Stem Cell-Adrenomedullin Therapy for Cancer Linked Lymphedema

摘要 淋巴水肿影响着美国数百万乳腺癌(BCA)幸存者中的38%到89%， 它是研究最少、相对低估、最不了解的并发症之一。 癌症或其治疗。BCA相关淋巴水肿是淋巴管破坏的结果 切除淋巴结或放射治疗，会导致虚弱的肢体肿胀，慢性炎症， 组织纤维化，以及对感染的易感性增加。这项研究解决的主要挑战是 攻克与BCA相关的淋巴水肿问题。我们提议的再生能源的成功开发 治疗将显著降低与癌症相关的淋巴水肿的发病率。这样做的理由是 建议是基于我们的新发现，其中两种肽激素-肾上腺髓质素(ADM)和中介 (IMD)-及其同源受体，降钙素受体样受体(CLR)和受体活性调节 蛋白质(RAMP)，是淋巴管发育所必需的，并且将是充分和必要的 将患者的成人组织干细胞重新编程为功能性淋巴管内皮细胞(LECs)。这些LEC 能形成新的淋巴管，恢复淋巴水肿患者的淋巴循环。然而，狂野- (Wt)型ADM和IMD在体内的半衰期较短；因此，我们假设新发明的 CLR-RAMP激动剂(指定为sCLR激动剂)是稳定的ADM和IMD类似物，将更有效 在对抗淋巴浮肿方面优于wt激素肽。因为BCA幸存者可能会被剥夺内皮细胞 对于淋巴管再生至关重要的祖细胞或干细胞，我们进一步假设 整合sCLR激动剂和干细胞的联合疗法将是最有效的方法 防止淋巴水肿的发生，或减轻淋巴水肿的衰弱作用。这个项目的目标是 是通过应用新型的sCLR激动剂和成人脂肪来克服BCA相关的淋巴水肿的问题。 组织干细胞(ASCs)或诱导多能干细胞(IPSCs)再生LECs以恢复淋巴管 体内循环，并了解sCLR激动剂介导的晶状体上皮细胞重编程的机制 来自ASCs或IPSCs，用于恢复正常的淋巴系统。我们计划通过研究 SCLR激动剂与ASCs或IPSCs联合治疗小鼠淋巴水肿的疗效 模特们。为了解决研究假设，我们将集中在两个具体目标：(1)确定疗效 用sCLR激动剂和ASCs或IPSCs联合治疗体内重建淋巴循环；以及 (2)阐明sCLR激动剂调节晶状体上皮细胞重编程的信号机制。 ASCs或IPSCs。总之，这项研究需要验证新的联合疗法的实用性。另外， 这一机制研究的结果将对潜在的信号机制提供重要的洞察。 SCLR激动剂促进ASCs或IPSCs的LECs重编程以再生功能淋巴管 系统，并有助于优化sCLR激动剂用于治疗淋巴水肿的治疗应用。

ABSTRACT Lymphedema affects between 38% and 89% of millions of breast cancer (BCa) survivors in the United States, and it is one of the least researched, relatively underestimated, and most poorly understood complications of cancer or its treatment. BCa-linked lymphedema occurs as a result of lymphatic vessel destruction during the removal of lymph nodes or radiation therapy, and it can lead to debilitating limb swelling, chronic inflammation, tissue fibrosis, and increased susceptibility to infection. This research addresses the overarching challenge of conquering the problems of BCa-linked lymphedema. Successful development of our proposed regenerative therapy will significantly reduce the morbidity associated with cancer-linked lymphedema. The rationale for this proposal is based on our new findings where two peptide hormones—adrenomedullin (ADM) and intermedin (IMD)—and their cognate receptors, the calcitonin receptor-like receptor (CLR) and receptor activity-modifying proteins (RAMPs), are required for lymphatic vessel development and will be sufficient and necessary to reprogram adult tissue stem cells from patients to functional lymphatic endothelial cells (LECs). These LECs can form new lymphatic vessels for restoring lymphatic circulation in patients with lymphedema. However, wild- type (wt) ADM and IMD have shorter half-lives in vivo; thus, we hypothesize that the newly invented stable CLR-RAMP agonists (designated sCLR agonists), which are stable ADM and IMD analogs, will be more potent than wt hormone peptides in combating lymphedema. Because BCa survivors may be deprived of endothelial progenitor or stem cells that are essential for the regeneration of lymphatic vessels, we further hypothesize that the combination therapy that integrates sCLR agonists and stem cells will be the most efficient approach to prevent the occurrence of, or to reduce the debilitating effects of lymphedema. The objectives of this project are to overcome the problems of BCa-linked lymphedema by applying novel sCLR agonists and adult adipose- tissue stem cells (ASCs) or induced pluripotent stem cells (iPSCs) to regenerate LECs to restore lymphatic circulation in vivo, and to understand the mechanisms for sCLR agonists-mediated reprogramming of LECs from ASCs or iPSCs for reviving a functional lymphatic system. We plan to achieve our Aims by studying the efficacy of a combination therapy that combines sCLR agonists and ASCs or iPSCs in the mouse lymphedema models. To address the study hypothesis, we will focus on two Specific Aims: (1) to determine the efficacy of combined treatment with sCLR agonists and ASCs or iPSCs in reconstructing lymphatic circulation in vivo; and (2) to elucidate the signaling mechanisms whereby sCLR agonists regulate reprogramming of LECs from ASCs or iPSCs. In summary, this study is required to validate the utility of novel combination therapies. Also, the results of this mechanistic study will provide significant insight into the signaling mechanisms underlying sCLR agonists-promoted reprogramming of LECs from ASCs or iPSCs for regenerating a functional lymphatic system, and to contribute to optimizing therapeutic applications of sCLR agonists for healing lymphedema.