Abstract
Chronic neurodegenerative and neuropsychiatric diseases are becoming more prevalent. Unfortunately, treatment of such diseases often gets complicated by the inability of many drugs of therapeutic relevance to cross the blood brain barrier (BBB), formed by non-fenestrated brain capillary endothelial cells (BCECs) and their intermingling tight junctions. Polypeptides like brain derived neurotropic factor (BDNF), erythropoietin (EPO), and FGL peptide (part of Neuronal adhesion molecule (NCAM)) are acknowledged for their neuroprotective and neurogenerative effects. Generally, however, these macromolecules are unable to cross the BBB. A promising approach to overcome the restricted properties of the BBB is gene therapy based on delivery of genetic material into BCECs. Theoretically this would result in protein expression and secretion from the BCEC and further into the brain.
The aim of this study has been to investigate the usage of BCEC as factories for recombinant protein production. A non-viral gene carrier was prepared from pullulan-spermine conjugated with plasmid DNA (Thomsen et al., 2011). In vitro transfection of Rat Brain Endothelial Cells (RBE4) and Human Brain Microvascular Endothelial cells (HBMECs) were conducted with three plasmids bearing cDNA encoding human BDNF, EPO or the FGL peptide. Results revealed a high expression of BDNF, EPO and FGL transcripts in transfected cells compared to the non-transfected cells, which strongly suggest that transfection were successful in both cells lines. To prove that the cells not only synthesize these proteins but also secret them, analysis of the cell media are ongoing. Additionally in vivo studies on rats subjected to injection of transfecting agents into the carotic artery are also being conducted.
- Thomsen LB, Lichota J, Kim KS, Moos T. Gene delivery by pullulan derivatives in brain capillary endothelial cells for protein secretion. J Control Release. 2011 Jan 18.
The aim of this study has been to investigate the usage of BCEC as factories for recombinant protein production. A non-viral gene carrier was prepared from pullulan-spermine conjugated with plasmid DNA (Thomsen et al., 2011). In vitro transfection of Rat Brain Endothelial Cells (RBE4) and Human Brain Microvascular Endothelial cells (HBMECs) were conducted with three plasmids bearing cDNA encoding human BDNF, EPO or the FGL peptide. Results revealed a high expression of BDNF, EPO and FGL transcripts in transfected cells compared to the non-transfected cells, which strongly suggest that transfection were successful in both cells lines. To prove that the cells not only synthesize these proteins but also secret them, analysis of the cell media are ongoing. Additionally in vivo studies on rats subjected to injection of transfecting agents into the carotic artery are also being conducted.
- Thomsen LB, Lichota J, Kim KS, Moos T. Gene delivery by pullulan derivatives in brain capillary endothelial cells for protein secretion. J Control Release. 2011 Jan 18.
| Originalsprog | Engelsk |
|---|---|
| Publikationsdato | 2011 |
| Status | Udgivet - 2011 |
| Begivenhed | 9th International conference on cerebral vascular biology - Leiden, Holland Varighed: 21 jun. 2011 → 25 jun. 2011 |
Konference
| Konference | 9th International conference on cerebral vascular biology |
|---|---|
| Land/Område | Holland |
| By | Leiden |
| Periode | 21/06/2011 → 25/06/2011 |