Figure S5. Body weight of mice xenografted with human primary GBM cells following treatment with combinations of radiotherapy and VX680 treatment.

Figure S5: JAK1/2 inhibitors block a transient STAT3 activation following IR in cultured murine proneural HGG cells.

Figure S4: Murine and human proneural HGG cells exposed to IR in vitro display reduced cell stiffness, reduced proliferation and increased TMZ resistance.

Compression-induced changes in the transcriptional GBM phenotype. Differentially expressed genes in tumorspheres derived from human GBMs (GBM5, GBM14, GBM34, and GBM43) when comparing compression to other conditions.

<p>Figure S1: Expression of mesenchymal genes, Pdpn and Chi3l1, in proneural and mesenchymal murine glioma models.</p>

Fig. S2. Interstitial fluid pressure (IFP) pressure sensors do not increase reactive gliosis or microglia activation during tumor progression. (A, B) GFAP+ reactive astrocytes (DAB; brown) and (C, D) Iba1+ tumor-associated macrophages (DAB; brown) in progressing GBM43 tumors in the absence or presence of IFP sensor. Nuclei are counterstained with hematoxylin (blue). Scale bar: 40 μm.

Figure S2. Expression of potential targets of VX680 in human GBM cultures.

Fig. S3. AF treatment increases drug uptake resulting in increased survival of GBM43 xenografted mice. (A, B) SPC diet increased uptake and reduced clearance of tail-vein injected gadolinium in mice xenografted with GBM43 cells. (C) Treatment regimen with SPC diet and/or Erlotinib for uptake and survival studies in GBM43 xenografts. (D) SPC treatment increased Erlotinib levels in GBM43 tumors 60 min following a single administration of Erlotinib. (n=4 for control and SPC, *P < 0.05; Student's t-test). (E) Kaplan-Meier (n=8 for all groups, **P < 0.01; Log-rank Mantel-Cox test or One way ANOVA) curve increased survival following SPC and Erlotinib treatment versus Erlotinib alone.

Figure S4. Sequential treatment with radiotherapy and the pan-AURK inhibitor VX680 to target TPCs in human primary GBM43 cultures.

Fig. S1. AF treatment increases ADC in tumor tissues. MR imaging showing an increase of (A) ADC in xenografted mice treated with nasal AF or SPC-diet with corresponding (B) T2w images.

<p>Figure S3: IR induces a sustained cell-intrinsic mesenchymal transition in HGG cell cultures.</p>

Fig. S6. AF peptide have no proliferative effect on pNKCC1 expressing human GBM cells under atmospheric pressure. (A) Adherent human GBM cells show immunoreactivity against an antibody recognizing Thr-phosphorylation site on human NKCC1. (B) AF treatment had no proliferative effects on human GBM5 cells grown under atmospheric pressure.

Fig. S7. AF reversed compression-induced transcriptional phenotype of GBM tumorspheres. Both (A-B) smaller and (C-D) larger GBM43 tumorspheres expanded in response to 48h compression. (***P < 0.05, Student's t-test). (E) Principle components analysis of all expressed genes. The first principle component is associated with GBM subtype, while the second principle component is associated with the presence of compression. (F) Heat map of genes differentially expressed when comparing compression to other conditions (see supplementary table S1 for gene list).

Fig. S5. AF prevents restoration of cell volume of GC22 cells under hyperosmolar conditions. (A) AF peptide (5 μg/ml) and bumetanide (BMT, 10 μM) reduced fluorescence of excited calcein-AM dye when GC22 cells were transferred to hypertonic conditions (432 mOsm). Scale bar: 10 μm. (B) Cell shrinkage was induced with hypertonic exposure in both BMT and AF peptide treated cells with no regulatory volume increase recorded within 20 minutes of hypertonic exposure. (C) Quantification of regulatory volume increase data. (n=3, *P < 0.05, one-way ANOVA).

Figure S3. Inhibition of phosphorylated histone H3 following incubation with the pan-AURK inhibitor VX680 in human primary GBM43 cultures.

Fig. S4. AF treatment results in no changes in body weight for mice xenografted with GBM43 cells and resulted in complete tumor regression in an EGFRvIII-expressing GBM allograft model. (A, B) SPC or Salovum treatment had no effect on body weight in GBM43 xenografts (n=7 for control, n=10 for SPC, and n=5 for Salovum). (C) SPC treatment resulted in complete tumor regression in 5/6 FVBn mice allografted with EGFRvIII and mCherry-expressing mouse GBM cells (dotted line; tumor border). Scale bar 2 mm.

