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Department of Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, New York
I. Introduction 1084
II. Characterization of the Sodium/Iodide Symporter Protein 1086
A. Generation of anti-NIS antibodies 1086
B. N-linked glycosylation of NIS: implications for the NIS secondary structure model 1087
De La Vieja, Antonio, Orsolya Dohan, Orlie Levy, and Nancy Carrasco. Molecular Analysis of the Sodium/
Iodide Symporter: Impact on Thyroid and Extrathyroid Pathophysiology. Physiol Rev 80: 1083–1105, 2000.—The
Na⫹/I⫺ symporter (NIS) is an intrinsic membrane protein that mediates the active transport of iodide into the thyroid
and other tissues, such as salivary glands, gastric mucosa, and lactating mammary gland. NIS plays key roles in
thyroid pathophysiology as the route by which iodide reaches the gland for thyroid hormone biosynthesis and as a
means for diagnostic scintigraphic imaging and for radioiodide therapy in hyperthyroidism and thyroid cancer. The
molecular characterization of NIS started with the 1996 isolation of a cDNA encoding rat NIS and has since
continued at a rapid pace. Anti-NIS antibodies have been prepared and used to study NIS topology and its secondary
structure. The biogenesis and posttranslational modifications of NIS have been examined, a thorough electrophys-
iological analysis of NIS has been conducted, the cDNA encoding human NIS (hNIS) has been isolated, the genomic
organization of hNIS has been elucidated, the regulation of NIS by thyrotropin and I⫺ has been analyzed, the
regulation of NIS transcription has been studied, spontaneous NIS mutations have been identified as causes of
congenital iodide transport defect resulting in hypothyroidism, the roles of NIS in thyroid cancer and thyroid
autoimmune disease have been examined, and the expression and regulation of NIS in extrathyroidal tissues have
been investigated. In gene therapy experiments, the rat NIS gene has been transduced into various types of human
cells, which then exhibited active iodide transport and became susceptible to destruction with radioiodide. The
continued molecular analysis of NIS clearly holds the potential of an even greater impact on a wide spectrum of
fields, ranging from structure/function of transport proteins to the diagnosis and treatment of cancer, both in the
thyroid and beyond.
www.physrev.physiology.org 0031-9333/00 $15.00 Copyright © 2000 the American Physiological Society 1083
1084 DE LA VIEJA, DOHAN, LEVY, AND CARRASCO Volume 80
approaches and techniques, leading to numerous re- transcription has been studied; spontaneous NIS muta-
ports in just the last 3 years (61, 63, 98, 107). This tions have been identified as causes of congenital hy-
review provides an overview of the most recent devel- pothyroidism; and the roles of NIS in thyroid cancer
opments on the molecular analysis of NIS, among and thyroid autoimmune disease have been examined.
which are the following: anti-NIS antibodies have been In addition, it has been shown that I⫺ transport in
prepared and used to study the topology of NIS and to at least some extrathyroidal tissues, such as breast and
experimentally test the proposed NIS secondary model gastric mucosa, is also mediated by NIS expressed in
so that some aspects of the above-described model these tissues, in which NIS is differently regulated and
have been experimentally confirmed and others re- subjected to distinct posttranslational modifications.
vised. The most recent revised secondary structure This notion effectively invalidates the previously held
model for NIS proposes 13 (Fig. 2B) rather than 12 view that NIS was a major thyroid-specific protein, like
transmembrane segments (Fig. 2A). The biogenesis and thyroglobulin (Tg) and thyroid peroxidase (TPO), pre-
posttranslational modifications of NIS have been exam- sumably not expressed in any other tissues. Gene ther-
ined; a thorough electrophysiological analysis of NIS apy experiments have recently been reported in which
has been conducted, in which NIS specificity and stoi- the rat NIS gene was transduced into human melanoma,
chiometry were studied and a mechanistic model was murine liver, murine colon, and human carcinoma cells,
proposed; the cDNA encoding human NIS (hNIS) has all of which then exhibited active I⫺ transport and
been isolated; the genomic organization of hNIS has became susceptible to destruction with radioiodide.
