Stem Cell Therapy for Pulmonary Hypertension

肺动脉高压的干细胞疗法

• 批准号: 6922436
• 负责人: Philip J Kadowitz
• 金额: $ 37.13万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-04-01 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6922436
• 关键词: calcitonin gene related peptide; cardiovascular disorder therapy; cell differentiation; drug administration rate /duration; enzyme linked immunosorbent assay; fluorescent in situ hybridization; genetic manipulation; genetic transcription; heart catheterization; histochemistry /cytochemistry; histology; immunocytochemistry; laboratory rat; lung; mesenchyme; nonhuman therapy evaluation; nonsurgical revascularization; polymerase chain reaction; pulmonary hypertension; respiratory function; stem cell transplantation; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): The broad long-term objectives of the proposed research are to develop a new mesenchymal stem-cell (MSC) based therapy for the treatment of pulmonary hypertension. Pulmonary hypertension is a serious, often fatal disease characterized by increased pulmonary arterial pressure, right-heart failure, and remodeling of the pulmonary vascular bed. Although current therapy has prolonged survival, the long-term outcome is not favorable. Gene therapy shows promise, however, in vivo administration of non-viral vectors or viral vectors leads to low level gene transfer, random gene expression, and adverse effects, including inflammation. MSCs are self-renewing adult stem cells from bone marrow, are easily isolated and expanded in culture, and have the potential to differentiate into many cell types. It is our hypothesis that gene-modified MSCs may have a beneficial effect in the treatment of pulmonary hypertension. The results of preliminary experiments in our laboratory show that the administration of gene-modified MSCs and wild-type or unmodified MSCs improve erectile function in the aged rat. The results of preliminary studies also show that intratracheal administration of wild-type MSCs attenuated monocrotaline-induced pulmonary hypertension and restored pulmonary vasodilator responses to acetylcholine, which were impaired by monocrotaline treatment. The experiments proposed in this application will test the hypothesis that intratracheal administration of MSCs or MSCs modified with the eNOS gene or the CGRP gene will have a beneficial effect in monocrotaline-induced pulmonary hypertension in the rat. In order to test this hypothesis, MSCs must be prepared, characterized, gene-modified, and injected into the lung of control rats and rats with monocrotaline-induced pulmonary hypertension. In addition to determining the effect of MSCs and gene-modified MSCs on baseline pulmonary pressures and blood flow, the effects of the MSCs on pulmonary vascular responses to ventilatory hypoxia and vasoactive agonists, including acetylcholine, and the fate of the injected MSCs in the lung will be investigated using histologic and histochemical techniques. Therefore, the first specific aim is to isolate, characterize and gene-modify MSCs from the rat. The second specific aim is to investigate the effect of injected MSCs on pulmonary vascular function and on monocrotaline-induced pulmonary hypertension using newly developed right-heart catheterization techniques. The third specific aim is to study the fate of injected MSCs in the lung using histochemical techniques, the deconvoluted microscope, and localization of the male Y chromosome in the lung of female rats using PCR and FISH methodologies. The results of these studies may lead to new MSC-based therapy for the treatment of pulmonary hypertension.

描述（由申请人提供）：拟议研究的广泛长期目标是开发一种新的基于间充质干细胞（MSC）的治疗肺动脉高压的疗法。肺动脉高压是一种严重的，通常是致命的疾病，其特征是肺动脉压升高，右心衰竭和肺血管床重塑。虽然目前的治疗延长了生存期，但长期结果并不理想。基因治疗显示出前景，然而，体内施用非病毒载体或病毒载体导致低水平的基因转移、随机基因表达和不良反应，包括炎症。间充质干细胞是来自骨髓的自我更新的成体干细胞，易于分离和培养扩增，并具有分化为多种细胞类型的潜力。我们的假设是，基因修饰的MSC可能在肺动脉高压的治疗中具有有益的效果。我们实验室的初步实验结果表明，基因修饰的MSC和野生型或未修饰的MSC的管理，提高勃起功能的老年大鼠。初步研究的结果还表明，肺内施用野生型MSC可减弱野百合碱诱导的肺动脉高压，并恢复肺血管扩张剂对乙酰胆碱的反应，这是野百合碱治疗所损害的。在本申请中提出的实验将检验以下假设：动脉内施用MSC或用eNOS基因或CGRP基因修饰的MSC将在大鼠中的野百合碱诱导的肺动脉高压中具有有益效果。为了验证这一假设，必须制备MSC，表征，基因修饰，并注射到对照大鼠和野百合碱诱导的肺动脉高压大鼠的肺中。除了确定MSC和基因修饰的MSC对基线肺压和血流量的影响之外，还将使用组织学和组织化学技术研究MSC对呼吸性缺氧和血管活性激动剂（包括乙酰胆碱）的肺血管反应的影响以及注射的MSC在肺中的命运。因此，第一个具体目标是从大鼠中分离、表征和基因修饰MSC。第二个具体的目的是调查的影响，注射骨髓间充质干细胞对肺血管功能和野百合碱诱导的肺动脉高压使用新开发的右心导管技术。第三个具体的目的是研究的命运，注射的MSC在肺组织化学技术，去卷积显微镜，和本地化的雄性Y染色体在雌性大鼠的肺中使用PCR和FISH方法。这些研究的结果可能会导致新的基于MSC的治疗肺动脉高压的疗法。