Novel therapies for obesity- or diabetes-related lymphatic dysfunction

翻译后摘要：内皮功能障碍，如淋巴功能障碍（LD）影响许多患者在美国。 根据美国国立卫生研究院的一份报告，LD是最不了解，相对低估，最少的疾病之一。 在美国研究疾病的并发症或其治疗。继发性LD是由内皮细胞引起的， 淋巴系统的功能障碍或获得性缺陷，并且通常与肥胖，糖尿病， 感染、肿瘤、创伤和治疗方式。虽然没有像乳房切除术后那样经常报道- 除了诱发LD外，肥胖也是当今美国实践中看到的LD的最常见原因之一。在 美国，超过20%的人口（约4000万）被认为是肥胖的。一般肥胖者LD的患病率 人群相对较低;然而，在病态肥胖患者中估计有74%的LD患病率， 报道LD是一种在病态肥胖患者中尚未得到充分研究的共病状况， 人口逐年增加。LD也可能与静脉药物滥用有关。如果不是 如果管理得当，LD可导致疼痛的肢体肿胀，慢性炎症，组织纤维化， 易受严重感染和其他疾病的影响。因此，存在未满足但紧急的医疗需求， 建立新的抗LD治疗剂，用于减少或克服LD在扩张中的衰弱作用， 患有病态肥胖或糖尿病的LD患者人群。这项研究旨在解决这一问题 解决问题。基本原理是基于我们的研究结果表明，肾上腺髓质素， （ADM）-及其同源受体，降钙素受体样受体（CGRP）和受体活性修饰 RAMPs是淋巴管发育所必需的，是必要的，也是足够的， 重编程成体组织干细胞以产生功能性淋巴管内皮细胞（LEC）。LEC可以形成 新的淋巴管，以恢复淋巴循环的LD患者患有LD。野生型ADM具有 体内半衰期极短，限制了其临床应用。值得注意的是，ADM（22-52）作为一种有效的免疫调节剂发挥作用。 ADM-受体（AMPA/RAMP 2）拮抗剂（称为AMRA），用于阻断血管生成和肿瘤生长。 因此，我们建议创建高度稳定的专有白蛋白（Alb）-ADM和Fc-ADM融合蛋白作为临床应用。 抗LD治疗剂和作为专有无毒抗血管生成/抗肿瘤蛋白的Alb-AMRA和Fc-AMRA 毒品具体目的：（目的1）产生一组Alb-ADM和Fc-ADM融合蛋白，表征融合蛋白的结构。 使用体外和体内测定，获得最具活性的融合蛋白，并产生Alb-AMRA和Fc-AMRA蛋白 同样的。(Aim 2）评估用Alb-2000的组合方案对淋巴再生的功效和特异性。 ADM联合Alb-AMRA或Fc-ADM联合Fc-AMRA对小鼠后肢LD模型的影响，并观察其毒性作用 体内研究。意义：在这项研究之后，我们将确定有效的抗LD方案， 开发（例如，毒性研究和药代动力学），这将在II期研究中解决。这项工作 强烈支持第二阶段，使这项工作成为一个引人注目的和一流的抗LD治疗。

Abstract: Endothelial dysfunction such as lymphatic dysfunction (LD) affects many patients in the U.S. According to a report of the NIH, LD is one of the most poorly understood, relatively underestimated, and least researched complications of diseases or its treatment in the U.S. Secondary LD is caused by an endothelial dysfunction or an acquired defect in the lymphatic system and is commonly associated with obesity, diabetes, infection, neoplasm, trauma, and therapeutic modalities. Although not reported as often as postmastectomy- induced LD, obesity is also one of the most common causes of LD seen in practice in the U.S. today. In the U.S., over 20% of the population (~40 millions) is considered obese. The prevalence of LD in general obese population is relative low; however, an estimated 74% prevalence of LD in morbidly obese patients has been reported. LD is one comorbid condition that has not been well studied in morbidly obese patients, whose population has been increasing year after year. LD may also be associated with intravenous drug abuse. If not properly managed, LD can lead to painful limb swelling, chronic inflammation, tissue fibrosis, and increased susceptibility to serious infection and other diseases. Thus, there is an unmet yet urgent medical need to establish novel anti-LD therapeutics for reducing or overcoming the debilitating effects of LD in the expanding population of LD patients with morbid obesity or diabetes. This research is proposed to tackle this poorly addressed problem. The rationale is based on our findings demonstrating that hormone—adrenomedullin (ADM)—and its cognate receptors, the calcitonin receptor-like receptor (CLR) and receptor activity-modifying proteins (RAMPs), are required for lymphatic vessel development and are necessary and sufficient to reprogram adult tissue stem cells to generate functional lymphatic endothelial cells (LECs). LECs can form new lymphatic vessels to restore lymphatic circulation in LD patients suffering from LD. Wild-type ADM has an extremely short half-life in vivo, limiting its clinical application. Notably, ADM(22–52) functions as a potent ADM-receptor (CLR/RAMP2) antagonist (termed AMRA) for blocking vascular angiogenesis and tumor growth. Thus, we propose creating highly stable proprietary albumin (Alb)-ADM and Fc-ADM fusion proteins as clinical anti-LD therapeutics and Alb-AMRA and Fc-AMRA as proprietary nontoxic anti-angiogenesis/antitumor protein drugs. Specific Aims: (Aim 1) Generate a panel of Alb-ADM and Fc-ADM fusion proteins, characterize the most active fusion proteins using in vitro and in vivo assays, and create Alb-AMRA and Fc-AMRA proteins similarly. (Aim 2) Assess the efficacy and specificity for lymphatic regeneration with combined regimens of Alb- ADM plus Alb-AMRA or Fc-ADM plus Fc-AMRA in the murine models of hindlimb LD, and conduct their toxicity studies in vivo. Significance: After this study, we will identify potent anti-LD regimens ready for preclinical development (e.g., toxicity studies and pharmacokinetics), which will be addressed during Phase II. This work strongly supports Phase II, making this work a compelling and first-in-class anti-LD therapy.