HLA-DQ-derived RTLs for Treatment of Celiac Disease

描述（由申请人提供）：乳糜泻是一种炎性自身免疫性疾病，其中CD 4 + T细胞靶向、损伤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麸质，一种在小麦、黑麦和大麦中发现的蛋白质。这种使人衰弱的疾病影响每120-300人中的1人，仅在美国就转化为大约220万人的市场规模。拟议工作的目标是在体外对人类HLA-DQ限制性T细胞克隆测试先导化合物（在I期资金下开发），并量化其控制HLA-DQ转基因小鼠中激活的肠道T细胞的能力。所提出的方法基于专利RTL技术（美国专利#6，270，772），Virogenomics拥有该技术的独家许可。RTL技术的治疗效果首先在多发性硬化的动物模型EAE（实验性自身免疫性脑脊髓炎）中描述。在这种动物模型中，RTLs抑制了致病性T细胞的活化，可用于预防和治疗疾病。这些分子在治疗其他人类疾病中的潜力为开发用于治疗乳糜泻的HLA-DQ衍生的人类RTL提供了强有力的理论基础。在I期资助下，设计并纯化了HLA-DQ衍生的重组T细胞受体配体（RTL）。这些分子也进行了严格的生物化学和生物物理特性。最近，已经开发了谷蛋白敏感性的huCD 4/DQ 2和huCD 4/DQ 8转基因小鼠模型。在huCD 4/DQ 8转基因模型中，T细胞对麸质的反应是CD 4依赖性的，HLA-DQ限制性的，并导致细胞因子IL-6、TGF-β和IL-10的产生，来自麸质喂养小鼠的肠淋巴细胞显示出活化的表型。我们将使用这些动物模型，通过体内概念验证实验，获取DQ衍生的RTL先导化合物。在II期资助下，我们提出1）DQ衍生的RTL对从乳糜泻患者获得的人T细胞系的生物学作用的体外表征，使用这些研究来改进我们的先导化合物以使其生物学活性最大化;和2）在乳糜泻转基因小鼠模型中调节细胞因子产生和阻断肠T细胞活化的RTL剂量要求的表征。

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.