been elucidated (Fig. 7) and the hNIS gene has been Therefore, it is now clearer than ever before that the
mapped to chromosome 19p; the regulation of NIS by continued molecular analysis of NIS holds the potential
TSH and I⫺ has been analyzed; the regulation of NIS of an even greater impact on a wide spectrum of fields,
1086 DE LA VIEJA, DOHAN, LEVY, AND CARRASCO Volume 80
ranging from structure/function of transport proteins to COOH terminus of the protein. The Ab immunoreacts
the diagnosis and treatment of cancer, both in the with a mature ⬃87-kDa polypeptide (i.e., NIS) and a par-
thyroid and beyond. tially glycosylated (⬃56 kDa) polypeptide in a line of
highly functional thyroid rat cells (FRTL-5 cells). Immu-
noreactivity is also observed in X. laevis oocytes and COS
II. CHARACTERIZATION OF THE
cells expressing NIS, and it is competitively blocked by
SODIUM/IODIDE SYMPORTER PROTEIN
the presence of excess synthetic peptide. This anti-COOH-
terminal NIS Ab was the first available tool to experimen-
A. Generation of Anti-NIS Antibodies tally probe the NIS secondary structure model, and it was
used to confirm the model-predicted cytosolic-side loca-
A major development in the molecular characteriza- tion of the COOH terminus by indirect immunofluores-
tion of NIS has been the generation of anti-NIS antibodies cence experiments in permeabilized FRTL-5 cells (62).
(Ab). Levy et al. (62) generated a high-affinity [dissocia- Subsequently and independently, another site-directed Ab
tion constant (Kd) ⬃100 pM] site-directed polyclonal anti- against the same COOH-terminal segment of NIS was
NIS Ab against the last 16 amino acid residues of the generated by Paire et al. (81), which similarly immunore-
July 2000 MOLECULAR ANALYSIS OF THE SODIUM/IODIDE SYMPORTER 1087
acts with an ⬃80- to 90-kDa glycosylated NIS protein from loop containing N225 are predicted to be on the extracel-
FRTL-5 cells. lular side, and the COOH terminus facing the cytosol, as
confirmed previously (Fig. 2B).
prisingly found not to generate a current, strongly sug- Eskandari et al. (34).]
1090 DE LA VIEJA, DOHAN, LEVY, AND CARRASCO Volume 80
125) ostensibly showing that 36Cl-labeled perchlorate en- et al. (75) reported that the biosynthesis of thyroid hor-
ters the cell may have been misinterpreted. Because mones by sheep thyroid slices was inhibited by high doses
[36Cl]chlorate (ClO⫺3 ) is a
36
Cl-labeled by-product of the of I⫺. Wolff and Chaikoff reported in 1948 (128) that
reaction employed to chemically synthesize [36Cl]per- organic binding of iodide in the rat thyroid was blocked
chlorate (ClO⫺4 ) for these uptake studies, it seems likely when I⫺ thyroid levels reached a critical high threshold, a
that [36Cl]chlorate, rather than perchlorate, accounts for phenomenon known as the acute Wolff-Chaikoff effect.
the presence of label in the cytosol of thyrocytes, given These researchers observed further that ⬃2 days later, in
that chlorate is readily translocated via NIS into the cell the presence of continued high plasma I⫺ concentrations,
(34). Current electrophysiological data, in conclusion, an “escape” or adaptation from the acute effect is ob-
strongly indicate that perchlorate is not translocated via served so that the level of organification of I⫺ is restored
NIS into the cell. and normal hormone biosynthesis resumes (129).
