RTB 1

项目摘要 胰腺癌是一种致命的疾病，预后很差，远处转移的频率很高。一 胰腺癌转移频繁的原因可能是原发肿瘤中的恶性细胞的能力 侵入静脉，提供直接进入肝脏的途径。虽然这种血管内转移是转移的关键第一步，但几乎没有 已知这一过程背后的分子和细胞变化。癌细胞渗入灶 小而复杂，需要高分辨率的三维(3D)方法来研究它们。在这 我们中心的三维多尺度肿瘤成像研究试验台，我们将全面表征 人胰腺癌3D血管内病灶的形态、细胞和分子变化 样本。与技术开发单元1合作，我们将采用我们的新型深度学习3D 连续切片胰腺癌静脉侵犯标本重建的CODA方法 血管内渗出的形态和细胞特征。我们还将进行多维免疫图谱分析 这些血管内病灶使用成像质量细胞术进行3D成像，它可以在每个组织上标记40个抗体 一节。这些分析将使我们能够比较血管内病灶和血管内的肿瘤微环境。 癌症的其他区域和未受累的静脉。我们会与科技发展组第二组合作， 应用一种新开发的空间分析方法分析这些血管内病灶的基因表达变化 转录组学/蛋白质组学方法(DBit-Seq)用于先前重建的胰腺癌样本 尾声。这种方法将提供首个人胰腺癌静脉侵袭的3D多尺度模型， 提供了前所未有的形态，细胞和分子细节的血管内和 开始转移。此外，我们的研究试验台将提供实施、 验证并反复改进来自两个技术开发中心的新方法。我们会 与这些技术开发中心密切合作，以评估CODA和 我们的人胰腺组织样本中的DBit-Seq，允许验证细胞类型鉴定和分子 通过补充技术进行的改变。我们的研究试验台上的研究将对 改进CODA和DBit-Seq成像平台，同时提供重要的生物学见解 胰腺癌转移的启动。

Project Summary Pancreatic cancer is a deadly disease with a dismal prognosis and a high frequency of distant metastasis. One likely reason for the frequent metastases in pancreatic cancer is the ability of malignant cells in the primary tumor to invade veins, providing direct access to the liver. While this intravasation is a key first step in metastasis, little is known about the molecular and cellular alterations that underlie this process. Foci of cancer cell intravasation are small and complex, requiring high-resolution three-dimensional (3D) approaches to study them. In this Research Test Bed of our Center for 3D Multiscale Cancer Imaging, we will comprehensively characterize the morphological, cellular, and molecular alterations of foci of intravasation in 3D in human pancreatic cancer samples. In collaboration with Technology Development Unit 1, we will employ our novel deep learning 3D reconstruction approach CODA to serially sectioned pancreatic cancer samples with venous invasion to quantify morphological and cellular features of intravasation. We will also perform multi-dimensional immune profiling of these intravasation foci in 3D using imaging mass cytometry, which can label with 40 antibodies on each tissue section. These analyses will allow us to compare the tumor microenvironment in foci of intravasation to that in other areas of cancer and in uninvolved veins. In collaboration with Technology Development Unit 2, we will analyze the gene expression changes in these intravasation foci by applying a newly developed spatial transcriptomics/proteomics approach (DBit-Seq) to pancreatic cancer samples previously reconstructed by CODA. This approach will provide the first 3D multi-scale models of venous invasion in human pancreatic cancer, providing unprecedented morphological, cellular, and molecular detail of the process of intravasation and initiation of metastasis. In addition, our Research Test Bed will provide a critical opportunity to implement, validate, and iteratively improve the novel approaches from the two Technology Development Centers. We will work closely with these Technology Development Centers in order to evaluate the performance of CODA and DBit-Seq in our human pancreatic tissue samples, allowing validation of the cell type identification and molecular alterations by complementary techniques. The studies in our Research Test Bed will promote significant improvement of the CODA and DBit-Seq imaging platforms while also providing important biological insights into the initiation of metastasis in pancreatic cancer.