Single‑cell transcriptomic analysis revealed the tumor‑associated microenvironment of papillary thyroid carcinoma with metastasis

Zhang,Ni; Liu,Qingbin; Wang,Qian; Liu,Xiuli; Zhang,Suya; Tian,Xinchen; Li,Long; Wang,Shuanglong; Lv,Bin; Jiang,Shulong

doi:10.3892/ol.2025.14876

Single‑cell transcriptomic analysis revealed the tumor‑associated microenvironment of papillary thyroid carcinoma with metastasis

Authors:
- Ni Zhang
- Qingbin Liu
- Qian Wang
- Xiuli Liu
- Suya Zhang
- Xinchen Tian
- Long Li
- Shuanglong Wang
- Bin Lv
- Shulong Jiang
View Affiliations

Affiliations: Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250000, P.R. China, Clinical Medical Laboratory Center, Jining First People's Hospital, Shandong First Medical University, Jining, Shandong 272000, P.R. China, Department of Ultrasonic Medicine, Jining First People's Hospital, Shandong First Medical University, Jining, Shandong 272000, P.R. China, Department of Breast and Thyroid Surgery, Jining First People's Hospital, Shandong First Medical University, Jining, Shandong 272000, P.R. China, Department of Ultrasonic Medicine, Jining First People's Hospital, Shandong First Medical University, Jining, Shandong 272000, P.R. China, Clinical Medical Laboratory Center, Jining First People's Hospital, Shandong First Medical University, Jining, Shandong 272000, P.R. China, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250000, P.R. China
Published online on: January 7, 2025 https://doi.org/10.3892/ol.2025.14876
Article Number: 130
Copyright: © Zhang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Papillary thyroid cancer (PTC) is frequently associated with inflammation and lymph node metastasis. Single‑cell RNA sequencing (scRNA‑seq) is a powerful tool to uncover rare cellular subpopulations and investigate the diverse functions inside tissue microenvironments. In the present study, scRNA‑seq analysis was employed to analyze the differences in macrophages, dendritic cells (DCs) and T cells between a metastatic PTC (PTC‑M) and its adjacent normal tissues, as well as a PTC tumor without metastasis. The findings revealed significant heterogeneity in immune cell populations in PTC‑M, suggesting that immunosuppressive components contribute to the development and metastasis of PTC. The current study revealed that the presence of alternatively activated M2 macrophages, conventional type 2 DCs (DC2s) and regulatory T cells (Tregs) was associated with increased lymph node metastasis and a more advanced stage of cancer. On the other hand, monocytes and B cells may have a beneficial effect in fighting against tumors. A group of tumor‑associated DC2s expressing both LAMP3 and CCL22 were shown to have a variety of immune‑related ligands. These cells have the ability to attract CD4+ T cells through communication between cells in the microenvironment. In this study, the immunological composition was examined at the level of individual cells and new prospective treatment approaches for PTC‑M were identified. The results support the hypothesis that myeloid cells and Tregs significantly contribute to tumor progression and metastasis by shaping the tumor microenvironment.

