title: Lentiviral vectors efficiently transduce human gonadotroph and somatotroph adenomas in vitro. Targeted expression of transgene by pituitary hormone promoters.
abstract: Despite important advances in human therapeutics, no specific treatment for both non-functioning gonadotroph and resistant somatotroph adenomas is available. Gene transfer by viral vectors can be considered as a promising way to achieve a specific and efficient treatment. Here we show the possibility of efficient gene transfer in human pituitary adenoma cells in vitro using a human immunodeficiency virus (HIV)-type 1-derived vector. Using enhanced green fluorescent protein (eGFP) gene as a marker placed under the phosphoglycerate kinase (PGK) promoter, gonadotroph and somatotroph adenomas were transduced even with moderate viral loads. The expression started at day 2, reached a peak at day 5, and it was still present at day 90. For targeting somatotroph and gonadotroph adenomas, human growth hormone (GH) promoter (GH -481, +54 bp) and two fragments of the human glycoprotein hormone alpha-subunit promoter (alpha-subunit 1 -520, +33 bp, and alpha-subunit 2 -907, +33 bp) were tested. In gonadotroph adenomas, the percentage of identified fluorescent cells and the fluorescence intensity analyzed by fluorescence-activated cell sorting indicated that the strength of the alpha-subunit 1 and alpha-subunit 2 promoters were comparable to that of the PGK promoter. Primary cultures of rat pituitary cells showed that alpha-subunit 1 is more selective to thyreotroph and gonadotroph phenotypes than alpha-subunit 2. GH promoter activity appeared weak in somatotroph adenomas. The human GH enhancer did not increase the GH promoter activity at all but the human prolactin promoter (-250 bp) allowed 4-fold more fluorescent cells to be obtained than the GH promoter. Several cell lines appeared too permissive to test cell-specificity of pituitary promoters. However, on human non-pituitary cell cultures, the tested pituitary promoters seemed clearly selective to target endocrine pituitary phenotypes. This study gives a starting point for a gene-therapy program using lentiviral vectors to transfer therapeutic genes in human pituitary adenomas.
authors.affiliation: Interactions Cellulaires Neuroendocriniennes, UMR 6544-Centre National de la Recherche Scientifique, Université de la Méditerranée, Institut Jean-Roche, Faculté de Médecine Nord, Marseille, France.
authors.forename: Catherine
authors.initials: C
authors.lastname: Roche
authors.forename: Alfredo J
authors.initials: AJ
authors.lastname: Zamora
authors.forename: David
authors.initials: D
authors.lastname: Taïeb
authors.forename: Esteban
authors.initials: E
authors.lastname: Lavaque
authors.forename: Ramahefarizo
authors.initials: R
authors.lastname: Rasolonjanahary
authors.forename: Henri
authors.initials: H
authors.lastname: Dufour
authors.forename: Claude
authors.initials: C
authors.lastname: Bagnis
authors.forename: Alain
authors.initials: A
authors.lastname: Enjalbert
authors.forename: Anne
authors.initials: A
authors.lastname: Barlier
mesh_terms: D000236:Adenoma / Q000378:metabolism / Q000628:therapy* / Q000821:virology; D000328:Adult; D000368:Aged; D045744:Cell Line, Tumor; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000627:therapeutic use; D015292:Glycoprotein Hormones, alpha Subunit / Q000235:genetics*; D006065:Gonadotropins, Pituitary / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D013006:Growth Hormone / Q000235:genetics / Q000378:metabolism; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D008297:Male; D008856:Microscopy, Fluorescence; D008875:Middle Aged; D010911:Pituitary Neoplasms / Q000378:metabolism / Q000628:therapy* / Q000821:virology; D011388:Prolactin / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D015292:Glycoprotein Hormones, alpha Subunit; D006065:Gonadotropins, Pituitary; D049452:Green Fluorescent Proteins; D011388:Prolactin; D013006:Growth Hormone