Comprehensive survey of HRAS, KRAS, and NRAS mutations in proliferative thyroid lesions from an ethnically diverse population.

BACKGROUND: The distribution and kind of rat sarcoma viral oncogenes homolog (RAS) mutations, as well as their clinical impact on different types of thyroid lesions, vary widely among the different populations studied. We performed a comprehensive mutational survey in the highly related RAS genes HRAS, KRAS, and NRAS in a case series of proliferative thyroid lesions with known BRAF mutational status, originating from an ethnically diverse group. MATERIALS AND METHODS: Mutational hotspot regions encompassing codons 12, 13, and 61 of the RAS genes were directly sequenced in 381 cases of thyroid lesions. In addition, the putative NRAS hotspot region encompassing codon 97 was sequenced in 36 thyroid lesions. The case series included lesions of Hashimoto's thyroiditis (HT), nodular goiters, hyperplastic nodules, follicular adenomas (FAs), Hurthle cell variants of FA, papillary thyroid carcinomas (PTCs), follicular variants of PTC (FVPTCs), microcarcinomas of PTC (micro PTCs; tumor size ≤1 cm), follicular TCs (FTCs), Hurthle cell variants of FTC, and non-well-differentiated TCs (NWDTCs). RESULTS: We identified RAS mutations in 16 out of 57 (28.1%) FAs, 2 out of 8 (25%) NWDTCs, 8 out of 42 (19.0%) FVPTCs, 2 out of 10 (20.0%) FTCs, 1 out of 12 (8.3%) Hurthle cell variants of FA, 3 out of 46 (6.5%) goiters, 1 out of 18 (5.6%) hyperplastic nodules, 3 out of 56 (5.4%) micro PTCs, 2 out of 115 (1.7%) PTCs, 0 out of 7 (0%) Hurthle cell variants of FTC, and 0 out of 10 (0%) HT lesions. NRAS codon 61 mutation was the predominant form, followed by HRAS codon 61 mutation. Only three mutations affected RAS codons 12 and 13, two of which were identified in goiters. No codon 97 mutation was detected in the examined FVPTCs. An as yet undescribed deletion of KRAS codon 59 was identified in one FA. DISCUSSION: RAS mutations in our case series were commonly associated with follicular-patterned thyroid lesions. Our data suggest that FAs with a RAS mutation may constitute precursor lesions for TC with follicular histology. The newly-discovered KRAS codon 59 deletion is one of the first reported codon deletions in a RAS hotspot region.

A Novel mutation in the SLC26A4 gene in a Chinese family with Pendred syndrome.

OBJECTIVE: To investigate the mutations in the SLC26A4 gene in a Chinese patient with Pendred syndrome. METHODS: The diagnosis of Pendred syndrome was confirmed by the family history, pure tone audiogram, perchlorate discharge test (PDT), and computed tomography (CT) of the temporal bone. DNA extraction, PCR and DNA sequencing were performed according to standard procedures. Mutations in the SLC26A4 gene were compared with 100 unrelated subjects to exclude common polymorphism. Splice-site mutation was further confirmed by restriction enzyme length polymorphism (RFLP) with the specifically designed primers. RESULTS: The proband presented with typical features of bilateral sensorineural deafness since childhood and goiter development in the early adulthood. Thyroid studies disclosed euthyroidism with elevated thyroglobulin, but negative for PDT. Marked enlargement of bilateral vestibular aqueduct (>1.5 mm) was found by CT of the temporal bone. A novel SLC26A4 splice-site mutation c.1263+1G>A (IVS10+1G>A) was identified in compound heterozygosity with the missense mutation c.1079C>T (p.A360V) in the proband. Both mutations were not found in the 100 unrelated Chinese. CONCLUSIONS: Our results support previous findings that Pendred syndrome can be caused by compound heterozygous mutation in the SLC26A4 gene, in which IVS10+1G>A is a novel pathogenic mutation.

