PO.TB09.02 · 肿瘤生物学
跨正常、癌前和癌组织的蛋白质与染色体改变的整合空间分析揭示正常输卵管中存在非整倍体
Integrative spatial profiling of protein and chromosomal alterations across normal, precancer, and cancer revealed the presence of aneuploidy in the normal fallopian tube
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：在同一FFPE切片内同时测量蛋白质表达和染色体改变，能够直接分析从正常上皮经癌前到癌状态的克隆选择和疾病进展。我们开发了一套基于一步式Orion多重成像平台（Lin等，Nature Cancer 2023）的统一工作流程，将高维蛋白质成像与传统DNA-FISH检测相整合，用于原位检测染色体拷贝数改变（CNA）。传统DNA-FISH虽在诊断实践中广泛使用，但需要蛋白酶消化和透化步骤，这些步骤会破坏抗原表位并扭曲核形态，妨碍可靠的蛋白质检测并限制与免疫荧光的整合。这一挑战跨越正常前驱组织和恶性组织，其中密集堆积或重叠的细胞核使单细胞CNA评分复杂化——鉴于人们日益认识到非整倍体甚至可在形态学正常的上皮中出现，这一问题现在尤为相关。
方法：多重成像的最新进展现已能够对携带CNA的单个细胞进行表型刻画，这是传统免疫荧光此前无法实现的能力。为实现这一潜力，我们优化了固定、杂交和成像参数，以在保持稳健CNA检测的同时保留抗原性和核结构。所得的ORION-FISH工作流程能够在同一切片内同时可视化和定量蛋白质状态与染色体改变。
结果：我们发现正常输卵管上皮携带MYC和/或CCNE1，二者均为高级别浆液性卵巢癌（HGSOC）及其前驱病变浆液性输卵管上皮内癌（STIC）中常见的CNA。我们还发现这些非整倍体细胞可能一直处于强烈的负选择压力之下，部分归因于强烈的免疫监视。目前，我们正在深入分析前驱样本和HGSOC标本，将在会议上展示。结论：我们将ORION-FISH方法应用于HGSOC及其前驱病变，以生成跨正常-癌前-癌连续体的非整倍体和细胞状态空间图谱，为我们在HGSOC癌前图谱2.0中的研究奠定了基础。
查看英文原文 English abstract
Introduction: Simultaneous measurement of protein expression and chromosomal alterations within the same FFPE section enables direct analysis of clonal selection and disease progression from normal epithelium through precancerous to cancerous states. We developed a unified workflow based on the one-shot Orion multiplexed imaging platform (Lin et al., Nature Cancer 2023) that integrates high-dimensional protein imaging with a conventional DNA-FISH assay for in situ detection of chromosomal copy number alterations (CNAs). Traditional DNA-FISH, although widely used in diagnostic practice, requires protease digestion and permeabilization steps that destroy antigen epitopes and distort nuclear morphology, preventing reliable protein detection and limiting integration with immunofluorescence. The challenge spans both normal precursor and malignant tissues, where densely packed or overlapping nuclei complicate single-cell CNA scoring - an issue that is now particularly relevant, given the growing recognition that aneuploidy can arise even in morphologically normal epithelia.
Method: Recent advances in multiplexed imaging now enable phenotypic characterization of individual cells harboring CNAs, a capability previously unattainable with conventional immunofluorescence. To realize this potential, we optimized fixation, hybridization, and imaging parameters to preserve both antigenicity and nuclear architecture while maintaining robust CNA detection. The resulting ORION-FISH workflow enables simultaneous visualization and quantification of protein states and chromosomal alterations within the same section.
Results: We have found that normal fallopian tube epithelium harbors MYC and/or CCNE1, both of which are common CNAs in High Grade Serous Ovarian Cancer (HGSOC) and its precursor lesion, S erous T ubal I ntraepithelial C arcinoma (STIC). We have also found that these aneuploid cells may have been under strong negative selection pressure, partially due to strong immune surveillance. Currently, we are analyzing in depth precursor samples and HGSOC specimens that will be presented at the meeting. Conclusion: We applied ORION-FISH approach to HGSOC and its precursors to generate spatial maps of aneuploidy and cell states across the normal-precancer-cancer continuum, establishing a foundation for our studies in the HGSOC Pre-Cancer Atlas 2.0.
利益披露 Disclosure
T. Kader, None.