Synergistic Effect of the Long-Term Overexpression of Bcl-2 and BDNF Lentiviral in Cell Protecting against Death and Generating TH Positive and CHAT Positive Cells from MSC

Abstract

Mesenchymal stem cells (MSC) are potentially a good material for transplantation in many diseases, including neurodegenerative diseases. The main problem with using them is the low percentage of surviving cells after the transplant procedure and the naturally poor ability of MSC to spontaneously differentiate into certain types of cells, which results in their poor integration with the host cells. The aim and the novelty of this work consists in the synergistic overexpression of two genes, BCL2 and BDNF, using lentiviral vectors. According to our hypothesis, the overexpression of the BCL2 gene is aimed at increasing the resistance of cells to stressors and toxic factors. In turn, the overexpression of the BDNF gene is suspected to direct the MSC into the neural differentiation pathway. As a result, it was shown that the overexpression of both genes and the overproduction of proteins is permanent and persists for at least 60 days. The synergistically transduced MSC were significantly more resistant to the action of staurosporine; 12 days after transduction, the synergistically transduced MSC had a six-times greater survival rate. The overexpression of the Bcl-2 and BDNF proteins was sufficient to stimulate a significant overexpression of the CHAT gene, and under specific conditions, the TH, TPH1, and SYP genes were also overexpressed. Modified MSC are able to differentiate into cholinergic and dopaminergic neurons, and the release of acetylcholine and dopamine may indicate their functionality.

The Different Molecular Code in Generation of Dopaminergic Neurons from Astrocytes and Mesenchymal Stem Cells

“…In the present study, although we found that S/F8/F2 induced MSCs to DA neuron-like cells, it was worth noting that the induced cells remained the original morphology with moderate TH expression levels. A similar expression pattern was found in many other reports on MSC-induced DA neurons [11,24,49,55]. Even in the optimized in vitro conditions, the induced MSCs developed monopolar neuron-like cells without complex neurites, lacking the synaptic structure of the functional neurons.…”

“…Similar limitations also exist in cell therapy with neural stem cells [7] and pluripotent induced stem cells (iPSCs) [3,8,9]. In contrast, mesenchymal stem cells (MSCs) isolated from the umbilical cord (UC-MSCs), adipose tissue (AD-MSCs), bone marrow (BM-MSCs), and other tissue sources [10] have displayed considerable superiorities of lower immunogenicity, less tumorigenicity, and higher ethical acceptance [5,11]. A series of studies have been reported that MSCs were able to rescue nigrostriatal circuits and improve behavioral performance in the PD models [12], which were considered as one of the ideal donor cells for PD treatment [13].…”

Mesenchymal Stem Cells: Therapeutic Mechanisms for Stroke

“…Additionally, transplanted MSCs carrying the GDNF gene also reduced the infarct volume, and the effect was similar to MSCs carrying the BDNF gene [ 23 ]. In addition, the overexpression of the BDNF gene could direct the MSC into the neural differentiation pathway [ 7 ]. In other aspects, the function of NGF was to prevent neuron apoptosis and to increase neuron proliferation [ 24 ], while VEGF enhanced angiogenesis [ 25 ], and PDGF promoted cell migration, the growth of primary cortical neurons, angiogenesis and axon growth, as well as the inhibition of neuroinflammation [ 26 , 27 , 28 ].…”

“…To treat stroke, the transplantation of mesenchymal stem cells (MSCs) can be used because of their intrinsic ability to play a therapeutic role. MSCs are pluripotent stem cells, with high self-renewal ability and multidirectional differentiation potential, that are capable of differentiating into adipocytes, chondrocytes, osteoblasts, neurons, and glial cells under certain conditions [ 4 , 5 , 6 , 7 ]. Moreover, MSCs can be extracted from different parts of the human body, and the isolation and amplification of MSCs is inexpensive.…”

Effects of the Co-Overexpression of the BCL and BDNF Genes on the Gamma-Aminobutyric Acid-Ergic Differentiation of Wharton’s-Jelly-Derived Mesenchymal Stem Cells

“…To investigate this hypothesis, two genes were selected: one from the BCL (B-cell lymphoma) gene family, which was expected to increase cell survival and resistance to toxic factors, and the other—BDNF (brain-derived neurotrophic factor)—whose overexpression should increase their ability to differentiate into the progenitors of nerve cells or mature cells or different types of neurons. Previous studies have shown that the co-overexpression of the BCL2 and BDNF or BCLXL and BDNF genes in an analogous transduction system increases cell survival under toxic conditions in an in vitro culture [ 13 , 14 ] and increases their ability to differentiate into cells that exhibit a cholinergic and/or dopaminergic phenotype [ 13 ].…”