title: The role of LAT1 in (18)F-DOPA uptake in malignant gliomas.
abstract: Positron emission tomography (PET) imaging with the amino acid tracer 6-(18)F-fluoro-L-3,4-dihydroxy-phenylalanine ((18)F-DOPA) may provide better spatial and functional information in human gliomas than CT or MRI alone. The L-type amino acid transporter 1 (LAT1) is responsible for membrane transport of large neutral amino acids in normal cells. This study assessed the relationship between LAT1 expression and (18)F-DOPA uptake in human astrocytomas. Endogenous LAT1 expression was measured in established glioblastoma (GBM) cell lines and primary GBM xenografts using Western blotting and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Uptake of (18)F-DOPA was approximated in vitro using (3)H-L-DOPA as an analog. Uptake of (3)H-L-DOPA was assessed in cells expressing LAT1 shRNA or LAT1 siRNA and compared to non-targeted (NT) control shRNA or siRNA sequences, respectively. To demonstrate the clinical relevance of these findings, LAT1 immunofluorescence staining was compared with corresponding regions of (18)F-DOPA PET uptake in patients with newly diagnosed astrocytomas. LAT1 mRNA and protein expression varies in GBM, and the extent of (3)H-L-DOPA uptake was positively correlated with endogenous LAT1 expression. Stable shRNA-mediated LAT1 knockdown in T98 and GBM28 reduced (3)H-L-DOPA uptake relative to NT shRNA by 57 (P < 0.0001) and 52 % (P < 0.001), respectively. Transient siRNA-mediated LAT1 knockdown in T98 reduced (3)H-L-DOPA uptake relative to NT siRNA up to 68 % (P < 0.01). In clinical samples, LAT1 expression positively correlated with (18)F-DOPA PET uptake (P = 0.04). Expression of LAT1 is strongly associated with (3)H-L-DOPA uptake in vitro and (18)F-DOPA uptake in patient biopsy samples. These results define LAT1 as a key determinant of (18)F-DOPA accumulation in GBM.
authors.affiliation: College of Medicine, Mayo Clinic, 200 First Street SW, Rochester, MN 55905, USA. Youland.Ryan@Mayo.edu
authors.forename: Ryan S
authors.initials: RS
authors.lastname: Youland
authors.forename: Gaspar J
authors.initials: GJ
authors.lastname: Kitange
authors.forename: Timothy E
authors.initials: TE
authors.lastname: Peterson
authors.forename: Deanna H
authors.initials: DH
authors.lastname: Pafundi
authors.forename: Judi A
authors.initials: JA
authors.lastname: Ramiscal
authors.forename: Jenny L
authors.initials: JL
authors.lastname: Pokorny
authors.forename: Caterina
authors.initials: C
authors.lastname: Giannini
authors.forename: Nadia N
authors.initials: NN
authors.lastname: Laack
authors.forename: Ian F
authors.initials: IF
authors.lastname: Parney
authors.forename: Val J
authors.initials: VJ
authors.lastname: Lowe
authors.forename: Debra H
authors.initials: DH
authors.lastname: Brinkmann
authors.forename: Jann N
authors.initials: JN
authors.lastname: Sarkaria
mesh_terms: D000818:Animals; D001692:Biological Transport; D015153:Blotting, Western; D001932:Brain Neoplasms / Q000378:metabolism* / Q000473:pathology; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives* / Q000493:pharmacokinetics; D005455:Fluorescent Antibody Technique; D005462:Fluorine Radioisotopes; D005910:Glioma / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D007124:Immunoenzyme Techniques; D027283:Large Neutral Amino Acid-Transporter 1 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D060787:Neoplasm Grading; D049268:Positron-Emission Tomography; D012333:RNA, Messenger / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
chemical_list: D005462:Fluorine Radioisotopes; D027283:Large Neutral Amino Acid-Transporter 1; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C043437:fluorodopa F 18; D004295:Dihydroxyphenylalanine