Endocrinol Metab.  2024 Apr;39(2):324-333. 10.3803/EnM.2023.1885.

Clinicopathological Features and Molecular Signatures of Lateral Neck Lymph Node Metastasis in Papillary Thyroid Microcarcinoma

• Lim J
• Lee HS
• Heo JH
• Song YS

• KMID: 2554637
• DOI: http://doi.org/10.3803/EnM.2023.1885

Abstract

Background
The predictive factors for lateral neck lymph node metastasis (LLNM) in papillary thyroid microcarcinoma (PTMC) remain undetermined. This study investigated the clinicopathological characteristics, transcriptomes, and tumor microenvironment in PTMC according to the LLNM status. We aimed to identify the biomarkers associated with LLNM development.
Methods
We retrospectively reviewed the medical records of patients with PTMC from two independent institutions between 2018 and 2022 (n=597 and n=467). We compared clinicopathological features between patients without lymph node metastasis (N0) and those with LLNM (N1b). Additionally, laser capture microdissection and RNA sequencing were performed on primary tumors from both groups, including metastatic lymph nodes from the N1b group (n=30; 20 primary tumors and 10 paired LLNMs). We corroborated the findings using RNA sequencing data from 16 BRAF-like PTMCs from The Cancer Genome Atlas. Transcriptomic analyses were validated by immunohistochemical staining.
Results
Clinicopathological characteristics, such as male sex, multifocality, extrathyroidal extension, lymphatic invasion, and central node metastasis showed associations with LLNM in PTMCs. Transcriptomic profiles between the N0 and N1b PTMC groups were similar. However, tumor microenvironment deconvolution from RNA sequencing and immunohistochemistry revealed an increased abundance of tumor-associated macrophages, particularly M2 macrophages, in the N1b group.
Conclusion
Patients with PTMC who have a male sex, multifocality, extrathyroidal extension, lymphatic invasion, and central node metastasis exhibited an elevated risk for LLNM. Furthermore, infiltration of M2 macrophages in the tumor microenvironment potentially supports tumor progression and LLNM in PTMCs.

Keyword

Thyroid cancer, papillary; Lymphatic metastasis; Biomarkers, tumor; Tumor microenvironment

Figure

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  Fig. 1. M2 macrophage infiltration compared between N1b and N0 primary cancers (PCs) of papillary thyroid microcarcinomas (PTMCs). (A) The abundance of M2 macrophages, (B) the M2 score, and (C) the expression of M2 macrophage markers in the CHA Bundang Medical Center (CHA) database. (D) The abundance of M2 macrophages, (E) the M2 score, and (F) the expression of M2 macrophage markers in The Cancer Genome Atlas (TCGA) database. BL, BRAF-like.

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  Fig. 2. Immunohistochemistry analysis of CD68 (pan-macrophage marker) and CD163 (M2 macrophages marker) in papillary thyroid microcarcinomas (PTMCs). (A, B) CD68-positive and (C, D) CD163-positive macrophage densities in PTMCs. (A, C) Representative images of immunohistochemical staining (200× magnification). The scale bar is 100 µm. (B, D) Statistical analysis of the average positive area in the N0 and N1b PTMCs.

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