Use of Gene Therapy: A Tool to Study Reproductive Function

基因疗法的使用：研究生殖功能的工具

• 批准号: 7920812
• 负责人: Ursula B. Kaiser
• 金额: $ 48.08万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7920812
• 关键词: Adult; Amino Acids; Base Pairing; Behavior; Binding Sites; Biological Models; Cells; Central Nervous System Diseases; Competence; Development; Disease; Estrogens; Exhibits; Feedback; Female; Fertility; Follicle Stimulating Hormone; Funding; Gene Expression; Gene Expression Regulation; Genes; Genetic Models; Goals; Gonadal Steroid Hormones; Gonadal structure; Gonadotropin Hormone Releasing Hormone; Growth; Herpesvirus 1; Hormones; Hypothalamic structure; Infertility; Lead; Length; Light; Luteinizing Hormone; Maintenance; Messenger RNA; Modeling; Mus; Neuraxis; Neurologic; Neurons; Neurosciences; Output; Ovarian; Ovulation; Pattern; Peptides; Peripheral; Physiological; Physiology; Pituitary Gland; Population; Pregnancy; Process; Production; Recovery; Recreation; Reproduction; Reproductive Behavior; Reproductive Physiology; Research; Role; Sex Behavior; Sexual Maturation; Signal Transduction; Simplexvirus; Source; Spermatogenesis; System; Testing; Tissues; Transgenes; Translations; Viral; Viral Vector; Wild Type Mouse; folliculogenesis; gene replacement; gene replacement therapy; gene therapy; gonadotropin releasing hormone associated peptide; hormone regulation; human GNRH1 protein; kisspeptin; male; prohormone; promoter; reproductive; reproductive axis; reproductive function; reproductive hormone; response; secretion process; tool; vector

The overall goal of this project is to explore the regulatory mechanisms that control reproduction using a unique genetic model system, the hypogonadal, GnRH-deficient hpg mouse with replacement of GnRH by targeted gene therapy. The ultimate test for the ability of viral vector systems to successfully direct cell-specific, physiologically regulated GnRH expression and release is the full recovery of reproductive function in the hpg mice. One of the unique features of our viral transgene that is essential for the recreation of reproductive integrity is the use of the GnRH gene with its own cognate promoter. The use of the endogenous GnRH gene promoter may allow regulated, pulsatile GnRH production and release, and hence stimulate pituitary LH and FSH production and lead to activation of the reproductive axis and induction of fertility. Our preliminary studies in hpg females injected with HSV.GnRH.GFP amplicon demonstrate coordinated stimulation of pituitary LH and FSH secretion and ovarian and uterine growth. In the hpg male, we have demonstrated spermatogenesis after delivery of HSV.GnRH.GFP amplicon vector. We hypothesize that delivery and expression of the GnRH gene to hypothalamic neurons in adult female and male hpg mice will result in the full recapitulation of reproductive function. In the first aim, we propose to more fully characterize the extent of reproductive recovery achieved by GnRH gene therapy, including ovarian folliculogenesis and ovulation, spermatogenesis, sexual reproductive behavior and fertility. Successful activation of reproductive function in this model would also suggest that neuronal inputs to GnRH neurons remain intact in hpg mice, so that appropriate modulation of GnRH output can occur. In the second aim, the effects of GnRH rescue on the regulatory mechanisms that control reproduction will be explored, including peripheral modulators such as positive and negative feedback effects of sex steroids, and central modulators such as kisspeptin. The proposed studies provide a genetic model for selective and regulated gene replacement therapy, and thus may have important implications not only for reproductive disorders such as infertility, but also for applications of gene therapy in the neurosciences, for treatment of neurological and other disorders where normal gene regulation is critical.

本项目的总体目标是探索控制生殖的调节机制， 独特的遗传模型系统，性腺功能减退，GnRH缺陷的hpg小鼠，用GnRH替代 靶向基因治疗最终测试病毒载体系统的能力，成功地指导细胞特异性， 生理调节的GnRH表达和释放是hpg中生殖功能的完全恢复 小鼠我们的病毒转基因的独特功能之一，这是必不可少的繁殖重建， 完整性是使用GnRH基因及其自身同源启动子。内源性GnRH基因的应用 启动子可允许调节的、脉动的GnRH产生和释放，从而刺激垂体LH， FSH的产生并导致生殖轴的激活和诱导生育。我们的初步研究 在用HSV、GnRH、GFP扩增子注射hpg雌性中，证实了垂体LH的协调刺激， FSH分泌与卵巢和子宫生长。在hpg男性中，我们已经证明了精子发生后， HSV、GnRH、GFP扩增子载体的递送。我们假设GnRH基因的传递和表达 对成年雌性和雄性hpg小鼠的下丘脑神经元的刺激将导致生殖功能的完全重演。 功能在第一个目标中，我们提议更充分地说明生殖恢复的程度 通过GnRH基因治疗，包括卵巢卵泡发生和排卵、精子发生、性生殖 行为和生育能力。在该模型中成功激活生殖功能也表明， 在hpg小鼠中，对GnRH神经元的神经元输入保持完整，因此适当的GnRH输出调节 可能发生。在第二个目标中，GnRH拯救对控制 生殖将被探讨，包括外围调制器，如积极和消极的反馈效应 性类固醇和中枢调节剂如kisspeptin这些研究为以下疾病提供了遗传模型： 选择性和调节性基因替代疗法，因此可能不仅对 生殖系统疾病，如不孕症，但也用于基因治疗在神经科学中的应用， 治疗神经系统和其他疾病，其中正常的基因调控是至关重要的。