Neuronal lipofuscinosis caused by Kufs disease/CLN4 DNAJC5 mutations but not by a CSPα/DNAJC5 deficiency

Santiago López-Begines; Nozha Borjini; Ángela Lavado-Roldán; Cristina Mesa-Cruz; Fabiola Mavillard; Vera I Wiersma; Fátima Rubio-Pastor; Emanuela Tumini; Carmen Paradela-Leal; María L Chiclana-Valcárcel; Carolina Aguado; Rafael Luján; Wiep Scheper; José L Nieto-González; Rafael Fernández-Chacón

doi:10.1126/sciadv.ads3393

Neuronal lipofuscinosis caused by Kufs disease/CLN4 DNAJC5 mutations but not by a CSPα/DNAJC5 deficiency

Santiago López-Begines
, Nozha Borjini
, Ángela Lavado-Roldán
, Cristina Mesa-Cruz
, Fabiola Mavillard
, Vera I Wiersma
, Fátima Rubio-Pastor
, Emanuela Tumini
, Carmen Paradela-Leal
, María L Chiclana-Valcárcel
, Carolina Aguado
, Rafael Luján
, Wiep Scheper
, José L Nieto-González
, Rafael Fernández-Chacón

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

Kufs disease/CLN4 is an autosomal dominant neurodegenerative disorder caused by unknown mechanisms through Leu115Arg and Leu116Δ mutations in the DNAJC5 gene that encodes the synaptic vesicle co-chaperone cysteine string protein α (CSPα/DNAJC5). To investigate the disease mechanisms in vivo, we generated three independent mouse lines overexpressing different versions of CSPα/DNAJC5 under the neuron-specific Thy1 promoter: wild-type (WT), Leu115Arg, and Leu116Δ. Mice expressing mutant Leu115Arg CSPα/DNAJC5 are viable but develop motor deficits. As described in patients with Kufs disease, we observed the pathological lipofuscinosis and intracellular structures resembling granular osmiophilic deposits (GRODs) in the mutant but not in the WT transgenic lines. Microglia engulf lipofuscin and lipofuscin-containing neurons. Notably, conventional or conditional knockout mice lacking CSPα/DNAJC5 did not exhibit any signs of increased lipofuscinosis or GRODs. Our novel mouse models provide a valuable tool to investigate the molecular mechanisms underlying Kufs disease/CLN4. DNAJC5 mutations cause neuronal lipofuscinosis through a cell-autonomous gain of a pathological function of CSPα/DNAJC5.

Original language	English
Article number	eads3393
Pages (from-to)	eads3393
Journal	Science advances
Volume	11
Issue number	21
DOIs	https://doi.org/10.1126/sciadv.ads3393
Publication status	Published - 23 May 2025

