Surface-immobilized aptamers for cancer cell isolation and microscopic cytology.
Exposing rare but highly malignant tumor cells that migrate from the primary tumor mass into adjacent tissue(s) or circulate in the bloodstream is critical for early detection and effective intervention(s). Here, we report on an aptamer-based strategy directed against epidermal growth factor receptor (EGFR), the most common oncogene in glioblastoma (GBM), to detect these deadly tumor cells. GBMs are characterized by diffuse infiltration into normal brain regions, and the inability to detect GBM cells renders the disease surgically incurable with a median survival of just 14.2 months. To test the sensitivity and specificity of our platform, anti-EGFR RNA aptamers were immobilized on chemically modified glass surfaces. Cells tested included primary human GBM cells expressing high levels of the wild-type EGFR, as well as genetically engineered murine glioma cells overexpressing the most common EGFR mutant (EGFRvIII lacking exons 2-7) in Ink4a/Arf-deficient astrocytes. We found that surfaces functionalized with anti-EGFR aptamers could capture both the human and murine GBM cells with high sensitivity and specificity. Our findings show how novel aptamer substrates could be used to determine whether surgical resection margins are free of tumor cells, or more widely for detecting tumor cells circulating in peripheral blood to improve early detection and/or monitoring residual disease after treatment.
Department of Bioengineering, Nanotechnology Research and Teaching Facility, [corrected] University of Texas at Arlington, Arlington, Texas 76019, USA.
Yuan
Y
Wan
Young-tae
YT
Kim
Na
N
Li
Steve K
SK
Cho
Robert
R
Bachoo
Andrew D
AD
Ellington
Samir M
SM
Iqbal
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D052157:Aptamers, Nucleotide / Q000235:genetics* / Q000378:metabolism; D001253:Astrocytes / Q000378:metabolism; D001483:Base Sequence; D001667:Binding, Competitive; D001932:Brain Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D066246:ErbB Receptors / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005909:Glioblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D008856:Microscopy, Fluorescence / Q000379:methods; D008969:Molecular Sequence Data; D009154:Mutation; D009474:Neurons / Q000378:metabolism; D016372:RNA, Antisense / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051381:Rats; D013234:Stem Cells / Q000378:metabolism; D013499:Surface Properties; D014407:Tumor Cells, Cultured
D000906:Antibodies; D052157:Aptamers, Nucleotide; D019941:Cyclin-Dependent Kinase Inhibitor p16; D016372:RNA, Antisense; D066246:ErbB Receptors