@misc{_Modifications,
 copyright={Creative Commons Attribution BY 4.0 license},
 howpublished={online},
 abstract={I aimed to investigate the influence of neuregulin-1 (NRG-1) on the functional properties of glial-restricted progenitor cells (GRPs). The hypothesis I sought to verify was that overexpressing neuregulin-1 in GRPs would alter their functional characteristics, potentially enhancing their application potential in the therapy of demyelination-related diseases. The study focused on GRPs isolated from mouse fetal nervous tissue (referred to as mGRPs). To induce overexpression of neuregulin-1 type I (NRG-1 type I), genetic modifications were performed on mGRPs through the transduction of cells using lentiviral vectors encoding NRG-1 type I. The research involved analyzing the expression levels of NRG-1 in the modified mGRPs, examining their phenotype, and investigating the functional properties of mGRPs-NRG-1 in vitro. Evaluation of mGRPs transduced with the pLenti-GIII-CMV-NRG-1 type I-mCherry vector demonstrated that these cells exhibit varying levels of NRG-1 type I transcript synthesis in subsequent transduction attempts, and their continued culture leads to spontaneous detachment of cells from the surface of the culture vessel and subsequent cell death. The unfavorable results obtained from transducing mGRPs with the pLenti-GIII-CMV-NRG-1 type I-mCherry vector prompted me to use the HIV-SFFV-NRG-1 type I-IRES-mRFP vector for the next phase of the study. This vector encodes NRG-1 type I sequence and the mRFP fluorescent reporter protein, separated by the IRES (Internal Ribosome Entry Site) sequence. This arrangement allows for the coexpression of both sequences under the transcriptional control of a shared promoter, while their translation occurs independently. In the next stage of the study, I attempted to transduce mGRPs with lentiviral activating particles designed to overexpress the endogenous NRG-1 gene (NRG-1-LAPs). However, simultaneous transduction of mGRPs with the three lentiviral vectors comprising the NRG-1-LAPs system proved ineffective. Therefore, the attempt to induce endogenous NRG-1 overexpression in mGRPs through transduction with lentiviral activating particles was unsuccessful. In light of the setbacks associated with genetic modifications aimed at overexpressing NRG-1 in mGRPs, I decided to conduct additional research based on the stimulation of mGRPs with exogenous recombinant mouse neuregulin-1 peptide (rmNRG-1) through its supplementation in the standard culture medium. Supplementation of mGRPs with rmNRG-1 peptide had a diverse impact on the phenotype of the cells in vitro. Based on the conducted research, the following conclusions can be drawn: - overexpression of NRG-1 type I, achieved by transducing mGRPs with lentiviral vectors, resulted in a decrease in the growth rate of these cells and promoted their differentiation towards oligodendrocytes; however, I did not observe a significant impact of NRG-1 type I overexpression on the myelination properties of mGRPs - my preliminary investigations into the stimulation of mGRPs using exogenous recombinant neuregulin-1 peptide indicate an enhanced proliferation rate and reduced migratory capacity of supplemented mGRPs, accompanied by a diminished capability of these cells to differentiate into mature oligodendrocytes - the findings presented in my doctoral thesis highlight that the influence of neuregulin-1 on the functional properties of mGRPs is contingent upon the experimental setup.},
 title={Modifications of mice glial-restricted progenitor cells using neuregulin-1 to enhance thier regenerative potential in the treatment of demyelinating diseases.},
 type={Text},
 URL={http://www.rcin.org.pl/Content/240042/Piotr%20Rogujski_rozprawa%20doktorska_l.pdf},
 keywords={Demyelinating disease, Stem Cells, Neuregulin-1, Lentiviral vector},
}