TY - JOUR
T1 - Generation and characterization of a human neuronal in vitro model for Rett Syndrome using a direct reprogramming method
AU - Huber, Anna
AU - Sarne, Victoria
AU - Beribisky, Alexander V
AU - Ackerbauer, Daniela
AU - Derdak, Sophia
AU - Madritsch, Silvia
AU - Etzler, Julia
AU - Huck, Sigismund
AU - Scholze, Petra
AU - Gorgulu, Ilayda
AU - Christodoulou, John
AU - Studenik, Christian R
AU - Neuhaus, Winfried
AU - Connor, Bronwen
AU - Laccone, Franco
AU - Steinkellner, Hannes
PY - 2024/3/1
Y1 - 2024/3/1
N2 - Rett Syndrome (RTT) is a severe neurodevelopmental disorder, afflicting 1 in 10,000 female births. It is caused by mutations in the X-linked methyl-CpG-binding protein gene (MECP2), which encodes for the global transcriptional regulator methyl CpG binding protein 2 (MeCP2). As human brain samples of RTT patients are scarce and cannot be used for downstream studies, there is a pressing need for in vitro modeling of pathological neuronal changes. In this study, we use a direct reprogramming method for the generation of neuronal cells from MeCP2-deficient and wild-type human dermal fibroblasts using two episomal plasmids encoding the transcription factors SOX2 and PAX6. We demonstrated that the obtained neurons exhibit a typical neuronal morphology and express the appropriate marker proteins. RNA-sequencing confirmed neuronal identity of the obtained MeCP2-deficient and wild-type neurons. Furthermore, these MeCP2-deficient neurons reflect the pathophysiology of RTT in vitro, with diminished dendritic arborization and hyperacetylation of histone H3 and H4. Treatment with MeCP2, tethered to the cell penetrating peptide TAT, ameliorated hyperacetylation of H4K16 in MeCP2-deficient neurons, which strengthens the RTT relevance of this cell model. We generated a neuronal model based on direct reprogramming derived from patient fibroblasts, providing a powerful tool to study disease mechanisms and investigating novel treatment options for RTT.
AB - Rett Syndrome (RTT) is a severe neurodevelopmental disorder, afflicting 1 in 10,000 female births. It is caused by mutations in the X-linked methyl-CpG-binding protein gene (MECP2), which encodes for the global transcriptional regulator methyl CpG binding protein 2 (MeCP2). As human brain samples of RTT patients are scarce and cannot be used for downstream studies, there is a pressing need for in vitro modeling of pathological neuronal changes. In this study, we use a direct reprogramming method for the generation of neuronal cells from MeCP2-deficient and wild-type human dermal fibroblasts using two episomal plasmids encoding the transcription factors SOX2 and PAX6. We demonstrated that the obtained neurons exhibit a typical neuronal morphology and express the appropriate marker proteins. RNA-sequencing confirmed neuronal identity of the obtained MeCP2-deficient and wild-type neurons. Furthermore, these MeCP2-deficient neurons reflect the pathophysiology of RTT in vitro, with diminished dendritic arborization and hyperacetylation of histone H3 and H4. Treatment with MeCP2, tethered to the cell penetrating peptide TAT, ameliorated hyperacetylation of H4K16 in MeCP2-deficient neurons, which strengthens the RTT relevance of this cell model. We generated a neuronal model based on direct reprogramming derived from patient fibroblasts, providing a powerful tool to study disease mechanisms and investigating novel treatment options for RTT.
KW - direct reprogramming
KW - induced neuronal progenitor cells
KW - MeCP2
KW - neurodevelopmental disorder
KW - Rett syndrome
KW - TAT-MeCP2
UR - http://www.scopus.com/inward/record.url?scp=85186450429&partnerID=8YFLogxK
U2 - 10.1089/scd.2023.0233
DO - 10.1089/scd.2023.0233
M3 - Article
C2 - 38164119
SN - 1547-3287
VL - 33
SP - 128
EP - 142
JO - Stem cells and development
JF - Stem cells and development
IS - 5-6
ER -