TY - JOUR
T1 - Rho kinase inhibition is essential during in vitro neurogenesis and promotes phenotypic rescue of human induced pluripotent stem cell-derived neurons with oligophrenin-1 loss of function
AU - Compagnucci, Claudia
AU - Barresi, Sabina
AU - Petrini, Stefania
AU - Billuart, Pierre
AU - Piccini, Giorgia
AU - Chiurazzi, Pietro
AU - Alfieri, Paolo
AU - Bertini, Enrico
AU - Zanni, Ginevra
PY - 2016
Y1 - 2016
N2 - Rho-GTPases have relevant functions in various aspects of neuronal development, such as differentiation, migration, and synaptogenesis. Loss of function of the oligophrenin-1 gene (OPHN1) causes X-linked intellectual disability with cerebellar hypoplasia and leads to hyperactivation of the rho kinase (ROCK) pathway. ROCK mainly acts through phosphorylation of the myosin phosphatise targeting subunit 1, triggering actin-myosin contractility. We show that during in vitro neurogenesis, ROCK activity decreases from day 10 until terminal differentiation, whereas in OPHN1-deficient human induced pluripotent stem cells (h-iPSCs), the levels of ROCK are elevated throughout differentiation. ROCK inhibition favors neuronal-like appearance of h-iPSCs, in parallel with transcriptional up regulation of nuclear receptor NR4A1, which is known to induce neurite outgrowth. This study analyzed the morphological, biochemical, and functional features of OPHN1-deficient h-iPSCs and their rescue by treatment with the ROCK inhibitor fasudil, shedding light on the relevance of the ROCK pathway during neuronal differentiation and providing a neuronal model for human OPHN1 syndrome and its treatment.
AB - Rho-GTPases have relevant functions in various aspects of neuronal development, such as differentiation, migration, and synaptogenesis. Loss of function of the oligophrenin-1 gene (OPHN1) causes X-linked intellectual disability with cerebellar hypoplasia and leads to hyperactivation of the rho kinase (ROCK) pathway. ROCK mainly acts through phosphorylation of the myosin phosphatise targeting subunit 1, triggering actin-myosin contractility. We show that during in vitro neurogenesis, ROCK activity decreases from day 10 until terminal differentiation, whereas in OPHN1-deficient human induced pluripotent stem cells (h-iPSCs), the levels of ROCK are elevated throughout differentiation. ROCK inhibition favors neuronal-like appearance of h-iPSCs, in parallel with transcriptional up regulation of nuclear receptor NR4A1, which is known to induce neurite outgrowth. This study analyzed the morphological, biochemical, and functional features of OPHN1-deficient h-iPSCs and their rescue by treatment with the ROCK inhibitor fasudil, shedding light on the relevance of the ROCK pathway during neuronal differentiation and providing a neuronal model for human OPHN1 syndrome and its treatment.
KW - 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
KW - Cell Biology
KW - Cell Differentiation
KW - Cell Shape
KW - Cells, Cultured
KW - Cytoskeletal Proteins
KW - Developmental Biology
KW - GTPase-Activating Proteins
KW - Humans
KW - In vitro neurogenesis
KW - Induced Pluripotent Stem Cells
KW - Models, Biological
KW - Mutation
KW - Nervous System Diseases
KW - Neurogenesis
KW - Neurons
KW - Nuclear Proteins
KW - Oligophrenin-1
KW - Phenotype
KW - ROCK inhibitors (fasudil, Y-27632)
KW - Rho-kinase signaling
KW - Syndrome
KW - rho-Associated Kinases
KW - 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine
KW - Cell Biology
KW - Cell Differentiation
KW - Cell Shape
KW - Cells, Cultured
KW - Cytoskeletal Proteins
KW - Developmental Biology
KW - GTPase-Activating Proteins
KW - Humans
KW - In vitro neurogenesis
KW - Induced Pluripotent Stem Cells
KW - Models, Biological
KW - Mutation
KW - Nervous System Diseases
KW - Neurogenesis
KW - Neurons
KW - Nuclear Proteins
KW - Oligophrenin-1
KW - Phenotype
KW - ROCK inhibitors (fasudil, Y-27632)
KW - Rho-kinase signaling
KW - Syndrome
KW - rho-Associated Kinases
UR - http://hdl.handle.net/10807/95381
UR - http://stemcellstm.alphamedpress.org/content/5/7/860.full.pdf
U2 - 10.5966/sctm.2015-0303
DO - 10.5966/sctm.2015-0303
M3 - Article
SN - 2157-6564
VL - 5
SP - 860
EP - 869
JO - Stem cells translational medicine
JF - Stem cells translational medicine
ER -