View Figures

View References

1	Pizzato M, Li M, Vignat J, Laversanne M, Singh D, La Vecchia C and Vaccarella S: The epidemiological landscape of thyroid cancer worldwide: GLOBOCAN estimates for incidence and mortality rates in 2020. Lancet Diabetes Endocrinol. 10:264–272. 2022. View Article : Google Scholar : PubMed/NCBI
2	Cheng F, Xiao J, Shao C, Huang F, Wang L, Ju Y and Jia H: Burden of Thyroid Cancer From 1990 to 2019 and Projections of Incidence and Mortality Until 2039 in China: Findings From Global Burden of Disease Study. Front Endocrinol (Lausanne). 12:7382132021. View Article : Google Scholar : PubMed/NCBI
3	Cabanillas ME, McFadden DG and Durante C: Thyroid cancer. Lancet. 388:2783–2795. 2016. View Article : Google Scholar : PubMed/NCBI
4	Suh YJ, Kwon H, Kim SJ, Choi JY, Lee KE, Park YJ, Park DJ and Youn YK: Factors affecting the locoregional recurrence of conventional papillary thyroid carcinoma after surgery: A Retrospective Analysis of 3381 Patients. Ann Surg Oncol. 22:3543–3549. 2015. View Article : Google Scholar : PubMed/NCBI
5	Zhan L, Feng HF, Yu XZ, Li LR, Song JL, Tu Y, Yuan JP, Chen C and Sun SR: Clinical and prognosis value of the number of metastatic lymph nodes in patients with papillary thyroid carcinoma. BMC Surg. 22:2352022. View Article : Google Scholar : PubMed/NCBI
6	Pu W, Shi X, Yu P, Zhang M, Liu Z, Tan L, Han P, Wang Y, Ji D, Gan H, et al: Single-cell transcriptomic analysis of the tumor ecosystems underlying initiation and progression of papillary thyroid carcinoma. Nat Commun. 12:60582021. View Article : Google Scholar : PubMed/NCBI
7	Xie Z, Li X, He Y, Wu S, Wang S, Sun J, He Y, Lun Y and Zhang J: Immune cell confrontation in the papillary thyroid carcinoma microenvironment. Front Endocrinol (Lausanne). 11:5706042020. View Article : Google Scholar : PubMed/NCBI
8	Galdiero MR, Varricchi G and Marone G: The immune network in thyroid cancer. Oncoimmunology. 5:e11685562016. View Article : Google Scholar : PubMed/NCBI
9	Yan T, Qiu W, Weng H, Fan Y, Zhou G and Yang Z: Single-Cell transcriptomic analysis of ecosystems in papillary thyroid carcinoma progression. Front Endocrinol (Lausanne). 12:7295652021. View Article : Google Scholar : PubMed/NCBI
10	Bergdorf K, Ferguson DC, Mehrad M, Ely K, Stricker T and Weiss VL: Papillary thyroid carcinoma behavior: Clues in the tumor microenvironment. Endocr Relat Cancer. 26:601–614. 2019. View Article : Google Scholar : PubMed/NCBI
11	Bentz BG, Miller BT, Holden JA, Rowe LR and Bentz JS: B-RAF V600E mutational analysis of fine needle aspirates correlates with diagnosis of thyroid nodules. Otolaryngol Head Neck Surg. 140:709–714. 2009. View Article : Google Scholar : PubMed/NCBI
12	Dura B, Choi JY, Zhang K, Damsky W, Thakral D, Bosenberg M, Craft J and Fan R: scFTD-seq: Freeze-thaw lysis based, portable approach toward highly distributed single-cell 3′ mRNA profiling. Nucleic Acids Res. 47:e162019. View Article : Google Scholar : PubMed/NCBI
13	Martin M: Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet J. 17:10–12. 2011. View Article : Google Scholar
14	Dobin A, Davis CA, Schlesinger F, Drenkow J, Zaleski C, Jha S, Batut P, Chaisson M and Gingeras TR: STAR: Ultrafast universal RNA-seq aligner. Bioinformatics. 29:15–21. 2013. View Article : Google Scholar : PubMed/NCBI
15	Liao Y, Smyth GK and Shi W: featureCounts: An efficient general purpose program for assigning sequence reads to genomic features. Bioinformatics. 30:923–930. 2014. View Article : Google Scholar : PubMed/NCBI
