Stem Cell Therapy for Pulmonary Hypertension

肺动脉高压的干细胞疗法

• 批准号: 7034579
• 负责人: Philip J Kadowitz
• 金额: $ 36.25万
• 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/7034579
• 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)：拟议研究的广泛的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方法。肺动脉高压是一种严重的、往往是致命的疾病，其特征是肺动脉压升高、右心衰竭和肺血管床重构。虽然目前的治疗方法延长了患者的生存期，但长期效果并不乐观。基因治疗显示出了希望，然而，在体内使用非病毒载体或病毒载体会导致低水平的基因转移、随机的基因表达和包括炎症在内的不良反应。MSCs是一种来自骨髓的自我更新的成体干细胞，在培养中很容易分离和扩增，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MSCs可能在治疗肺动脉高压方面有有益的作用。我们实验室的初步实验结果表明，基因修饰的MSCs和野生型或未修饰的MSCs可以改善老年大鼠的勃起功能。初步研究结果还表明，气管内注射野生型MSCs可减轻野百合碱所致的肺动脉高压，并恢复乙酰胆碱所致的肺血管扩张反应，而野百合碱治疗可使其受损。本申请中提出的实验将检验这样一种假设，即气管内注射MSCs或eNOS基因或CGRP基因修饰的MSCs将对野百合碱诱导的大鼠肺动脉高压产生有益影响。为了验证这一假设，必须制备、鉴定、基因修饰MSCs，并将其注射到对照组大鼠和野百合碱诱导的肺动脉高压大鼠的肺中。除了确定MSCs和基因修饰的MSCs对基础肺压力和血流的影响外，还将利用组织学和组织化学技术研究MSCs对呼吸性低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MSCs在肺中的去向。因此，第一个特定的目标是分离、鉴定和基因修饰大鼠骨髓间充质干细胞。第二个具体目的是利用新开发的右心插管技术研究注射MSCs对肺血管功能和野百合碱诱导的肺动脉高压的影响。第三个特定目的是利用组织化学技术、去卷曲显微镜以及用PCR和FISH方法对雄性Y染色体在雌性大鼠肺中的定位，研究注射MSCs在肺中的去向。这些研究的结果可能导致基于MSC的新的治疗肺动脉高压的方法。