Neurosurgical delivery of chemotherapeutics, targeted toxins, genetic and viral therapies in neuro-oncology.

Local delivery of biologic agents, such as gene and viruses, has been tested preclinically with encouraging success, and in some instances clinical trials have also been performed. In addition, the positive pressure infusion of various therapeutic agents is undergoing human testing and approval has already been granted for routine clinical use of biodegradable implants that diffuse a chemotherapeutic agent into peritumoral regions. Safety in glioma patients has been shown, but anticancer efficacy needs additional refinements in the technologies employed. In this review, we will describe these modalities and provide a perspective on needed improvements that should render them more successful.

PML-nuclear bodies accumulate DNA in response to polyomavirus BK and simian virus 40 replication.

Promyelocytic nuclear bodies (PML-NBs) are distinct nuclear structures that are involved in apoptosis, differentiation, transcriptional regulation and DNA damage response. These bodies have also been shown to associate with nuclear sites of viral DNA replication. In the present study, we used BrdU pulse labeling to demonstrate that PML-NBs accumulate newly synthesized DNA in cells infected by the polyomaviruses simian virus 40 (SV40) or polyomavirus BK (BKV). Sequestration of DNA molecules in these structures depended on active viral DNA replication, and was observed exclusively in cells that contained prominent viral replication domains. Furthermore, a significant portion of the accumulated DNA was found to be single-stranded, indicating that the sequestered DNA had been subjected to processing by nuclease or DNA unwinding activities. siRNA-mediated suppression of the PML protein prevented the recruitment of single-stranded DNA into nuclear foci, but did not significantly affect the overall efficiency of viral DNA replication. These results indicate a role of PML and PML-NBs in post-replication DNA processing, and suggest that PML-NBs become linked to sites of viral DNA synthesis due to a role of these structures in DNA metabolism.

Prospects for delivery of recombinant angiostatin by cell-encapsulation therapy.

Implantation of encapsulated nonautologous cells that have been genetically modified to secrete proteins with tumor suppressor properties represents an alternative nonviral strategy to cancer gene therapy. We report an approach to raise the yield of recombinant proteins from encapsulated cells substantially. We hypothesized that by optimizing the encapsulation procedure, the production efficacy from the encapsulated cells could be increased. HEK 293 EBNA cells were genetically engineered to produce angiostatin. Encapsulation was performed by varying bead size, cellular density, homogeneity, and ion composition of the gel. The morphology and viability of the cells and the release of angiostatin were studied. Computer software was developed for three-dimensional imaging and quantification of cell viability. Angiostatin production was assessed at 3, 6, and 11 weeks using enzyme-linked immunosorbent assay (ELISA). Inhomogeneous gels facilitated cell growth and viability. The most efficient inhomogeneous microcapsules were generated by reducing the size and cellular density of the beads. The viability and the production of angiostatin were 3 to 5 times higher than in the homogeneous capsules. Significant amounts of viable cells were present in both homogeneous and inhomogeneous beads after 6 months of culture. The stability of the alginate matrix was greatly enhanced by gelling in the presence of barium. In conclusion, the viability and production efficacy of recombinant angiostatin from alginate-encapsulated cells can be increased considerably by optimizing the encapsulation procedure. The development of such optimized microcapsules brings cell-encapsulation therapy further towards clinical use in cancer therapy.

Progress and challenges for cell encapsulation in brain tumour therapy.

Cell encapsulation provides a method to circumvent the host immune system by encapsulating cells or tissues in immunoisolating, semipermeable membranes before implantation. The technology has been widely studied with an aim of developing bio-organs transplantable into patients without the need of immunosuppression, and in cancer therapy, the principle of cell encapsulation may be further exploited. Encapsulated recombinant cells represent factories or bioreactors for the production of therapeutic proteins. By implanting the bioreactors in the vicinity of the tumour, long-term local de novo delivery of the therapeutic proteins may be achieved. Malignant brain tumours such as glioblastoma multiforme (GBM) remain highly lethal neoplasms, refractory to current therapies. Researchers and medical professionals are working on developing translational therapies to combat these aggressive tumours. Numerous clinical trials on gene therapy for glioma patients have been conducted over the last decade, but the results have largely been disappointing. Cell encapsulation represents an alternative method for local delivery of therapeutic proteins with antineoplastic properties to glioma patients. The concept has not yet reached clinical trials, but encouraging results have been achieved in rats bearing gliomas when implanting endostatin-secreting encapsulated cells into the rat brain. This review primarily focuses on the recent progress that has been made with cell encapsulation technology. In addition, the challenges this field faces before clinical application in brain tumour patients is discussed.

Mechanisms of tumor cell invasion and angiogenesis in the central nervous system.

Despite extensive progress in characterizing the genetic events involved in the development of gliomas, the cellular origin and the defined molecular mechanisms that lead to their occurrence are still unclear. It is known that tumours are of monoclonal origin. This is contrasted by the fact that gliomas frequently express features of different glial cell lineages. With the identification of pluripotent neural stem cells and the growth factors that control neural cell development, we are now making early inroads towards understanding glial cell migration as well as the neural cell plasticity within the adult central nervous system (CNS). Gliomas share several fetal antigens with immature brain cells. It is therefore tempting to speculate that the migration of neural precursor cells actually represents the normal counterpart of glioma cell migration. The migratory behavior of gliomas may be due to a predetermined interplay between normal brain tissue and the migrating cells, where the brain represents a permissive tissue for guiding cells with certain phenotypic traits to migrate along specific anatomical structures. Malignant progression is also accompanied by extensive angiogenesis which is especially prominent in glioblastoma multiforme (GBM). For cell proliferation to take place, several cell signaling cues mediated by specific growth factors are shared between the glioma cells and the endothelial cells while others are unique for endothelial cells. Therefore the endothelial cell compartment represents a promising target for novel therapeutic strategies including gene therapy and cell-based therapies.