Patient characteristics.

<p>Patient characteristics.</p

MRI scans before (a-e) and under (f-j) therapy.

<p><b>a, f:</b> Reduced leptomeningeal enhancement (white arrows) after 8 weeks of therapy with bevacizumab and lomustine in patient 3. <b>b, g:</b> Regression of leptomeningeal contrast-enhancing nodule (white arrow) on the septum pellucidum on T1-weighted images after eight weeks of therapy with bevacizumab and temozolomide in patient 4. <b>c, h:</b> This regression (black arrow) in patient 3 was also visible on T2-weighted images, which makes pure pseudoresponse unlikely. <b>d, i:</b> Regression of leptomeningeal contrast-enhancing nodules (white arrowheads) on the surface of the medullar conus and the lumbar nerve roots on T1 weighted images (Th10-L2) in patient 9 before and after radiotherapy plus eight weeks of therapy with bevacizumab and lomustine. <b>e, j:</b> This regression of contrast-enhancement (white arrowheads) in patient 9 was also apparent in the thoracic spine (Th5-Th9) which was not treated with radiotherapy.</p

Treatment and outcome.

<p>Treatment and outcome.</p

Model spectra.

<p>Graphical user interface on T2-weighted MR image with two small white boxes indicating an example voxel for tumor (left) and control tissue (right) within the measured area (large white box) (A). Corresponding T1-weighted contrast enhanced MR image (B).Corresponding spectra for the marked tumor (C) and control tissue voxels (D). The original signal is shown in black, the LCModel fit in red and the isolated signal for myoinsitol in blue. A * marks the creatine peaks.</p

Kaplan-Meier-Curves based on delta MI.

<p>Kaplan-Meier-Curves for patients with delta MI values > 1.817 (OS 275 days, color: red) compared to patients with values < 1.817 (OS 164 days, color: brown). The figure shows an extract with an OS of 1829 days for the remaining patient of the brown cohort and a censored survival of 1424 days for the one remaining patient in the red cohort. MI = Myoinositol; OS = overall survival; delta MI = MI control tissue minus tumor concentrations at baseline.</p

Box-Whisker-plots.

<p>(A) Box-Whisker-plot showing the median (baseline CCNU/VM26 = 2.73, BVZ = 2.47; post treatment CCNU/VM26 = 2.38, BVZ = 3.61) and quartiles for tumor at baseline and post treatment for BVZ (orange) and CCNU/VM26 (blue). (B) Box-Whisker-plot showing the median (baseline CCNU/VM26 = 4.83, BVZ = 4.97; post treatment CCNU/VM26 = 4.83, BVZ = 6.08) and quartiles for the control tissue at baseline and post treatment for BVZ (orange) and CCNU/VM26 (blue). Each * marks a significant increase from baseline to post treatment.</p

Kaplan-Meier-Curves based on control tissue MI.

<p>Kaplan-Meier-Curves for patients with a MI concentration in the control tissue at baseline > 4.696 (OS 270 days, color: red) compared to patients with a MI concentration < 4.696 (median overall survival (OS 206 days, color: brown).The figure shows an extract with an OS of 1829 days for the remaining patient of the brown cohort and a censored survival of 1424 days for the remaining patient in the red cohort. MI = Myoinositol; OS = overall survival; delta MI = MI control tissue minus tumor concentrations at baseline.</p

Patient characteristics.

<p>Patient characteristics.</p

Overview of essential intracellular members of the mTORC1-pathway and respective targeted therapies.

<p>Signaling from growth factor receptors such as EGFR or a nutrient rich environment leads to an activation of the mTOR-pathway, while nutrient depletion inhibits signaling. The mTORC1-complex consists of the mTOR-protein itself and the subunits raptor (regulatory associated protein of mTOR), mLST8 (mammalian lethal with sec thirteen 8) and PRAS40 (proline-rich Akt1 substrate 1). Activation of the pathway leads to phosphorylation of the ribosomal protein S6 at phospho-sites Ser235/236 and Ser240/244 by the S6-kinase (S6K). Additionally, the translational repressor 4EBP1 (eukaryotic initiation factor 4E binding protein-1) is phosporylated at Thr37/Thr46, thereby relieving its translational inhibition and promoting translation especially of mRNAs with a TOP motif [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127123#pone.0127123.ref014" target="_blank">14</a>]. First generation mTORC1-inhibitors are rapamycin and its derivatives temsirolimus and everolimus. Mechanistically these compounds bind the intracellular adaptor protein FKBP12 to form an allosteric mTORC1 inhibitory complex. 4EBP1 phosphorylation though mTORC1-dependent is largely resistant to the mTORC1 inhibitor rapamycin and its derivatives. Recently, ATP-competitive mTORC (2^nd generation) inhibitors like torin1 and torin2 which are efficient in dephosphorylating 4EBP1 have been developed [<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0127123#pone.0127123.ref015" target="_blank">15</a>].</p

Blocking lysosomal acidification or inhibiting cathepsin B rescues glioma cells from B10 cytotoxicity under hypoxia.

<p>To block lysosomal acidification we used bafilomycin A1 (BafA1). A, LN-308 or LNT-229 cells were co-treated with 10 nM BafA1 and 10 µM B10 under hypoxic conditions (1% O₂) for 48 h. Cell density was assessed by CV (n = 4, SD, *p<0.05). B, cell death was quantified by LDH-release after treating LN-308 or LNT-229 cells with BafA1 and B10 under hypoxic conditions (0.1% O₂) for 19 h (LN-308) or 27 h (LNT-229) (n = 4, SD, * = p<0.05). To inhibit cathepsin B we used the cathepsin inhibitor CA074-Me. C, LN-308 or LNT-229 cells were co-treated with 10 µM CA074-Me and 10 µM B10 under hypoxic conditions (1% O₂) for 48 h. Cell density was assessed by CV (n = 4, SD, *p<0.05). D, cell death was quantified by LDH release after treating LN-308 or LNT-229 cells with CA074-Me and B10 under hypoxic conditions (0.1% O₂) for 19 h (LN-308) or 25 h (LNT-229) (n = 4, SD, *p<0.05).</p