Mutual influence of tumor cells, extracellular matrix and macrophages in intraocular murine melanoma

葡萄膜黑色素瘤是成人高加索人群中最常见的原发眼内肿瘤。虽然原发肿瘤的局部治疗非常有希望，但随着肝转移的发生，预后急剧恶化。一些研究表明巨噬细胞参与葡萄膜黑色素瘤和其他恶性肿瘤的发病机制。在我们的小鼠肿瘤模型中，眼内注射的肿瘤细胞形成具有侵袭性黑色素瘤组织学特征(如血管生成、血管生成抑制和巨噬细胞浸润)的肿瘤块，类似于人类葡萄膜黑色素瘤和其他肿瘤。由于眼睛是一个半封闭的隔室，可以获得持续的血液供应，这些眼内肿瘤代表了一般肿瘤生物学中孤立参数研究的模型。我们研究的目的是分析巨噬细胞和肿瘤细胞在巨噬细胞极化和预后相关的血管生成拟态方面的相互影响。我们假设促血管生成M2巨噬细胞通过刺激肿瘤细胞建立血管生成拟态来促进肿瘤生长。我们的目的是确定这一过程中涉及的因素，特别是关于血管生成拟态和巨噬细胞极化的因素。我们还想为血管生成拟态建立一个可靠的标记。设计了人和小鼠肿瘤细胞以及各自的巨噬细胞在跨膜细胞培养系统中的体外实验。我们的小鼠模型用于体外结果的形态控制。我们还想建立一种3D细胞培养，它可能会取代我们的老鼠模型来解决特定的问题。在第二步中，将识别并选择性地敲除参与血管生成拟态和巨噬细胞极化产生的因素，以研究特定的信号通路和潜在的治疗策略。在CCR2和CX3CR1基因敲除小鼠中，研究了肿瘤相关巨噬细胞的起源(CCR2依赖与CX3CR1依赖的招募)以及巨噬细胞在肿瘤生长和血管结构发育中的特定作用。综上所述，这些研究将有助于了解肿瘤和巨噬细胞之间的相互作用，以及肿瘤相关巨噬细胞在巨噬细胞极化和血管生成拟态产生方面的起源，这与预后相关，因此预后较差。

Uveal melanoma is the most common primary intraocular tumor in the adult Caucasian population. While local treatment of the primary tumor is very promising, prognosis worsens dramatically with the occurrence of liver metastasis. Several studies were able to show that macrophages are involved in the pathogenesis of uveal melanoma and other malignancies.In our murine tumor model, intraocularly injected tumor cells develop tumor masses with histologic characteristics of aggressive melanoma (such as angiogenesis, vasculogenic mimcry and infiltration with macrophages) similar to human uveal melanoma and other tumors. Since the eye represents a semi-closed compartment with access to constant blood supply, these intraocular tumors represent a model for studies of isolated parameters in general tumor biology.The purpose of our studies is the analysis of the mutual influence of macrophages and tumor cells with respect to macrophage polarisation and prognostically relevant vasculogenic mimicry. We hypothesize that proangiogenic M2 macrophages contribute to tumor growth via the stimulation of tumor cells to build vasculogenic mimicry. Our aim is to identify the factors involved in this process, in particular with regard to vasculogenic mimicry and macrophage polarisation. We further want to establish a reliable marker for vasculogenic mimicry. The designed in vitro experiments with human and murine tumor cells and respective macrophages in a transwell-membrane cell culture system. Our mouse model is used for a morphological control of the in vitro findings. We further want to establish a 3D cell culture, which might replace our mouse model for specific questions. In a second step, factors involved in the production of vasculogenic mimicry and macrophage polarisation will be identified and selectively knocked out in order to study specific signalling pathways and potentially therapeutic strategies.The origin of tumor-associated macrophages (CCR2-dependent versus CX3CR1-dependent recruitment) and the specific role of macrophages on tumor growth and the development of vascular structures are investigated in CCR2- and CX3CR1-knockout mice. In summary, these studies will help to understand the interaction between the tumor and macrophages as well as the origin of tumor-associated macrophages with respect to macrophage polarisation and producation of vasculogenic mimicry which are prognostically relevant for metastases and thus a worse prognosis.