Glioblastoma Lines as the Disease Model

Glioblastoma (GBM) is the most aggressive type of brain tumor arising from glial cells accounting for 52% of all parenchymal brain cancer cases and 20% of all intracranial tumors. GBM has pronounced mitotic activity, substantial tendency toward neoangiogenesis (microvascular proliferation), necrosis, and high proliferative rates. Because of their intrinsic infiltrative nature, GBM has a highly aggressive malignant clinical course. The adjuvant chemo-radiotherapy (RT) with temozolomide (TMZ) after maximal safe resection remains the standard of care.

Brain cancer is a unique because of the "blood-brain barrier", which severely restricts the bloodstream of the brain. While the blood brain barrier (BBB) is great for protecting the brain from danger, when the brain has cancer cells, the BBB can be a problem. Therefore, it is important to find new drug targets. The mechanisms underlying radio- and chemo-resistance are poorly understood. Recent studies suggest that up-regulation of the molecular target of rapamycin (mTOR) plays a pivotal role in determining resistance to treatment. The upregulation of mTOR in GBM has been reported by multiple experimental and pathological findings. Moreover, the up-regulation of mTOR is also the key for cell growth and cell proliferation as demonstrated by in vivo studies. In fact, specific factors derived from brain endothelial cells maintain glioblastoma stem-like cell expansion through the mTOR pathway. The key to successful treatment of glioblastoma will be no doubt in the realization that this clinic is an entity in biologic terms, more than one disease, and is likely that specific targeted therapies will be effective in molecularly defined subsets. In this way, the molecular classification of these tumors will be defined in clinically relevant terms based on the identification of markers that define subsets and are predictive of response to promising agents. Additional investigations and identifications of new biomarkers will help to better define the clinical and biologic subtypes of glioblastoma and an improved disease control. In short, the brain tumor has peculiar ad personam mutations. This is why the investigators have decided to set up primary lines starting from the patient's biopsy.

Expected results of the scientific research project:

In the first instance, the primary outcome will be to establish cell cultures and stem cells that faithfully reproduce in vitro the physiology of the tumor maintaining the same characteristics of patient's neoplasm

1. Evaluate the effect of new target drugs on the proliferation of primary and continuous human glioblastoma cell lines by setting up growth curves and methyl thiazolyl tetrazolium (MTT) toxicity assays.
2. Screening of natural and synthetic drugs using patient-derived primary glioblastoma cell lines
3. Characterize the mechanisms and proteins involved in the apoptotic and / or autophagic pathway with immunohistochemistry and western blot assays in control and treated cells.
4. Validation of previously identified molecular targets in preclinical models of brain cancers. The investigators are able to identify novel molecular determinants that can be targeted by pharmacological intervention to decrease or block the tumor growth.