Mice harbouring a SCA28 patient mutation in AFG3L2 develop late-onset ataxia associated with enhanced mitochondrial proteotoxicity

Cecilia Mancini, Eriola Hoxha, Luisa Iommarini, Alessandro Brussino, Uwe Richter, Francesca Montarolo, Claudia Cagnoli, Roberta Parolisi, Diana Iulia Gondor Morosini, Valentina Nicolò, Francesca Maltecca, Luisa Muratori, Giulia Ronchi, Stefano Geuna, Francesca Arnaboldi, Elena Donetti, Elisa Giorgio, Simona Cavalieri, Eleonora Di Gregorio, Elisa Pozzi & 11 andra Marta Ferrero, Evelise Riberi, Giorgio Casari, Fiorella Altruda, Emilia Turco, Giuseppe Gasparre, Brendan J Battersby, Anna Maria Porcelli, Enza Ferrero, Alfredo Brusco, Filippo Tempia

Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

Sammanfattning

Spinocerebellar ataxia 28 is an autosomal dominant neurodegenerative disorder caused by missense mutations affecting the proteolytic domain of AFG3L2, a major component of the mitochondrial m-AAA protease. However, little is known of the underlying pathogenetic mechanisms or how to treat patients with SCA28. Currently available Afg3l2 mutant mice harbour deletions that lead to severe, early-onset neurological phenotypes that do not faithfully reproduce the late-onset and slowly progressing SCA28 phenotype. Here we describe production and detailed analysis of a new knock-in murine model harbouring an Afg3l2 allele carrying the p.Met665Arg patient-derived mutation. Heterozygous mutant mice developed normally but adult mice showed signs of cerebellar ataxia detectable by beam test. Although cerebellar pathology was negative, electrophysiological analysis showed a trend towards increased spontaneous firing in Purkinje cells from heterozygous mutants with respect to wild-type controls. As homozygous mutants died perinatally with evidence of cardiac atrophy, for each genotype we generated mouse embryonic fibroblasts (MEFs) to investigate mitochondrial function. MEFs from mutant mice showed altered mitochondrial bioenergetics, with decreased basal oxygen consumption rate, ATP synthesis and mitochondrial membrane potential. Mitochondrial network formation and morphology was altered, with greatly reduced expression of fusogenic Opa1 isoforms. Mitochondrial alterations were also detected in cerebella of 18-month-old heterozygous mutants and may be a hallmark of disease. Pharmacological inhibition of de novo mitochondrial protein translation with chloramphenicol caused reversal of mitochondrial morphology in homozygous mutant MEFs, supporting the relevance of mitochondrial proteotoxicity for SCA28 pathogenesis and therapy development.
Originalspråkengelska
TidskriftNeurobiology of Disease
Volym124
Sidor (från-till)14-28
Antal sidor15
ISSN0969-9961
DOI
StatusPublicerad - 1 okt 2018
MoE-publikationstypA1 Tidskriftsartikel-refererad

