Lang Fabian, Contreras-Gerenas María F, Gelléri Márton, Neumann Jan, Kröger Ole, Sadlo Filip, Berniak Krzysztof, Marx Alexander, Cremer Christoph, Wagenknecht Hans-Achim, Allgayer Heike
Institute of Organic Chemistry, Karlsruhe Institute of Technology (KIT), Fritz-Haber-Weg 6, Campus Süd, 76131 Karlsruhe, Germany.
Department of Experimental Surgery-Cancer Metastasis, Mannheim Medical Faculty, Ruprecht-Karls University of Heidelberg, Ludolf-Krehl-Straße 13-17, 68167 Mannheim, Germany.
Cancers (Basel). 2021 Jul 22;13(15):3692. doi: 10.3390/cancers13153692.
Tumour cell heterogeneity, and its early individual diagnosis, is one of the most fundamental problems in cancer diagnosis and therapy. Single molecule localisation microscopy (SMLM) resolves subcellular features but has been limited to cultured cell lines only. Since nuclear chromatin architecture and microRNAs are critical in metastasis, we introduce a first-in-field approach for quantitative SMLM-analysis of chromatin nanostructure in individual cells in resected, routine-pathology colorectal carcinoma (CRC) patient tissue sections. Chromatin density profiles proved to differ for cells in normal and carcinoma colorectal tissues. In tumour sections, nuclear size and chromatin compaction percentages were significantly different in carcinoma versus normal epithelial and other cells of colorectal tissue. SMLM analysis in nuclei from normal colorectal tissue revealed abrupt changes in chromatin density profiles at the nanoscale, features not detected by conventional widefield microscopy. SMLM for microRNAs relevant for metastasis was achieved in colorectal cancer tissue at the nuclear level. Super-resolution microscopy with quantitative image evaluation algorithms provide powerful tools to analyse chromatin nanostructure and microRNAs of individual cells from normal and tumour tissue at the nanoscale. Our new perspectives improve the differential diagnosis of normal and (metastatically relevant) tumour cells at the single-cell level within the heterogeneity of primary tumours of patients.
肿瘤细胞异质性及其早期个体诊断是癌症诊断和治疗中最基本的问题之一。单分子定位显微镜（SMLM）能够分辨亚细胞特征，但仅限于培养的细胞系。由于核染色质结构和微小RNA在转移过程中至关重要，我们引入了一种首次用于在切除的常规病理结直肠癌（CRC）患者组织切片中对单个细胞的染色质纳米结构进行定量SMLM分析的方法。染色质密度分布图显示，正常和癌性结直肠组织中的细胞存在差异。在肿瘤切片中，癌组织与正常上皮细胞及结直肠组织的其他细胞相比，细胞核大小和染色质压缩百分比存在显著差异。对正常结直肠组织细胞核的SMLM分析揭示了纳米尺度下染色质密度分布图的突然变化，这是传统宽视场显微镜无法检测到的特征。在结直肠癌组织的细胞核水平上实现了对与转移相关的微小RNA的SMLM分析。具有定量图像评估算法的超分辨率显微镜提供了强大的工具，可在纳米尺度上分析正常和肿瘤组织中单个细胞的染色质纳米结构和微小RNA。我们的新观点改善了在患者原发性肿瘤异质性范围内，在单细胞水平上对正常和（与转移相关的）肿瘤细胞的鉴别诊断。
