FLOAT System to Study Salivary Gland Cancer Invasion

这款R21的目标是使用我们最新开发的双色荧光示踪矫形器 同种异体移植(FLOAT)系统研究唾液腺的分子机制 癌症(SGC)进展。人类SGC的生物信息学研究已经确定了可能的分子途径 调控肿瘤侵袭，但后续分析被证明是具有挑战性的。细胞培养模型受到以下限制 缺乏天然的肿瘤微环境和转基因动物模型受到资源密集型的限制 区分肿瘤细胞和周围间质的方法和困难。因此，新的方法 这使得研究人员能够快速测试分子假说，同时保留 肿瘤的微环境，是需要的。此外，这些模型需要考虑其他因素的影响 偏远地区的共病，如肥胖症。我们的提案将使用浮动系统来满足这一需求， 简单且易于实验处理的原位移植模型，使研究人员能够可视化 植入唾液肿瘤细胞及邻近微环境，可准确模拟体内多个方面 SGC进展。该系统利用DsRed荧光标记唾液肿瘤细胞，并对其进行移植 进入表达绿色荧光蛋白的小鼠的唾液腺。双色荧光允许 研究人员快速评估相对于邻近间质的肿瘤细胞。我们的初步数据 证明浮动系统可用于显示固有的肿瘤自噬能力 调节荷瘤腺体的结构和存活率。我们现在将使用浮动系统来 评估肥胖如何促进体内原发SGC病变的局部侵袭。需要局部入侵才能 SGC进展，但这一多方面的过程还没有被很好地理解。另外，尽管两种肿瘤的变化 细胞和周围的基质已经被报道在癌症进展过程中，这是一种直接的机制联系 饮食诱导的肥胖和加速的SGC进展之间的关系尚未确定。我们假设 饮食诱导的肥胖通过诱导肿瘤的变化来加剧SGC的局部侵袭 允许或加强局部入侵的微环境。鉴于肿瘤的关键作用 在肿瘤进展的微环境中，我们进一步预测了饮食诱导肥胖对类型的影响 肿瘤微环境中的细胞活动促进了SGC的侵袭。在目标1中，我们将使用浮动 系统，以测试饮食诱导肥胖对SGC局部侵袭、基因表达和血脂谱的影响。 该目标包括3D基质辅助激光解吸/电离(MALDI)-组织成像质谱学 (TIMS)分析侵袭性肿瘤的脂谱异质性。在目标2中，我们将使用浮动系统来 测试饮食诱导的肥胖对微环境中纤维化和炎症的影响。彩车 系统代表了可视化和区分异类组件的强大的第一步 在局部侵袭过程中宿主-肿瘤相互作用的研究，以及有效测试新疗法的新系统。

The goal of this R21 is to use our newly developed dual-color Fluorescence-tracing Orthotopic AlloTransplantation (FLOAT) system to investigate the molecular mechanisms underlying salivary gland cancer (SGC) progression. Bioinformatics studies of human SGC have identified putative molecular pathways that regulate tumor invasion, but follow-up analysis has proved challenging. Cell culture models are limited by the lack of a native tumor microenvironment and transgenic animal models are limited by resource-intensive methods and difficulties distinguishing tumor cells from the surrounding stroma. Therefore, new approaches that allow researchers to rapidly test molecular hypotheses, while preserving the native 3D structure of the tumor microenvironment, are needed. Furthermore, these models need to consider the impact of other comorbidities in distant sites, such as obesity. Our proposal will address this need using the FLOAT system, a simple and experimentally tractable orthotopic transplantation model, which allows researchers to visualize implanted salivary tumor cells and the adjacent microenvironment, to accurately model many aspects of in vivo SGC progression. This system utilizes DsRed fluorescence to label salivary tumor cells, which are transplanted into the salivary glands of green fluorescent protein-expressing mice. Dual-color fluorescence allows researchers to rapidly evaluate the tumor cells relative to the adjacent stroma. Our preliminary data demonstrate that the FLOAT system was useful for demonstrating that intrinsic tumor autophagy capacity modulates the architecture of tumor-bearing glands and survival. We will now use the FLOAT system to evaluate how obesity promotes local invasion of primary SGC lesions in vivo. Local invasion is required for SGC progression, but this multifaceted process is not well understood. Also, although changes in both tumor cells and the surrounding matrix have been reported during cancer progression, a direct mechanistic link between diet-induced obesity and accelerated SGC progression has not been established. We hypothesize that diet-induced obesity exacerbates SGC local invasion by inducing changes in the tumor microenvironment that permit or enhance local invasion. Given the crucial role of the tumor microenvironment in cancer progression, we further predict that the effect of diet-induced obesity on the type and activity of cells in the tumor microenvironment facilitates SGC invasion. In Aim 1, we will use the FLOAT system to test the effects of diet-induced obesity on SGC local invasion, gene expression, and lipid profiles. This Aim includes 3D matrix-assisted laser desorption/ionization (MALDI)-tissue-imaging mass spectrometry (TIMS) analysis of lipid profile heterogeneity in the invading tumor. In Aim 2, we will use the FLOAT system to test the effects of diet-induced obesity on fibrosis and inflammation in the microenvironment. The FLOAT system represents a powerful first step for both visualizing and distinguishing the heterogeneous components of host-tumor interaction during local invasion and a new system for efficiently testing novel therapeutics.