Vetenskapsgrenar

  • 1184 Genetik, utvecklingsbiologi, fysiologi

Citera det här

Mancini, Cecilia ; Hoxha, Eriola ; Iommarini, Luisa ; Brussino, Alessandro ; Richter, Uwe ; Montarolo, Francesca ; Cagnoli, Claudia ; Parolisi, Roberta ; Gondor Morosini, Diana Iulia ; Nicolò, Valentina ; Maltecca, Francesca ; Muratori, Luisa ; Ronchi, Giulia ; Geuna, Stefano ; Arnaboldi, Francesca ; Donetti, Elena ; Giorgio, Elisa ; Cavalieri, Simona ; Di Gregorio, Eleonora ; Pozzi, Elisa ; Ferrero, Marta ; Riberi, Evelise ; Casari, Giorgio ; Altruda, Fiorella ; Turco, Emilia ; Gasparre, Giuseppe ; Battersby, Brendan J ; Porcelli, Anna Maria ; Ferrero, Enza ; Brusco, Alfredo ; Tempia, Filippo. / Mice harbouring a SCA28 patient mutation in AFG3L2 develop late-onset ataxia associated with enhanced mitochondrial proteotoxicity. I: Neurobiology of Disease. 2018 ; Vol. 124. s. 14-28.
@article{81d29801aa5f48a1ba82e584b22398b2,
title = "Mice harbouring a SCA28 patient mutation in AFG3L2 develop late-onset ataxia associated with enhanced mitochondrial proteotoxicity",
abstract = "Spinocerebellar ataxia 28 is an autosomal dominant neurodegenerative disorder caused by missense mutations affecting the proteolytic domain of AFG3L2, a major component of the mitochondrial m-AAA protease. However, little is known of the underlying pathogenetic mechanisms or how to treat patients with SCA28. Currently available Afg3l2 mutant mice harbour deletions that lead to severe, early-onset neurological phenotypes that do not faithfully reproduce the late-onset and slowly progressing SCA28 phenotype. Here we describe production and detailed analysis of a new knock-in murine model harbouring an Afg3l2 allele carrying the p.Met665Arg patient-derived mutation. Heterozygous mutant mice developed normally but adult mice showed signs of cerebellar ataxia detectable by beam test. Although cerebellar pathology was negative, electrophysiological analysis showed a trend towards increased spontaneous firing in Purkinje cells from heterozygous mutants with respect to wild-type controls. As homozygous mutants died perinatally with evidence of cardiac atrophy, for each genotype we generated mouse embryonic fibroblasts (MEFs) to investigate mitochondrial function. MEFs from mutant mice showed altered mitochondrial bioenergetics, with decreased basal oxygen consumption rate, ATP synthesis and mitochondrial membrane potential. Mitochondrial network formation and morphology was altered, with greatly reduced expression of fusogenic Opa1 isoforms. Mitochondrial alterations were also detected in cerebella of 18-month-old heterozygous mutants and may be a hallmark of disease. Pharmacological inhibition of de novo mitochondrial protein translation with chloramphenicol caused reversal of mitochondrial morphology in homozygous mutant MEFs, supporting the relevance of mitochondrial proteotoxicity for SCA28 pathogenesis and therapy development.",
keywords = "AFG3L2, Mitochondrial dynamics, Mouse knock-in, Proteotoxicity, SCA28, 1184 Genetics, developmental biology, physiology",
author = "Cecilia Mancini and Eriola Hoxha and Luisa Iommarini and Alessandro Brussino and Uwe Richter and Francesca Montarolo and Claudia Cagnoli and Roberta Parolisi and {Gondor Morosini}, {Diana Iulia} and Valentina Nicol{\`o} and Francesca Maltecca and Luisa Muratori and Giulia Ronchi and Stefano Geuna and Francesca Arnaboldi and Elena Donetti and Elisa Giorgio and Simona Cavalieri and {Di Gregorio}, Eleonora and Elisa Pozzi and Marta Ferrero and Evelise Riberi and Giorgio Casari and Fiorella Altruda and Emilia Turco and Giuseppe Gasparre and Battersby, {Brendan J} and Porcelli, {Anna Maria} and Enza Ferrero and Alfredo Brusco and Filippo Tempia",
year = "2018",
month = "10",
day = "1",
doi = "10.1016/j.nbd.2018.10.018",
language = "English",
volume = "124",
pages = "14--28",
journal = "Neurobiology of Disease",
issn = "0969-9961",
publisher = "ACADEMIC PRESS INC ELSEVIER SCIENCE",

}

Mancini, C, Hoxha, E, Iommarini, L, Brussino, A, Richter, U, Montarolo, F, Cagnoli, C, Parolisi, R, Gondor Morosini, DI, Nicolò, V, Maltecca, F, Muratori, L, Ronchi, G, Geuna, S, Arnaboldi, F, Donetti, E, Giorgio, E, Cavalieri, S, Di Gregorio, E, Pozzi, E, Ferrero, M, Riberi, E, Casari, G, Altruda, F, Turco, E, Gasparre, G, Battersby, BJ, Porcelli, AM, Ferrero, E, Brusco, A & Tempia, F 2018, 'Mice harbouring a SCA28 patient mutation in AFG3L2 develop late-onset ataxia associated with enhanced mitochondrial proteotoxicity', Neurobiology of Disease, vol. 124, s. 14-28. https://doi.org/10.1016/j.nbd.2018.10.018

Mice harbouring a SCA28 patient mutation in AFG3L2 develop late-onset ataxia associated with enhanced mitochondrial proteotoxicity. / Mancini, Cecilia; Hoxha, Eriola; Iommarini, Luisa; Brussino, Alessandro; Richter, Uwe; Montarolo, Francesca; Cagnoli, Claudia; Parolisi, Roberta; Gondor Morosini, Diana Iulia; Nicolò, Valentina; Maltecca, Francesca; Muratori, Luisa; Ronchi, Giulia; Geuna, Stefano; Arnaboldi, Francesca; Donetti, Elena; Giorgio, Elisa; Cavalieri, Simona; Di Gregorio, Eleonora; Pozzi, Elisa; Ferrero, Marta; Riberi, Evelise; Casari, Giorgio; Altruda, Fiorella; Turco, Emilia; Gasparre, Giuseppe; Battersby, Brendan J; Porcelli, Anna Maria; Ferrero, Enza; Brusco, Alfredo; Tempia, Filippo.

I: Neurobiology of Disease, Vol. 124, 01.10.2018, s. 14-28.

