TY - JOUR
T1 - Synapse alterations precede neuronal damage and storage pathology in a human cerebral organoid model of CLN3-juvenile neuronal ceroid lipofuscinosis
AU - Gomez-Giro, Gemma
AU - Arias-Fuenzalida, Jonathan
AU - Jarazo, Javier
AU - Zeuschner, Dagmar
AU - Ali, Muhammad
AU - Possemis, Nina
AU - Bolognin, Silvia
AU - Halder, Rashi
AU - Jager, Christian
AU - Kuper, Willemijn F. E.
AU - van Hasselt, Peter M.
AU - Zaehres, Holm
AU - del Sol, Antonio
AU - van der Putten, Herman
AU - Scholer, Hans R.
AU - Schwamborn, Jens C.
N1 - Funding Information:
J. C. S.’s lab is supported by Fonds National de la Recherche (FNR) Luxembourg (Proof of Concept, FNR/PoC16/11559169). We would like to thank the LCSB bio-imaging platform for flow cytometry support and the stem cells, gene editing and organoid platform for support. Zdenka Hodak of the LCSB Metabolomics Platform for providing technical assistance. NCL-Stiftung and Batten Research Fonds (BRF) supported G.GG. J.J. was supported by FNR (AFR, Aides à la Formation-Recherche).
Publisher Copyright:
© 2019 The Author(s).
PY - 2019/12/30
Y1 - 2019/12/30
N2 - The juvenile form of neuronal ceroid Lipofuscinosis (JNCL) is the most common form within this group of rare lysosomal storage disorders, causing pediatric neurodegeneration. The genetic disorder, which is caused by recessive mutations affecting the CLN3 gene, features progressive vision loss, cognitive and motor decline and other psychiatric conditions, seizure episodes, leading to premature death. Animal models have traditionally aid the understanding of the disease mechanisms and pathology and are very relevant for biomarker research and therapeutic testing. Nevertheless, there is a need for establishing reliable and predictive human cellular models to study the disease. Since patient material, particularly from children, is scarce and difficult to obtain, we generated an engineered a CLN3-mutant isogenic human induced pluripotent stem cell (hiPSC) line carrying the c.1054C -> T pathologic variant, using state of the art CRISPR/Cas9 technology. To prove the suitability of the isogenic pair to model JNCL, we screened for disease-specific phenotypes in non-neuronal two-dimensional cell culture models as well as in cerebral brain organoids. Our data demonstrates that the sole introduction of the pathogenic variant gives rise to classical hallmarks of JNCL in vitro. Additionally, we discovered an alteration of the splicing caused by this particular mutation. Next, we derived cerebral organoids and used them as a neurodevelopmental model to study the particular effects of the CLN3(Q352x) mutation during brain formation in the disease context. About half of the mutation -carrying cerebral organoids completely failed to develop normally. The other half, which escaped this severe defect were used for the analysis of more subtle alterations. In these escapers, whole-transcriptome analysis demonstrated early disease signatures, affecting pathways related to development, corticogenesis and synapses. Complementary metabolomics analysis confirmed decreased levels of cerebral tissue metabolites, some particularly relevant for synapse formation and neurotransmission, such as gamma-amino butyric acid (GABA). Our data suggests that a mutation in CLN3 severely affects brain development. Furthermore, before disease onset, disease associated neurodevelopmental changes, particular concerning synapse formation and function, occur.
AB - The juvenile form of neuronal ceroid Lipofuscinosis (JNCL) is the most common form within this group of rare lysosomal storage disorders, causing pediatric neurodegeneration. The genetic disorder, which is caused by recessive mutations affecting the CLN3 gene, features progressive vision loss, cognitive and motor decline and other psychiatric conditions, seizure episodes, leading to premature death. Animal models have traditionally aid the understanding of the disease mechanisms and pathology and are very relevant for biomarker research and therapeutic testing. Nevertheless, there is a need for establishing reliable and predictive human cellular models to study the disease. Since patient material, particularly from children, is scarce and difficult to obtain, we generated an engineered a CLN3-mutant isogenic human induced pluripotent stem cell (hiPSC) line carrying the c.1054C -> T pathologic variant, using state of the art CRISPR/Cas9 technology. To prove the suitability of the isogenic pair to model JNCL, we screened for disease-specific phenotypes in non-neuronal two-dimensional cell culture models as well as in cerebral brain organoids. Our data demonstrates that the sole introduction of the pathogenic variant gives rise to classical hallmarks of JNCL in vitro. Additionally, we discovered an alteration of the splicing caused by this particular mutation. Next, we derived cerebral organoids and used them as a neurodevelopmental model to study the particular effects of the CLN3(Q352x) mutation during brain formation in the disease context. About half of the mutation -carrying cerebral organoids completely failed to develop normally. The other half, which escaped this severe defect were used for the analysis of more subtle alterations. In these escapers, whole-transcriptome analysis demonstrated early disease signatures, affecting pathways related to development, corticogenesis and synapses. Complementary metabolomics analysis confirmed decreased levels of cerebral tissue metabolites, some particularly relevant for synapse formation and neurotransmission, such as gamma-amino butyric acid (GABA). Our data suggests that a mutation in CLN3 severely affects brain development. Furthermore, before disease onset, disease associated neurodevelopmental changes, particular concerning synapse formation and function, occur.
KW - JNCL
KW - CLN3 disease
KW - CRISPR/Cas9
KW - Cerebral organoids
KW - Neurodevelopment
KW - Synapses
KW - PLURIPOTENT STEM-CELLS
KW - MOUSE MODEL
KW - CATHEPSIN-D
KW - PARKINSONS-DISEASE
KW - LYSOSOMAL STORAGE
KW - PROTEIN
KW - CLN3
KW - MUTATIONS
KW - GENE
KW - GENERATION
U2 - 10.1186/s40478-019-0871-7
DO - 10.1186/s40478-019-0871-7
M3 - Article
C2 - 31888773
SN - 2051-5960
VL - 7
JO - Acta neuropathologica communications
JF - Acta neuropathologica communications
IS - 1
M1 - 222
ER -