Novel immunosuppressive mechanism of human bone marrow mesenchymal stem cells in experimental Crohn’s disease

克罗恩病(CD)是一种慢性肠道炎症性疾病，影响超过70万人 在美国的个人，并在世界各地变得越来越普遍。抗肿瘤坏死因子等生物制品的使用 抗黏附分子药物显著改善了CD的生活质量 然而，生物制品往往会随着时间的推移而失效，并有明显的副作用。新的 利用成人间充质干细胞免疫抑制能力的细胞治疗 针对CD等免疫介导性疾病的(MSC)目前正在进行临床试验。累积的正文 包括我们自己的工作在内的数据表明，局部注射MSC对患者来说是一种有前途的治疗方法 有对抗肿瘤坏死因子无效的肛周瘘。然而，系统的MSC治疗腔性CD 疗效较低，这可能是由于许多原因没有得到充分的调查。所有的 进行的小鼠研究表明，MSC治疗对需要 手法以发展炎症为主，主要用于治疗大肠炎症。 然而，近三分之二的人类CD患者有小肠受累。因此，有一个 迫切需要研究疾病相关和临床前CD小鼠模型中的MSC治疗。在这 本研究旨在研究人骨髓间充质干细胞(HMSC)对SAMP-1/YitFc(SAMP)小鼠的治疗作用。 Samp菌株是一种独特的模型，可自发发展为类CD小肠(SI)。 炎症与人类疾病有着惊人的相似之处。我强劲的初步数据显示 结果表明，SAMP小鼠建立的SI炎症用一剂骨髓来源的治疗 人骨髓间充质干细胞(HMSC)炎症反应明显减轻，粘膜愈合良好。因此，我们的 假设hMSC治疗SAMP小鼠的SI炎症是通过一种涉及调控的机制 从宿主组织的转录组到抗炎和组织再生途径。这就做 在两个具体目标中测试这一假设。在特定的目标1中，我们将使用我们的新SAMP小鼠 医疗复发模型，包括3D立体显微镜(用于粘膜愈合)等新技术 检验hMSC可治疗、预防SI炎症并确定最佳途径的假设 HMSC管理部门。此外，使用一种针对免疫抑制进行优化的新型hMSC，我们将 确定是否可以提高MSC疗法的治疗效果。在目标2中，使用人和老鼠 MSCs采用三重报告、最先进的成像技术、激光捕获 显微解剖，宿主细胞和间充质干细胞的单细胞RNA测序 骨髓间充质干细胞治疗SAMP的愈合机制(S)。我们的临床前实验设计 试验1)已确诊疾病的治疗，2)缓解的维持，3)骨髓间充质干细胞的最佳途径 给药，4)使用增强的免疫抑制hMSC 5)hMSC对粘膜愈合的作用，以及6) 比较生物学方法理解愈合机制(S)具有很高的相关性 人类IBD。这项拨款产生的数据将导致对人类的更机械化的方法 治疗CD患者的临床试验，并可能发现促进组织生长的新分子 再生和治疗，这将构成未来机械式R01的基础。因此，我们有 通过产生新的科学想法，在我们初步发现的基础上提出了一项建议 不仅与CD有关，而且可能与其他免疫介导性疾病有关，其中MSCs 目前正处于临床前或早期临床试验阶段。

Crohn’s disease (CD) is chronic intestinal inflammatory disease that affects more than 700,000 individuals in the US and is becoming more common worldwide. The use of biologics such as anti-TNF and anti-adhesion molecule medications has significantly improved the quality of life of CD patients, however biologics tend to lose efficacy with time and have significant side effects. New cell based therapies that utilize the immunosuppressive capacity of adult mesenchymal stem cells (MSC) for immune mediated diseases like CD are currently in clinical trials. A cumulative body of data, including our own work shows that locally injected MSC are a promising therapy for patients with perianal fistulae refractory to anti-TNF’s. However, systemic MSC therapy for luminal CD has lower efficacy, which may be due to myriad reasons with many inadequately investigated. All the murine studies performed demonstrate the benefit of MSC therapy in mouse models that require manipulation to develop inflammation and are focused on treating large intestinal inflammation. Though, nearly two thirds of human CD patients have small intestinal involvement. Thus, there is a critical need to study MSC therapy in disease relevant and preclinical murine models of CD. In this study, we propose to study the SAMP-1/YitFc (SAMP) mouse for treatment with human MSC (hMSC).The SAMP strain is a unique model that spontaneously develops CD-like small intestinal (SI) inflammation which has impressive similarities to the human disease. My strong preliminary data has shown that SAMP mice with established SI inflammation treated with one dose of bone marrow derived human MSC (hMSC) had significantly lower inflammation and had mucosal healing. Therefore, our hypothesis is that hMSC treat SI inflammation in SAMP mice by a mechanism involving the modulation of transcriptome of host tissue to an anti-inflammatory and tissue regeneration pathway. I will test this hypothesis in 2 specific aims. In specific aim 1, we will use the SAMP mice, our new SAMP medical recurrence model, novel techniques including 3D stereomicroscopy (for mucosal healing) to test the subhypothesis that hMSC can treat, prevent SI inflammation and determine the optimal route of hMSC administration. In addition, using a novel hMSC optimized for immunosuppression, we will determine if we can enhance the treatment efficacy of MSC therapy. In aim 2, using human and mouse MSCs transduced with triple reporter, state-of-the-art imaging techniques, laser capture microdissection, single cell RNA sequencing of host cells and MSCs we will investigate the mechanism(s) of healing in SAMP after treatment with MSC. Our preclinical experimental design that tests 1) treatment of established disease, 2) maintenance of remission, 3) the optimal route of MSC administration, 4) use of enhanced immunosuppressive hMSC 5) hMSC effect on mucosal healing, and 6) comparative biology approach to understand the mechanism (s) of healing has high relevance for human IBD. The data generated from this grant will lead to a more mechanistic approach to human clinical trials for treating CD patients and may identify novel molecules that promote tissue regeneration and healing that will form the basis for mechanistic R01s in the future. Thus, we have generated a proposal that builds upon our preliminary findings by generating new scientific ideas that have relevance not only to CD, but potentially to other immune mediated diseases where MSCs are currently in preclinical or early-stage clinical trials.