Age and BRAF^V600E Mutation Stratified Patients with Cytologically Benign Thyroid Nodules

Figures & data

Figure 1 The diagram of the study. The sample used for analysis and the analysis diagram of the study.

Table 1 Correlation Between Clinical Characteristics and BRAF^600E Mutation in All Thyroid Nodules

Characteristics	BRAF^V600E		P-value
	Positive (89)	Negative (128)
Gender (Males)	23/89(25.8%)	33/89(25.7%)	0.992
Age (mean ± S.D.)	46.01±12.640	48.11±11.783	0.233
Drinking	5/89 (5.6%)	6/128 (4.7%)	0.496
Smoking	7/89 (7.9%)	14/128 (10.9%)	0.451
TBSRTC II	17/89 (19.1%)	86/128 (67.2%)	<0.001***
TBSRTC III–IV	31/89 (34.8%)	24/128 (18.8%)	0.007**
TBSRTC V–VI	41/89 (46.1%)	18/128 (14.1%)	<0.001***
C-TIRADS 4b-4c	37/89 (41.6%)	18/128 (14.1%)	<0.001***

Notes: ***P<0.001, **P<0.01.

Abbreviations: mean±S.D., mean ± standard deviation; TBSRTC, the Bethesda System for Reporting Thyroid Cytopathology; C-TIRADS, Chinese Thyroid imaging reporting and data system.

Figure 2 Associations of BRAF^V600E mutation with thyroid nodule classifications and clinicopathological stages. (A) Alluvial diagram of the BRAF^V600E mutation with different classification of US classification C-TIRADS, TBSRTC categories, and Pathological diagnosis; (B–E) Correlation between BRAF^V600E mutation status and (B) tumor stages, (C) T stages, (D) N stages, (E) M stages in TCGA-THCA cohort, red: positive BRAF^V600E mutation, blue: negative BRAF^V600E mutation; (F) The correlation between BRAF^V600E mutation status and the expression of BRAF in TCGA-THCA cohort.

Table 2 Clinical Characteristics Associated with Malignant in Thyroid Nodules

Characteristics	Malignant (39)	Benign (4)	P-value
Gender (Male)	14/39(35.9%)	0/4(0%)	0.192
Age (mean±S.D.)	45.92±12.55	48.75±8.26	0.664
Drinking	6/39(15.4%)	0/4(0%)	0.535
Smoking	8/39(20.5%)	0/4(0%)	0.315
Positive BRAF^V600E	29/39 (74.4%)	0/4 (0%)	0.008**
TBSRTC II	7/39(17.9%)	1/4(25%)	0.576
TBSRTC III–IV	12/39(30.8%)	3/4(75%)	0.114
TBSRTC V–VI	20/39(51.3%)	0(0%)	0.072
C-TIRADS 3–4a	18/39(46.2%)	3/4(75%)	0.286

Notes: **P<0.01.

Abbreviations: mean±S.D., mean ± standard deviation; TBSRTC, the Bethesda System for Reporting Thyroid Cytopathology; C-TIRADS, Chinese Thyroid imaging reporting and data system.

Table 3 Diagnostic Efficacy of Different Methods for Discrimination Malignant in Thyroid Nodules

Diagnosis Methods	AUC	Sensitivity	Specificity
BRAF^V600E	0.872	74.4%	100.0%
TBSRTC	0.747	51.3%	100.0%
C-TIRADS	0.721	53.8%	75.0%
BRAF^V600E + TBSRTC	0.923	84.6%	100.0%
BRAF^V600E +C-TIRADS	0.824	89.7%	75.0%
TBSRTC +C-TIRADS	0.747	74.4%	75.0%
Three methods combined	0.837	92.3%	75.0%

Abbreviations: AUC, area under the curve; TBSRTC, the Bethesda System for Reporting Thyroid Cytopathology; C-TIRADS, Chinese Thyroid imaging reporting and data system.

Table 4 Correlations of BSRTC Categories with BRAF^V600E Mutation and Pathological Results

TBSRTC	all	BRAF^V600E Mutation	The Rate of BRAF^V600E Mutation	Malignant Rate
II	8	Positive	3(37.5%)	3/3(100%)
		Negative	5(62.5%)	4/5(80%)
III–IV	15	Positive	10(66.7%)	10/10(100%)
		Negative	5(33.3%)	2/5(25%)
V–VI	20	Positive	16(80%)	16/16(100%)
		Negative	4(20%)	4/4(100%)

Abbreviation: TBSRTC, the Bethesda System for Reporting Thyroid Cytopathology.

Table 5 The Prediction of BRAF^V600E Mutation on Malignancy in Thyroid Nodules with Benign Cytology

Studies	Number of Cases in Benign FNA	Malignant in BRAF^V600E Mutation
Kim et al, 2014^Citation18	4028	15/17
Zhang et al, 2022^Citation19	49	20/21
Chen et al, 2018^Citation17	292	31/36

Abbreviation: FNA, fine needle aspiration.