Whereas the mechanism responsible for the acute Wolff-
Chaikoff effect has yet to be fully elucidated, it has been
III. REGULATION OF SODIUM/IODIDE proposed to be the result of organic iodocompounds act-
SYMPORTER PROTEIN EXPRESSION ing as mediators. The iodolipid ␣-iodohexadecanal has
been suggested to be one such mediator, on account of its
TSH is the primary hormonal regulator of thyroid ability to inhibit NADPH oxidase, TPO, and TSH-induced
roid, lung, forebrain, and pituitary and in the adult thyroid (95) have recently shown that both TTF-2 and HNF-3,
and lung; 2) TTF-2, a forkhead protein detected in devel- another forkhead transcription factor also present in the
oping thyroid and anterior pituitary and in the adult thy- thyroid, bind to the same site and thus participate in the
roid; and 3) Pax8, a nuclear protein member of the murine regulation of the TPO but not the Tg promoter. In addi-
family of paired-domain (PD) containing genes, present in tion, upstream enhancers have also been identified in
the developing thyroid, kidney, and midbrain boundary both promoters. Three TTF-1 binding sites for the bovine
and in the adult thyroid and kidney. Specific combinations Tg promoter are present 2 kb upstream. Two TTF-1 and
of these factors have been proposed to regulate transcrip- one Pax8 binding site, recently reported (35), are found in
tion of the thyroid-specific proteins Tg and TPO and also a 230-bp enhancer located 5.5 kb upstream of the TPO
of the TSHr. initiation codon (22).
The Tg and TPO minimal promoter structures are The TSHr promoter is very different from the Tg and
very similar to each other but different from the TSHr TPO promoters (Fig. 6). The TSHr minimal promoter
promoter (22) (Fig. 6). The Tg and TPO promoters from region is localized in the 5⬘-upstream region between
FIG. 6. Schematic representation of the structures of the Tg, TPO, TSH receptor (TSHr), and NIS promoters. Symbols
refer to binding sites mapped by DNase I footprinting. Numbers indicate the distances from the corresponding
transcriptional start sites. For specific details, see Reference 22.
1092 DE LA VIEJA, DOHAN, LEVY, AND CARRASCO Volume 80
neither TTF-2 nor Pax8 binding sites have been identified could result from the presence of an upstream tissue-
in the TSHr promoter. The TSHr transcript has also been specific enhancer or from the influence of chromatin
found in retro-orbital fibroblasts and in adipose tissue (32, structure and/or methylation, as there are numerous CpG
60). Rat adipose tissue expresses TSHr mRNA levels sim- dinucleotides.
ilar to those found in the thyroid (100). The promoter
regulation is different in adipocytes from thyroid cells,
but both display multiple start sites and important regu- E. Analysis of the Rat NIS Promoter
lation by CREB (99).
TPO and Tg expression are both upregulated by TSH Tong et al. (112) isolated a 16.4-kb fragment of
(41, 116) but by different regulatory mechanisms. TPO genomic rNIS DNA. They localized the start site at ⫺98 bp
and TSHr regulation takes place faster than Tg regulation. and one TATA box between ⫺124 and ⫺118 (AATAAAT),
Although Tg transcriptional activation kinetics depend on with a minimal promoter localized between ⫺199 and
the experimental model used, i.e., it is rapid (⬃1 h) in ⫺110 bp. Surprisingly, however, they concluded that 8 kb
thyroid slides and slow (⬃8 h) in primary cultures, TPO of the 5⬘-flanking region of the NIS gene is not sufficient to
induction is rapid in both experimental models (41). All confer thyroid-specific transcription. Neither TTF-2 nor
three genes are controlled in the thyroid at the transcrip- Pax8 putative binding sites were localized within 2 kb
tional level mainly by a cAMP-dependent mechanism, upstream of the initiation codon. However, the conclu-
-hydroxylase (111), prohormone convertase 1 (47), and ability to organify accumulated I⫺; therefore, they behave
proenkephalin genes (19). In NUE, both Pax8 and the like PTU-treated thyroid tissue; 2) TSH exerts no regula-
unidentified CRE-like binding factor act synergistically to tory influence on nonthyroid I⫺ accumulation; and 3) at
obtain full TSH/cAMP-dependent transcription. However, least salivary glands and gastric mucosa concentrate thio-
this enhancer is also able to mediate cAMP-dependent cyanate, unlike the thyroid where it is metabolized (oxi-
transcription by a novel PKA-independent mechanism dized). Despite these differences, several reports of pa-
(78). tients suffering the simultaneous genetic absence of I⫺
The present picture of the NIS promoter shows two transport in the thyroid, the salivary glands, and the gas-