Funding

This work was supported by Spanish Agencia Estatal de Investigación and Ministerio de Ciencia, Innovación y Universidades and European Regional Development Fund (ERDF), (MICIU/AEI/10.13039/501100011033, FEDER, UE): BFU2016-76050-P (R.F.-C.), PID2019-105530GB-I00 (R.F.-C.), PID2022-138957NBI00 (R.F.-C.), PID2021-125875OB-I00 (R.L.), BES-2011-046029 (Á.L.-R.), BES-2017-082324 (C.M.-C.), PRE2020/095166 (C.P.-L.), and FPU18/01700 (F.R.-P.); Andalusian Consejería de Universidad, Investigación e Innovación, CUII, P12-CTS-2232, P18FR-2144, CTS-600 (R.F.-C.); Andalusian Consejería de Salud y Consumo, RH-0064-2021 (N.B.); INVESTIGO Program funded by the European Union through the Recovery, Transformation, and Resilience Plan for Andalusia (M.L.C.-V.); Institute of Health Carlos III (ISCIII) and MICIU: FORTALECE (FORT23/00008) and CIBERNED (R.F.-C.); Junta de Comunidades de Castilla-La Mancha, SBPLY/21/180501/000064 (to R.L.); ZonmMW Memorabel/Alzheimer Nederland, no. 733050101 (W.S.). center (dKFZ) and the institute of Genetics and Molecular and cellular Biology [Gie-ceRBM (iGBMc)] for providing caMKii-cre-eRt2 transgenic mice, to J. vitorica for providing antibodies, and to A. Pascual and M. e. Saez for technical advice and support related to genomic sequences. thanks to A. Arroyo and M. c. Rivero for previous technical assistance with genotyping. We are indebted to O. Pintado for his legacy and pioneering work in the generation of genetically modified mice, including mice reported in this study, during more than two decades at the University of Seville. Funding: this work was supported by Spanish Agencia estatal de investigación and Ministerio de ciencia, innovación y Universidades and european Regional development Fund (eRdF), (MiciU/Aei/10.13039/501100011033, FedeR, Ue): BFU2016-76050-P (R.F.-c.), Pid2019-105530GB-i00 (R.F.-c.), Pid2022-138957nBi00 (R.F.-c.), Pid2021-125875OB-i00 (R.L.), BeS-2011-046029 (Á.L.-R.), BeS-2017-082324 (c.M.-c.), PRe2020/095166 (c.P.-L.), and FPU18/01700 (F.R.-P.); Andalusian consejería de Universidad, investigación e innovación, cUii, P12-ctS-2232, P18FR-2144, ctS-600 (R.F.-c.); Andalusian consejería de Salud y consumo, Rh-0064-2021 (n.B.); inveStiGO Program funded by the european Union through the Recovery, transformation, and Resilience Plan for Andalusia (M.L.c.-v.); institute of health carlos iii (iSciii) and MiciU: FORtALece (FORt23/00008) and ciBeRned (R.F.-c.); Junta de comunidades de castilla-La Mancha, SBPLY/21/180501/000064 (to R.L.); ZonmMW Memorabel/Alzheimer nederland, no. 733050101 (W.S.). Author contributions: S.L.-B. generated and characterized mouse lines, designed and performed experiments, and analyzed data; n.B. designed experiments and analyzed data pertaining to microglia and 3d imaging reconstructions and behavior; Á.L.-R. and F.M. designed and carried out plasmid constructions for transgenesis; Á.L.-R. carried out generation and characterization of transgenic mouse lines; c.M.-c. generated a conditional KO mouse line and performed experiments and analyzed data; v.i.W. and W.S. performed experiments and analyzed data pertaining to GvB; S.L.-B., c.A., and R.L. performed experiments and analyzed data pertaining electron microscopy data; S.L.-B., e.t., M.L.c.-v. performed experiments and analysis related to genome sequencing; e.t. performed cathepsin d–related experiments; c.P.-L. and F.R.-P. performed experiments and analysis related to immunolabeling of brain sections; J.L.n.-G. set up protocols for electron microscopy and co-supervised c.M.-c.’s experiments; R.F.-c. conceived the study, designed experiments, analyzed data, and wrote the paper with input from all authors. Competing interests: the authors declare that they have no competing interests. Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper, in the Supplementary Materials, and at https://datadryad.org/ stash (dOi: 10.5061/dryad.dfn2z35c4). Genomic sequences are available at BioProject accession number PRJnA1174377. custom macros for image analysis based on Fiji (imageJ) are available at https://github.com/SLopezBegines/imageJ-Macros, and scripts for behavioral analysis are available at https://github.com/SLopezBegines/R_code and at https://datadryad. org/stash (dOi: 10.5061/dryad.dfn2z35c4). the transgenic mouse models (thy1-GFP-cSPα-Wt, thy1-GFP-cSPα-L115R, and thy1-GFP-cSPα-L116Δ) can be provided by R.F.-c.’s pending scientific review and a completed material transfer agreement. Requests for transgenic mice should be submitted to R.F.-c. ([email protected]).

Funders	Funder number
Instituto de Salud Carlos III
Resilience Plan for Andalusia
Andalusian Consejería de Universidad, Investigación e Innovación
Andalusian consejería de Salud y consumo
European Commission
Centro de Investigación Biomédica en Red sobre Enfermedades Neurodegenerativas
Agencia Estatal de Investigación
ZonmMW Memorabel
Andalusian Consejería de Salud y Consumo	RH-0064-2021
European Regional Development Fund	MICIU/AEI/10.13039/501100011033
Ministerio de Ciencia, Innovación y Universidades	Pid2022-138957nBi00, Pid2019-105530GB-i00, MiciU/Aei/10.13039/501100011033, FORT23/00008, BFU2016-76050-P
Junta de Comunidades de Castilla-La Mancha	SBPLY/21/180501/000064
CUII	P12-CTS-2232, CTS-600, P18FR-2144
Alzheimer Nederland	733050101
Federación Española de Enfermedades Raras	BES-2011-046029, PID2022-138957NBI00, BES-2017-082324, FPU18/01700, PID2021-125875OB-I00, PRE2020/095166, PID2019-105530GB-I00

