Tech Core 1

项目摘要-技术1 确定推动转移性乳腺癌进展的关键分子和细胞事件， 胰腺癌可能会导致改善机制的理解和诊断工具和治疗。 然而，人类胰腺/乳腺肿瘤的显微解剖学-它们的空间组织和相关的组织结构， 细胞和非细胞成分中的含量-由于其本质上是三维的， 非对称和高度异质。结合联合收割机结构和空间的三维成像工具 需要大量的组学信息来定位癌组织中的这些转移驱动事件。 具体来说，我们的目的是调查静脉侵犯，这是一个常见的途径转移的乳腺癌， 胰腺癌细胞在Tech 1中，我们提出开发一种新的3D多尺度成像方法CODA， 将使我们能够探测肿瘤的表型异质性，从几厘米到微米尺度， 复用串行成像和配准/深度学习算法。我们建议的CODA成像 方法解决了现有技术的组织清除和3D成像方法的缺陷，包括不一致的 透明和染色、缺乏验证、抗体渗透性差和有限的多重能力。CODA 绕过了光学显微镜的限制，允许大体积成像，同时实现微米 无论成像组织的体积如何，都可以提高分辨率。CODA的另一个关键优势是， 连续切割H&E切片的优点在于，它可以很容易地结合其他成像方式，以提取高质量的图像。 3D样本的细胞/分子含量。这些包括免疫细胞化学（CODA+IHC）， 免疫荧光（CODA+IF）和成像质谱细胞术（CODA+IMC）。这一点尤其重要，因为 内渗/外渗和癌细胞侵入的许多潜在的分子和细胞贡献者 静脉，包括免疫细胞和癌症相关的成纤维细胞，需要无法检测到的特异性标记 在H&E幻灯片中。这些建议的CODA扩展版本提供了一个独特的机会来制作新的3D 接近和远离血管的人PDAC和乳腺肿瘤的多重图谱。CODA及其集成 CODA+X版本将在中心的RTB单元中进行测试。

Project Summary – Tech 1 The identification of key molecular and cellular events that drive the progression of metastatic breast and pancreatic cancer could lead to improved mechanistic understanding and diagnostic tools and treatments. However, the micro-anatomy of human pancreatic/breast tumors – their spatial organization and associated content in cellular and non-cellular components – is poorly understood as it is intrinsically three-dimensional, non-symmetric, and highly heterogeneous. Three-dimensional imaging tools that combine structural and spatial omic information in large volumes are required to locate these metastatic driver events in cancerous tissue. Specifically, we aim to investigate venous invasion, which is a common route of metastasis of breast and pancreatic cancer cells. In Tech1, we propose to develop a new 3D multiscale imaging method, CODA, which will allow us to probe the phenotypic heterogeneity of tumors from the multi-cm to the micron scale via multiplexing serial imaging and registration/deep-learning algorithms. Our proposed repertoire of CODA imaging methods addresses deficiencies of state-of-art tissue clearing and 3D imaging methods, including inconsistent clearing and staining, lack of validation, poor antibody penetration and limited multiplexing capabilities. CODA bypasses the limitations of optical microscopy, allowing for large volumetric imaging while achieving micron resolution regardless of the volume of the imaged tissue. Another key advantage of CODA, which is based on serially cut H&E sections, is that it can readily incorporate other imaging modalities to extract high cellular/molecular content from the 3D samples. These include immunocytochemistry (CODA+IHC), immunofluorescence (CODA+IF), and imaging mass cytometry (CODA+IMC). This is particularly important as many potential molecular and cellular contributors of intravasation/extravasation and cancer cell invasion into veins, including immune cells and cancer-associated fibroblasts, require specific labels that cannot be detected in H&E slides. These proposed expanded versions of CODA offer a unique opportunity to produce new 3D multiplex maps of human PDAC and breast tumors near and far from blood vessels. CODA and its integrated versions CODA+X will be tested in the RTB units of the Center.