The effect of unrestricted cycling KD on patient-derived IDH1^wt and IDH1^mut flank xenograft growth.

Tumor volumes for mice subcutaneously engrafted with IDH1mut GBM164 at 21 days, IDH1mut GBM 196 at 14 days, and IDH1wt GBM12 at 14 days, either on SD or cycling KD (initiated 3 days after engraftment). Time points differed due to differing rates of tumor growth. Bars = means, P values calculated by unpaired t-tests.</p

Effect of ketones on patient-derived IDH1^wt and IDH1^mut tumor cell proliferation <i>in vitro</i>.

Absolute cell counts over 14 days of culture in either normal glucose (NG) or low glucose (LG), with or without 10 mM β-hydroxybutyrate (BHB). Each data point is shown as mean ±SEM. Two-way ANOVA analyses of the final time points are described in the Results text and Table 3.</p

Two-way ANOVA analyses of <i>in vitro</i> proliferation of IDH^wt and IDH^mut tumor cells in varying glucose and ketogenic culture conditions.

Two-way ANOVA analyses of in vitro proliferation of IDHwt and IDHmut tumor cells in varying glucose and ketogenic culture conditions.</p

Summary of cell lines used.

Summary of cell lines used.</p

Evaluation of brain delivery of Evans Blue (EB) by various peptides (A) and by different concentration of peptide K16ApoE (B).

<p>67.5 picomoles of each of the peptides (K16, ApoE and K16ApoE) was either injected first followed in 10 min by injection of EB (40 ul of a 2% solution) or the dye and the peptides were mixed together and then injected. Mice were perfused with saline 2 h after injection and then brains were collected for visualization.</p

Basic metabolic parameters of mice on an unrestricted cycling KD.

Saphenous vein blood samples tested for glucose (A) and ketones (B) at the end of each week. Mouse body weights are shown in (C). Green bars indicate weeks in which mice were on KD.</p

K16ApoE-mediated brain delivery of blue (EB), red (Crocein Scarlet) and green (Light Green SF) dyes to the brain.

<p>Three different approaches were assessed for dye delivery: 1. K16ApoE was injected first then a given dye was injected 10 min after (second columns of brain specimens); 2. K16ApoE was mixed with 300 ug of cetuximab and injected followed by injection of a given dye 10 min after 3^rd column of brain specimens), and 3. K16ApoE and the dyes were mixed and injected (fourth column of brain specimens). The first column of brain specimens represents animals receiving injection of a given dye alone. Mice were perfused with saline 2 h after injection and then brains were collected for visualization. 67.5 picomole of K16ApoE was used in each experiment. 40 ul of a 2% solution of each of the dyes were used for injection into a 20 g mouse (amount of dye injected varied accordingly with weight of mice).</p

Summary of macronutrient composition of standard and experimental diets presented as proportion of total kilocalories.

Summary of macronutrient composition of standard and experimental diets presented as proportion of total kilocalories.</p

Quantification of K16ApoE-mediated brain-uptake of cisplatin (Cp) and methotrexate (MTX).

<p>300 ug of the carrier peptide K16ApoE, 300 ug of cetuximab and 300 ug of cisplatin (Cp) were used in this experiment. Group 1- these animals received only Cp or MTX. Group 2- these animals received injection of K16ApoE then injection of either Cp or MTX. Group 3- these animals received an injection of K16ApoE mixed with cetuximab, followed by an injection of Cp or MTX. Group 4- these animals received an injection of K16ApoE mixed with Cp or MTX. Post-perfused brains were collected after 1 h of final injection and processed for respective assays. Fold change for Group 2 has been obtained by dividing the mean value for Group 2 by the mean value for group 1; fold change for Group 3 has been obtained by dividing the mean value for this group by the mean value of Group 1, and so on. ‘% delivery’ indicates the fraction of Cp or MTX in brain compared to the injected dose. Six animals in each group have been used.</p

Brain imaging and quantification (via microSPECT) of brain-uptake of I-125 via insulin injection.

<p>I-125 was injected 10 min after injection of 250 ug, 500 ug and 1000 ug of insulin, respectively, and 200 ug of K16ApoE. Quantification of I-125 in the brain was done after cardiac perfusion. Six mice were evaluated for each group.</p