The Neurofilament-Derived Peptide NFL-TBS.40-63 Targets Neural Stem Cells and Affects Their Properties

Abstract

Targeting neural stem cells (NSCs) in the adult brain represents a promising approach for developing new regenerative strategies, because these cells can proliferate, self-renew, and differentiate into new neurons, astrocytes, and oligodendrocytes. Previous work showed that the NFL-TBS.40-63 peptide, corresponding to the sequence of a tubulin-binding site on neurofilaments, can target glioblastoma cells, where it disrupts their microtubules and inhibits their proliferation. We show that this peptide targets NSCs in vitro and in vivo when injected into the cerebrospinal fluid. Although neurosphere formation was not altered by the peptide, the NSC self-renewal capacity and proliferation were reduced and were associated with increased adhesion and differentiation. These results indicate that the NFL-TBS.40-63 peptide represents a new molecular tool to target NSCs to develop new strategies for regenerative medicine and the treatment of brain tumors. STEM CELLS TRANSLATIONAL MEDICINE 2016;5:901-913 SIGNIFICANCEIn the present study, the NFL-TBS.40-63 peptide targeted neural stem cells in vitro when isolated from the subventricular zone and in vivo when injected into the cerebrospinal fluid present in the lateral ventricle. The in vitro formation of neurospheres was not altered by the peptide; however, at a high concentration of the peptide, the neural stem cell (NSC) self-renewal capacity and proliferation were reduced and associated with increased adhesion and differentiation. These results indicate that the NFL-TBS.40-63 peptide represents a new molecular tool to target NSCs to develop new strategies for regenerative medicine and the treatment of brain tumors.

The neurofilament derived-peptide NFL-TBS.40-63 enters in-vitro in human neural stem cells and increases their differentiation

“…The peptide NFL-TBS.40-63 is able to enter in NSCs from human fetus in a dose-dependent manner as previously shown for NSCs from newborn and adult rats [ 12 ] or glioblastoma cells [ 22 , 23 ]. It was also internalized by oligodendrocytes [ 24 ] and certain human tumor cell lines (HeLa, MCF7, LNCaP) [ 9 , 22 ], while its internalization was very low in healthy astrocytes or neurons [ 11 , 22 ].…”

“…The possibility of manipulating NSCs directly in the brain for neurodegenerative therapies would be a major breakthrough in the context of neurological disorders. As previously shown, the NFL-TBS.40-63 peptide is a promising tool to target newborn or adult rat NSCs by modifing their behavior [ 12 ]. It is however essential to first evaluate the potential of this peptide to modify human NSCs before considering this strategy as a therapeutic approach.…”

“…This peptide not only increases oligodendrocyte differentiation and maturation, but also protects oligodendrocytes in a demyelination model [ 11 ]. Recently, we showed that the peptide can target newborn and adult rat NSCs (rNSCs), and modify rNSCs properties in vitro , in particular differentiation [ 12 ].…”

Enhanced and preferential internalization of lipid nanocapsules into human glioblastoma cells: effect of a surface-functionalizing NFL peptide

“…brain neural stem cells. 18,22,41 Previously, Paillard et al reported that the internalization of untargeted LNCs was not preferentially targeted into GBM cells and entered also healthy astrocytes. 38 Additionally, surface-functionalization with the fluoNFL peptide significantly enhanced the uptake of LNCs in NHA, compared to control LNCs (LNC-DiA), NFL being a cell penetrating peptide.…”

CRISPR: express delivery to any DNA address

“…Common experimental strategies exploited to deliver components to the cell interior include microinjection, infection by viruses engineered to express desired products and diverse chemically mediated transfection strategies. However, delivery of therapeutics to specific cell types, the silver bullet critical for many applications, is often difficult to achieve (e.g., Lépinoux‐Chambaud et al , ). For the widest application of potential CRISPR‐based therapeutic strategies, overcoming this current limitation remains an important objective.…”