Molecular and Proteomic Profiles of Radioiodine Refractory Papillary Thyroid Cancer

Background Despite the generally favorable prognosis of papillary thyroid cancers (PTCs) following surgery with/without radioactive iodine (RAI) therapy, approximately one-third of patients experiencing recurrence and metastasis eventually develop resistance to RAI, leading to poor outcomes. However, the mechanisms underlying RAI-refractoriness remain elusive. This study aimed to assess the molecular and proteomic characteristics of RAI-refractory PTC (RR-PTC) for deeper insights.

The medical records were reviewed for the selection and grouping of RR-PTC patients and RAI-sensitive controls. RR-PTC patients were divided into three subgroups: continuous RAI uptake (ID), loss of uptake at the first I-131 treatment (iDF) and lost gradually (iDG). Proteomic profiling and targeted next-generation sequencing were performed on primary lesions. The incidence of gene mutations and fusions was compared across groups. Bioinformatic analysis was subsequently conducted to identify the differentially expressed proteins and enriched pathways.

Forty-eight PTC patients with recurrence and/or metastasis were included. The expression profiles of the RR-PTC and control groups were similar. In the subgroup comparison, enriched pathways related to MAPK and TNF signaling were associated with negative I-131 uptake and tumor tolerance with positive I-131 uptake. The BRAFV600E mutation was less common in the ID group, whereas the TERT promoter mutation was more common in the iDF group. NCOA4-RET fusion was more common in the ID group.

The present study constructed the first proteomic profile of RR-PTC. The identified proteins and pathways may be promising biomarkers and drug targets. Gene alterations can aid in the early diagnosis of RR-PTC. Competing Interest Statement The authors have declared no competing interest. Funding Statement This research was supported by grants from the Interdisciplinary Innovative Talents Foundation from Renmin Hospital of Wuhan University (JCRCFZ-2022-015), the Fundamental Research Funds for the Central Universities (2042019kf0229), the Natural Science Foundation of Hubei Province, China (2023AFB701), and the Thyroid Research Project for Young and Middle-aged Doctors from Bethune Charitable Foundation (JKM2022-B12). Author Declarations I confirm all relevant ethical guidelines have been followed, and any necessary IRB and/or ethics committee approvals have been obtained. Yes The details of the IRB/oversight body that provided approval or exemption for the research described are given below: The Institutional Ethical Committee of Renmin Hospital of Wuhan University gave ethical approval for this study (No. WDRY2021-K032). The requirement for obtaining informed consent from the involved patients was waived. I confirm that all necessary patient/participant consent has been obtained and the appropriate institutional forms have been archived, and that any patient/participant/sample identifiers included were not known to anyone (e.g., hospital staff, patients or participants themselves) outside the research group so cannot be used to identify individuals. Yes I understand that all clinical trials and any other prospective interventional studies must be registered with an ICMJE-approved registry, such as ClinicalTrials.gov. I confirm that any such study reported in the manuscript has been registered and the trial registration ID is provided (note: if posting a prospective study registered retrospectively, please provide a statement in the trial ID field explaining why the study was not registered in advance). Yes I have followed all appropriate research reporting guidelines, such as any relevant EQUATOR Network research reporting checklist(s) and other pertinent material, if applicable. Yes Footnotes Author list and data availability were modified. Data Availability The proteomic data have been deposited in the iProX database with the project ID IPX0009103001. Calculation files and additional data are available in the Mendeley database (DOI: 10.17632/yfpfvktrxn). No custom code was used in the current study. Abbreviations - ANOVA - analysis of variance - ATA - American Thyroid Association - AUC - area under the curve (of ROC) - BRAF - B-Raf proto-oncogene, serine/threonine kinase - BRS - BRAF-RAS score - CC - correlation coefficient - COSMIC - the Catalogue Of Somatic Mutations In Cancer - CT - computed tomography - CYT - immune cytolytic activity score - DEG - differentially expressed gene - DEP - differentially expressed protein - DIA - data-independent acquisition - DTC - differentiated thyroid cancer - ECP - eosinophil cationic protein - ES - enrichment score - ERK - extracellular signal-regulated kinase - ETE - extrathyroidal extension - FC - fold change - FFPE - formalin-fixed paraffin-embedded - F-18-FDG - 2-[18F]fluoro-2-deoxy-D-glucose - GEO - the Gene Expression Omnibus database - GO - gene ontology - GS - gene significance - GSEA - gene set enrichment analysis - GSVA - gene set variation analysis - HR - hazard ratio - ID - positive RAI uptake and disease persistence - Id - RAI uptake positive and disease remission - iDF - negative RAI uptake at the first time of RAI treatment with disease persistence - iDG - RAI uptake lost gradually after previous RAI treatments with disease persistence - IIS - immune infiltration score - IP - inositol phosphate - IPA - ingenuine pathway analysis - KEGG - Kyoto Encyclopedia of Genes and Genomes - KNN - K-nearest neighbor - LASSO - least absolute shrinkage and selection operator - LC - liquid chromatography - LNM - lymph node metastatic lesion - MAPK - mitogen-activated protein kinase - MET - MET proto-oncogene - MKI - multi-kinase inhibitor - MM - module membership - MS - mass spectrometry - MRI - magnetic resonance imaging - NIS - sodium iodide symporter - PCA - principal component analysis - PCT - pressure cycling technology - PET - positron emission tomography - RAI - radioactive iodine - RAIR - RAI-refractoriness - RF - random forest - RFS - recurrence-free survival - RMA - robust multiarray average - ROC - receiver operating characteristic - RR-PTC - radioactive iodine refractory papillary thyroid cancer - SPECT - single photon emission computerized tomography - ssGSEA - single sample GSEA - SVM-RFE - support vector machine-recursive feature elimination - TAK1 - transforming growth factor-β activated kinase 1 - TCGA - The Cancer Genome Atlas Program - TERT - telomerase reverse transcriptase - TERTp - TERT promoter - TDS - thyroid differentiation score - TIS - T cell infiltration score - TNF - tumor necrosis factor - TNGS - targeted next-generation sequencing - TOM - topological overlap matrix - WBS - whole-body scanning - WGCNA - weighted correlation network analysis