HLA-DQ-derived RTLs for Treatment of Celiac Disease

描述(申请人提供)：乳糜泻是一种炎症性自身免疫性疾病，在这种疾病中，CD4+T细胞靶向、损害并最终破坏小肠的绒毛组织结构，干扰从食物中吸收营养。患有乳糜泻的人不能耐受面筋，面筋是一种存在于小麦、黑麦和大麦中的蛋白质。这种令人衰弱的疾病每120-300人中就有1人受到影响，仅在美国就意味着大约220万人的市场规模。这项拟议工作的目标是在体外测试先导化合物(在第一阶段资金支持下开发)在人类HLA-DQ限制性T细胞克隆上，并量化它们在HLA-DQ转基因小鼠中控制激活的肠道T细胞的能力。提出的方法是基于病毒基因组学拥有独家许可证的专利RTL技术(美国专利#6,270,772)。RTL技术的治疗效果首次在多发性硬化症的动物模型EAE(实验性自身免疫性脑脊髓炎)中被描述。在该动物模型中，RTLS抑制了致病T细胞的激活，可用于预防和治疗疾病。这些分子在治疗其他人类疾病方面的潜力为开发基于人类白细胞抗原DQ的人类RTL用于治疗乳糜泻提供了强有力的理论基础。在第一阶段的资助下，设计并纯化了人类白细胞抗原-DQ衍生的重组T细胞受体配体(RTL)。这些分子还经过了严格的生化和生物物理表征。近年来，已经建立了对面筋敏感的huCD4/DQ2和huCD4/DQ8转基因小鼠模型。在huCD4/DQ8转基因模型中，T细胞对面筋的反应是依赖于面筋的，人类白细胞抗原DQ受到限制，并导致细胞因子IL-6、转化生长因子-β和白介素10的产生，而摄入面筋的小鼠的肠道淋巴细胞表现为活化的表型。我们将使用这些动物模型通过体内概念验证实验来获取DQ衍生的RTL先导化合物。在第二阶段资助中，我们建议1)体外表征从乳糜泻患者获得的DQ来源的RTL对人类T细胞系的生物学效应，利用这些研究来改进我们的先导化合物，以最大限度地提高它们的生物活性；以及2)表征RTL的剂量需求，以调节细胞因子的产生，并在乳糜泻转基因小鼠模型中阻断肠道T细胞的激活。

DESCRIPTION (provided by applicant): Celiac disease is an inflammatory autoimmune disease in which CD4+ T cells target, damage and eventually destroy the villous tissue structures of the small intestine, interfering with the absorption of nutrients from food. People who have celiac disease cannot tolerate gluten, a protein found in wheat, rye and barley. This debilitating disease affects 1 of every 120-300 people, which in the United States alone translates to a market size of roughly 2.2 million people. The goal of the proposed work is to test lead compounds (developed under Phase I funding) in vitro on human HLA-DQ-restricted T cell clones, and quantify their ability to control activated intestinal T cells in HLA-DQ transgenic mice. The approach presented is based on patented RTL technology (US Patent #6,270,772) for which Virogenomics holds an exclusive license. The therapeutic efficacy of the RTL technology was first described in EAE (experimental autoimmune encephalomyelitis), an animal model of multiple sclerosis. In this animal model, RTLs inhibited activation of pathogenic T cells and could be used to prevent and treat disease. The potential of these molecules in the treatment of other human diseases provided a strong rationale to develop HLA-DQ-derived human RTLs for treatment of celiac disease. Under Phase I funding, HLA-DQ-derived Recombinant T cell receptor Ligands (RTLs) were designed and purified. These molecules also underwent rigorous biochemical and biophysical characterization. Recently, huCD4/DQ2 and huCD4/DQ8 transgenic mouse models of gluten sensitivity have been developed. In the huCD4/DQ8 transgenic model, the T cell response to gluten was CD4 dependent, HLA-DQ restricted and led to the production of cytokines IL-6, TGF-beta and IL-10, with intestinal lymphocytes from gluten-fed mice displaying an activated phenotype. We will use these animal models to take DQ-derived RTL lead compounds through proof-of-concept experiments in vivo. Under Phase II funding we propose 1) In vitro characterization of the biological effects of DQ-derived RTLs on human T cell lines obtained from patients with celiac disease, using these studies to refine our lead compounds to maximize their biological activity; and 2) Characterization of RTL dosage requirements to modulate cytokine production and block activation of intestinal T cells in transgenic mouse models of celiac disease.