Improvement in reproductive parameters in hypogonadal female mice by regulated gene replacement therapy in the central nervous system.
One of the challenges of gene targeting is to achieve regulated transgene expression in specific target cells. The hypogonadal (hpg) mice are genetically deficient in hypothalamic gonadotropin-releasing hormone (GnRH) production due to a deletion in the GnRH gene, resulting in hypogonadotropic hypogonadism. Here we show an improvement in reproductive parameters of adult female homozygous hpg mice by direct infusion into the hypothalamic preoptic area (POA) of a herpes simplex virus (HSV)-based amplicon vector containing a 13.5 kb genomic fragment encoding the GnRH gene together with its cognate promoter and regulatory elements. Following vector injection, GnRH-expressing neurons were detected in the POA, and pituitary and plasma gonadotropin levels as well as ovarian and uterine weights increased. In addition, a subset of injected hpg mice demonstrated cyclic estrous changes, consistent with regulated control of GnRH production. Administration of kisspeptin-10 resulted in an increase in plasma luteinizing hormone levels, further supporting appropriate regulation of the introduced GnRH transgene. These findings indicate that delivery of the GnRH gene resulted in selective neuronal expression of GnRH and regulated hypothalamic GnRH release. To our knowledge, this is the first example of the correct targeting of a gene under its cognate promoter to neurons resulting in selective and regulated synthesis of a biologically active peptide, and thus may have a wide range of applications in the treatment of human disorders.
Jeong
K-H
KH
Division of Endocrinology, Diabetes, and Hypertension, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
Bakowska
J C
JC
Song
I O
IO
Fu
N
N
Breakefield
X O
XO
Kaiser
U B
UB
D000818:Animals; D005260:Female; D005640:Follicle Stimulating Hormone / Q000097:blood; D005786:Gene Expression Regulation / Q000187:drug effects; D018390:Gene Targeting; D005818:Genetic Engineering; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000235:genetics; D007987:Gonadotropin-Releasing Hormone / Q000032:analysis / Q000235:genetics* / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics; D018259:Herpesvirus 1, Human / Q000235:genetics*; D007006:Hypogonadism / Q000378:metabolism / Q000628:therapy*; D007031:Hypothalamus / Q000378:metabolism; D007150:Immunohistochemistry; D059648:Kisspeptins; D007986:Luteinizing Hormone / Q000097:blood / Q000378:metabolism; D051379:Mice; D008817:Mice, Mutant Strains; D009474:Neurons / Q000737:chemistry / Q000378:metabolism; D009842:Oligopeptides / Q000494:pharmacology; D011301:Preoptic Area / Q000737:chemistry / Q000378:metabolism; D011401:Promoter Regions, Genetic; D019076:Transgenes
C106628:KISS1 protein, human; D059648:Kisspeptins; D009842:Oligopeptides; D049452:Green Fluorescent Proteins; D007987:Gonadotropin-Releasing Hormone; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone