Imaging lymph function in humans and in a murine model of lymphedema

描述(申请人提供)：淋巴管和淋巴功能障碍与转移性癌症、哮喘、糖尿病和肥胖症密切相关。然而，尽管淋巴管在这些疾病中很重要，但非侵入性监测淋巴功能的诊断成像方法还不存在。最近，我们报道了使用非特异性近红外荧光团IC-Green以及针对淋巴血管内皮受体LYVE-1的淋巴内皮分子染色，以非侵入性和动态的方式成像完整猪和人类淋巴功能的能力。此外，在开发出分子靶向“激活”淋巴管内皮细胞的显像剂后，我们假设我们可以成像淋巴管生成。我们的结果证明了监测淋巴流量及其分子标志物在诊断和治疗患有原发和继发性淋巴水肿的人群中淋巴疾病的可行性。在这项研究应用中，我们假设：近红外(NIR)光学方法可以用来成像淋巴流动、功能和重塑，以提供诊断和关于治疗反应的信息。具体地说，我们寻求：(1)改进成像硬件并开发用于临床前和临床成像研究中淋巴功能的自动成像和分析的软件方法；(2)开发、合成和验证针对LYVE-1、透明质酸酶和α-9整合素的分子靶向显像剂，这些显像剂参与淋巴水肿和淋巴管生成过程；(3)在免疫活性的SKH-1小鼠获得性淋巴水肿模型中建立功能性淋巴成像方法，以使用微量的非特异性IC-Green和IRDye800CW来评估淋巴水肿的时间进展，然后使用针对LYVE-1、透明质酸降解和增殖的LEC和BEC增殖上表达的α-9、α-v整合素的分子显像剂来跟踪进展和治疗反应；最后，(4)对受试者志愿者进行成像，以建立基线淋巴功能成像参数，并对淋巴水肿患者进行成像，以评估淋巴功能成像参数随疾病进展和对常规治疗的反应/无反应而发生的变化。这项拟议的工作利用了新的开发仪器和显像剂，建立了淋巴水肿的小鼠模型，并建立了一个临床试验团队，将淋巴成像方面的发现转化为患者。 项目简介：淋巴管和淋巴功能障碍与转移性癌症、哮喘、糖尿病和肥胖症密切相关。然而，尽管淋巴管在这些疾病中很重要，但非侵入性监测淋巴功能的诊断成像方法还不存在。在这项工作中，我们开发了新的成像技术来评估淋巴疾病和健康中的淋巴功能和分子标志物。

DESCRIPTION (provided by applicant): Malfunctioning lymphatics and lymphatic function are strongly implicated in metastatic cancer, asthma, diabetes, and obesity. Yet despite the importance of the lymphatics in these diseases, diagnostic imaging approaches to non-invasively monitor lymph function are non-existent. Recently, we reported the ability to non-invasively and dynamically image lymph function in the intact swine as well as in humans using a non- specific near-infrared fluorophore, IC-Green, as well as a molecular stain of lymph endothelium that targets the lymph vascular endothelium receptor, LYVE-1. In addition, upon developing imaging agents that molecularly target "activated" lymph endothelium, we hypothesize that we can image lymphangiogenesis. Our results demonstrate the feasibility of monitoring lymph flow and its molecular markers for the diagnosis and management of lymph diseases in a population of patients who suffer from primary and secondary lymphedema. In this research application, we hypothesize that: near infrared (NIR) optical methods can be used to image lymph flow, function, and remodeling to provide diagnostics and information on response to therapy. Specifically, we seek to: (1) Refine imaging hardware and develop software methodologies for automated imaging and analysis of lymph function in preclinical and clinical imaging studies; (2) Develop, synthesize, and validate molecularly targeting imaging agents that are specific to LYVE-1, hyaluronidase, and alpha-9 integrins that are involved in the process of lymphedema and lymphangiogenesis; (3) Develop functional lymph imaging methodologies in an immunocompetent SKH-1 mouse model of acquired lymphedema in order to assess the temporal progression of lymphedema using trace amounts of non-specific IC-Green and IRDye800CW to assess lymphatic function, and then to use molecular imaging agents that target LYVE-1, hyaluronan degradation, and alpha-9, alpha-v integrins expressed on proliferating LEC and BEC proliferation to track the progression and response to therapy; and finally to (4) Conduct imaging on human subject volunteers to establish baseline lymph function imaging parameters as well as on lymphedema patients to assess alteration in lymph function imaging parameters with disease progression and response/non-response to conventional therapy. The proposed work utilizes new developmental instrumentation and imaging agents, established mouse models of lymphedema, and a clinical trial team to translate discoveries in lymph imaging to patients. Project Narrative: Malfunctioning lymphatics and lymphatic function are strongly implicated in metastatic cancer, asthma, diabetes, and obesity. Yet despite the importance of the lymphatics in these diseases, diagnostic imaging approaches to non-invasively monitor lymph function are non-existent. In this work we develop new imaging techniques to assess lymph function and molecular markers in lymph disease and health.