眼附属器皮脂腺癌的单细胞RNA分析揭示了复杂的肿瘤微环境并鉴定出新的生物标志物。
Single-Cell RNA Profiling of Ocular Adnexal Sebaceous Carcinoma Reveals a Complex Tumor Microenvironment and Identifies New Biomarkers.
作者信息
Zhang Michelle G, Gallo Ryan A, Tan Charissa H, Camacho Matthew, Fasih-Ahmad Sohaib, Moeyersoms Acadia H M, Sayegh Yoseph, Dubovy Sander R, Pelaez Daniel, Rong Andrew J
机构信息
From the Dr. Nasser Ibrahim Al-Rashid Orbital Vision Research Center (M.G.Z., R.A.G., A.H.M., D.P., and A.J.R.), Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, USA; Sylvester Comprehensive Cancer Center (M.G.Z., R.A.G., D.P., and A.J.R.), University of Miami Miller School of Medicine, Miami, Florida, USA.
Department of Ophthalmology (C.H.T., M.C., S.F.A., Y.S., and S.R.D.), Florida Lions Ocular Pathology Laboratory, Bascom Palmer Eye Institute, University of Miami Miller School of Medicine, Miami, Florida, USA.
出版信息
Am J Ophthalmol. 2025 Feb;270:8-18. doi: 10.1016/j.ajo.2024.10.001. Epub 2024 Oct 10.
PURPOSE
Ocular adnexal sebaceous carcinoma (OaSC) is an aggressive malignancy that often necessitates orbital exenteration. Its tumor composition and transcriptional profile remain largely unknown, which poses a significant barrier to medical advances. Here, we report the first in-depth transcriptomic analysis of OaSC at the single-cell resolution and discern mechanisms underlying cancer progression for the discovery of potential globe-sparing immunotherapies, targeted therapies, and biomarkers to guide clinical management.
DESIGN
Laboratory investigation with a retrospective observational case series.
METHODS
Single-cell RNA sequencing was performed on six patient specimens: three primary tumors, two tumors with pagetoid spread, and a normal tarsus sample. Cellular components were identified via gene signatures. Molecular pathways underlying tumorigenesis and pagetoid spread were discerned via gene ontology analysis of the differentially expressed genes between specimens. CALML5 immunohistochemistry was performed on an archival cohort of OaSC, squamous cell carcinoma, ocular surface squamous neoplasia (OSSN), and basal cell carcinoma cases.
RESULTS
Analysis of 29,219 cells from OaSC specimens revealed tumor, immune, and stromal cells. Tumor-infiltrating immune cells include a diversity of cell types, including exhausted T-cell populations. In primary OaSC tumors, mitotic nuclear division and oxidative phosphorylation pathways are upregulated, while lipid biosynthesis and metabolism pathways are downregulated. Epithelial tissue migration pathways are upregulated in tumor cells undergoing pagetoid spread. Single-cell RNA sequencing analyses also revealed that CALML5 is upregulated in OaSC tumor cells. Diffuse nuclear and cytoplasmic CALML5 staining was present in 28 of 28 (100%) OaSC cases. Diffuse nuclear and membranous CALML5 staining was present in 5 of 25 (20%) squamous cell carcinoma and OSSN cases, while diffuse nuclear staining was present in 1 of 12 (8%) basal cell carcinoma cases.
CONCLUSIONS
This study reveals a complex OaSC tumor microenvironment and confirms that the CALML5 immunohistochemical stain is a sensitive diagnostic marker.
目的
眼附属器皮脂腺癌（OaSC）是一种侵袭性恶性肿瘤，常需行眼眶内容物剜除术。其肿瘤组成和转录谱在很大程度上仍不清楚，这对医学进展构成了重大障碍。在此，我们报告了首例以单细胞分辨率对OaSC进行的深入转录组分析，并识别癌症进展的潜在机制，以发现潜在的保眼球免疫疗法、靶向疗法以及指导临床管理的生物标志物。
设计
采用回顾性观察病例系列进行实验室研究。
方法
对6例患者标本进行单细胞RNA测序：3例原发性肿瘤、2例呈派杰样扩散的肿瘤以及1例正常睑板样本。通过基因特征识别细胞成分。通过对标本间差异表达基因进行基因本体分析，识别肿瘤发生和派杰样扩散的分子途径。对OaSC、鳞状细胞癌、眼表鳞状上皮内瘤（OSSN）和基底细胞癌病例的存档队列进行钙调蛋白样蛋白5（CALML5）免疫组织化学检测。
结果
对来自OaSC标本的29219个细胞进行分析，发现了肿瘤细胞、免疫细胞和基质细胞。肿瘤浸润免疫细胞包括多种细胞类型，包括耗竭的T细胞群体。在原发性OaSC肿瘤中，有丝分裂核分裂和氧化磷酸化途径上调，而脂质生物合成和代谢途径下调。在发生派杰样扩散的肿瘤细胞中，上皮组织迁移途径上调。单细胞RNA测序分析还显示，CALML5在OaSC肿瘤细胞中上调。28例（100%）OaSC病例中均出现弥漫性核及胞质CALML5染色。25例鳞状细胞癌和OSSN病例中有5例（20%）出现弥漫性核及膜性CALML5染色，而12例基底细胞癌病例中有1例（8%）出现弥漫性核染色。
结论
本研究揭示了复杂的OaSC肿瘤微环境，并证实CALML5免疫组织化学染色是一种敏感的诊断标志物。
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