Revertierung von Neurofibromatose Typ 1 Haploinsuffizienzeffekten durch Proteintransduktion in vitro
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Enzyme replacement therapies have been performed since the 1970s to treat the lack of proteins in human blood or in lysosomes of human cells. Up to now, there are no comparable treatments to replace lacking proteins in the cytosol of cells of patients suffering from genetic diseases. Here, this approach is exemplarily studied in fibroblasts of neurofibromatosis type 1 (NF1) patients. In these cells, the amount of the cytosolic neurofibromin is reduced. This protein reduction leads to a hyperactivation of the mitogen activated protein kinase pathway and to an altered capability of the fibroblasts to orientate themselves on nanostructured surfaces. In cell cultures it could be shown that the missing amounts of neurofibromin in NF1 haploinsufficient fibroblasts can be replaced using protein transduction methods. Two test systems were used to illustrate that the transduced neurofibromin can assume the functions of the lacking proteins in these cells and revert cellular effects based on NF1 haploinsufficiency. The endolysosomal entrapment of transduced proteins is the bottleneck of cytosolic protein transduction. For the cell culture experiments, this problem was solved by performing photochemical internalization (PCI) treatments. PCI was used to translocate endosomal entrapped proteins to the cytosol of the transduced fibroblasts. For that systemic PCI treatments are not realizable in patients, future studies will have to examine protein modifications that allow a cytosolic delivery of transduced proteins without an interfering from an external source. Experiments using a neurofibromin fused to the protein transduction domain of HIV-TAT revealed promising evidence that modifications can replace secondary treatments like PCI. Hence, the cytosolic replacement of lacking intracellular proteins seems to be a possible treatment for patients with monogenic diseases like NF1.
Subject HeadingsEnzymtherapie [GND]
Typ 1 [GND]
Cell-penetrating peptides [MeSH]
Enzyme replacement therapy; Methods [MeSH]