Protein Conjugation For Antigen-Specific Treatment Of Thyroid Autoimmune Diseases

摘要 每6个美国人中就有1人患有自身免疫性疾病，给美国医疗体系造成的损失超过1000亿美元 每年一次。最常见的自身免疫性疾病之一是格雷夫斯病，3%的女性和 0.5%的男性。Graves病的主要原因是形成针对甲状腺的抗体。 刺激素受体(TSHR)。TSHR是一种G蛋白偶联受体，具有可溶性的胞外受体 A亚基。据了解，这个A亚基是TSHR自身免疫识别的触发因素，并且 由此产生的抗体。这些甲状腺刺激性抗体(TSAb)会导致细胞过度激活 通过模拟甲状腺激素对TSHR的影响。过度刺激甲状腺会导致 甲状腺机能亢进症，或甲状腺激素甲状腺激素分泌过多，导致焦虑，体重减轻， 最终导致骨质疏松症和甲状腺癌。Graves病的一种常见并存是Graves病 眼病(GO)，由免疫攻击视神经成纤维细胞引起，可导致视神经病变 纤维化，最终导致失明。与大多数自身免疫性疾病一样，目前还没有治疗方法 解决Graves病的根本原因--TSHR的免疫识别。相反，现代的方法 Graves病的治疗依赖于症状管理，主要是通过联合放射治疗 碘(RAI)和甲状腺抑制剂，如他巴唑。在RAI治疗中，碘131被给予患有 其目的是消除甲状腺功能，从而减轻甲亢症状。不幸的是，RAI经常导致 在完全甲状腺切除术中，引发慢性甲状腺功能减退症，并需要终身依赖 补充甲状腺激素。RAI还与严重GO增加20%有关，这是一种非常有问题的 推定治疗的结果。相比之下，他咪唑等甲状腺抑制剂可以一次服用数年。 时间作为日常治疗，但往往会导致复发。Graves病患者跨治疗策略分析 已经显示出生活质量的显著下降，表明需要改进治疗策略。 需要的是一种治疗策略，解决自身免疫性疾病的根本原因，在 本案是对TSHR的识别和瞄准。最近的研究表明，抗原特异性免疫 当抗原可以偶联到某些产生耐受的受体、纳米粒和 蛋白质。然而，与大多数蛋白质抗原一样，TSHR作为一种基因融合生产是具有挑战性的。我们 已经开发出一种新的蛋白质偶联技术，能够仅使用天然的蛋白质将完整的蛋白质融合在一起 氨基酸。我们建议将这项技术应用于抗原特异性疗法的创建，方法是融合 TSHR对耐受性龋病的治疗，并将其应用于Graves‘s小鼠模型以评估其对血清的影响 甲状腺激素和TSAb水平。

Abstract Autoimmune disorders effect 1 in 6 Americans and cost the American Healthcare system over $100 billion annually. One of the most common autoimmune disorders is Graves’ disease, which effects 3% of women and 0.5% of men. The primary cause of Graves’ disease is the formation of antibodies against the Thyroid Stimulating Hormone Receptor (TSHR). The TSHR is a G-protein coupled receptor with a soluble extracellular A-subunit. It is understood that this A-subunit is the trigger for autoimmune recognition of the TSHR and resultant production of antibodies. These Thyroid Stimulating Antibodies (TSAb) cause over activation of the thyroid by mimicking the effect of thyroid hormones on the TSHR. Overstimulation of the thyroid leads to hyperthyroidism, or an overproduction of the thyroid hormone thyroxine which results in anxiety, weight loss, and ultimately osteoporosis and thyroid cancer. A common co-morbidity of Graves’ disease is Graves’ opthalmopathy (GO) which is caused by immunological attack on optical fibroblasts and can lead to optical fibrosis and eventually loss of vision. As with most autoimmune disorders, there is not treatment available to address the root cause of Graves’ disease –the immune recognition of the TSHR. Instead, modern approaches to treatment of Graves’ disease rely on symptom management, primarily through a combination of radioactive iodine (RAI) and thyroid inhibitors such as methimazole. In RAI therapy, iodine 131 is given to the patient with the goal of ablating thyroid function, thus diminishing hyperthyroid symptoms. Unfortunately, RAI often results in complete thyroid ablation, triggering chronic hypothyroidism and necessitation lifelong dependence on thyroxine supplements. RAI is also associated with a 20% increase in severe GO, a highly problematic outcome for a putative therapy. Thyroid inhibitors such as methimazole in contrast can be given for years at a time as a daily treatment, but often lead to relapse. Analysis of Graves’ patients across treatment strategies has shown significant decrease in quality of life, suggesting that improved therapeutic strategies are required. What is needed is a therapeutic strategy that addresses the fundamental cause of autoimmune disorders, in this case the recognition and targeting of the TSHR. Recent work has shown that antigen specific immune tolerance is possible when antigens can be coupled to certain tolerogenic receptors, nanoparticles, and proteins. However, as with most protein antigens, the TSHR is challenging to produce as a genetic fusion. We have developed a novel protein coupling technique capable of fusing intact proteins together using only native amino acids. We propose to apply this technique to the creation of antigen specific therapeutics by fusing the TSHR to tolerogenic cariers and delivering these to mouse models of Graves’ to assess the impact on serum thyroxine and TSAb levels.