title: Selective Targeting to Glioma with Nucleic Acid Aptamers.
abstract: Malignant glioma is characterised by a rapid growth rate and high capacity for invasive infiltration to surrounding brain tissue; hence, diagnosis and treatment is difficult and patient survival is poor. Aptamers contribute a promising and unique technology for the in vitro imaging of live cells and tissues, with a potentially bright future in clinical diagnostics and therapeutics for malignant glioma. The binding selectivity, uptake capacity and binding target of two DNA aptamers, SA43 and SA44, were investigated in glioma cells and patient tissues. The binding assay showed that SA43 and SA44 bound with strong affinity (Kd, 21.56 ± 4.60 nM and Kd, 21.11 ± 3.30 nM respectively) to the target U87MG cells. Quantitative analysis by flow cytometry showed that the aptamers were able to actively internalise in U87MG and 1321N1 glioma cells compared to the non-cancerous and non-glioma cell types. Confocal microscopy confirmed staining in the cytoplasm, and co-localisation studies with endoplasmic reticulum, Golgi apparatus and lysosomal markers suggested internalisation and compartmentalisation within the endomembrane system. Both aptamers selectively bound to Ku 70 and Ku 80 DNA repair proteins as determined by aptoprecipitation (AP) followed by mass spectrometry analysis and confirmation by Western blot. In addition, aptohistochemical (AHC) staining on paraffin embedded, formalin fixed patient tissues revealed that the binding selectivity was significantly higher for SA43 aptamer in glioma tissues (grade I, II, III and IV) compared to the non-cancerous tissues, whereas SA44 did not show selectivity towards glioma tissues. The results indicate that SA43 aptamer can differentiate between glioma and non-cancerous cells and tissues and therefore, shows promise for histological diagnosis of glioma.
authors.affiliation: School of Pharmacy and Biomedical Sciences, College of Clinical and Biomedical Sciences, University of Central Lancashire, Preston, Lancashire, PR1 2HE, United Kingdom.
authors.forename: Shraddha
authors.initials: S
authors.lastname: Aptekar
authors.affiliation: School of Pharmacy and Biomedical Sciences, College of Clinical and Biomedical Sciences, University of Central Lancashire, Preston, Lancashire, PR1 2HE, United Kingdom; Department of Neuropathology, Lancashire Teaching Hospitals (NHS trust), Preston, PR2 9HT, United Kingdom.
authors.forename: Mohit
authors.initials: M
authors.lastname: Arora
authors.affiliation: School of Pharmacy and Biomedical Sciences, College of Clinical and Biomedical Sciences, University of Central Lancashire, Preston, Lancashire, PR1 2HE, United Kingdom.
authors.forename: Clare Louise
authors.initials: CL
authors.lastname: Lawrence
authors.affiliation: School of Pharmacy and Biomedical Sciences, College of Clinical and Biomedical Sciences, University of Central Lancashire, Preston, Lancashire, PR1 2HE, United Kingdom.
authors.forename: Robert William
authors.initials: RW
authors.lastname: Lea
authors.affiliation: Department of Neuropathology, Lancashire Teaching Hospitals (NHS trust), Preston, PR2 9HT, United Kingdom.
authors.forename: Katherine
authors.initials: K
authors.lastname: Ashton
authors.affiliation: Department of Neuropathology, Lancashire Teaching Hospitals (NHS trust), Preston, PR2 9HT, United Kingdom.
authors.forename: Tim
authors.initials: T
authors.lastname: Dawson
authors.affiliation: School of Pharmacy and Biomedical Sciences, College of Clinical and Biomedical Sciences, University of Central Lancashire, Preston, Lancashire, PR1 2HE, United Kingdom.
authors.forename: Jane Elizabeth
authors.initials: JE
authors.lastname: Alder
authors.affiliation: School of Pharmacy and Biomedical Sciences, College of Clinical and Biomedical Sciences, University of Central Lancashire, Preston, Lancashire, PR1 2HE, United Kingdom.
authors.forename: Lisa
authors.initials: L
authors.lastname: Shaw
mesh_terms: D052157:Aptamers, Nucleotide / Q000737:chemistry / Q000378:metabolism*; D019921:Biotinylation; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D002470:Cell Survival; D011232:Chemical Precipitation; D005434:Flow Cytometry; D005910:Glioma / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007150:Immunohistochemistry; D018613:Microscopy, Confocal; D009363:Neoplasm Proteins / Q000378:metabolism; D009690:Nucleic Acid Conformation; D013347:Subcellular Fractions / Q000378:metabolism; D013696:Temperature
chemical_list: D052157:Aptamers, Nucleotide; D009363:Neoplasm Proteins