A small synthetic cripto blocking Peptide improves neural induction, dopaminergic differentiation, and functional integration of mouse embryonic stem cells in a rat model of Parkinson's disease.
Cripto is a glycosylphosphatidylinositol-anchored coreceptor that binds Nodal and the activin type I (ALK)-4 receptor, and is involved in cardiac differentiation of mouse embryonic stem cells (mESCs). Interestingly, genetic ablation of cripto results in increased neuralization and midbrain dopaminergic (DA) differentiation of mESCs, as well as improved DA cell replacement therapy (CRT) in a model of Parkinson's disease (PD). In this study, we developed a Cripto specific blocking tool that would mimic the deletion of cripto, but could be easily applied to embryonic stem cell (ESC) lines without the need of genetic manipulation. We thus screened a combinatorial peptide library and identified a tetrameric tripeptide, Cripto blocking peptide (BP), which prevents Cripto/ALK-4 receptor interaction and interferes with Cripto signaling. Cripto BP treatment favored neuroectoderm formation and promoted midbrain DA neuron differentiation of mESCs in vitro and in vivo. Remarkably, Cripto BP-treated ESCs, when transplanted into the striatum of PD rats, enhanced functional recovery and reduced tumor formation, mimicking the effect of genetic ablation of cripto. We therefore suggest that specific blockers such as Cripto BP may be used to improve the differentiation of ESC-derived DA neurons in vitro and their engraftment in vivo, bringing us closer towards an application of ESCs in CRT.
Lonardo
Enza
E
Stem Cell Fate Laboratory, Institute of Genetics and Biophysics, Naples, Italy.
Parish
Clare L
CL
Ponticelli
Salvatore
S
Marasco
Daniela
D
Ribeiro
Diogo
D
Ruvo
Menotti
M
De Falco
Sandro
S
Arenas
Ernest
E
Minchiotti
Gabriella
G
D030201:Activin Receptors, Type I / Q000378:metabolism; D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D004195:Disease Models, Animal; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism; D051379:Mice; D009363:Neoplasm Proteins / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects*; D009842:Oligopeptides / Q000494:pharmacology* / Q000627:therapeutic use*; D010300:Parkinson Disease / Q000188:drug therapy* / Q000628:therapy; D011485:Protein Binding / Q000187:drug effects; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D033581:Stem Cell Transplantation
D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; D009842:Oligopeptides; C489450:Tdgf1 protein, mouse; D004815:Epidermal Growth Factor; D030201:Activin Receptors, Type I; C498933:Acvr1b protein, mouse