High-content screening identifies a small molecule that restores AP-4-dependent protein trafficking in neuronal models of AP-4-associated hereditary spastic paraplegia

  • Nat Commun. 2024 Jan 17;15(1):584. doi: 10.1038/s41467-023-44264-1.
Afshin Saffari  1  2 ,  Barbara Brechmann  1 ,  Cedric Böger  1 ,  Wardiya Afshar Saber  1 ,  Hellen Jumo  1  3 ,  Dosh Whye  3 ,  Delaney Wood  3 ,  Lara Wahlster  4 ,  Julian E Alecu  1 ,  Marvin Ziegler  1 ,  Marlene Scheffold  1 ,  Kellen Winden  1 ,  Jed Hubbs  3 ,  Elizabeth D Buttermore  3 ,  Lee Barrett  3 ,  Georg H H Borner  5 ,  Alexandra K Davies  5  6 ,  Darius Ebrahimi-Fakhari  7  8 ,  Mustafa Sahin  1  3
Affiliations
  • 1. Department of Neurology & F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
  • 2. Division of Child Neurology and Inherited Metabolic Diseases, Heidelberg University Hospital, Heidelberg, Germany.
  • 3. Rosamund Stone Zander Translational Neuroscience Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, 02115, USA.
  • 4. Department of Hematology & Oncology, Boston Children's Hospital & Dana-Farber Cancer Institute, Harvard Medical School, Boston, MA, 02115, USA.
  • 5. Department of Proteomics and Signal Transduction, Max-Planck-Institute of Biochemistry, Martinsried, 82152, Germany.
  • 6. School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, University of Manchester, Manchester, M13 9PT, UK.
  • 7. Department of Neurology & F.M. Kirby Neurobiology Center, Boston Children's Hospital, Harvard Medical School, Boston, MA, 02115, USA. [email protected].
  • 8. Movement Disorders Program, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, MA, 02115, USA. [email protected].
PMID: 38233389 DOI: 10.1038/s41467-023-44264-1
Abstract

Unbiased phenotypic screens in patient-relevant disease models offer the potential to detect therapeutic targets for rare diseases. In this study, we developed a high-throughput screening assay to identify molecules that correct aberrant protein trafficking in adapter protein complex 4 (AP-4) deficiency, a rare but prototypical form of childhood-onset hereditary spastic paraplegia characterized by mislocalization of the Autophagy protein ATG9A. Using high-content microscopy and an automated image analysis pipeline, we screened a diversity library of 28,864 small molecules and identified a lead compound, BCH-HSP-C01, that restored ATG9A pathology in multiple disease models, including patient-derived fibroblasts and induced pluripotent stem cell-derived neurons. We used multiparametric orthogonal strategies and integrated transcriptomic and proteomic approaches to delineate potential mechanisms of action of BCH-HSP-C01. Our results define molecular regulators of intracellular ATG9A trafficking and characterize a lead compound for the treatment of AP-4 deficiency, providing important proof-of-concept data for future studies.

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