Forskningsoutput: TidskriftsbidragArtikelVetenskapligPeer review

TY - JOUR

T1 - Mice harbouring a SCA28 patient mutation in AFG3L2 develop late-onset ataxia associated with enhanced mitochondrial proteotoxicity

AU - Mancini, Cecilia

AU - Hoxha, Eriola

AU - Iommarini, Luisa

AU - Brussino, Alessandro

AU - Richter, Uwe

AU - Montarolo, Francesca

AU - Cagnoli, Claudia

AU - Parolisi, Roberta

AU - Gondor Morosini, Diana Iulia

AU - Nicolò, Valentina

AU - Maltecca, Francesca

AU - Muratori, Luisa

AU - Ronchi, Giulia

AU - Geuna, Stefano

AU - Arnaboldi, Francesca

AU - Donetti, Elena

AU - Giorgio, Elisa

AU - Cavalieri, Simona

AU - Di Gregorio, Eleonora

AU - Pozzi, Elisa

AU - Ferrero, Marta

AU - Riberi, Evelise

AU - Casari, Giorgio

AU - Altruda, Fiorella

AU - Turco, Emilia

AU - Gasparre, Giuseppe

AU - Battersby, Brendan J

AU - Porcelli, Anna Maria

AU - Ferrero, Enza

AU - Brusco, Alfredo

AU - Tempia, Filippo

PY - 2018/10/1

Y1 - 2018/10/1

N2 - Spinocerebellar ataxia 28 is an autosomal dominant neurodegenerative disorder caused by missense mutations affecting the proteolytic domain of AFG3L2, a major component of the mitochondrial m-AAA protease. However, little is known of the underlying pathogenetic mechanisms or how to treat patients with SCA28. Currently available Afg3l2 mutant mice harbour deletions that lead to severe, early-onset neurological phenotypes that do not faithfully reproduce the late-onset and slowly progressing SCA28 phenotype. Here we describe production and detailed analysis of a new knock-in murine model harbouring an Afg3l2 allele carrying the p.Met665Arg patient-derived mutation. Heterozygous mutant mice developed normally but adult mice showed signs of cerebellar ataxia detectable by beam test. Although cerebellar pathology was negative, electrophysiological analysis showed a trend towards increased spontaneous firing in Purkinje cells from heterozygous mutants with respect to wild-type controls. As homozygous mutants died perinatally with evidence of cardiac atrophy, for each genotype we generated mouse embryonic fibroblasts (MEFs) to investigate mitochondrial function. MEFs from mutant mice showed altered mitochondrial bioenergetics, with decreased basal oxygen consumption rate, ATP synthesis and mitochondrial membrane potential. Mitochondrial network formation and morphology was altered, with greatly reduced expression of fusogenic Opa1 isoforms. Mitochondrial alterations were also detected in cerebella of 18-month-old heterozygous mutants and may be a hallmark of disease. Pharmacological inhibition of de novo mitochondrial protein translation with chloramphenicol caused reversal of mitochondrial morphology in homozygous mutant MEFs, supporting the relevance of mitochondrial proteotoxicity for SCA28 pathogenesis and therapy development.

AB - Spinocerebellar ataxia 28 is an autosomal dominant neurodegenerative disorder caused by missense mutations affecting the proteolytic domain of AFG3L2, a major component of the mitochondrial m-AAA protease. However, little is known of the underlying pathogenetic mechanisms or how to treat patients with SCA28. Currently available Afg3l2 mutant mice harbour deletions that lead to severe, early-onset neurological phenotypes that do not faithfully reproduce the late-onset and slowly progressing SCA28 phenotype. Here we describe production and detailed analysis of a new knock-in murine model harbouring an Afg3l2 allele carrying the p.Met665Arg patient-derived mutation. Heterozygous mutant mice developed normally but adult mice showed signs of cerebellar ataxia detectable by beam test. Although cerebellar pathology was negative, electrophysiological analysis showed a trend towards increased spontaneous firing in Purkinje cells from heterozygous mutants with respect to wild-type controls. As homozygous mutants died perinatally with evidence of cardiac atrophy, for each genotype we generated mouse embryonic fibroblasts (MEFs) to investigate mitochondrial function. MEFs from mutant mice showed altered mitochondrial bioenergetics, with decreased basal oxygen consumption rate, ATP synthesis and mitochondrial membrane potential. Mitochondrial network formation and morphology was altered, with greatly reduced expression of fusogenic Opa1 isoforms. Mitochondrial alterations were also detected in cerebella of 18-month-old heterozygous mutants and may be a hallmark of disease. Pharmacological inhibition of de novo mitochondrial protein translation with chloramphenicol caused reversal of mitochondrial morphology in homozygous mutant MEFs, supporting the relevance of mitochondrial proteotoxicity for SCA28 pathogenesis and therapy development.

KW - AFG3L2

KW - Mitochondrial dynamics

KW - Mouse knock-in

KW - Proteotoxicity

KW - SCA28

KW - 1184 Genetics, developmental biology, physiology

U2 - 10.1016/j.nbd.2018.10.018

DO - 10.1016/j.nbd.2018.10.018

M3 - Article

VL - 124

SP - 14

EP - 28

JO - Neurobiology of Disease

JF - Neurobiology of Disease

SN - 0969-9961

ER -