Figure 3 ROC analysis of diagnosis methods for malignancy. The ROC analysis of TBSRTC, BRAF^V600E mutation status, and C-TIRADS for malignancy in thyroid nodules.

Table 6 Correlation Between BRAF^V600E Mutation and Clinical Characteristics in Thyroid Nodules with Benign Cytology

Characteristics	BRAF^V600E		P-value
	Positive (%)	Negative (%)
Gender (Males)	4(23.5%)	22(25.6%)	0.564
Age (mean±S.D.)	47.67±9.38	56.35±11.89	0.0012**
Drinking	1(5.9%)	3(3.5%)	0.52
Smoking	1(5.9%)	7(8.1%)	0.607
C-TIRADS 4b-4c	6(7%)	2(11.8%)	0.393
Multifocality	13(76.5%)	45(53.2%)	0.056
Right Location	10(58.8)	34(39.5)	0.142
Irregular Morphology	14(82.4)	56(65.1)	0.164
Hypoechoic	16(94.1%)	80(93%)	0.675
Unclear Boundary	12(70.6)	55(64%)	0.41
Size (mm) median (IQR)	8(4.6–14.5)	8(6–12.25)	0.879

Note: **P<0.01.

Abbreviations: mean±S.D., mean ± standard deviation; C-TIRADS, Chinese Thyroid imaging reporting and data system; IQR, interquartile range.

Table 7 The Relationship Between BRAF^V600E Mutation and Clinical Characteristics in C-TIRADS 3–4a Patients with Benign Cytology

Characteristics	BRAF^V600E		P-value
	Negative	Positive
Gender (Male)	3(20%)	21(26%)	0.441
Age (mean±S.D.)	56.87±12.55	47.28±9.31	0.001**
Drinking	1(6.7%)	3(3.8%)	0.503
Smoking	1(6.7%)	6(7.5%)	0.695
Multifocality	12(80%)	40(50%)	0.032*
Location (Right)	10(66.7%)	30(37.5%)	0.036*
Irregular Morphology	12(80%)	52(65%)	0.254
Hypoechoic	14(93.3%)	74(92.5%)	0.695
Unclear Boundary	10(66.7%)	51(63.7%)	0.829
Size (mm) median (IQR)	7.0(4.4–15.0)	8.0(6.0–13.0)	0.786

Notes: *P<0.05, **P<0.01.

Abbreviations: C-TIRADS, Chinese Thyroid imaging reporting and data system; mean±S.D., mean ± standard deviation; IQR, interquartile range.

Figure 4 The correlation of BRAF^V600E mutation status and patient age. (A–E) Comparison of patient age between positive and negative BRAF^V600E mutation (A) in benign cytology TBSRTC II, (B) in C-TIRADS 3–4a with benign cytology, (C) in all the patients of our cohort, (D) in TCGA-THCA cohort, and (E) in TCGA-THCA-Asian cohort (E). (F) Liner correlation between patient age and BRAF^V600E mutation rate in different cohorts. Green: TCGA-THCA Asian cohort, red: FHPT cohort in TBSRTC II, blue: FHPT cohort in C-TIRADS 3–4a with TBSRTC II.

Figure 5 The association in immune response with BRAF^V600E and patients age. (A) The volcano plot of differentially expressed genes (DEGs) between older and younger patients in TCGA-THCA-Asian, younger: ≤50 years vs.older: >50 years. (B) KEGG analysis of DEGs between younger and older patients in TCGA-THCA-Asian. (C) The volcano plot of DEGs between patients with positive and negative BRAF^V600E mutation in TCGA-THCA-Asian. (D) KEGG analysis intersection DEGs between BRAF^V600E mutations groups and age groups. (E) The proportion of immunosimulators in younger and older patients, positive and negative BRAF^V600E mutation in TCGA-THCA-Asian. Younger: ≤50 years vs.older: >50 years *p < 0.05, **p < 0.01.

Kim SY, Kim EK, Kwak JY, Moon HJ, Yoon JH. What to do with thyroid nodules showing benign cytology and BRAF(V600E) mutation? A study based on clinical and radiologic features using a highly sensitive analytic method. Surgery. 2015;157(2):354–361. doi:10.1016/j.surg.2014.09.003

 PubMed Web of Science ®Google Scholar

Zhang Y, Lu F, Shi H, et al. Predicting malignancy in thyroid nodules with benign cytology results: the role of Conventional Ultrasound, Shear Wave Elastography and BRAF V600E. Clin Hemorheol Microcirc. 2022;81(1):33–45. doi:10.3233/CH-211337

 PubMed Web of Science ®Google Scholar

Chen X, Zhou Q, Wang F, et al. Value of BRAF V600E in high-risk thyroid nodules with benign cytology results. AJNR Am J Neuroradiol. 2018;39(12):2360–2365. doi:10.3174/ajnr.A5898

 PubMed Web of Science ®Google Scholar