Keywords

Animals
HSP40 Heat-Shock Proteins/genetics
Mice
Membrane Proteins/genetics
Mutation
Disease Models, Animal
Neuronal Ceroid-Lipofuscinoses/genetics
Mice, Knockout
Neurons/metabolism
Humans
Mice, Transgenic

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10.1126/sciadv.ads3393

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López-Begines, S., Borjini, N., Lavado-Roldán, Á., Mesa-Cruz, C., Mavillard, F., Wiersma, V. I., Rubio-Pastor, F., Tumini, E., Paradela-Leal, C., Chiclana-Valcárcel, M. L., Aguado, C., Luján, R., Scheper, W., Nieto-González, J. L., & Fernández-Chacón, R. (2025). Neuronal lipofuscinosis caused by Kufs disease/CLN4 DNAJC5 mutations but not by a CSPα/DNAJC5 deficiency. Science advances, 11(21), eads3393. Article eads3393. https://doi.org/10.1126/sciadv.ads3393

@article{48f4d30ea4cb4a5caac8eb858d3626c3,

title = "Neuronal lipofuscinosis caused by Kufs disease/CLN4 DNAJC5 mutations but not by a CSPα/DNAJC5 deficiency",

abstract = "Kufs disease/CLN4 is an autosomal dominant neurodegenerative disorder caused by unknown mechanisms through Leu115Arg and Leu116Δ mutations in the DNAJC5 gene that encodes the synaptic vesicle co-chaperone cysteine string protein α (CSPα/DNAJC5). To investigate the disease mechanisms in vivo, we generated three independent mouse lines overexpressing different versions of CSPα/DNAJC5 under the neuron-specific Thy1 promoter: wild-type (WT), Leu115Arg, and Leu116Δ. Mice expressing mutant Leu115Arg CSPα/DNAJC5 are viable but develop motor deficits. As described in patients with Kufs disease, we observed the pathological lipofuscinosis and intracellular structures resembling granular osmiophilic deposits (GRODs) in the mutant but not in the WT transgenic lines. Microglia engulf lipofuscin and lipofuscin-containing neurons. Notably, conventional or conditional knockout mice lacking CSPα/DNAJC5 did not exhibit any signs of increased lipofuscinosis or GRODs. Our novel mouse models provide a valuable tool to investigate the molecular mechanisms underlying Kufs disease/CLN4. DNAJC5 mutations cause neuronal lipofuscinosis through a cell-autonomous gain of a pathological function of CSPα/DNAJC5.",

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author = "Santiago L{\'o}pez-Begines and Nozha Borjini and {\'A}ngela Lavado-Rold{\'a}n and Cristina Mesa-Cruz and Fabiola Mavillard and Wiersma, \{Vera I\} and F{\'a}tima Rubio-Pastor and Emanuela Tumini and Carmen Paradela-Leal and Chiclana-Valc{\'a}rcel, \{Mar{\'i}a L\} and Carolina Aguado and Rafael Luj{\'a}n and Wiep Scheper and Nieto-Gonz{\'a}lez, \{Jos{\'e} L\} and Rafael Fern{\'a}ndez-Chac{\'o}n",

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month = may,

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doi = "10.1126/sciadv.ads3393",

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López-Begines, S, Borjini, N, Lavado-Roldán, Á, Mesa-Cruz, C, Mavillard, F, Wiersma, VI, Rubio-Pastor, F, Tumini, E, Paradela-Leal, C, Chiclana-Valcárcel, ML, Aguado, C, Luján, R, Scheper, W, Nieto-González, JL & Fernández-Chacón, R 2025, 'Neuronal lipofuscinosis caused by Kufs disease/CLN4 DNAJC5 mutations but not by a CSPα/DNAJC5 deficiency', Science advances, vol. 11, no. 21, eads3393, pp. eads3393. https://doi.org/10.1126/sciadv.ads3393

