Targeting Adenovirus for Gene Therapy of Uterine Leiomyoma

靶向腺病毒用于子宫平滑肌瘤的基因治疗

基本信息

批准号：
8335561
负责人：
Ayman Al-Hendy
金额：
$ 6.35万
依托单位：
MEHARRY MEDICAL COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-24 至 2015-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8335561
关键词：
Ablation Adenovirus Vector Adenoviruses Affect African American Aftercare Animal Model Apoptosis Apoptosis Promoter Apoptotic Applications Grants Attenuated Biological Assay Biological Models Bioluminescence Bromodeoxyuridine CAV2 gene Caring Cells Chronic Common Neoplasm Cyclin D1 Data Delayed Childbearing Dependency Development Disease Distant Dominant-Negative Mutation Economic Burden Endometrium Estrogen Receptors Estrogens Evaluation Face Fatigue Female Female Genital Neoplasms Fertility Fibroid Tumor Future Ganciclovir Generations Genes Genetic Transcription HSV-Tk Gene Healthcare Systems Hormonal Human Hysterectomy Image Immune Immune response In Situ Nick-End Labeling Incidence Injection of therapeutic agent Iron deficiency anemia Leiomyoma Lesion Literature Liver Luc Gene Luciferases MSLN gene Mammary gland Medical Methods Minority Modeling Modification Mus Normal tissue morphology Operative Surgical Procedures Ovulation PECAM1 gene Parents Pathogenesis Patients Positron-Emission Tomography Pregnancy Premenopause Progesterone Progesterone Receptors Randomized Rattus Receptor Gene Reproductive Health Resolution Role SCID Mice SLPI gene Safety Sampling Signal Transduction Site Symptoms TK Gene Testing Therapeutic Thymidine Kinase Time Tissues Toxic effect Ultrasonography Uterine Fibroids Vascular Endothelial Growth Factors Vascular blood supply Woman Women&apos s Health Work disorder control gene therapy health disparity improved in vivo in vivo Model indexing insight interest mouse model myometrium novel pre-clinical public health relevance reproductive reproductive function research study response response marker single photon emission computed tomography social steroid hormone therapeutic gene treatment response tumor vector

项目摘要

DESCRIPTION (provided by applicant): Uterine leiomyoma (fibroids) are the most common gynecologic tumors in premenopausal women and the leading indication for hysterectomy. Uterine fibroids are also a major health disparity issue being 3-4 folds more frequent in African American compared to white women. We have generated most of the literature on the utility of fibroid gene therapy, demonstrating its potential as a localized treatment for uterine leiomyoma that can control the disease while preserving fertility potential. In this proposal we are aiming to screen a wide range of improved adenoviral vector transductional and transcription modifications, to develop an optimal uterine fibroids ON-normal tissue OFF targeted adenoviral vector for effective and safe localized treatment of uterine fibroids. In this revised competing renewal application, we propose three specific aims; Specific aim one: we will assess adenovirus targeting strategies towards uterine leiomyoma in the Eker rat model. Four targeted adenoviral vectors expressing luciferase gene as a marker (Ad5-RGD-luc, Ad5-CAV2- luc, Ad5-SLPI-luc, and Ad5-MSLN-CRAD-luc) will be directly delivered into the uterine fibroid tumors of the Eker rat, and their efficiency and safety profile will be compared to the parent Ad5-luc first generation unmodified adenoviral vector using bioluminescence imaging and tissue luciferase assays. Specific aim 2: we will construct a human leiomyoma-targeted adenovirus vector using best targeting strategy identified above, and then proceed to subclone and express the fibroid therapeutic genes. Three therapeutic genes will be used that we have shown to be effective in local ablation of fibroid lesions; dominant negative estrogen receptor (DN-ER), dominant negative progesterone receptor (DN- PR) or thymidine kinase/ganciclovir (TK/GCV) approach. Specific aim 3: In this aim we will test the therapeutic utility of direct intratumor delivery of the three targeted adenoviral vectors (Ad-T-DN-ER, Ad-T-DN-PR and Ad-T-TK) in two animal models; female Eker rats, the only immune competent model for uterine fibroids, and Memy 1, our novel SCID mouse model harboring engrafted human fibroid tissues. The tumors will be sized biweekly by high resolution ultrasound. Functional in vivo PET and SPECT imaging will be used for real time evaluation of tumor proliferation and apoptosis. Detailed evaluation of tumor response as well as additional toxicity and safety assays will be performed. These experiments will provide a valuable model system to further our understanding of the role of estrogen, progesterone and apoptosis in the pathogenesis of uterine fibroids in a unique in vivo setting. This will have major positive effects on minority women health, and reproductive health in general. PUBLIC HEALTH RELEVANCE: For women with symptomatic fibroids who want to preserve fertility, there are currently limited conservative treatment options that will treat their fibroids without compromising subsequent chances of achieving a healthy pregnancy. In this proposal we are aiming to develop an improved gene therapy approach for effective and safe localized treatment of uterine fibroids. Additionally it will enhance our understanding of the role of steroid hormones in pathogenesis of uterine fibroids using unique in vivo model systems.