<p>Figure S2: Fractionated doses of radiation extend survival and induce a PMT in proneural HGG murine model.</p>

Kesebolite-(Ce), ideal formula CeCa2Mn(AsO4)[SiO3]3, is a new mineral (IMA No. 2019-097) recovered from mine dumps at the Kesebol Mn-(Fe-Cu) deposit in Västra Götaland, Sweden. It occurs with rhodonite, baryte, quartz, calcite, talc, andradite, rhodochrosite, K-feldspar, hematite, gasparite-(Ce), chernovite-(Y) and ferriakasakaite-(Ce). It forms mostly euhedral crystals, with lengthwise striation. The mineral is dark grayish-brown to brown, translucent, with light brown streak. It is optically biaxial (+), with weak pleochroism, and ncalc = 1.74. H = 5–6 and VHN100 = 825. Fair cleavage is observed on {100}. The calculated density is 3.998(5) g·cm−3. Kesebolite-(Ce) is monoclinic, P21/c, with unit-cell parameters from X-ray single-crystal diffraction data: a = 6.7382(3), b = 13.0368(6), c = 12.0958(6) Å, β = 98.578(2)°, and V = 1050.66(9) Å3, with Z = 4. Strongest Bragg peaks in the X-ray powder pattern are: [I(%), d(Å) (hkl)] 100, 3.114 (20-2); 92, 2.924 (140); 84, 3.138 (041); 72, 2.908 (014); 57, 3.228 (210); 48, 2.856 (042); 48, 3.002 (132). The unique crystal structure was solved and refined to R1 = 4.6%. It consists of 6-periodic single silicate chains along (001); these are interconnected to infinite (010) strings of alternating, corner-sharing MnO6 and AsO4 polyhedra, altogether forming a trellis-like framework parallel to (100).

Abstract Background: Patients with type 1 diabetes mellitus (T1DM) have an increased risk of Addison's disease (AD) (1), and recognizing those at risk would be of great value. Objective: To determine if there are early clinical indicators that may denote the development AD in adult patients with T1DM. Methods: Nationwide, matched, observational cohort study linking data from Swedish national registries [among others National Diabetes Register (NDR), Inpatient Register and Prescribed Drug Register] with a coverage of >97%. Patients with T1DM who developed AD (cases) were matched to 5 control subjects with T1DM that did not develop AD. Clinical data (including co-morbidities) and drug prescriptions were assessed: a) prior to baseline (inclusion into the study), and b) 2 years prior to AD diagnosis. Analysis of covariance and estimated group proportions, with corresponding 95% confidence intervals (CI), were used for comparisons between groups. Results: Between 1998-2013 and among 36,514 adult patients with T1DM in NDR, 66 patients were diagnosed with concomitant AD. These cases were matched to 330 controls with T1DM. Prior to baseline, cases had higher proportion of prescription of thyroid/anti-thyroid drugs than controls (9.1% vs 1.8%). Prior to AD diagnosis, cases compared with controls had higher proportion of diabetic retinopathy (12.1% vs 2.1%), infections requiring hospital admission (16.7% vs 2.1%), higher frequency in prescription of thyroid/anti-thyroid drugs (28.8% vs 7.0%), and prescription of glucagon (18.2% vs 6.4%). No difference in HbA1c (glycated haemoglobin) was seen between the groups prior to baseline or prior to AD diagnosis. Conclusions: These population-based, nation-wide, real-world data suggest that medical treatment for thyroid disease, a severe infection and prescription of glucagon for hypoglycemia should raise the awareness of AD in adult patients with T1DM. Our data also suggest that the development of diabetic retinopathy could also be associated with glucocorticoid deficiency prior to the development of AD. Reference: (1) Chantzichristos D et al. Eur J Endocrinol. 2018;178(1):115-122. Disclosure statement: The authors have nothing to disclose. Funding: The National Diabetes Registry receives financial support from the Swedish Association of Local Authorities and Regions and the Region Västra Götaland. The study was financed by grants from the Swedish state under the agreement between the Swedish government and the county councils, the ALF-agreement (ALFGBG-719531) and the Swedish Research Council (2015-02561). The funders of the study had no role in study design, data collection, data analysis, data interpretation, or writing of the report.