Tumour-associated glial host cells display a stem-like phenotype with a distinct gene expression profile and promote growth of GBM xenografts.

BACKGROUND: Little is known about the role of glial host cells in brain tumours. However, supporting stromal cells have been shown to foster tumour growth in other cancers. METHODS: We isolated stromal cells from patient-derived glioblastoma (GBM) xenografts established in GFP-NOD/scid mice. With simultaneous removal of CD11b+ immune and CD31+ endothelial cells by fluorescence activated cell sorting (FACS), we obtained a population of tumour-associated glial cells, TAGs, expressing markers of terminally differentiaed glial cell types or glial progenitors. This cell population was subsequently characterised using gene expression analyses and immunocytochemistry. Furthermore, sphere formation was assessed in vitro and their glioma growth-promoting ability was examined in vivo. Finally, the expression of TAG related markers was validated in human GBMs. RESULTS: TAGs were highly enriched for the expression of glial cell proteins including GFAP and myelin basic protein (MBP), and immature markers such as Nestin and O4. A fraction of TAGs displayed sphere formation in stem cell medium. Moreover, TAGs promoted brain tumour growth in vivo when co-implanted with glioma cells, compared to implanting only glioma cells, or glioma cells and unconditioned glial cells from mice without tumours. Genome-wide microarray analysis of TAGs showed an expression profile distinct from glial cells from healthy mice brains. Notably, TAGs upregulated genes associated with immature cell types and self-renewal, including Pou3f2 and Sox2. In addition, TAGs from highly angiogenic tumours showed upregulation of angiogenic factors, including Vegf and Angiopoietin 2. Immunohistochemistry of three GBMs, two patient biopsies and one GBM xenograft, confirmed that the expression of these genes was mainly confined to TAGs in the tumour bed. Furthermore, their expression profiles displayed a significant overlap with gene clusters defining prognostic subclasses of human GBMs. CONCLUSIONS: Our data demonstrate that glial host cells in brain tumours are functionally distinct from glial cells of healthy mice brains. Furthermore, TAGs display a gene expression profile with enrichment for genes related to stem cells, immature cell types and developmental processes. Future studies are needed to delineate the biological mechanisms regulating the brain tumour-host interplay.

Establishment of a novel dsRed NOD/Scid mouse strain to investigate the host and tumor cell compartments.

Here we describe a NOD/Scid mouse strain expressing the dsRed transgene. The strain is maintained by inbreeding of homozygous dsRed NOD/Scid siblings, and expresses red fluorescence from various organs. The model allows engraftment of human tumor tissue, and engrafted tumors were separated into stromal and malignant cell compartments. Furthermore, we compared tumor-associated and normal fibroblast for expression of fibroblast-associated markers, and identified a marker panel that was upregulated in the tumor-associated fibroblasts. In conclusion, we propose that this model may be used in a variety of studies of tumor progression and to elucidate the role of the tumor microenvironment.

Neopterin: a promising marker for the inflammation in polycystic ovary syndrome.

BACKGROUND: Several markers of low-grade chronic inflammation are altered in women with polycystic ovary syndrome (PCOS). Neopterin (NEO) is a marker of celullar immunity, and oxidative stress, mainly produced by activated macrophages. We aimed to evaluate the NEO levels in PCOS patients and correlate them with antropometric and biochemical parameters. METHODS: The study groups consisted of 69 women with PCOS and 46 healthy controls. Both groups were divided into two subgroups according to their body mass index (BMI): <25 = normoweight, >25 = overweight. The clinical and biochemical parameters and serum NEO levels were analyzed. RESULTS: Circulating levels of NEO were significantly (p < 0.001) higher in women with PCOS (normoweight: 15.9 ± 4.7 nmol/l; overweight 13.3 ± 8.1 nmol/l) compared to controls (normoweight: 8.6 ± 2.0 nmol/l; overweight 9.2 ± 1.8 nmol/l) regardless of their weight classes. Waist-to-hip ratio (WHR) (p < 0.05), free and total testosterone (p < 0.001) were significantly elevated in women with PCOS compared to controls after controlling for the effect of obesity. CONCLUSION: Circulating NEO level s are elevated in PCOS independent of body mass index supporting the suggestion of PCOS is a low-grade chronic inflammatory state.

The outcome of adding peripheral neuromodulation (Stoller afferent neuro-stimulation) to anti-muscarinic therapy in women with severe overactive bladder.

OBJECTIVE: Anti-muscarinic treatment alone and peripheral neuromodulation with concomitant anti-muscarinic treatment were compared in patients with severe overactive bladder. METHODS: In this prospective study, 40 women with severe overactive bladder according to the 7-day voiding diary without any prior treatment completed the Incontinence Impact Questionnaire (IIQ-7) and were randomised into anti-muscarinic-alone and combination treatment groups. Twenty women received daily 4 mgs of tolterodine orally and in 20 women Stoller afferent neuro-stimulation (SANS) therapy was performed concomitantly for 12 weeks to the same anti-muscarinic regimen. After 12 weeks of therapy, two of the patients drop out of the study and remaining patients filled out the IIQ-7 questionnaire and the 7-day voiding diary again. Pretreatment and post-treatment QoL scores and the 7-day voiding diaries were compared. Mann-Whitney U, Wilcoxon and two sided significance tests were used. RESULTS: Thirty-eight women fulfilling the criteria were included in the study. Severity of overactive bladder symptoms decreased significantly in both treatment groups. However, the decrease in combination treatment group was more significant than the anti-muscarinic-alone group. Adverse events were similar between the two groups. CONCLUSION: Combining SANS and anti-muscarinic therapy resulted in significantly better clinical outcomes and IIQ-7 scores as compared with anti-muscarinic treatment alone in patients with severe overactive bladder.

Membrane-Depolarizing Channel Blockers Induce Selective Glioma Cell Death by Impairing Nutrient Transport and Unfolded Protein/Amino Acid Responses.

Glioma-initiating cells (GIC) are considered the underlying cause of recurrences of aggressive glioblastomas, replenishing the tumor population and undermining the efficacy of conventional chemotherapy. Here we report the discovery that inhibiting T-type voltage-gated Ca2+ and KCa channels can effectively induce selective cell death of GIC and increase host survival in an orthotopic mouse model of human glioma. At present, the precise cellular pathways affected by the drugs affecting these channels are unknown. However, using cell-based assays and integrated proteomics, phosphoproteomics, and transcriptomics analyses, we identified the downstream signaling events these drugs affect. Changes in plasma membrane depolarization and elevated intracellular Na+, which compromised Na+-dependent nutrient transport, were documented. Deficits in nutrient deficit acted in turn to trigger the unfolded protein response and the amino acid response, leading ultimately to nutrient starvation and GIC cell death. Our results suggest new therapeutic targets to attack aggressive gliomas. Cancer Res; 77(7); 1741-52. ©2017 AACR.