Cancer and glioma : an integrated approach of gene therapy and bioluminescence imaging

Glioblastoma multiforme (GBM) is the most malignant variant of glioma. This tumor does not only display an extremely aggressive, invasive growth pattern, but is also very difficult to treat. With a two-year survival rate of 40% and a median survival of 12-18 months after treatment, prognosis is poor. Current treatment options are not successful in halting tumor progression and GBM tumors are highly heterogeneous, display all kinds of anti-apoptotic escape routes, suppress the immune system, invade the surrounding parenchyma with unmatched aggressiveness and possess a whole array of tools to rearrange the extra tumoral environment to their advantage. The aim of this thesis is to combine the strengths of gene-therapy and bioluminescence Imaging (BLI) for the development of novel reporter systems in order to study glioma tumor biology and its response to therapeutic compounds. We optimized the currently available BLI luciferases (Gaussia luciferase, Vargula hilgendorfi) and assays (Gluc blood assay, Mycoplasma detection assay). We explored a new multimodal targeted liposome formulation with increased relaxivity for the treatment and imaging of cancer. Finally we combined the newly developed and enhanced reporters to test a new therapeutic combination for the treatment of Glioma (TRAIL, Lanatoside C).UBL - phd migration 201

Cell-based Immunotherapy Against Gliomas: From Bench to Bedside

Scientific Assessment and Innovation in Neurosurgical Treatment Strategie

Enhanced Gaussia Luciferase Blood Assay for Monitoring of in Vivo Biological Processes

Scientific Assessment and Innovation in Neurosurgical Treatment Strategie

Advances in stem cell therapy against gliomas

Scientific Assessment and Innovation in Neurosurgical Treatment Strategie

Sensitive Assay for Mycoplasma Detection in Mammalian Cell Culture

Scientific Assessment and Innovation in Neurosurgical Treatment Strategie

Codon-Optimized Luciola italica Luciferase Variants for Mammalian Gene Expression in Culture and In Vivo

Luciferases have proven to be useful tools in advancing our understanding of biologic processes. Having a multitude of bioluminescent reporters with different properties is highly desirable. We characterized codon-optimized thermostable green- and red-emitting luciferase variants from the Italian firefly Luciola italica for mammalian gene expression in culture and in vivo. Using lentivirus vectors to deliver and stably express these luciferases in mammalian cells, we showed that both variants displayed similar levels of activity and protein half-lives as well as similar light emission kinetics and higher stability compared to the North American firefly luciferase. Further, we characterized the red-shifted variant for in vivo bioluminescence imaging. Intramuscular injection of tumor cells stably expressing this variant into nude mice yielded a robust luciferase activity. Light emission peaked at 10 minutes post-D-luciferin injection and retained > 60% of signal at 1 hour. Similarly, luciferase activity from intracranially injected glioma cells expressing the red-shifted variant was readily detected and used as a marker to monitor tumor growth over time. Overall, our characterization of these codon-optimized luciferases lays the groundwork for their further use as bioluminescent reporters in mammalian cells

Codon-Optimized Luciola italica Luciferase Variants for Mammalian Gene Expression in Culture and In Vivo

Scientific Assessment and Innovation in Neurosurgical Treatment Strategie

Type grouping in rat skeletal muscle after crush injury

Object. Accuracy of reinnervation is an important factor that determines outcome after nerve injury and repair. Type grouping-the clustering of muscle fibers of the same type after reinnervation-can be used to investigate the accuracy of reinnervation. In this study, the degree of type grouping after crush injury in rats was compared with the clustering of muscle fibers after autografting or single-lumen nerve grafting. Methods. Twelve weeks after sciatic nerve crush injury in rats, clustering of Type I muscle fibers was analyzed in the target muscle with adenosine 5'-triphosphatase staining. In addition, the number of regenerated axons was determined in the nerve distal to the crush injury. Results were compared with that of the authors' previous study. Results. Type grouping was more abundant after crush injury than after autograft or single-lumen nerve graft repair. Conclusions. Crush injury leads to more clustered innervation of muscle fibers, probably because the Schwann cell basal lamina tubes are not interrupted as they are in autograft or artificial nerve graft repair. This finding adds to understanding the processes playing a role in nerve regeneration. (DOI: 10.3171/2010.9.JNS091656)Stem cells & developmental biolog

Directed Molecular Evolution Reveals Gaussia Luciferase Variants with Enhanced Light Output Stability

Scientific Assessment and Innovation in Neurosurgical Treatment Strategie