Dual Reporter Gene Imaging
The human and rodent sodium iodide symporters ( NIS ) have recently been cloned and are being investigated as potential therapeutic and reporter genes. We have extended this effort by constructing an internal ribosomal entry site (IRES)-linked human NIS (hNIS) -enhanced green fluorescent protein ( e...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
SAGE Publishing
2005-04-01
|
| Series: | Molecular Imaging |
| Online Access: | https://doi.org/10.1162/15353500200504193 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1841563103195561984 |
|---|---|
| author | Jiantu Che Mikhail Doubrovin Inna Serganova Ludmila Ageyeva Pat Zanzonico Ronald Blasberg |
| author_facet | Jiantu Che Mikhail Doubrovin Inna Serganova Ludmila Ageyeva Pat Zanzonico Ronald Blasberg |
| author_sort | Jiantu Che |
| collection | DOAJ |
| description | The human and rodent sodium iodide symporters ( NIS ) have recently been cloned and are being investigated as potential therapeutic and reporter genes. We have extended this effort by constructing an internal ribosomal entry site (IRES)-linked human NIS (hNIS) -enhanced green fluorescent protein ( eGFP ) hybrid reporter gene for both nuclear and optical imaging. A self-inactivating retroviral vector, termed pQCNIG, containing hNIS-IRES-eGFP dual reporter gene, driven by a constitutive CMV promoter, was constructed and used to generate RG2-pQCNIG cells and RG2-pQCNIG tumors. 131 I-iodide and 99m TcO 4 -pertechnetate accumulation studies plus fluorescence microscopy and intensity assays were performed in vitro, and gamma camera imaging studies in RG2-pQCNIG and RG2 tumor-bearing athymic rats were performed. RG2-pQCNIG cells expressed high levels of hNIS protein and showed high intensity of eGFP fluorescence compared with RG2 wild-type cells. RG2-pQCNIG cells accumulated Na 131 I and 99m TcO 4 – to a 50:1 and a 170:1 tissue/medium ratio at 10 min, compared with 0.8:1.2 tissue/medium ratio in wild-type RG2 cells. A significant correlation between radiotracer accumulation and eGFP fluorescence intensity was demonstrated. RG2-pQCNIG and RG2 tumors were readily differentiated by in vivo gamma camera imaging; radiotracer uptake increased in RG2-pQCNIG but declined in RG2 tumors over the 50-min imaging period. Stomach and thyroid were the major organs of radionuclide accumulation. The IRES-linked hNIS-eGFP dual reporter gene is functional and stable in transduced RG2-pQCNIG cells. Optical and nuclear imaging of tumors produced from these cell lines provides the opportunity to monitor tumor growth and response to therapy. These studies indicate the potential for a wider application of hNIS reporter imaging and translation into patient studies using radioisotopes that are currently available for human use for both SPECT and PET imaging. |
| format | Article |
| id | doaj-art-871ab4eaee1f4505b50a5628f05c0e3c |
| institution | Kabale University |
| issn | 1536-0121 |
| language | English |
| publishDate | 2005-04-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Molecular Imaging |
| spelling | doaj-art-871ab4eaee1f4505b50a5628f05c0e3c2025-01-03T00:12:12ZengSAGE PublishingMolecular Imaging1536-01212005-04-01410.1162/1535350020050419310.1162_15353500200504193Dual Reporter Gene ImagingJiantu CheMikhail DoubrovinInna SerganovaLudmila AgeyevaPat ZanzonicoRonald BlasbergThe human and rodent sodium iodide symporters ( NIS ) have recently been cloned and are being investigated as potential therapeutic and reporter genes. We have extended this effort by constructing an internal ribosomal entry site (IRES)-linked human NIS (hNIS) -enhanced green fluorescent protein ( eGFP ) hybrid reporter gene for both nuclear and optical imaging. A self-inactivating retroviral vector, termed pQCNIG, containing hNIS-IRES-eGFP dual reporter gene, driven by a constitutive CMV promoter, was constructed and used to generate RG2-pQCNIG cells and RG2-pQCNIG tumors. 131 I-iodide and 99m TcO 4 -pertechnetate accumulation studies plus fluorescence microscopy and intensity assays were performed in vitro, and gamma camera imaging studies in RG2-pQCNIG and RG2 tumor-bearing athymic rats were performed. RG2-pQCNIG cells expressed high levels of hNIS protein and showed high intensity of eGFP fluorescence compared with RG2 wild-type cells. RG2-pQCNIG cells accumulated Na 131 I and 99m TcO 4 – to a 50:1 and a 170:1 tissue/medium ratio at 10 min, compared with 0.8:1.2 tissue/medium ratio in wild-type RG2 cells. A significant correlation between radiotracer accumulation and eGFP fluorescence intensity was demonstrated. RG2-pQCNIG and RG2 tumors were readily differentiated by in vivo gamma camera imaging; radiotracer uptake increased in RG2-pQCNIG but declined in RG2 tumors over the 50-min imaging period. Stomach and thyroid were the major organs of radionuclide accumulation. The IRES-linked hNIS-eGFP dual reporter gene is functional and stable in transduced RG2-pQCNIG cells. Optical and nuclear imaging of tumors produced from these cell lines provides the opportunity to monitor tumor growth and response to therapy. These studies indicate the potential for a wider application of hNIS reporter imaging and translation into patient studies using radioisotopes that are currently available for human use for both SPECT and PET imaging.https://doi.org/10.1162/15353500200504193 |
| spellingShingle | Jiantu Che Mikhail Doubrovin Inna Serganova Ludmila Ageyeva Pat Zanzonico Ronald Blasberg Dual Reporter Gene Imaging Molecular Imaging |
| title | Dual Reporter Gene Imaging |
| title_full | Dual Reporter Gene Imaging |
| title_fullStr | Dual Reporter Gene Imaging |
| title_full_unstemmed | Dual Reporter Gene Imaging |
| title_short | Dual Reporter Gene Imaging |
| title_sort | dual reporter gene imaging |
| url | https://doi.org/10.1162/15353500200504193 |
| work_keys_str_mv | AT jiantuche dualreportergeneimaging AT mikhaildoubrovin dualreportergeneimaging AT innaserganova dualreportergeneimaging AT ludmilaageyeva dualreportergeneimaging AT patzanzonico dualreportergeneimaging AT ronaldblasberg dualreportergeneimaging |