Stem Cell Therapy for Pulmonary Hypertension

这项研究的广泛的长期目标是开发一种新的间充质干细胞（MSC） 肺动脉高压的治疗方法肺动脉高压是一种严重的， 以肺动脉压升高、右心衰竭和肺动脉重构为特征的疾病 肺血管床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。MSC是自我更新的成体 来自骨髓的干细胞容易分离并在培养物中扩增，并且具有分化成 许多细胞类型。我们的假设是，基因修饰的MSC可能在治疗 肺动脉高压在本实验室进行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改善老年大鼠的勃起功能。的结果 初步研究还表明，腹膜内给予野生型MSC可减弱野百合碱诱导的 肺动脉高压和恢复肺血管舒张反应乙酰胆碱，这是受损的 野百合碱治疗在本申请中提出的实验将测试以下假设： 施用MSC或用eNOS基因或CGRP基因修饰的MSC将在以下方面具有有益效果： 野百合碱诱导的大鼠肺动脉高压。为了检验这一假设，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大鼠的肺中。 肺动脉高压除了确定MSC和基因修饰的MSC对基线的影响之外， 肺动脉压和血流量，MSC对肺血管反应的影响， 缺氧和血管活性激动剂，包括乙酰胆碱，以及肺中注射的MSC的命运将被确定。 使用组织学和组织化学技术进行研究。因此，第一个具体目标是分离， 表征和基因修饰来自大鼠的MSC。第二个具体目标是研究注射的效果。 骨髓间充质干细胞对肺血管功能及肺动脉高压的影响 开发了右心导管插入技术。第三个具体目的是研究注射的MSC在小鼠中的命运。 肺使用组织化学技术，去卷积显微镜，和定位的男性Y染色体， 采用PCR和FISH技术对雌性大鼠肺组织进行了检测。这些研究的结果可能会导致新的MSC- 肺动脉高压的治疗方法

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.