The Cancer Genome Atlas Analysis Predicts MicroRNA for Targeting Cancer Growth and Vascularization in Glioblastoma

Abstract

Using in silico analysis of The Cancer Genome Atlas (TCGA), we identified microRNAs associated with glioblastoma (GBM) survival, and predicted their functions in glioma growth and progression. Inhibition of two "risky" miRNAs, miR-148a and miR-31, in orthotopic xenograft GBM mouse models suppressed tumor growth and thereby prolonged animal survival. Intracranial tumors treated with uncomplexed miR-148a and miR-31 antagomirs exhibited reduced proliferation, stem cell depletion, and normalized tumor vasculature. Growth-promoting functions of these two miRNAs were, in part, mediated by the common target, the factor inhibiting hypoxia-inducible factor 1 (FIH1), and the downstream pathways involving hypoxia-inducible factor HIF1α and Notch signaling. Therefore, miR-31 and miR-148a regulate glioma growth by maintaining tumor stem cells and their niche, and providing the tumor a way to activate angiogenesis even in a normoxic environment. This is the first study that demonstrates intratumoral uptake and growth-inhibiting effects of uncomplexed antagomirs in orthotopic glioma.

Genome Editing Reveals Glioblastoma Addiction to MicroRNA-10b

“…RNA isolation, qRT-PCR, and protein analysis by western blotting were performed as previously described. 34 Cell viability was assessed using WST1 (Roche), according to the manufacturer's instructions, 2 days post-transfection for the monolayer cultures and 5 days post-transfection for neurospheres. Wound healing assay was performed as described.…”

“…34 When bioluminescence reached the exponential phase with signal of 10 6 photons/s (10 days after LN229 and 19 days after GBM8 implantation), the lentiviral CRISPR-Cas9 constructs (3 Â 10 5 TU)…”

The role of micro-RNA in the regulation of signal pathways in gliomas

“…Глобальный экспрессионный анализ показывает, что активация EGFR и инактивация PTEN (phosphatase and tensin homolog) индуцируют экспрессию miR-146a, представляя собой механизм отрицательной обратной связи, сдерживающий опухолевый рост [38]. Для глиобластом, обработанных ингибиторами miR-148a и miR-31, отмечено снижение темпов пролиферации, стволовых свойств опухолевых клеток, нормализация сосудистой системы, что частично опосредовано общей мишенью данных микро-РНК -FIH1 (factor inhibiting hypoxiainducible factor 1) и последующего сигналинга HIF1a (hypoxia inducible factor 1 alpha subunit) и Notch [39]. Показано увеличение экспрессии miR-148a при глиобластоме [37].…”

MicroRNAs in glioblastoma multiforme pathogenesis and therapeutics

“…66 found that miRNAs can also regulate GBM growth through maintenance of tumor stem cells and stem cell niches. Inhibition of miR‐148a and miR‐31 using antisense oligonucleotides was able to reduce cancer cell proliferation, deplete stem cells, and normalize tumor vasculature.…”

“…Inhibition of miR‐148a and miR‐31 using antisense oligonucleotides was able to reduce cancer cell proliferation, deplete stem cells, and normalize tumor vasculature. These effects were mediated in part through miR‐148a and miR‐31 repression of factor‐inhibiting hypoxia‐inducible factor 1 (FIH1), which can influence angiogenesis and tumor stemness through HIF1 α and Notch signaling 66. Thus, their inhibition was able to effectively suppress tumor growth and prolong survival time in orthotopic xenograft GBM mouse models.…”