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  

Abstract

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. STEM

Wnt5a cooperates with canonical Wnts to generate midbrain dopaminergic neurons in vivo and in stem cells

“…Shh and Fgf8 have been described previously as being expressed at the cross-section between the FP and isthmus and have been demonstrated to regulate the development of VM DA neurons in a coordinated manner (47,48) and to promote the differentiation of stem cells into midbrain DA neurons (28,32,44,(49)(50)(51). Similarly, Wnt1 and Wnt5a are coexpressed and intersect in defined spatial and temporal patterns in the developing midbrain: Wnt1 is expressed in the midbrain as early as E8 (52) and is found in two lateral bands flanking the FP from E10.5-12.5 (22), and Wnt5a is expressed in the VM, including the FP, from E9.5-13.5 (17).…”

Cripto-1: At the Crossroads of Embryonic Stem Cells and Cancer

“…The Cripto-1 blocking peptide was able to induce formation of neuroectoderm and increased midbrain dopaminergic neuron differentiation of mouse ES cells in vitro and in vivo (Lonardo et al, 2010). Moreover, mouse ES cells treated with Cripto-1 blocking peptide enhanced functional recovery and reduced tumor formation in Parkinsonian rats (Lonardo et al, 2010). Overexpression of Cripto-1 has also been observed in the cerebral cortical tissues of macaques that had been infected with a chimeric simian human immunodeficiency virus (SHIV) by cDNA array analysis (Stephens et al, 2006).…”

“…In a rat animal model of Parkinson's disease, mouse ES cells that are null for Cripto-1 expression (Cripto-1 -/ -ES cells), when grafted at low density into rats, differentiated into neuronal cells in the brain and were able to restore normal behavior without producing teratomas (Parish et al, 2005). Recently, a tetrameric tripeptide, which prevents Cripto-1/ALK4 interaction and interferes with Cripto-1 signaling was identified (Lonardo et al, 2010). The Cripto-1 blocking peptide was able to induce formation of neuroectoderm and increased midbrain dopaminergic neuron differentiation of mouse ES cells in vitro and in vivo (Lonardo et al, 2010).…”

“…Recently, a tetrameric tripeptide, which prevents Cripto-1/ALK4 interaction and interferes with Cripto-1 signaling was identified (Lonardo et al, 2010). The Cripto-1 blocking peptide was able to induce formation of neuroectoderm and increased midbrain dopaminergic neuron differentiation of mouse ES cells in vitro and in vivo (Lonardo et al, 2010). Moreover, mouse ES cells treated with Cripto-1 blocking peptide enhanced functional recovery and reduced tumor formation in Parkinsonian rats (Lonardo et al, 2010).…”

Control of Immune Response to Allogeneic Embryonic Stem Cells by CD3 Antibody–Mediated Operational Tolerance Induction

“…The differentiated products of embryonic stem cells (ESCs) have been used successfully in animal models to treat diverse diseases such as myocardial infarction (1,2), heart ischemia-reperfusion injury (3) and Parkinson's disease (4,5). Challenging the initially prevailing dogma, their implantation in allogeneic hosts was shown to trigger an immune response (6,7) that, despite being described as less intense than that of differentiated tissues (8,9), required the use of immunosuppressive treatments and thus represented an obstacle to widespread clinical application (7,10,11).…”