Single-Cell RNA Profiling of Ocular Adnexal Sebaceous Carcinoma Reveals a Complex Tumor Microenvironment and Identifies New Biomarkers

IF 4.1 1区医学 Q1 OPHTHALMOLOGY

American Journal of Ophthalmology Pub Date : 2024-10-10 DOI:10.1016/j.ajo.2024.10.001

Michelle G. Zhang , Ryan A. Gallo , Charissa H. Tan , Matthew Camacho , Sohaib Fasih-Ahmad , Acadia H.M. Moeyersoms , Yoseph Sayegh , Sander R. Dubovy , Daniel Pelaez , Andrew J. Rong

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引用次数: 0

Abstract

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.

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眼附属皮脂腺癌的单细胞 RNA 图谱揭示了复杂的肿瘤微环境，并确定了新的生物标记物。

目的：眼附属皮脂腺癌（OaSC）是一种侵袭性恶性肿瘤，通常需要进行眼眶外切除术。它的肿瘤组成和转录特征在很大程度上仍不为人所知，这对医学进步构成了重大障碍。在此，我们报告了首次以单细胞分辨率对眼眶外肿瘤进行的深入转录组学分析，并揭示了癌症进展的内在机制，从而发现潜在的保全免疫疗法、靶向疗法和生物标志物，以指导临床治疗：设计：实验室调查与回顾性观察病例系列：方法：对六例患者标本进行单细胞 RNA 测序：三例原发性肿瘤、两例蝶形扩散肿瘤和一例正常跗骨样本。通过基因特征确定了细胞成分。通过对标本间差异表达基因的基因本体分析，确定了肿瘤发生和鳞状细胞扩散的分子途径。对OaSC、鳞状细胞癌（SCC）、眼表鳞状细胞瘤（OSSN）和基底细胞癌（BCC）病例的档案队列进行了CALML5免疫组化：结果：对来自 OaSC 标本的 29,219 个细胞进行了分析，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，包括衰竭的T细胞群。在原发性 OaSC 肿瘤中，有丝分裂核分裂和氧化磷酸化通路上调，而脂质生物合成和代谢通路下调。scRNA-seq分析还发现，CALML5在OaSC肿瘤细胞中上调。28例OaSC病例中有28例（100%）出现弥漫性核和胞质CALML5染色。25例SCC和OSSN病例中有5例（20%）出现弥漫性核和膜CALML5染色，而12例BCC病例中有1例（8%）出现弥漫性核染色：本研究揭示了复杂的 OaSC 肿瘤微环境，并证实 CALML5 免疫组化染色是一种敏感的诊断标志物。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

American Journal of Ophthalmology 医学-眼科学

CiteScore

9.20

自引率

7.10%

发文量

406

审稿时长

36 days

期刊介绍： The American Journal of Ophthalmology is a peer-reviewed, scientific publication that welcomes the submission of original, previously unpublished manuscripts directed to ophthalmologists and visual science specialists describing clinical investigations, clinical observations, and clinically relevant laboratory investigations. Published monthly since 1884, the full text of the American Journal of Ophthalmology and supplementary material are also presented online at www.AJO.com and on ScienceDirect. The American Journal of Ophthalmology publishes Full-Length Articles, Perspectives, Editorials, Correspondences, Books Reports and Announcements. Brief Reports and Case Reports are no longer published. We recommend submitting Brief Reports and Case Reports to our companion publication, the American Journal of Ophthalmology Case Reports. Manuscripts are accepted with the understanding that they have not been and will not be published elsewhere substantially in any format, and that there are no ethical problems with the content or data collection. Authors may be requested to produce the data upon which the manuscript is based and to answer expeditiously any questions about the manuscript or its authors.