Salivary Gland-Based Gene Therapy for Pulmonary Arterial Hypertension

基于唾液腺的肺动脉高压基因治疗

• 批准号: 8608587
• 负责人: RAYMOND Louis Benza
• 金额: $ 18.01万
• 依托单位: ALLEGHENY-SINGER RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-02-01 至 2016-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8608587
• 关键词: Address; Adverse effects; Affect; Aftercare; Anabolism; Animal Model; Area; Biological Response Modifier Therapy; Bioreactors; Blood Circulation; Cannulations; Caregivers; Cell membrane; Cells; Chimeric Proteins; Chronic; Complex; Continuous Infusion; DNA; Diagnosis; Disease; Dose; Drug Formulations; Encapsulated; Endocrine; Endothelin Receptor Antagonist; Enzymes; Epoprostenol; Gene Delivery; Gene Expression; Gene Transfer; Goals; Heart; Human; Immune response; In Vitro; Individual; Institutes; Labyrinth; Left; Life; Lipids; Longevity; Lung; Mammalian Cell; Mammals; Measures; Mechanics; Mediating; Microbubbles; Modality; Modeling; Oral cavity; Organ; Outpatients; Pancreas; Pathway interactions; Patients; Pneumonectomy; Procedures; Process; Production; Progressive Disease; Property; Prostacyclin synthase; Prostaglandins; Prostaglandins I; Proteins; Pulmonary Vascular Resistance; Rattus; Regimen; Reliance; Research; Salivary Glands; Serum; Signal Pathway; Site; Solutions; Structure of submaxillary duct; Submandibular gland; Survival Rate; Symptoms; System; Techniques; Technology; Testing; Therapeutic; Thromboxane A2; Transgenes; Translating; Ultrasonic Therapy; Ultrasonography; Vascular Endothelium; Viral Vector; advanced disease; base; cyclooxygenase 1; design; effective therapy; gene delivery system; gene therapy; gene transfer vector; in vivo; indexing; inhibitor/antagonist; innovation; non-viral gene delivery; novel; pressure; public health relevance; pulmonary arterial hypertension; research study; restoration; therapeutic transgene; urinary; vector; viral gene delivery

DESCRIPTION (provided by applicant): The overall goal of this proposal is to design and test a gene therapy strategy for Pulmonary Arterial Hypertension (PAH). PAH is a rare, deadly and incurable disease with a mean survival of 2.8 years from onset of symptoms if left untreated. Of the three classes of approved therapeutics, endothelin receptor antagonists, phospodiesterase inhibitors, and prostacyclins, prostacyclin is the most effective therapy. However, complicated delivery systems and potential side effects associated with the present formulation of prostanoids (e.g. prostacyclin) have deterred some patients and caregivers from instituting this highly effective class of agents. The challenge to be addressed by this proposal is the need for a therapeutic regimen that allows for endogenous production of prostacyclin therapy within the patient's own body, throughout the entire lifetime of the patient. Because prostacyclin can be produced endogenously through expression of the enzyme Prostacyclin Synthase (PGIS), gene therapy has previously shown proof-of-principle efficacy in animal models by enabling endogenous production of prostacyclin and reversal of experimental PAH. This proposal will build upon this concept, but will execute this strategy using newly developed gene transfer technology that obviates the viral gene delivery vectors that have been used in prior studies. Viral vectors have been extremely useful in earlier studies, but have limited duration of expression, and due to host immune response, cannot affect life- long therapy nor can they be re-dosed. Our gene delivery system, which used ultrasound-induced microbubble cavitation to allow entry of non-viral DNA vectors into cells, is thought to evade host immune responses, theoretically allowing re-dosing of the Prostacyclin Synthase therapeutic transgene as a periodic boosters throughout the entire lifespan of the patient. An additional innovation of this proposal toward achieving the field-wide goal of endogenous prostacyclin production is the choice of the salivary glands as the therapeutic biosynthesis site. The salivary glands can be accessed through a bloodless, outpatient procedure, and contain a robust endocrine secretory pathway capable of secreting transgene products into the intravascular space. The encapsulated, fixed volume of the intraductal labyrinth of the salivary glands also allow precise control of the delivery system, making ultrasound-assisted gene transfer (UAGT) far more practical and consistent than has been observed in other organs (e.g. heart or pancreas). A final innovation of this proposal is the first in vivo application of a Cox-1/PGIS fusion protein that produces dramatically higher levels of prostacyclin than PGIS alone. In summary, this project seeks to innovate the enticing idea of PGIS- based gene therapy for PAH aggregating three enabling technologies: 1) a practical, re-dosable gene delivery technique, 2) a novel biosynthesis site (salivary glands) that can be accessed through an outpatient procedure, and 3) a Cox-1/PGIS fusion protein transgene that produces superior levels of prostacyclin therapeutic. The efficacy of this gene therapy strategy will be tested in a highly relevant rat model of severe PAH.

描述（由申请人提供）：本提案的总体目标是设计和测试肺动脉高压（PAH）的基因治疗策略。PAH是一种罕见、致命和无法治愈的疾病，如果不进行治疗，从症状发作起的平均生存期为2.8年。在三类批准的治疗药物（内皮素受体拮抗剂、磷酸二酯酶抑制剂和前列环素）中，前列环素是最有效的治疗药物。然而，与本发明的前列腺素类制剂（例如前列环素）相关的复杂的递送系统和潜在的副作用已经阻止了一些患者和护理人员使用这类高效的药剂。该提议要解决的挑战是需要一种治疗方案，其允许在患者的整个生命周期内在患者自身体内内源性产生前列环素治疗。由于前列环素可以通过前列环素合酶（PGIS）的表达内源性产生，因此基因治疗先前已在动物模型中通过实现前列环素的内源性产生和实验性PAH的逆转而显示出原理性疗效。该提案将建立在这一概念的基础上，但将使用新开发的基因转移技术来执行这一策略，该技术消除了先前研究中使用的病毒基因递送载体。病毒载体在早期研究中非常有用，但表达持续时间有限，并且由于宿主免疫应答，不能影响终身治疗，也不能重新给药。我们的基因递送系统使用超声诱导的微泡空化来允许非病毒DNA载体进入细胞，被认为可以逃避宿主免疫反应，理论上允许在患者的整个生命周期中作为周期性增强剂重新给药前列环素合酶治疗转基因。该提议朝向实现内源性前列环素产生的全领域目标的另一个创新是选择唾液腺作为治疗性生物合成位点。唾液腺可以通过无血的门诊手术进入，并且含有能够将转基因产物分泌到血管内空间中的稳健的内分泌途径。唾液腺导管内迷路的封装、固定体积也允许精确控制递送系统，使得超声辅助基因转移（UAGT）比在其他器官（例如心脏或胰腺）中观察到的更实用和一致。该提议的最后一个创新是首次在体内应用考克斯-1/PGIS融合蛋白，其产生比单独的PGIS显著更高水平的前列环素。总之，该项目旨在创新基于PGIS的PAH基因治疗的诱人理念，聚合三种使能技术：1）实用的、可重复给药的基因递送技术，2）可通过门诊手术进入的新型生物合成位点（唾液腺），以及3）产生上级前列环素治疗剂水平的考克斯-1/PGIS融合蛋白转基因。将在高度相关的重度PAH大鼠模型中检测该基因治疗策略的疗效。