Transcriptional targeting to anterior pituitary lactotrophic cells using recombinant adenovirus vectors in vitro and in vivo in normal and estrogen/sulpiride-induced hyperplastic anterior pituitaries.
The use of pituitary cell type-specific promoters is a powerful molecular tool to achieve pituitary cell type-specific transcriptional targeting of transgenes encoded by viral vectors. It has recently been proposed that transcriptional targeting of therapeutic genes could be harnessed as a gene therapy strategy for the treatment of pituitary disease. We describe the successful use of the human PRL promoter (hPrl) encoded within recombinant adenovirus vectors to target transgene expression of Herpes Simplex Virus Type 1-Thymidine Kinase (HSV1-TK) or beta-galactosidase to lactotrophic cells in vitro and in vivo. Functionally, the restriction of expression of HSV1-TK to lactotrophic tumor cells, using the hPrl promoter, resulted in the cell type-specific induction of apoptosis in the lactotrophic GH3 tumor cell line, in the presence of ganciclovir (GCV). In the corticotrophic AtT20 cell line, we detected neither HSV1-TK expression, nor apoptosis in the presence of GCV. The hPrl promoter encoded within a recombinant adenoviral vector also restricted transgene expression to lactotrophic cells in primary anterior pituitary (AP) cultures, and importantly, within the anterior pituitary gland in vivo. When the HSV1-TK driven by hPrl promoter was used in an in vivo model ofestrogen/sulpiride lactotroph induced hyperplasia within the AP in situ, the treatment was not effective in either reducing the weight of the gland, the number of lactotrophic cells within the transduced area in vivo, or the circulating PRL levels. This is in contrast to the human cytomegalovirus promoter (hCMV) driving expression of HSV1-TK in the same experimental paradigm, which was effective in reducing pituitary weight and circulating PRL levels. Our results have important implications in the design of gene therapy strategies for pituitary tumors. We demonstrate that both the choice of the in vivo animal model, i.e. adenoma in the AP gland in situ, and the particular gene therapy strategy chosen, i.e. use of strong ubiquitous promoters vs. weaker but cell type-specific promoters, determine the experimental therapeutic outcome.
Southgate
T D
TD
Molecular Medicine and Gene Therapy Unit, School of Medicine, University of Manchester, United Kingdom.
Windeatt
S
S
Smith-Arica
J
J
Gerdes
C A
CA
Perone
M J
MJ
Morris
I
I
Davis
J R
JR
Klatzmann
D
D
Löwenstein
P R
PR
Castro
M G
MG
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D014150:Antipsychotic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D002460:Cell Line; D004967:Estrogens / Q000494:pharmacology*; D005696:Galactosidases / Q000235:genetics; D018390:Gene Targeting / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics*; D018259:Herpesvirus 1, Human / Q000201:enzymology; D006965:Hyperplasia / Q000139:chemically induced / Q000473:pathology; D007150:Immunohistochemistry; D007202:Indicators and Reagents; D010903:Pituitary Gland, Anterior / Q000166:cytology* / Q000473:pathology; D010908:Pituitary Hormones, Anterior / Q000097:blood; D051381:Rats; D011915:Rats, Inbred BUF; D013469:Sulpiride / Q000494:pharmacology*; D014158:Transcription, Genetic / Q000235:genetics*; D019076:Transgenes / Q000235:genetics
D014150:Antipsychotic Agents; D004967:Estrogens; D007202:Indicators and Reagents; D010908:Pituitary Hormones, Anterior; D013469:Sulpiride; D005696:Galactosidases