Pathogenic substitution of IVS15 + 5G > A in SLC26A4 in patients of Okinawa Islands with enlarged vestibular aqueduct syndrome or Pendred syndrome.

BACKGROUND: Pendred syndrome (PS) and nonsyndromic hearing loss associated with enlarged vestibular aqueduct (EVA) are caused by SLC26A4 mutations. The Okinawa Islands are the southwestern-most islands of the Japanese archipelago. And ancestral differences have been reported between people from Okinawa Island and those from the main islands of Japan. To confirm the ethnic variation of the spectrum of SLC26A4 mutations, we investigated the frequencies of SLC26A4 mutations and clinical manifestations of patients with EVA or PS living in the Okinawa Islands. METHODS: We examined 22 patients with EVA or PS from 21 unrelated families in Okinawa Islands. The patient's clinical history, findings of physical and otoscopic examinations, hearing test, and computed tomography (CT) scan of the temporal bones were recorded. To detect mutations, all 21 exons and the exon-intron junctions of SLC26A4 were sequenced for all subjects. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) for SLC26A4 and calculations using the comparative CT (2(-ΔΔCT)) method were used to determine the pathogenicity associated with gene substitutions. RESULTS: SLC26A4 mutations were identified in 21 of the 22 patients. We found a compound heterozygous mutation for IVS15 + 5G > A/H723R in nine patients (41%), a homozygous substitution of IVS15 + 5G > A in six patients (27%), and homozygous mutation for H723R in five patients (23%). The most prevalent types of SLC26A4 alleles were IVS15 + 5G > A and H723R, which both accounted for 15/22 (68%) of the patients. There were no significant correlations between the types of SLC26A4 mutation and clinical manifestations. Based on qRT-PCR results, expression of SLC26A4 was not identified in patients with the homozygous substitution of IVS15 + 5G > A. CONCLUSIONS: The substitution of IVS15 + 5G > A in SLC26A4 was the most common mutation in uniquely found in patients with PS and EVA in Okinawa Islands. This suggested that the spectrum of SLC26A4 mutation differed from main islands of Japan and other East Asian countries. The substitution of IVS15 + 5G > A leads to a loss of SLC26A expression and results in a phenotype of PS and EVA.

Identification of two heterozygous deafness mutations in SLC26A4 (PDS) in a Chinese family with two siblings.

OBJECTIVE: To detect genetic cause of two Chinese siblings (patient 1 and 2) with Pendred syndrome. DESIGN: Patients and their parents underwent clinical and genetic evaluations. To identify genetic mutations, sequencing of SLC26A4 was carried out. STUDY SAMPLE: Two siblings and their parents. RESULTS: Clinical evaluations showed that patient 1 suffered from bilateral postlingual progressive sensorineural hearing loss with enlarged vestibular aqueduct and slight diffuse multinodular goiter with euthyroid, and patient 2 suffered from bilateral prelingual progressive sensorineural hearing loss with enlarged vestibular aqueduct and no goiter with euthyroid. Furthermore, the sequence analysis of SLC26A4 indicated that either of the two siblings presented a compound heterozygote for the c.919A>G mutation in the splice site of intron 7 and for the c.1548insC mutation in exon 14. Their mother was a heterozygous carrier of the splice site mutation in intron 7, and their father was a heterozygous carrier of the insertion mutation in exon 14. CONCLUSIONS: Mutation analysis identified a compound heterozygous mutation (c.919A>G/c.1548insC) in SLC26A4 in two Chinese siblings with Pendred syndrome. Also, c.1548insC was first reported in the Chinese population. Although the two siblings from the same family carried the same genotype, they presented different phenotypes.

An investigation of epidemiologic factors associated with large nodular goiter.

