Abstract
The adult human olfactory bulb neural stem/progenitor cells (OBNC/PC) are promising candidate for cell-based therapy for traumatic and neurodegenerative insults. Exogenous application of NGF was suggested as a promising therapeutic strategy for traumatic and neurodegenerative diseases, however effective delivery of NGF into the CNS parenchyma is still challenging due mainly to its limited ability to cross the blood–brain barrier, and intolerable side effects if administered into the brain ventricular system. An effective method to ensure delivery of NGF into the parenchyma of CNS is the genetic modification of NSC to overexpress NGF gene. Overexpression of NGF in adult human OBNSC is expected to alter their proliferation and differentiation nature, and thus might enhance their therapeutic potential. In this study, we genetically modified adult human OBNS/PC to overexpress human NGF (hNGF) and green fluorescent protein (GFP) genes to provide insight about the effects of hNGF and GFP genes overexpression in adult human OBNS/PC on their in vitro multipotentiality using DNA microarray, immunophenotyping, and Western blot (WB) protocols. Our analysis revealed that OBNS/PC-GFP and OBNS/PC-GFP-hNGF differentiation is a multifaceted process involving changes in major biological processes as reflected in alteration of the gene expression levels of crucial markers such as cell cycle and survival markers, stemness markers, and differentiation markers. The differentiation of both cell classes was also associated with modulations of key signaling pathways such MAPK signaling pathway, ErbB signaling pathway, and neuroactive ligand-receptor interaction pathway for OBNS/PC-GFP, and axon guidance, calcium channel, voltage-dependent, gamma subunit 7 for OBNS/PC-GFP-hNGF as revealed by GO and KEGG. Differentiated OBNS/PC-GFP-hNGF displayed extensively branched cytoplasmic processes, a significant faster growth rate and up modulated the expression of oligodendroglia precursor cells markers (PDGFRα, NG2 and CNPase) respect to OBNS/PC-GFP counterparts. These findings suggest an enhanced proliferation and oligodendrocytic differentiation potential for OBNS/PC-GFP-hNGF as compared to OBNS/PC-GFP.
Highlights
Exogenous application of nerve growth factor (NGF) for the treatment of traumatic and neurodegenerative insults is a promising therapeutic strategy
Understanding mechanisms involved in modulation of proliferation and differentiation potential of adult human OBNSC is crucial for enhancement of their therapeutic potentials against a wide range of traumatic and neurodegenerative diseases such as Alzheimer’s disease (AD), Parkinson’s disease (PD) among others
NGF release was repressed in both human NGF (hNGF)+ and Whole Transcript (WT) cells, higher level of hNGF could still be detected in olfactory bulb neural stem/progenitor cells (OBNS/PC)-GFPhNGF (Figure 1B)
Summary
Exogenous application of nerve growth factor (NGF) for the treatment of traumatic and neurodegenerative insults is a promising therapeutic strategy. NGF enhances the survival of cholinergic neurons in basal forebrain in rats [1,2,3] and primates [4,5,6,7], and phase-I clinical trial of NGF gene therapy for Alzheimer’s disease (AD) provided promising data [8,9]. An effective method to ensure delivery of NGF into the parenchyma of CNS is the genetic modification of cells to overexpress NGF gene(s). In this regard, engraftments of cells that secrete NGF promote the growth of host spinal axons after injury [15] and protect cholinergic neurons from degeneration in chemical lesions [16,17] or aged brain [18,19,20]. Transplantation of fibroblasts encoding NGF gene in the primate brain rescued degenerating cholinergic neurons, and reduce degree of cognitive decline [20]
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
