A KCNC3 mutation causes a neurodevelopmental, non-progressive SCA13 subtype associated with dominant negative effects and aberrant EGFR trafficking

Swati Khare, [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Fernandez-Funez, Pedro; Waters, Michael F.] Univ Florida, Dept Neurol, Gainesville, FL 32611 USA; [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Butler, Brittany; Khoshbouei, Habibeh; Smithson, Lisa; Golde, Todd E.; Rincon-Limas, Diego E.; Lewis, Jada; Fernandez-Funez, Pedro; Nick, Harry S.; Waters, Michael F.] Univ Florida, McKnight Brain Inst, Gainesville, FL 32611 USA; [Khare, Swati; Waters, Michael F.] Univ Florida, Dept Biomed Engn, Gainesville, FL 32611 USA; [Zhang, Yalan; Kaczmarek, Leonard K.] Yale Univ, Dept Pharmacol, New Haven, CT USA; [Butler, Brittany; Khoshbouei, Habibeh; Rayaprolu, Sruti; Smithson, Lisa; Golde, Todd E.; Waters, Michael F.] Univ Florida, Dept Neurosci, Gainesville, FL 32610 USA; [Hathorn, Tyisha; Ranum, Laura P. W.] Univ Florida, Dept Mol Genet & Microbiol, Gainesville, FL USA; [Paucar, Martin] Karolinska Univ Hosp, Dept Neurol, Stockholm, Sweden; [Paucar, Martin] Karolinska Inst, Dept Clin Neurosci, Stockholm, Sweden; [Morse, Richard] Dartmouth Hitchcock Med Ctr, Dept Neurol, Lebanon, NH 03766 USA; [Raff, Michael; Simon, Julie] Multicare Hlth Syst, Genom Inst, Tacoma, WA USA; [Ld, Magnus Nordenskjo E.] Karolinska Univ Hosp, Dept Genet, Stockholm, Sweden; [Ld, Magnus Nordenskjo E.] Karolinska Inst, Dept Mol Med & Surg, Ctr Mol Med, Stockholm, Sweden; [Wirdefeldt, Karin] Karolinska Inst, Dept Med Epidemiol & Biostat, Stockholm, Sweden; [Waters, Michael F.] St Josephs Hosp, Dept Neurol, Barrow Neurol Inst, Phoenix, AZ 85013 USA
Jerelyn A. Nick, [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Fernandez-Funez, Pedro; Waters, Michael F.] Univ Florida, Dept Neurol, Gainesville, FL 32611 USA; [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Butler, Brittany; Khoshbouei, Habibeh; Smithson, Lisa; Golde, Todd E.; Rincon-Limas, Diego E.; Lewis, Jada; Fernandez-Funez, Pedro; Nick, Harry S.; Waters, Michael F.] Univ Florida, McKnight Brain Inst, Gainesville, FL 32611 USA; [Khare, Swati; Waters, Michael F.] Univ Florida, Dept Biomed Engn, Gainesville, FL 32611 USA; [Zhang, Yalan; Kaczmarek, Leonard K.] Yale Univ, Dept Pharmacol, New Haven, CT USA; [Butler, Brittany; Khoshbouei, Habibeh; Rayaprolu, Sruti; Smithson, Lisa; Golde, Todd E.; Waters, Michael F.] Univ Florida, Dept Neurosci, Gainesville, FL 32610 USA; [Hathorn, Tyisha; Ranum, Laura P. W.] Univ Florida, Dept Mol Genet & Microbiol, Gainesville, FL USA; [Paucar, Martin] Karolinska Univ Hosp, Dept Neurol, Stockholm, Sweden; [Paucar, Martin] Karolinska Inst, Dept Clin Neurosci, Stockholm, Sweden; [Morse, Richard] Dartmouth Hitchcock Med Ctr, Dept Neurol, Lebanon, NH 03766 USA; [Raff, Michael; Simon, Julie] Multicare Hlth Syst, Genom Inst, Tacoma, WA USA; [Ld, Magnus Nordenskjo E.] Karolinska Univ Hosp, Dept Genet, Stockholm, Sweden; [Ld, Magnus Nordenskjo E.] Karolinska Inst, Dept Mol Med & Surg, Ctr Mol Med, Stockholm, Sweden; [Wirdefeldt, Karin] Karolinska Inst, Dept Med Epidemiol & Biostat, Stockholm, Sweden; [Waters, Michael F.] St Josephs Hosp, Dept Neurol, Barrow Neurol Inst, Phoenix, AZ 85013 USA
