Paracrine interactions between primary human macrophages and human fibroblasts enhance murine mammary gland humanization in vivo.
Paracrine interactions between primary human macrophages and human fibroblasts enhance murine mammary gland humanization in vivo.
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DOI：
10.1186/bcr3215
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发表时间：
2012-06-25
期刊：
影响因子：
--
通讯作者：
Vonderhaar BK
中科院分区：
文献类型：
--
作者：
Fleming JM;Miller TC;Kidacki M;Ginsburg E;Stuelten CH;Stewart DA;Troester MA;Vonderhaar BK
巨噬细胞包括正常腺体发育所必需的乳腺微环境的基本组分。然而，没有可行的体内模型来研究它们在正常人类乳腺功能中的作用。我们假设，将原代人巨噬细胞添加到小鼠乳腺中将增强并提供一种新的方法来检查人源化过程中的免疫-基质细胞相互作用。在存在或不存在异位雌激素刺激的情况下，原代人巨噬细胞用于人源化小鼠乳腺。通过细胞因子/趋化因子ELISA、酶谱法、蛋白质印迹分析、侵袭和增殖测定来鉴定增强的人源化机制;用免疫组织学分析证实结果。巨噬细胞和雌激素刺激的联合治疗显着提高了总腺体人源化的百分比和植入/生长成功率。时程分析显示，注射后两周，人巨噬细胞消失，表明成纤维细胞的整体生长和侵袭力的改善为上皮细胞增殖和结构形成提供了更大的基质床。雌激素刺激的巨噬细胞在体外显著增强成纤维细胞的增殖和侵袭，并显著增加人源化腺体中增殖细胞核抗原（PCNA）阳性细胞，这表明它们促进成纤维细胞人源化。细胞因子/趋化因子ELISA、酶谱和Western分析确定TNFα和MMP 9是雌激素刺激的巨噬细胞增强人源化的潜在机制。TNFα和MMP 9的特异性抑制剂验证了这些分子对体外成纤维细胞行为的影响，以及通过人源化腺体的人特异性MMP 9表达的免疫组织化学分析。最后，与仅用成纤维细胞人源化的腺体相比，用巨噬细胞人源化的腺体具有增强的植入和肿瘤生长。在此，我们证明了复杂的免疫和基质细胞旁分泌的相互作用，在人源化的体内模型系统。我们用体外分析证实了我们的体内结果，突出了该模型可互换地证实体外和体内结果的价值。理解驱动旁分泌细胞相互作用的信号网络是至关重要的，因为肿瘤细胞利用这些信号机制来支持其生长和侵袭特性。本报告提出了一个动态的体内模型，研究初级人类免疫/成纤维细胞/上皮细胞的相互作用，并推进我们的知识，基质衍生的信号，促进肿瘤发生。
Macrophages comprise an essential component of the mammary microenvironment necessary for normal gland development. However, there is no viable in vivo model to study their role in normal human breast function. We hypothesized that adding primary human macrophages to the murine mammary gland would enhance and provide a novel approach to examine immune-stromal cell interactions during the humanization process. Primary human macrophages, in the presence or absence of ectopic estrogen stimulation, were used to humanize mouse mammary glands. Mechanisms of enhanced humanization were identified by cytokine/chemokine ELISAs, zymography, western analysis, invasion and proliferation assays; results were confirmed with immunohistological analysis. The combined treatment of macrophages and estrogen stimulation significantly enhanced the percentage of the total gland humanized and the engraftment/outgrowth success rate. Timecourse analysis revealed the disappearance of the human macrophages by two weeks post-injection, suggesting that the improved overall growth and invasiveness of the fibroblasts provided a larger stromal bed for epithelial cell proliferation and structure formation. Confirming their promotion of fibroblasts humanization, estrogen-stimulated macrophages significantly enhanced fibroblast proliferation and invasion in vitro, as well as significantly increased proliferating cell nuclear antigen (PCNA) positive cells in humanized glands. Cytokine/chemokine ELISAs, zymography and western analyses identified TNFα and MMP9 as potential mechanisms by which estrogen-stimulated macrophages enhanced humanization. Specific inhibitors to TNFα and MMP9 validated the effects of these molecules on fibroblast behavior in vitro, as well as by immunohistochemical analysis of humanized glands for human-specific MMP9 expression. Lastly, glands humanized with macrophages had enhanced engraftment and tumor growth compared to glands humanized with fibroblasts alone. Herein, we demonstrate intricate immune and stromal cell paracrine interactions in a humanized in vivo model system. We confirmed our in vivo results with in vitro analyses, highlighting the value of this model to interchangeably substantiate in vitro and in vivo results. It is critical to understand the signaling networks that drive paracrine cell interactions, for tumor cells exploit these signaling mechanisms to support their growth and invasive properties. This report presents a dynamic in vivo model to study primary human immune/fibroblast/epithelial interactions and to advance our knowledge of the stromal-derived signals that promote tumorigenesis.
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影响因子：
8.4
作者：
Curran, S;Murray, GI
通讯作者：
Murray, GI
影响因子：
64.8
作者：
Asselin-Labat, Marie-Liesse;Vaillant, Francois;Visvader, Jane E.
通讯作者：
Visvader, Jane E.
影响因子：
2.5
作者：
Goldman, Armond S.;Chheda, Sadhana;Schmalstieg, Frank C.
通讯作者：
Schmalstieg, Frank C.
影响因子：
3.6
作者：
Cohen, PE;Chisholm, O;Pollard, JW
通讯作者：
Pollard, JW
影响因子：
4.8
作者：
Adib-Conquy, M;Cavaillon, JM
通讯作者：
Cavaillon, JM