title: A quantitative method for measuring gene expression magnitude and volume delivered by gene therapy vectors.
abstract: This study describes a quantitative method to measure the magnitude and distribution of gene expression following local delivery of an adenoviral vector containing the human sodium iodide symporter (hNIS) reporter gene into the canine prostate. Following systemic administration of Na(99m)TcO(4), autoradiographs of prostate sections depicting hNIS-dependent (99m)TcO(4)(-) uptake were digitized and stacked to produce a three-dimensional reconstruction of gene expression. Frequency histograms reflecting hNIS gene expression magnitude and volume were used to quantify hNIS function. The method demonstrated submillimeter resolution allowing for precise measurements of gene expression magnitude and volume in vivo. The method developed here could be applied to other reporter gene systems in which the readout can be digitized from thin tissue sections.
authors.affiliation: Department of Radiation Oncology, Henry Ford Health System, Detroit, MI 48202, USA.
authors.forename: Kenneth N
authors.initials: KN
authors.lastname: Barton
authors.forename: Xueqing
authors.initials: X
authors.lastname: Xia
authors.forename: Hui
authors.initials: H
authors.lastname: Yan
authors.forename: Hans
authors.initials: H
authors.lastname: Stricker
authors.forename: Gregory
authors.initials: G
authors.lastname: Heisey
authors.forename: Fang-Fang
authors.initials: FF
authors.lastname: Yin
authors.forename: Tavarekere N
authors.initials: TN
authors.lastname: Nagaraja
authors.forename: Guopei
authors.initials: G
authors.lastname: Zhu
authors.forename: Andrew
authors.initials: A
authors.lastname: Kolozsvary
authors.forename: Joseph D
authors.initials: JD
authors.lastname: Fenstermacher
authors.forename: Mei
authors.initials: M
authors.lastname: Lu
authors.forename: Jae Ho
authors.initials: JH
authors.lastname: Kim
authors.forename: Svend O
authors.initials: SO
authors.lastname: Freytag
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Brown
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001345:Autoradiography; D004285:Dogs; D004305:Dose-Response Relationship, Drug; D015870:Gene Expression; D017930:Genes, Reporter; D005821:Genetic Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D017403:In Situ Hybridization; D008297:Male; D011467:Prostate / Q000378:metabolism; D012974:Sodium Iodide / Q000494:pharmacology; D013667:Technetium / Q000494:pharmacology
chemical_list: D013667:Technetium; D012974:Sodium Iodide