Neuronal Differentiation of Neural Progenitor Cells by Intracellular Delivery of Synthetic Oligopeptide Derived from Von Hippel-Lindau Protein

Abstract

Intracellular delivery of synthetic oligopeptides has the potential to promote the occurrence of various cellular events such as cell death, proliferation, growth inhibition, metabolic changes, and morphological changes. However, the regulation of cellular differentiation by intracellular delivery of synthetic oligopeptides has been little studied. Von Hippel-Lindau protein (pVHL) is one of the proteins that functions to induce the differentiation of neural progenitor cells (NPCs). To function in these cells, pVHL forms a complex composed of itself, elongin BC, Clu-2, and Rbx-1. It is suggested that the binding site of elongin BC in pVHL plays a critical role in pVHL function, i.e., ubiquitination, which is related to neuronal differentiation. So, we synthesized an oligopeptide corresponding to the elongin BC binding site, and delivered the oligopeptide into NPCs by using a mixture of trifluoroacetylated lipopolyamine and diloeoyl phosphatidylethanolamine (BioPorter) to form a peptide-lipid complex. After intracellular delivery of the oligopeptide, induction of differentiation of NPCs was shown in terms of neurite outgrowth and by immunocytochemical and electrophysiological means. The intracellular delivery of the synthetic oligopeptide derived from pVHL may provide a safe and valuable approach for the neuronal differentiation of NPCs.

Effect of Cdc42 domains on filopodia sensing, cell orientation, and haptotaxis

“…Peptides were delivered by BioPORTER reagent (Genlantis, San Diego, CA). It was previously found that BioPORTER (trifluoroacetylated lipopolyamine and dioleoyl phosphatidylethanolamine) was non-toxic, and the structure and function of the peptides were preserved in the cell [22]. Peptides were made up in sterile phosphate-buffered saline (PBS, pH 7.4) to a concentration of 50 μg/ml and combined with BioPORTER reagent according to the manufacturer's instructions.…”

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

“…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.…”

Transplantation of Von Hippel–Lindau peptide delivered neural stem cells promotes recovery in the injured rat spinal cord

“…By 9-fluorenylmethyloxycarbonyl solid-phase method, we chemically synthesized a peptide corresponding to the 157-171 amino acid sequence of the amino acid region of the pVHL which was shown to be related to neuronal differentiation [8]. The synthesized peptide was linked with the PTD of HIV-TAT protein (TATVHL-peptide, YGRKKRRQRRRDTLKERCLQVVRSLVK), thereby facilitating peptide entry into the cells.…”

“…However, as the virus vectors are used in the VHL gene transfer, it might have problems in infectious risk, toxicity, and bioethics. Recently we showed neuronal differentiation of neural progenitor cells with intracellular delivery of a synthetic oligopeptide derived from pVHL [8]. A novel approach, alternative to the gene transfer technology, has been developed for the delivery of a broad spectrum of exogenous proteins and chemically synthesized peptides into living cells [9].…”