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，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(MSCs)正在进行几项临床试验，用于治疗免疫介导性炎症性疾病。然而，缺乏相关的动物模型来研究MSC在这些疾病中的治疗仍然限制了这种治疗方案的发展。这些疾病通常表现为口腔粘膜的损害。口腔黏膜丘疹和口疮样溃疡是克罗恩病的共同特征，移植物抗宿主病(GVHD)患者中有25%-70%的患者存在自身免疫性Vesci溃烂口腔病变，寻常型天疱疮除了影响患者的皮肤外，还会影响患者的口腔黏膜。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的粘膜疾病都是由不适当的 对自身抗原的免疫反应。猫慢性牙周炎(FCGS)是一种自然发生的、疼痛和虚弱的免疫介导性炎症性疾病，其病因可能是多因素的。疾病症状也是对抗原的不适当免疫反应的结果，这种免疫反应表现在严重的口腔溃烂和增生性粘膜病变以及全身炎症迹象中。令人振奋的初步数据表明，在患有FCGS的猫中，全身应用MSC可以缓解疾病并使全身炎症生物标志物正常化。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/系统免疫系统都很容易成像和采样。我们的主要假设是，这种新的、自然发生的动物疾病可以作为研究MSC治疗免疫介导性炎症性疾病的模型。这项拟议的研究将加强我们对基于MSC的免疫调节、MSC治疗在自然发生的免疫介导性炎症性疾病背景下的全身影响的基本理解，并产生新的再生疗法。为了建立这种动物模型，我们将使用流式细胞术来表征MSCs的表面蛋白表型。然后，我们将通过核素扫描，全面阐明氚标记的MSCs在移植到患有CGS的猫体内后的迁移和归巢。在跟踪MSCs的同时，我们将监测血白细胞、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实践。除了生物标志物外，盲人、委员会认证的兽医定期检查将监测口腔粘膜炎症的临床症状的变化。我们还将比较人类和猫科动物口腔损伤的病理，以确定它们在组织学和免疫组织化学上的相似性。最后，我们将确定患有慢性口腔炎症性疾病的人类是否也有可测量的生物标记物的变化，包括循环淋巴细胞亚群和血清蛋白，在患有FCGS的猫中注意到。

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.