16	Satija R, Farrell JA, Gennert D, Schier AF and Regev A: Spatial reconstruction of single-cell gene expression data. Nat Biotechnol. 33:495–502. 2015. View Article : Google Scholar : PubMed/NCBI
17	Yu G, Wang LG, Han Y and He QY: clusterProfiler: An R package for comparing biological themes among gene clusters. OMICS. 16:284–287. 2012. View Article : Google Scholar : PubMed/NCBI
18	Hänzelmann S, Castelo R and Guinney J: GSVA: Gene set variation analysis for microarray and RNA-seq data. BMC Bioinformatics. 14:72013. View Article : Google Scholar : PubMed/NCBI
19	Qiu X, Hill A, Packer J, Lin D, Ma YA and Trapnell C: Single-cell mRNA quantification and differential analysis with Census. Nat Methods. 14:309–315. 2017. View Article : Google Scholar : PubMed/NCBI
20	Gulati GS, Sikandar SS, Wesche DJ, Manjunath A, Bharadwaj A, Berger MJ, Ilagan F, Kuo AH, Hsieh RW, Cai S, et al: Single-cell transcriptional diversity is a hallmark of developmental potential. Science. 367:405–411. 2020. View Article : Google Scholar : PubMed/NCBI
21	Efremova M, Vento-Tormo M, Teichmann SA and Vento-Tormo R: CellPhoneDB: Inferring cell-cell communication from combined expression of multi-subunit ligand-receptor complexes. Nat Protoc. 15:1484–1506. 2020. View Article : Google Scholar : PubMed/NCBI
22	Carstens JL, Correa de Sampaio P, Yang D, Barua S, Wang H, Rao A, Allison JP, LeBleu VS and Kalluri R: Spatial computation of intratumoral T cells correlates with survival of patients with pancreatic cancer. Nat Commun. 8:150952017. View Article : Google Scholar : PubMed/NCBI
23	Bankhead P, Loughrey MB, Fernández JA, Dombrowski Y, McArt DG, Dunne PD, McQuaid S, Gray RT, Murray LJ, Coleman HG, et al: QuPath: Open source software for digital pathology image analysis. Sci Rep. 7:168782017. View Article : Google Scholar : PubMed/NCBI
24	Skytthe MK, Graversen JH and Moestrup SK: Targeting of CD163+ Macrophages in Inflammatory and Malignant Diseases. Int J Mol Sci. 21:54972020. View Article : Google Scholar : PubMed/NCBI
25	Wu Y, Yang S, Ma J, Chen Z, Song G, Rao D, Cheng Y, Huang S, Liu Y, Jiang S, et al: Spatiotemporal immune landscape of colorectal cancer liver metastasis at single-cell level. Cancer Discov. 12:134–153. 2022. View Article : Google Scholar : PubMed/NCBI
26	Huang X, Li Y, Fu M and Xin HB: Polarizing macrophages in vitro. Methods Mol Biol. 1784:119–126. 2018. View Article : Google Scholar : PubMed/NCBI
27	Lv SL, Zeng ZF, Gan WQ, Wang WQ, Li TG, Hou YF, Yan Z, Zhang RX and Yang M: Lp-PLA2 inhibition prevents Ang II-induced cardiac inflammation and fibrosis by blocking macrophage NLRP3 inflammasome activation. Acta Pharmacol Sin. 42:2016–2032. 2021. View Article : Google Scholar : PubMed/NCBI
28	Tang TT, Lv LL, Pan MM, Wen Y, Wang B, Li ZL, Wu M, Wang FM, Crowley SD and Liu BC: Hydroxychloroquine attenuates renal ischemia/reperfusion injury by inhibiting cathepsin mediated NLRP3 inflammasome activation. Cell Death Dis. 9:3512018. View Article : Google Scholar : PubMed/NCBI
29	Rebeck GW: The role of APOE on lipid homeostasis and inflammation in normal brains. J Lipid Res. 58:1493–1499. 2017. View Article : Google Scholar : PubMed/NCBI
30	Zhou L, Zhuo H, Ouyang H, Liu Y, Yuan F, Sun L, Liu F and Liu H: Glycoprotein non-metastatic melanoma protein b (Gpnmb) is highly expressed in macrophages of acute injured kidney and promotes M2 macrophages polarization. Cell Immunol. 316:53–60. 2017. View Article : Google Scholar : PubMed/NCBI
