ROLE OF HUMORAL IMMUNITY IN METASTASIS

体液免疫在转移中的作用

• 批准号: 8171585
• 负责人: EDWARD F PATZ
• 金额: $ 0.55万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171585
• 关键词: Affinity; Alpha-glucosidase; Animal Model; Animals; Antibodies; Autoantibodies; Blood specimen; CD34 gene; Cancer Patient; Cancer cell line; Cell Line; Cells; Computer Retrieval of Information on Scientific Projects Database; Conditioned Culture Media; Data; Development; Disease; Early Diagnosis; Funding; Future; Glucosidase II; Grant; Hematopoietic stem cells; Human; Humoral Immunities; Image; Imaging Techniques; Immune response; Immunocompromised Host; In Vitro; Injection of therapeutic agent; Institution; Lung; Malignant Neoplasms; Malignant neoplasm of lung; Membrane Glycoproteins; Metastatic Adenocarcinoma; Metastatic Lesion; Monitor; Mus; Neoplasm Metastasis; Outcome; Passive Immunity; Patients; Phenotype; Play; Process; Proteins; Recurrence; Research; Research Personnel; Resources; Role; Serum; Serum-Free Culture Media; Source; Staging; Tail; Time; United States National Institutes of Health; Veins; Western Blotting; base; cancer cell; cell motility; design; extracellular; host neoplasm interaction; in vitro Assay; migration; mouse model; neoplastic cell; research study; response; theories; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We propose the theory that the host response is intimately involved in the process of metastasis, and that a better understanding of the tumor-host interaction is essential if there is to be an improvement in outcomes. We have recently discovered autoantibodies in lung cancer patients that have a strong association to the non-metastatic phenotype. Based on these findings, we have formulated the hypothesis that the humoral response in some patients with cancer plays a determining role in whether or not metastasis occurs. We propose initially to investigate an autoantibody prevalent in non-metastatic patients and determine if it is protective against metastasis, and if so, by what mechanism. We have found that conditioned medium (CM) from H1568 cells (a highly metastatic human lung cancer cell line) is capable of initiating CD34+ hematopoietic progenitor cell (HPC) migration in an in vitro assay that mimics one of the initial steps in the development of the metastatic niche. Sera from patients with stage I non-recurrent or late stage metastatic adenocarcinomas were used to probe western blots of proteins from CM from this cell line in order to identify autoantibodies to proteins with possible migration promoting functions. We identified antibodies to alpha-glucosidase II in 70% of patients with non-recurrent lung cancer and in none of the patients with metastatic lung cancer. Addition of an anti-alpha-glucosidase II antibody to the CM decreased CD34+ cell migration in vitro by 31-63%. This suggests that H1568 cells secrete an extracellular form of glucosidase II that is functional on cell surface glycoproteins of CD34+ cells. We want to determine first if priming animals with conditioned medium will accelerate metastatic disease, and then determine if inhibition of glucosidase II will reduce metastasis. We will use imaging (microCT) to follow the development and progression of lung metastasis. This will allow us to use a minimal number of animals to determine differences in rates, size, and distribution of metastatic lesions. We will begin by using a well established mouse model of metastasis through tail vein injection. 1. Eight immunocompromised mice will be used. Four mice will be primed with condition medium from H1568 lung cancer cells for 3 days. Four mice will be primed with serum free medium. We will then inject all mice with approximately one million tumor cells, and the animals will be imaged two times per week for three weeks (previous studies have suggested that the metastasis will be well established by 21 days). As above we will determine if priming the animals will increase metastasis. 2. We will then plan to perform a second experiment and pre-treat 4 mice with affinity purified human anti-alpha-glucosidase II antibody at the time of CM injection to see if this blocks HPC migration (monitored from a small blood sample taken at each imaging procedure) and number or size of metastases (passive immunity). Four control mice will not receive the antibody. These two initial imaging studies will provide us with preliminary data to efficiently design future studies. As we develop a new paradigm for early detection and the process of metastasis, the capability for non-invasive imaging will become an invaluable research tool to evaluate our animal models.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 我们提出的理论，宿主反应密切参与转移的过程中，更好地了解肿瘤-宿主的相互作用是必不可少的，如果要有一个改善的结果。我们最近在肺癌患者中发现了与非转移性表型有很强关联的自身抗体。基于这些发现，我们提出了一个假设，即某些癌症患者的体液反应在是否发生转移中起着决定性作用。我们建议首先调查非转移性患者中普遍存在的自身抗体，并确定它是否对转移具有保护作用，如果是，则通过何种机制。 我们已经发现，来自H1568细胞（一种高转移性人肺癌细胞系）的条件培养基（CM）能够在体外测定中启动CD 34+造血祖细胞（HPC）迁移，该测定模拟了转移性小生境发展的初始步骤之一。来自I期非复发性或晚期转移性腺癌患者的血清用于探测来自该细胞系的CM的蛋白质的蛋白质印迹，以鉴定针对具有可能的迁移促进功能的蛋白质的自身抗体。我们在70%的非复发性肺癌患者中发现了α-葡萄糖苷酶II抗体，而在转移性肺癌患者中没有发现。向CM中添加抗α-葡糖苷酶II抗体可使体外CD 34+细胞迁移降低31- 63%。这表明H1568细胞分泌对CD 34+细胞的细胞表面糖蛋白起作用的胞外形式的葡糖苷酶II。我们想首先确定用条件培养基引发动物是否会加速转移性疾病，然后确定抑制葡萄糖苷酶II是否会减少转移。 我们将使用成像（microCT）来跟踪肺转移的发展和进展。这将使我们能够使用最少数量的动物来确定转移性病变的发生率、大小和分布的差异。我们将开始通过使用一个良好建立的小鼠转移模型，通过尾静脉注射。 1.将使用8只免疫功能低下的小鼠。将用来自H1568肺癌细胞的条件培养基致敏四只小鼠3天。4只小鼠将用无血清培养基预处理。然后，我们将向所有小鼠注射大约一百万个肿瘤细胞，每周对动物进行两次成像，持续三周（以前的研究表明，转移将在21天内充分建立）。如上所述，我们将确定引发动物是否会增加转移。 2.然后，我们将计划进行第二次实验，并在CM注射时用亲和纯化的人抗α-葡糖苷酶II抗体预处理4只小鼠，以观察这是否阻断HPC迁移（从每次成像程序采集的少量血液样本中监测）和转移的数量或大小（被动免疫）。四只对照小鼠将不接受抗体。 这两个初步的成像研究将为我们提供初步的数据，以有效地设计未来的研究。随着我们开发一种新的早期检测和转移过程的模式，非侵入性成像的能力将成为评估我们的动物模型的宝贵研究工具。