Yalan Zhang, [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Fernandez-Funez, Pedro; Waters, Michael F.] Univ Florida, Dept Neurol, Gainesville, FL 32611 USA; [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Butler, Brittany; Khoshbouei, Habibeh; Smithson, Lisa; Golde, Todd E.; Rincon-Limas, Diego E.; Lewis, Jada; Fernandez-Funez, Pedro; Nick, Harry S.; Waters, Michael F.] Univ Florida, McKnight Brain Inst, Gainesville, FL 32611 USA; [Khare, Swati; Waters, Michael F.] Univ Florida, Dept Biomed Engn, Gainesville, FL 32611 USA; [Zhang, Yalan; Kaczmarek, Leonard K.] Yale Univ, Dept Pharmacol, New Haven, CT USA; [Butler, Brittany; Khoshbouei, Habibeh; Rayaprolu, Sruti; Smithson, Lisa; Golde, Todd E.; Waters, Michael F.] Univ Florida, Dept Neurosci, Gainesville, FL 32610 USA; [Hathorn, Tyisha; Ranum, Laura P. W.] Univ Florida, Dept Mol Genet & Microbiol, Gainesville, FL USA; [Paucar, Martin] Karolinska Univ Hosp, Dept Neurol, Stockholm, Sweden; [Paucar, Martin] Karolinska Inst, Dept Clin Neurosci, Stockholm, Sweden; [Morse, Richard] Dartmouth Hitchcock Med Ctr, Dept Neurol, Lebanon, NH 03766 USA; [Raff, Michael; Simon, Julie] Multicare Hlth Syst, Genom Inst, Tacoma, WA USA; [Ld, Magnus Nordenskjo E.] Karolinska Univ Hosp, Dept Genet, Stockholm, Sweden; [Ld, Magnus Nordenskjo E.] Karolinska Inst, Dept Mol Med & Surg, Ctr Mol Med, Stockholm, Sweden; [Wirdefeldt, Karin] Karolinska Inst, Dept Med Epidemiol & Biostat, Stockholm, Sweden; [Waters, Michael F.] St Josephs Hosp, Dept Neurol, Barrow Neurol Inst, Phoenix, AZ 85013 USA
Kira Galeano, [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Fernandez-Funez, Pedro; Waters, Michael F.] Univ Florida, Dept Neurol, Gainesville, FL 32611 USA; [Khare, Swati; Nick, Jerelyn A.; Galeano, Kira; Butler, Brittany; Khoshbouei, Habibeh; Smithson, Lisa; Golde, Todd E.; Rincon-Limas, Diego E.; Lewis, Jada; Fernandez-Funez, Pedro; Nick, Harry S.; Waters, Michael F.] Univ Florida, McKnight Brain Inst, Gainesville, FL 32611 USA; [Khare, Swati; Waters, Michael F.] Univ Florida, Dept Biomed Engn, Gainesville, FL 32611 USA; [Zhang, Yalan; Kaczmarek, Leonard K.] Yale Univ, Dept Pharmacol, New Haven, CT USA; [Butler, Brittany; Khoshbouei, Habibeh; Rayaprolu, Sruti; Smithson, Lisa; Golde, Todd E.; Waters, Michael F.] Univ Florida, Dept Neurosci, Gainesville, FL 32610 USA; [Hathorn, Tyisha; Ranum, Laura P. W.] Univ Florida, Dept Mol Genet & Microbiol, Gainesville, FL USA; [Paucar, Martin] Karolinska Univ Hosp, Dept Neurol, Stockholm, Sweden; [Paucar, Martin] Karolinska Inst, Dept Clin Neurosci, Stockholm, Sweden; [Morse, Richard] Dartmouth Hitchcock Med Ctr, Dept Neurol, Lebanon, NH 03766 USA; [Raff, Michael; Simon, Julie] Multicare Hlth Syst, Genom Inst, Tacoma, WA USA; [Ld, Magnus Nordenskjo E.] Karolinska Univ Hosp, Dept Genet, Stockholm, Sweden; [Ld, Magnus Nordenskjo E.] Karolinska Inst, Dept Mol Med & Surg, Ctr Mol Med, Stockholm, Sweden; [Wirdefeldt, Karin] Karolinska Inst, Dept Med Epidemiol & Biostat, Stockholm, Sweden; [Waters, Michael F.] St Josephs Hosp, Dept Neurol, Barrow Neurol Inst, Phoenix, AZ 85013 USA