31	Guilliams M, Ginhoux F, Jakubzick C, Naik SH, Onai N, Schraml BU, Segura E, Tussiwand R and Yona S: Dendritic cells, monocytes and macrophages: A unified nomenclature based on ontogeny. Nat Rev Immunol. 14:571–578. 2014. View Article : Google Scholar : PubMed/NCBI
32	Maier B, Leader AM, Chen ST, Tung N, Chang C, LeBerichel J, Chudnovskiy A, Maskey S, Walker L, Finnigan JP, et al: A conserved dendritic-cell regulatory program limits antitumour immunity. Nature. 580:257–262. 2020. View Article : Google Scholar : PubMed/NCBI
33	Zhang Q, He Y, Luo N, Patel SJ, Han Y, Gao R, Modak M, Carotta S, Haslinger C, Kind D, et al: Landscape and dynamics of single immune cells in hepatocellular carcinoma. Cell. 179:829–845.e20. 2019. View Article : Google Scholar : PubMed/NCBI
34	Liu W, Hu H, Shao Z, Lv X, Zhang Z, Deng X, Song Q, Han Y, Guo T, Xiong L, et al: Characterizing the tumor microenvironment at the single-cell level reveals a novel immune evasion mechanism in osteosarcoma. Bone Res. 11:42023. View Article : Google Scholar : PubMed/NCBI
35	Rowshanravan B, Halliday N and Sansom DM: CTLA-4: A moving target in immunotherapy. Blood. 131:58–67. 2018. View Article : Google Scholar : PubMed/NCBI
36	Rapp M, Wintergerst MWM, Kunz WG, Vetter VK, Knott MML, Lisowski D, Haubner S, Moder S, Thaler R, Eiber S, et al: CCL22 controls immunity by promoting regulatory T cell communication with dendritic cells in lymph nodes. J Exp Med. 216:1170–1181. 2019. View Article : Google Scholar : PubMed/NCBI
37	Yin H, Tang Y, Guo Y and Wen S: Immune microenvironment of thyroid cancer. J Cancer. 11:4884–4896. 2020. View Article : Google Scholar : PubMed/NCBI
38	Yu H, Huang X, Liu X, Jin H, Zhang G, Zhang Q and Yu J: Regulatory T cells and plasmacytoid dendritic cells contribute to the immune escape of papillary thyroid cancer coexisting with multinodular non-toxic goiter. Endocrine. 44:172–181. 2013. View Article : Google Scholar : PubMed/NCBI
39	He D, Wang D, Lu P, Yang N, Xue Z, Zhu X, Zhang P and Fan G: Single-cell RNA sequencing reveals heterogeneous tumor and immune cell populations in early-stage lung adenocarcinomas harboring EGFR mutations. Oncogene. 40:355–368. 2021. View Article : Google Scholar : PubMed/NCBI
40	Choi J, Gyamfi J, Jang H and Koo JS: The role of tumor-associated macrophage in breast cancer biology. Histol Histopathol. 33:133–145. 2018.PubMed/NCBI
41	Chung W, Eum HH, Lee HO, Lee KM, Lee HB, Kim KT, Ryu HS, Kim S, Lee JE, Park YH, et al: Single-cell RNA-seq enables comprehensive tumour and immune cell profiling in primary breast cancer. Nat Commun. 8:150812017. View Article : Google Scholar : PubMed/NCBI
42	Fan C, Wu J, Shen Y, Hu H, Wang Q, Mao Y, Ye B and Xiang M: Hypoxia promotes the tolerogenic phenotype of plasmacytoid dendritic cells in head and neck squamous cell carcinoma. Cancer Med. 11:922–930. 2022. View Article : Google Scholar : PubMed/NCBI
43	Zhang QY, Ye XP, Zhou Z, Zhu CF, Li R, Fang Y, Zhang RJ, Li L, Liu W, Wang Z, et al: Lymphocyte infiltration and thyrocyte destruction are driven by stromal and immune cell components in Hashimoto's thyroiditis. Nat Commun. 13:7752022. View Article : Google Scholar : PubMed/NCBI
44	Propper DJ and Balkwill FR: Harnessing cytokines and chemokines for cancer therapy. Nat Rev Clin Oncol. 19:237–253. 2022. View Article : Google Scholar : PubMed/NCBI
45	Berlato C, Khan MN, Schioppa T, Thompson R, Maniati E, Montfort A, Jangani M, Canosa M, Kulbe H, Hagemann UB, et al: A CCR4 antagonist reverses the tumor-promoting microenvironment of renal cancer. J Clin Invest. 127:801–813. 2017. View Article : Google Scholar : PubMed/NCBI