TY - JOUR

T1 - Neuronal lipofuscinosis caused by Kufs disease/CLN4 DNAJC5 mutations but not by a CSPα/DNAJC5 deficiency

AU - López-Begines, Santiago

AU - Borjini, Nozha

AU - Lavado-Roldán, Ángela

AU - Mesa-Cruz, Cristina

AU - Mavillard, Fabiola

AU - Wiersma, Vera I

AU - Rubio-Pastor, Fátima

AU - Tumini, Emanuela

AU - Paradela-Leal, Carmen

AU - Chiclana-Valcárcel, María L

AU - Aguado, Carolina

AU - Luján, Rafael

AU - Scheper, Wiep

AU - Nieto-González, José L

AU - Fernández-Chacón, Rafael

PY - 2025/5/23

Y1 - 2025/5/23

N2 - Kufs disease/CLN4 is an autosomal dominant neurodegenerative disorder caused by unknown mechanisms through Leu115Arg and Leu116Δ mutations in the DNAJC5 gene that encodes the synaptic vesicle co-chaperone cysteine string protein α (CSPα/DNAJC5). To investigate the disease mechanisms in vivo, we generated three independent mouse lines overexpressing different versions of CSPα/DNAJC5 under the neuron-specific Thy1 promoter: wild-type (WT), Leu115Arg, and Leu116Δ. Mice expressing mutant Leu115Arg CSPα/DNAJC5 are viable but develop motor deficits. As described in patients with Kufs disease, we observed the pathological lipofuscinosis and intracellular structures resembling granular osmiophilic deposits (GRODs) in the mutant but not in the WT transgenic lines. Microglia engulf lipofuscin and lipofuscin-containing neurons. Notably, conventional or conditional knockout mice lacking CSPα/DNAJC5 did not exhibit any signs of increased lipofuscinosis or GRODs. Our novel mouse models provide a valuable tool to investigate the molecular mechanisms underlying Kufs disease/CLN4. DNAJC5 mutations cause neuronal lipofuscinosis through a cell-autonomous gain of a pathological function of CSPα/DNAJC5.

AB - Kufs disease/CLN4 is an autosomal dominant neurodegenerative disorder caused by unknown mechanisms through Leu115Arg and Leu116Δ mutations in the DNAJC5 gene that encodes the synaptic vesicle co-chaperone cysteine string protein α (CSPα/DNAJC5). To investigate the disease mechanisms in vivo, we generated three independent mouse lines overexpressing different versions of CSPα/DNAJC5 under the neuron-specific Thy1 promoter: wild-type (WT), Leu115Arg, and Leu116Δ. Mice expressing mutant Leu115Arg CSPα/DNAJC5 are viable but develop motor deficits. As described in patients with Kufs disease, we observed the pathological lipofuscinosis and intracellular structures resembling granular osmiophilic deposits (GRODs) in the mutant but not in the WT transgenic lines. Microglia engulf lipofuscin and lipofuscin-containing neurons. Notably, conventional or conditional knockout mice lacking CSPα/DNAJC5 did not exhibit any signs of increased lipofuscinosis or GRODs. Our novel mouse models provide a valuable tool to investigate the molecular mechanisms underlying Kufs disease/CLN4. DNAJC5 mutations cause neuronal lipofuscinosis through a cell-autonomous gain of a pathological function of CSPα/DNAJC5.

KW - Animals

KW - HSP40 Heat-Shock Proteins/genetics

KW - Mice

KW - Membrane Proteins/genetics

KW - Mutation

KW - Disease Models, Animal

KW - Neuronal Ceroid-Lipofuscinoses/genetics

KW - Mice, Knockout

KW - Neurons/metabolism

KW - Humans

KW - Mice, Transgenic

U2 - 10.1126/sciadv.ads3393

DO - 10.1126/sciadv.ads3393

M3 - Article

C2 - 40397740

SN - 2375-2548

VL - 11

SP - eads3393

JO - Science advances

JF - Science advances

IS - 21

M1 - eads3393

ER -

Neuronal lipofuscinosis caused by Kufs disease/CLN4 DNAJC5 mutations but not by a CSPα/DNAJC5 deficiency

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