A quantitative method for measuring gene expression magnitude and volume delivered by gene therapy vectors.
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.
Barton
Kenneth N
KN
Department of Radiation Oncology, Henry Ford Health System, Detroit, MI 48202, USA.
Xia
Xueqing
X
Yan
Hui
H
Stricker
Hans
H
Heisey
Gregory
G
Yin
Fang-Fang
FF
Nagaraja
Tavarekere N
TN
Zhu
Guopei
G
Kolozsvary
Andrew
A
Fenstermacher
Joseph D
JD
Lu
Mei
M
Kim
Jae Ho
JH
Freytag
Svend O
SO
Brown
Stephen L
SL
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
D013667:Technetium; D012974:Sodium Iodide