描述（由申请人提供）：子宫平滑肌瘤（肌瘤）是绝经前妇女中最常见的妇科肿瘤，也是子宫切除术的领先指示。与白人妇女相比，子宫肌瘤也是非裔美国人的主要健康差异问题的主要频率更高。我们已经生成了有关肌瘤基因疗法实用性的大多数文献，证明了它作为子宫平滑肌瘤的局部治疗的潜力，可以控制该疾病，同时保持生育能力的潜力。在此提案中，我们旨在筛选出广泛改进的腺病毒载体的转导和转录修饰，以开发出靶向的腺病毒载体的正常组织，以有效且安全的局部化子宫构局部治疗子宫构件。在经过修订的竞争续约应用中，我们提出了三个具体目标。具体目标：我们将评估Eker大鼠模型中针对子宫平滑肌瘤的腺病毒靶向策略。表达荧光素酶基因作为标记的四个有针对性的腺病毒载体（AD5-RGD-LUC，AD5-CAV2- LUC，AD5-SLPI-LUC和AD5-MSLN-CRAD-LUC）将直接传递到使用Eker Rat的子宫纤维肿瘤中，并将其使用eker的效率和安全性与父母的效率相比，并将其与父母的效率进行比较。生物发光成像和组织荧光素酶测定。具体目的2：我们将使用上面确定的最佳靶向策略来构建一个靶向人类的腺病毒载体，然后继续进行亚克隆并表达肌瘤治疗基因。将使用三种治疗基因，这些基因在肌瘤病变的局部消融中有效。显性雌激素受体（DN-ER），显性负孕酮受体（DN-PR）或胸苷激酶/Ganciclovir（TK/GCV）方法。特定目的3：在此目标中，我们将在两个动物模型中测试三个靶向腺病毒载体（AD-T-DN-ER，AD-T-DN-PR和AD-TK）的直接肿瘤内递送的治疗效用；雌性Eker大鼠是子宫肌瘤的唯一免疫胜任模型，以及Memy 1，这是我们具有植入人类肌瘤组织的新型SCID小鼠模型。肿瘤将通过高分辨率超声进行双周尺寸。体内PET和SPECT成像的功能将用于实时评估肿瘤增殖和凋亡。将对肿瘤反应以及其他毒性和安全测定法进行详细评估。这些实验将提供一个有价值的模型系统，以进一步了解雌激素，孕酮和凋亡在独特的体内环境中子宫肌瘤发病机理中的作用。这将对少数族裔妇女健康和生殖健康产生重大积极影响。公共卫生相关性：对于有症状性肌瘤的女性，想要保留生育能力的肌瘤，目前有限的保守治疗选择将治疗其肌瘤，而不会损害随后获得健康怀孕的机会。在此提案中，我们旨在开发一种改进的基因治疗方法，以有效且安全的子宫肌瘤局部治疗。此外，它将增强我们对使用独特的体内模型系统中类固醇激素在子宫肌瘤发病机理中的作用的理解。