An Animal Model of Chronic Oral Inflammation for Stem Cell-Based Therapy

描述（由申请人提供）：我们建议开发一种自然发生的猫模型，用于通过给予基于干细胞的再生医学来解决慢性粘膜炎症。间充质干细胞（MSC）正在用于治疗免疫介导的炎性疾病的若干临床试验中进行研究。然而，缺乏相关的动物模型研究MSC治疗这些疾病继续限制这种治疗选择的发展。这些疾病经常表现为口腔粘膜的病变。口腔粘膜中的丘疹样皱褶和口疮样溃疡是克罗恩病的常见特征，25-70%的移植物抗宿主病（GVHD）患者存在自身免疫性水疱溃疡性口腔病变，并且除了皮肤外，寻常天疱疮还可影响患者的口腔粘膜。虽然这些疾病是由不同的病因引起的，但所有这些慢性和衰弱性粘膜疾病都是不适当的治疗的结果。 对自身抗原的免疫反应。猫慢性牙龈口炎（FCGS）是一种自然发生的，疼痛和衰弱，免疫介导的炎症性疾病与可能的多因素病因。疾病症状也是对抗原的不适当免疫应答的结果，其表现为严重的口腔溃疡和增生性粘膜病变以及炎症的全身体征。令人兴奋的初步数据表明，在患有FCGS的猫中全身MSC施用导致疾病缓解和炎症的全身生物标志物的正常化。这种疾病模型是研究MSC-免疫系统相互作用的理想模型，因为口腔和局部/全身免疫系统都很容易成像和采样。我们的总体假设是，这种新的，自然发生的动物疾病可以作为一个模型，研究免疫介导的炎症性疾病的MSC治疗。这项拟议的研究将增强我们对基于MSC的免疫调节的基本理解，MSC治疗在自然发生的免疫介导的炎症性疾病背景下的全身影响，并产生新的再生疗法。为了开发这种动物模型，我们将使用流式细胞术表征MSC的表面蛋白表型。然后，我们将充分阐明锝标记的间充质干细胞移植到患有CGS的猫后，通过荧光造影的迁移和归巢。在跟踪MSC的同时，我们将监测血液白细胞、淋巴结细胞亚群、血清蛋白和唾液蛋白的变化，以确定治疗粘膜病变的最佳方法。除生物标志物外，由设盲、委员会认证的兽医牙医进行的定期检查将监测口腔粘膜炎症临床症状的变化。我们还将比较人类和猫科动物口腔病变的病理学，以确定其组织学和免疫组化的相似性。最后，我们将确定是否患有慢性口腔炎性疾病的人共享可测量的生物标志物的改变，包括循环淋巴细胞亚群和血清蛋白，在FCGS猫中注意到。

DESCRIPTION (provided by applicant): We propose to develop a naturally-occurring feline model for the resolution of chronic mucosal inflammation through the administration of stem cell-based regenerative medicine. Mesenchymal stem cells (MSCs) are being investigated in several clinical trials for the treatment of immune- mediated inflammatory diseases. However, a lack of relevant animal models for studying MSC therapy in these disorders continues to limit the development of this therapeutic option. These disorders frequently manifest themselves as lesions of the oral mucosa. Papular folds and aphthous-like ulcers in the oral mucosa are a common feature of Crohn's disease, auto-immune vesci ulo-ulcerative oral lesions are present in 25-70% of patients with graft-versus-host disease (GVHD) and pemphigus vulgaris can affect the patient's oral mucosa in addition to their skin. While these diseases arise from a diverse etiology, all of these chronic and debilitating mucosal diseases are the result of an inappropriate immune response to auto-antigens. Feline chronic gingivostomatitis (FCGS) is a naturally occurring, painful and debilitating, immune-mediated inflammatory disorder with a possible multi-factorial etiology. The disease symptoms are also the result of an inappropriate immune response to antigens that manifests itself in severe oral ulcerative and proliferative mucosal lesions and in systemic signs of inflammation. Exciting preliminary data demonstrate that systemic MSC administration in cats with FCGS results in disease remission and normalization of systemic biomarkers of inflammation. This disease model is ideal to study MSC-immune system interactions as both the oral cavity and local/systemic immune system are readily imaged and sampled. Our overarching hypothesis is that this novel, naturally occurring animal disease can be used as a model to study MSC treatment of immune-mediated inflammatory disorders. The proposed study will enhance our fundamental understanding of MSC-based immunomodulation, the systemic impact of MSC therapy in the context of a naturally occurring immune- mediated inflammatory disease, and generate novel regenerative therapies. To develop this animal model we will characterize the surface protein phenotype of the MSCs using flow cytometry. We will then fully elucidate the migration and homing of technetium-labeled MSCs following transplantation into cats afflicted with CGS via scintigraphy. In conjunction to tracking the MSCs we will monitor changes in blood leukocytes, lymph node cell subsets, serum proteins, and salivary proteins to determine the best practices to treat mucosal lesions. In addition to biomarkers, regular examinations by blinded, board certified veterinary dentists will monitor changes in clinical symptoms of inflammation in the oral mucosa. We will also compare the pathology of human and feline oral lesions to determine their histological and immunohistochemical similarities. Finally, we will determine if humans with chronic oral inflammatory diseases share the alterations in measurable biomarkers, including circulating lymphocyte subsets and serum proteins, noted in cats with FCGS.