Multidisciplinary Sciences; Butler, Brittany; Khoshbouei, Habibeh; Rayaprolu, Sruti; Hathorn, Tyisha; Ranum, Laura P. W.; Smithson, Lisa; Golde, Todd E.; Paucar, Martin; Morse, Richard; Raff, Michael; Simon, Julie; Ld, Magnus Nordenskjo E.; Wirdefeldt, Karin; Rincon-Limas, Diego E.; Lewis, Jada; Kaczmarek, Leonard K.; Fernandez-Funez, Pedro; Nick, Harry S.; Waters, Michael F.

Abstract

The autosomal dominant spinocerebellar ataxias (SCAs) are a diverse group of neurological disorders anchored by the phenotypes of motor incoordination and cerebellar atrophy. Disease heterogeneity is appreciated through varying comorbidities: dysarthria, dysphagia, oculomotor and/or retinal abnormalities, motor neuron pathology, epilepsy, cognitive impairment, autonomic dysfunction, and psychiatric manifestations. Our study focuses on SCA13, which is caused by several allelic variants in the voltage-gated potassium channel KCNC3 (Kv3.3). We detail the clinical phenotype of four SCA13 kindreds that confirm causation of the KCNC3(R423H) allele. The heralding features demonstrate congenital onset with non-progressive, neurodevelopmental cerebellar hypoplasia and lifetime improvement in motor and cognitive function that implicate compensatory neural mechanisms. Targeted expression of human KCNC3(R423H) in Drosophila triggers aberrant wing veins, maldeveloped eyes, and fused ommatidia consistent with the neurodevelopmental presentation of patients. Furthermore, human KCNC3(R423H) expression in mammalian cells results in altered glycosylation and aberrant retention of the channel in anterograde and/or endosomal vesicles. Confirmation of the absence of plasma membrane targeting was based on the loss of current conductance in cells expressing the mutant channel. Mechanistically, genetic studies in Drosophila, along with cellular and biophysical studies in mammalian systems, demonstrate the dominant negative effect exerted by the mutant on the wild-type (WT) protein, which explains dominant inheritance. We demonstrate that ocular co-expression of KCNC3(R423H) with Drosophila epidermal growth factor receptor (dEgfr) results in striking rescue of the eye phenotype, whereas KCNC3(R423H) expression in mammalian cells results in aberrant intracellular retention of human epidermal growth factor receptor (EGFR). Together, these results indicate that the neurodevelopmental consequences of KCNC3(R423H) may be mediated through indirect effects on EGFR signaling in the developing cerebellum. Our results therefore confirm the KCNC3(R423H) allele as causative for SCA13, through a dominant negative effect on KCNC3 WT and links with EGFR that account for dominant inheritance, congenital onset, and disease pathology.