Thyroid Cancer Screening Using Tumor-Associated DN T Cells as Immunogenomic Markers

Shahnawaz Imam; Rodis D Paparodis; Shafiya Imtiaz Rafiqi; Sophia Ali; Azra Niaz; Abed Kanzy; Yara E Tovar; Mohammed A Madkhali; Ahmed Elsherif; Feras Khogeer; Zeeshan A Zahid; Haider Sarwar; Tamanna Karim; Nancy Salim; Juan C Jaume

doi:10.3389/fonc.2022.891002

Thyroid Cancer Screening Using Tumor-Associated DN T Cells as Immunogenomic Markers

Front Oncol. 2022 May 27:12:891002. doi: 10.3389/fonc.2022.891002. eCollection 2022.

Authors

Shahnawaz Imam^{1

2}, Rodis D Paparodis^{1

2

3}, Shafiya Imtiaz Rafiqi^{1

2}, Sophia Ali^{1

2}, Azra Niaz^{1

2}, Abed Kanzy^{1

2}, Yara E Tovar^{1

2}, Mohammed A Madkhali^{1

2}, Ahmed Elsherif^{1

2}, Feras Khogeer^{1

2}, Zeeshan A Zahid^{1

2}, Haider Sarwar^{1

2

4}, Tamanna Karim^{1

2}, Nancy Salim^{1

2}, Juan C Jaume^{1

2}

Affiliations

¹ Division of Endocrinology, Diabetes and Metabolism, Department of Medicine, College of Medicine and Life Sciences, University of Toledo, Toledo, OH, United States.
² Center for Diabetes and Endocrine Research (CeDER), University of Toledo, Toledo, OH, United States.
³ Private Practitioner, Patras, Greece.
⁴ Windsor University School of Medicine, Cayon St. Kitts West Indies, Saint Kitts and Nevis.

Abstract

Background: Thyroid nodules are an extremely common entity, and surgery is considered the ultimate diagnostic strategy in those with unclear malignant potential. Unfortunately, strategies aiming to predict the risk of malignancy have inadequate specificity. Our group recently found that the microenvironment of thyroid cancer is characterized by an enhanced immune invasion and activated immune response mediated by double-negative T lymphocytes (DN T) (CD3⁺CD4^-CD8^-), which are believed to enable or promote tumorigenesis. In the present work, we try to use the DN T cells' proportion in thyroid fine-needle aspiration (FNA) material as a predictor of the risk of malignancy.

Methods: We recruited 127 patients and obtained ultrasound-guided FNA samples from subjects with cytology-positive or suspicious for malignancy and from those with benign nodular goiter associated with compressive symptoms (such as dysphagia, shortness of breath, or hoarseness), Hashimoto thyroiditis, and Graves' disease. Out of 127, we investigated 46 FNA samples of patients who underwent total thyroidectomy and for which postoperative histological diagnosis by the academic pathologists was available. We specifically measured the number of cells expressing CD3⁺CD4^-CD8^- (DN T) as a function of total CD3⁺ cells in FNA samples using flow cytometry. We correlated their FNA DN T-cell proportions with the pathological findings.

Results: The DN T cells were significantly more abundant in lymphocytic infiltrates of thyroid cancer cases compared to benign nodule controls (p < 0.0001). When the DN T-cell population exceeded a threshold of 9.14%, of total CD3⁺ cells, the negative likelihood ratio of being cancer-free was 0.034 (96.6% sensitivity, 95% CI, 0.915-1.000, p < 0.0001). DN T cells at <9.14% were not found in any subject with benign disease (specificity 100%). The high specificity of the test is promising, since it abolishes a false-positive diagnosis and in turn unnecessary surgical procedures.

Conclusion: The present study proposes DN T cells' proportion as a preoperative diagnostic signature for thyroid cancer that with integration of RNA transcriptomics can provide a simplified technology based on the PCR assay for the ease of operation.

Keywords: DN T cells; PTC; active surveillance; immune editing; immunogenomic marker; thyroid cancer.