BACKGROUND: Sporadic nodular goiter is a common problem in the United States and significant compressive symptoms may occur with progression to a critical size. METHODS: Potential epidemiological variables associated with the development of large unilateral (> or = 50 g) and bilateral (> or = 100 g) nodular goiter were investigated including: age, gender, race, body mass index (BMI), family history of thyroid disease, pregnancy at time of diagnosis, insurance status, and tobacco or alcohol use. Data were obtained from an IRB-approved thyroid database and retrospective chart review of consecutive patients operated on for nodular goiter from 1990 through 2005. A univariate and multivariate analysis of epidemiological variables in patients with "large" versus "small" nodular goiter was completed. RESULTS: Of the 488 patients operated on for nodular goiter, 113 (23%) were classified as "large," 43 with unilateral (mean 106 +/- 72 g) and 70 with bilateral enlargement (mean 173 +/- 92 g) and 375 (77%) were classified as "small," 179 with unilateral (18 +/- 10 g) and 196 with bilateral (37 +/- 24 g) enlargement. Based on univariate analysis, African-American race, age > or = 40 years, BMI > or = 30 kg/m2, and lack of insurance were associated with an increased risk of large nodular goiter (P < or = 0.001), whereas alcohol use was protective (P = 0.002). A multivariate analysis revealed that African-American race [adjusted odds ratio (adj. OR) 3.3, 95% CI = 2.0-5.4], age > or = 40 years (adj. OR 2.1, 95% CI = 1.2-3.8), and BMI > or = 30 kg/m2 (adj. OR 2.5, 95% CI = 1.5-4.0) were independently associated with large nodular goiter. No significant differences were observed in gender, family history of thyroid disease, pregnancy, or tobacco use (P > 0.1). CONCLUSIONS: African-American race, obesity, and increasing age are independent risk factors for the development of large nodular goiter. These results may be helpful in determining how best to monitor patients with nodular goiter, with earlier intervention to help prevent progressive enlargement and its sequelae.

Goitre prevalence in indigenous population of Evenkia.

Representatives of indigenous northern peoples (the Evenks and the Kets) living in a large Siberian territory named Evenkia were studied. There were children 6-14 years of age and adults aged 18-45 years. Thyroid volumes were determined by ultrasound. Iodine excretion was estimated. Normal thyroid volumes were found in 27.5% of the children. A high prevalence of goitre was detected in the adults. 55.4% of the women and 29.1% of the men had thyroid volumes above normal upper limits. Nodular goitres were found in 10.8% of the adults. 58.8% of the subjects had urinary iodine excretion below 10 micrograms/dl. Data show that the territory of Evenkia, settled by northern aborigines, is iodine deficient. The usage of the iodinated salt is ineffective. The preservation of the iodine content in salt remains the main problem in this area. In addition, some other goitrogenic agents may contribute to the high goitre prevalence in the indigenous population in Evenkia.

Antibodies to the thyroid gland and to the thyrotrophin receptor in African and Indian thyrotoxic patients.

Sixty two thyrotoxic patients, 34 African and 28 Indian, were studied in order to assess the prevalence of thyroid antibodies and TSH binding inhibitory activity (TBI): 45 had Graves' disease and 17 had toxic nodular goitres. Microsomal and thyroglobulin antibodies were positive more often in Indian than in African patients with Graves' disease (microsomal 52% vs 37.4%, P less than 0.05; thyroglobulin 38.1% vs 4.2%, P less than 0.001). Patients with toxic nodular goitres had a lower prevalence of positive microsomal antibodies (P less than 0.01), but not of thyroglobulin antibodies (P = 0.1) when compared with patients with Graves' disease. TBI activity measured by a radioreceptor assay was positive in 43 of the 45 (95%) patients with Graves' disease and only 1 of the 17 patients (5.9%) with toxic nodular goitre. It thus appears that TBI activity is a sensitive marker in the diagnosis of Graves' disease and that there is a lower prevalence of thyroglobulin and microsomal antibodies in African patients compared with Indian patients.