Gliomas are a common type of brain tumour and for the treatment of a patient it is important to determine the tumour’s grade of malignancy. This is done today by a biopsy, a histopathological analysis of the tumourous tissue, that is classified by the World Health Organization on a malignancy scale from I to IV. Recent studies have shown that the local image variance (LIV) and the intratumoural susceptibility signal (ITSS) in susceptibility-weighted MR images correlate to the tumour grade. This thesis project aims to develop a software program as aid for the radiologists when grading a glioma. The software should by image analysis be able to separate the gliomas into low grade (I-II) and high grade (III-IV). The result is a graphical user interface written in Python 3.4.3. The user chooses an image, draws a region of interest and starts the analysis. The analyses implemented in the program are LIV and ITSS mentioned above, and the code can be extended to contain other types of analyses as research progresses. To validate the image analysis, 16 patients with glioma grades confirmed by biopsy are included in the study. Their susceptibility-weighted MR images were analysed with respect to LIV and ITSS, and the outcome of those image analyses was tested versus the known grades of the patients. No statistically significant difference could be seen between the high and the low grade group, in the case of LIV. This was probably due to hemorrhage and calcification, characteristic for some tumours and interpreted as blood vessels. Concerning ITSS a statistically significant difference could be seen between the high and the low grade group (p < 0.02). The sensitivity and specificity was 80% and 100% respec- tively. Among these 16 gliomas, 11 were astrocytic tumours and between low and high grade astrocytomas a statistically significant difference was shown. The degree of LIV was significantly different between the two groups (p < 0.03) and the sensitivity and specificity were 86% and 100% respectively. The degree of ITSS was significantly different between the two groups (p < 0.04) and the sensitivity and specificity were 86% and 100% respectively. Spearman correlation showed a correlation between LIV and tumour grade (for all gliomas r = 0.53 and p < 0.04, for astrocytomas r = 0.84 and p < 0.01). A correlation was also found between ITSS and tumour grade (for all gliomas r = 0.69 and p < 0.01, for astrocytomas r = 0.63 and p < 0.04). The results indicate that SWI is useful for distinguishing between high and low grade astrocytoma with 1.5T imaging within this cohort. It also seems possible to distinguish between high and low grade glioma with ITSS.

EGFR TKIs are standard therapy for advanced NSCLC. In order to define their role in early disease, we implemented a phase II trial of neoadjuvant gefitinib in clinical stage I NSCLC. Tumour shrinkage was seen in 43% of patients, with 11% achieving RECIST partial response (PR). Analysis of molecular markers showed EGFR TKD mutations in 17% of cases, being the only associated with PR. For the first time we defined the histopathological response of NSCLC to these agents, characterized by reduction in tumour cellularity and proliferative index as well as presence of non-mucinous BAC histology. Clinical PR tumours also presented large areas of stromal fibrosis with presence of focal residual tumour. In a characterization of intracellular signalling response, EGFR dephosphorylation in the residues Y1068 and Y1173 was not concordant and only the former was significantly reduced. pAkt Ser473/Akt and Thr308/Akt ratios were significantly reduced but observed among both, clinical responders and resistant patients. Interestingly, reduction in pEGFR Y1068 was significantly associated with increase in tumour cellularity (p=0.047), Ki-67 index (p=0.018) and tumour growth (p=0.019) with a residual perinuclear localization been detected, suggesting a novel mechanism of resistance involving receptor internalization. Finally, we determined that the EGFR protein remains stable up to one hour of post resection ischemia but two to three tumour samples are necessary for an adequate tumour representation. Furthermore, EGFR cytoplasmic compartment presented the best association with clinical response in our cohort. Taking all together, we were able to generate the first clinical trial exploring the use of an EGFR TKI in early NSCLC, characterizing for the first time the histopathological and signalling responses to these agents with an evidence of a potential novel mechanism of resistance. Finally, we observed that multiple samples collection for an adequate tumour representation, and assessment of the cytoplasmic compartment, are warrant.

EGFR TKIs are standard therapy for advanced NSCLC. In order to define their role in early disease, we implemented a phase II trial of neoadjuvant gefitinib in clinical stage I NSCLC. Tumour shrinkage was seen in 43% of patients, with 11% achieving RECIST partial response (PR). Analysis of molecular markers showed EGFR TKD mutations in 17% of cases, being the only associated with PR. For the first time we defined the histopathological response of NSCLC to these agents, characterized by reduction in tumour cellularity and proliferative index as well as presence of non-mucinous BAC histology. Clinical PR tumours also presented large areas of stromal fibrosis with presence of focal residual tumour. In a characterization of intracellular signalling response, EGFR dephosphorylation in the residues Y1068 and Y1173 was not concordant and only the former was significantly reduced. pAkt Ser473/Akt and Thr308/Akt ratios were significantly reduced but observed among both, clinical responders and resistant patients. Interestingly, reduction in pEGFR Y1068 was significantly associated with increase in tumour cellularity (p=0.047), Ki-67 index (p=0.018) and tumour growth (p=0.019) with a residual perinuclear localization been detected, suggesting a novel mechanism of resistance involving receptor internalization. Finally, we determined that the EGFR protein remains stable up to one hour of post resection ischemia but two to three tumour samples are necessary for an adequate tumour representation. Furthermore, EGFR cytoplasmic compartment presented the best association with clinical response in our cohort. Taking all together, we were able to generate the first clinical trial exploring the use of an EGFR TKI in early NSCLC, characterizing for the first time the histopathological and signalling responses to these agents with an evidence of a potential novel mechanism of resistance. Finally, we observed that multiple samples collection for an adequate tumour representation, and assessment of the cytoplasmic compartment, are warrant.