Glioblastomas shed large quantities of small, membrane-bound microvesicles into the circulation. Although these hold promise as potential biomarkers of therapeutic response, their identification and quantification remain challenging. Here, we describe a highly sensitive and rapid analytical technique for profiling circulating microvesicles directly from blood samples of patients with glioblastoma. Microvesicles, introduced onto a dedicated microfluidic chip, are labeled with target-specific magnetic nanoparticles and detected by a miniaturized nuclear magnetic resonance system. Compared with current methods, this integrated system has a much higher detection sensitivity and can differentiate glioblastoma multiforme (GBM) microvesicles from nontumor host cell–derived microvesicles. We also show that circulating GBM microvesicles can be used to analyze primary tumor mutations and as a predictive metric of treatment-induced changes. This platform could provide both an early indicator of drug efficacy and a potential molecular stratifier for human clinical trials.
At a glance
- Detection of mutations in EGFR in circulating lung-cancer cells. N. Engl. J. Med. 359, 366–377 (2008). et al.
- Integrating high-throughput technologies in the quest for effective biomarkers for ovarian cancer. Nat. Rev. Cancer 10, 371–378 (2010). , &
- Exosomes: proteomic insights and diagnostic potential. Expert Rev. Proteomics 6, 267–283 (2009). , , &
- Membrane vesicles as conveyors of immune responses. Nat. Rev. Immunol. 9, 581–593 (2009). , &
- Glioblastoma microvesicles transport RNA and proteins that promote tumour growth and provide diagnostic biomarkers. Nat. Cell Biol. 10, 1470–1476 (2008). et al.
- Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles derived from tumour cells. Nat. Cell Biol. 10, 619–624 (2008). et al.
- Exosomes—vesicular carriers for intercellular communication. Curr. Opin. Cell Biol. 21, 575–581 (2009). &
- Microvesicles as mediators of intercellular communication in cancer—the emerging science of cellular 'debris'. Semin. Immunopathol. 33, 455–467 (2011). et al.
- Shedding microvesicles: artefacts no more. Trends Cell Biol. 19, 43–51 (2009). , &
- Proteomic and immunologic analyses of brain tumor exosomes. FASEB J. 23, 1541–1557 (2009). et al.
- Generation of novel, secreted epidermal growth factor receptor (EGFR/ErbB1) isoforms via metalloprotease-dependent ectodomain shedding and exosome secretion. J. Cell. Biochem. 103, 1783–1797 (2008). et al.
- Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences. Nat. Commun. 2, 180 (2011). et al.
- Chip-NMR biosensor for detection and molecular analysis of cells. Nat. Med. 14, 869–874 (2008). , , &
- Miniature magnetic resonance system for point-of-care diagnostics. Lab Chip 11, 2282–2287 (2011). et al.
- Micro-NMR for rapid molecular analysis of human tumor samples. Sci. Transl. Med. 3, 71ra16 (2011). et al.
- Exosomes: composition, biogenesis and function. Nat. Rev. Immunol. 2, 569–579 (2002). , &
- Bioorthogonal chemistry amplifies nanoparticle binding and enhances the sensitivity of cell detection. Nat. Nanotechnol. 5, 660–665 (2010). , , , &
- Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr. Protoc. Cell Biol. 30, 3.22 (2006). , , &
- Amplification and/or overexpression of platelet-derived growth factor receptors and epidermal growth factor receptor in human glial tumors. Cancer Res. 52, 4550–4553 (1992). et al.
- Increased expression of podoplanin in malignant astrocytic tumors as a novel molecular marker of malignant progression. Acta Neuropathol. 111, 483–488 (2006). et al.
- EphA2 as a novel molecular marker and target in glioblastoma multiforme. Mol. Cancer Res. 3, 541–551 (2005). , , &
- An integrated genomic analysis of human glioblastoma multiforme. Science 321, 1807–1812 (2008). et al.
- Cancer-associated IDH1 mutations produce 2-hydroxyglutarate. Nature 462, 739–744 (2009). et al.
- Clinical relevance of microparticles from platelets and megakaryocytes. Curr. Opin. Hematol. 17, 578–584 (2010). , &
- Expression of epidermal growth factor receptor or ErbB3 facilitates geldanamycin-induced down-regulation of ErbB2. Mol. Cancer Res. 7, 275–284 (2009). et al.
- Targeting the dynamic HSP90 complex in cancer. Nat. Rev. Cancer 10, 537–549 (2010). , , &
- The novel Hsp90 inhibitor NXD30001 induces tumor regression in a genetically engineered mouse model of glioblastoma multiforme. Mol. Cancer Ther. 9, 2618–2626 (2010). et al.
- Oncogenic EGFR signaling cooperates with loss of tumor suppressor gene functions in gliomagenesis. Proc. Natl. Acad. Sci. USA 106, 2712–2716 (2009). et al.
- Cancer Genome Atlas Research Network. Comprehensive genomic characterization defines human glioblastoma genes and core pathways. Nature 455, 1061–1068 (2008).
- Integrated genomic analysis identifies clinically relevant subtypes of glioblastoma characterized by abnormalities in PDGFRA, IDH1, EGFR, and NF1. Cancer Cell 17, 98–110 (2010). et al.
- The type III epidermal growth factor receptor mutation. Biological significance and potential target for anti-cancer therapy. Ann. Oncol. 12, 745–760 (2001). , , &
- Molecular determinants of the response of glioblastomas to EGFR kinase inhibitors. N. Engl. J. Med. 353, 2012–2024 (2005). et al.
- The parametric response map is an imaging biomarker for early cancer treatment outcome. Nat. Med. 15, 572–576 (2009). et al.
- Prospective analysis of parametric MRI biomarkers: Identification of early and distinct glioma response patterns not predicted by standard radiographic assessment. Clin. Cancer Res. 17, 4751–4760 (2011). et al.
- Parametric response map as an imaging biomarker to distinguish progression from pseudoprogression in high-grade glioma. J. Clin. Oncol. 28, 2293–2299 (2010). et al.
- Magnetic resonance of 2-hydroxyglutarate in IDH1-Mutated low-grade gliomas. Sci. Transl. Med. 4, 116ra5 (2012). et al.
- Changes in tumor metabolism as readout for mammalian target of rapamycin kinase inhibition by rapamycin in glioblastoma. Clin. Cancer Res. 14, 3416–3426 (2008). et al.
- Highly magnetic core-shell nanoparticles with a unique magnetization mechanism. Angew. Chem. Int. Edn Engl. 50, 4663–4666 (2011). , , &
- Supplementary Text and Figures (3M)
Supplementary Methods, Supplementary Figures 1–8 and Supplementary Tables 1–4