Phenotypic characterization of Cdkl5-knockdown neurons establishes elongated cilia as a functional assay for CDKL5 Deficiency Disorder

Alessia Di Nardo; Alina Rühmkorf; Patricia Award; Ashton Brennecke; Michela Fagiolini; Mustafa Sahin

doi:10.1016/j.neures.2021.10.001

Phenotypic characterization of Cdkl5-knockdown neurons establishes elongated cilia as a functional assay for CDKL5 Deficiency Disorder

Neurosci Res. 2022 Mar:176:73-78. doi: 10.1016/j.neures.2021.10.001. Epub 2021 Oct 5.

Authors

Alessia Di Nardo¹, Alina Rühmkorf², Patricia Award³, Ashton Brennecke⁴, Michela Fagiolini⁵, Mustafa Sahin⁶

Affiliations

¹ F.M. Kirby Neurobiology Center, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Rosamund Stone Zander Translational Neuroscience Center, Boston Children's Hospital, Boston, MA 02115, USA; Genetic and Developmental Disorders Research Unit, Biogen 115 Broadway, Cambridge, MA 02142, USA(1).
² F.M. Kirby Neurobiology Center, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Rosamund Stone Zander Translational Neuroscience Center, Boston Children's Hospital, Boston, MA 02115, USA.
³ F.M. Kirby Neurobiology Center, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA.
⁴ Genetic and Developmental Disorders Research Unit, Biogen 115 Broadway, Cambridge, MA 02142, USA(1).
⁵ F.M. Kirby Neurobiology Center, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Hock E. Tan and K. Lisa Yang Center for Autism Research at Harvard University, Boston, MA 02115, USA.
⁶ F.M. Kirby Neurobiology Center, Department of Neurology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Rosamund Stone Zander Translational Neuroscience Center, Boston Children's Hospital, Boston, MA 02115, USA. Electronic address: Mustafa.sahin@childrens.harvard.edu.

Abstract

CDKL5 Deficiency Disorder (CDD) is a severe encephalopathy characterized by intractable epilepsy, infantile spasms, and cognitive disabilities. The detrimental CNS manifestations and lack of therapeutic interventions represent unmet needs, necessitating identification of CDD-dependent phenotypes for in vitro disease modeling and therapeutic testing. Here, we optimized a high-content assay to quantify cilia in CDKL5-deficient neurons. Our work shows that Cdkl5-knockdown neurons have elongated cilia and uncovers cilium lengthening in hippocampi of Cdkl5 knockout mice. Collectively, our findings identify cilia length alterations under CDKL5 activity loss in vitro and in vivo and reveal elongated cilia as a robust functional phenotype for CDD.

Keywords: Autism; CDKL5; Cilia; Neurodevelopmental disorder; Seizure.

MeSH terms

Animals
Cilia
Epileptic Syndromes* / genetics
Mice
Neurons
Phenotype
Protein Serine-Threonine Kinases* / genetics

Phenotypic characterization of Cdkl5-knockdown neurons establishes elongated cilia as a functional assay for CDKL5 Deficiency Disorder

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