Efficient Generation of Dopamine Neuron-Like Cells From Skin-Derived Precursors With a Synthetic Peptide Derived from von Hippel-Lindau Protein

Abstract

Skin-derived precursors (SKPs) from mammalian dermis represent neural crest-related stem cells capable of differentiating into both neural and mesodermal progency. SKPs are of clinical interest because they serve as accessible autologous donor cells for neuronal repair for neuronal intractable diseases. However, little is known about the efficient generation of neurons from SKPs, and phenotypes of neurons generated from SKPs have been restricted. In addition, the neuronal repair using their generated neurons as donor cells has not been achieved. The von Hippel-Lindau protein (pVHL) is one of the proteins that play an important role during neuronal differentiation, and recently neuronal differentiation of neural progenitor cells by intracellular delivery of a synthetic VHL peptide derived from elongin BC-binding site has been demonstrated. In the present study, a synthetic VHL peptide derived from elongin BC-binding site was conjugated to the protein transduction domain (PTD) of HIV-TAT protein (TATVHL peptide) to facilitate entry into cells, and we demonstrate the efficient generation of cells with dopaminergic phenotype from SKPs with the intracellular delivery of TATVHL peptide, and characterized the generated cells. The TATVHL peptide-treated SKPs expressed neuronal marker proteins, particularly dopamine neuron markers, and also up-regulated mRNA levels of proneural basic helix-loop-helix factors. After the TATVHL peptide treatment, transplanted SKPs into Parkinson's disease (PD) model rats sufficiently differentiated into dopamine neuron-like cells in PD model rats, and partially but significantly corrected behavior of PD model rats. The generated dopamine neuron-like cells are expected to serve as donor cells for neuronal repair for PD.

Engrafted VHL peptide-delivered bone marrow stromal cells promote spinal cord repair in rats

“…In this study, we proposed a novel method using intracellular delivery of VHL peptide for neuronal differentiation in MSCs. Compared with neural progenitor/stem cells [8,9] or skin-derived precursors [10], MSCs showed less neuronal differentiation by intracellular delivery of VHL peptide. Although the mechanism of neuronal differentiation by intracellular delivery of VHL peptide is unclear, the response to VHL peptide might be different in cell types.…”

“…As described earlier [8][9][10], we chemically synthesized a peptide corresponding to the 157-171 amino acid sequence in the amino acid region of the pVHL. [11].…”

“…We showed earlier that the VHL protein had the potential to transform neural progenitor cells into neurons in vitro [7]. Recently, intracellular delivery of a synthetic oligopeptide derived from the VHL protein (VHL peptide) showed neuronal differentiation in neural progenitor/stem cells and skin-derived precursors [8][9][10], and transplantation of VHL peptide-delivered neural stem cells promoted recovery in the injured rat spinal cord [9]. In this study, we showed that intracellular delivery of VHL peptide induced neuronal differentiation in MSCs.…”

Prospect of cell therapy for Parkinson's disease

“…The results showed efficient generation of cells with dopaminergic neuronal and proneural markers. Furthermore, transplanting these cells into Parkinson's disease model rats showed promising results, and that the cells generated dopaminergic neuron-like cells [34]. …”

Modeling neural differentiation on micropatterned substrates coated with neural matrix components

“…These precursor cells have the potential to generate progeny of the neuronal and glial lineages, amongst other cell types (Fernandes et al, 2008; Hunt et al, 2009). Importantly, dopaminergic neurons have been generated in vitro from SKPs and transplanted into the striata of a Parkinson disease (PD) rat model (Kubo et al, 2009; Higashida et al, 2010). Although these recent findings will need validation by independent research groups, they hint the potential to isolate autologous dermal precursor cells and, after in vitro differentiation to the desired lineage, apply the differentiated neurons for cell replacement therapies of Parkinson's disease.…”