnight Endowment for Neuroscience, the Ellison Medical Foundation, and the Gottlieb Family Foundation.

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Contributions

V.J.M. identified the retromer stabilizing site and carried out the in silico screens and designed and performed the in vitro retromer chaperone characterization; D.E.B. designed, coordinated and performed the in vivo retromer chaperone characterization; S.S. assisted with the immunohistochemical staining and genetic recombination in neurons; C.V. assisted with neuronal cultures; M.R.A. assisted with the in vitro thermal studies; V.M.P. assisted with the APP fragment analysis; R.T.S. helped with Aβ measurements; and G.A.P., D.R. and S.A.S. conceived of and supervised the studies and wrote the manuscript.

Corresponding authors

Correspondence to Gregory A Petsko, Dagmar Ringe or Scott A Small.

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The authors declare no competing financial interests.

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Supplementary Results, Supplementary Table 1, Supplementary Figures 1–7 and MedKoo certificate. (PDF 1821 kb)

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Mecozzi, V., Berman, D., Simoes, S. et al. Pharmacological chaperones stabilize retromer to limit APP processing. Nat Chem Biol 10, 443–449 (2014). https://doi.org/10.1038/nchembio.1508

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