title: SSTR2-based reporters for assessing gene transfer into non-small cell lung cancer: evaluation using an intrathoracic mouse model.
abstract: The most common cause of cancer-related deaths in North America is lung cancer, 85% of which is non-small cell lung cancer (NSCLC). Gene therapy is a promising approach, but has been hindered by lack of methods for localizing and quantifying gene expression in vivo. Human somatostatin receptor subtype-2 (SSTR2)-based reporters can be used to follow gene expression in vivo using ligands with greater affinity for this subtype. NSCLCs can express SSTR subtypes, which may interfere with SSTR2-based reporters. We assessed whether a SSTR2-based reporter can serve as a reporter of gene transfer into NSCLCs. SSTR subtype expression was assessed in NSCLC cell lines A549, H460, and H1299 using RT-PCR. After infection with an adenovirus containing hemagglutinin-A-tagged-SSTR2 (Ad-HA-SSTR2) or control insert, expression was assessed by immunologic techniques and binding to clinically-approved (111)In-octreotide. In vivo, after magnetic resonance (MR) imaging, intrathoracic H460 tumors were injected with Ad-HA-SSTR2 or control virus (n = 6 mice/group) under ultrasound guidance. Intravenous injection of (111)In-octreotide 2 days later was followed by planar and single-photon emission computed tomography (SPECT) imaging. Biodistribution into tumors was assessed in vivo using anatomic MR and functional gamma-camera images and ex vivo using excised organs/tumors. In human lung tumor samples (n = 70), SSTR2 expression was assessed using immunohistochemistry. All three NSCLC cell lines expressed different SSTR subtypes, but none expressed SSTR2. Upon Ad-HA-SSTR2 infection, HA-SSTR2 expression was seen in all three cell lines using antibodies targeting the HA domain or (111)In-octreotide targeting the receptor domain (p < 0.05). Intrathoracic tumors infected with Ad-HA-SSTR2 were clearly visible by gamma-camera imaging; expression was quantified by both in vivo and ex vivo biodistribution analysis and demonstrated greater uptake in tumors infected with Ad-HA-SSTR2 compared with control virus (p < 0.05). Immunohistochemistry found that 78% of NSCLCs are negative for and 13% have low levels of SSTR2 expression. It is concluded that SSTR2-based reporters can serve as reporters of gene transfer into NSCLCs.
authors.affiliation: Department of Experimental Diagnostic Imaging, University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA.
authors.forename: S P
authors.initials: SP
authors.lastname: Singh
authors.forename: L
authors.initials: L
authors.lastname: Han
authors.forename: R
authors.initials: R
authors.lastname: Murali
authors.forename: L
authors.initials: L
authors.lastname: Solis
authors.forename: J
authors.initials: J
authors.lastname: Roth
authors.forename: L
authors.initials: L
authors.lastname: Ji
authors.forename: I
authors.initials: I
authors.lastname: Wistuba
authors.forename: V
authors.initials: V
authors.lastname: Kundra
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000628:therapy; D000077192:Adenocarcinoma of Lung; D000256:Adenoviridae / Q000378:metabolism; D000818:Animals; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics* / Q000628:therapy; D045744:Cell Line, Tumor; D015902:Gamma Cameras; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics / Q000628:therapy; D008297:Male; D051379:Mice; D008819:Mice, Nude; D023421:Models, Animal; D017481:Receptors, Somatostatin / Q000235:genetics* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015899:Tomography, Emission-Computed, Single-Photon
chemical_list: D017481:Receptors, Somatostatin; C492049:Sstr2 protein, mouse