important regions (Fig. 6): 1) a proximal promoter, still tric mucosa strongly hinted at a “genetic link” among
questionably thyroid specific, reported to be TSH/cAMP these I⫺ transport systems, suggesting that extrathyroidal
regulated by a TRE binding sequence, mediated by a new I⫺ transport is catalyzed by plasma membrane proteins
NTF-1 protein (76). This upregulation, however, is lower that are very similar, if not identical, to thyroid NIS (13,
than the TSH-induced mRNA expression reported by the 16, 42, 125). Moreover, thyroidal and extrathyroidal I⫺
same group (53). 2) The NIS promoter also has an up- accumulation generate I⫺ concentration gradients of sim-
stream enhancer that is thyroid specific, with Pax8 and ilar magnitude (⬃20- to 40-fold under steady-state condi-
CRE-like binding factors. It seems likely that both regions tions). Hence, the isolation and characterization of the
in the promoter act synergistically to achieve full high- NIS cDNA from rat thyroid (21) made it possible to ex-
yields a large number of false positives due to the high Ref. 62), a finding consistent with the identity among
sensitivity of the RT-PCR technique (38). Therefore, the human thyroid NIS, mg-NIS, and g-NIS predicted by the
detection of the NIS amplified product by RT-PCR in a cloning of human NIS cDNA from gastric mucosa and
given tissue cannot be regarded as sufficient evidence that mammary glands by Spitzweg et al. (108).
NIS is functionally expressed in that tissue. A thorough
characterization of NIS protein expression is necessary to
C. Regulation of Mammary Gland NIS
properly evaluate the significance or results obtained by
RT-PCR and even Northern analyses, and once NIS pro-
Subsequently, Tazebay et al. (111a) investigated
tein expression has been demonstrated, a correlation with
whether a correlation existed between I⫺ accumulation
Na⫹-dependent, perchlorate-sensitive, active I⫺ accumu-
activity and mg-NIS expression in the various physiolog-
lation in that tissue must be established. By these criteria,
ical stages of the mammary gland. They found that mg-
and with the consideration of the above results, NIS ap-
NIS was absent in nubile and pregnant mammary gland,
pears to be expressed and active in extrathyroidal tissues
but clearly present in lactating mammary gland. To ascer-
previously known to exhibit NIS activity, such as salivary
tain whether weaning had an effect on mg-NIS expres-
glands and gastric mucosa. The significance of the detec-
sion, mother rats were removed from their litters. Twenty-
tion of the NIS-amplified product by RT-PCR in other
four hours after weaning, mg-NIS expression was
human and rat tissues still awaits further characteriza-
significantly decreased, and after 48 h, it was barely de-
ture of AITD is the expression of HLA class II molecules, CHO cells stably expressing recombinant rat NIS. The
induced by the T cell secreted interferon (IFN)-␥, on the investigators observed also that some normal sera and
follicular epithelial cells, which are normally class II neg- patients’ sera that did not immunoreact on immunoblots
ative. Another characteristic feature of AITD is the pres- nevertheless caused ⬃90% inhibition of NIS activity. Be-
ence in patients’ sera of autoAb that are directed against cause this inhibitory effect was lost after dialyzing these
specific thyroid antigens (Tg, TPO, and TSHr) (121). Io- sera, the authors concluded that some sera contained a
dine is an important susceptibility and modulating factor dialyzable inhibitor, independent of the presence or ab-
in the development of thyroid autoimmunity. High levels sence of anti-NIS autoAb, and thus suggested that purified
of iodine can cause thyroid damage and thyroiditis, par- IgG be used to accurately evaluate the putative autoAb
ticularly against the background of a previously low in- activity. The investigators also concluded that the pres-
take of iodine. Significantly, the incidence of AITD is ence of anti-NIS autoAb may play a role in the pathogen-
higher in areas of iodine sufficiency than in areas of iodine esis of Hashimoto’s thyroiditis and thus may modulate
deficiency (37). thyroid function. All of these preliminary observations
Given that thyroid-associated proteins such as Tg, call for further study and confirmation.
TPO, and TSHr are known targets for autoAb in AITD, the Morris et al. (74) synthesized 21 peptides correspond-
molecular identification of NIS has made it possible for ing to putative extracellular segments of rat NIS, based on
the first time to experimentally assess whether NIS is also the first 12 transmembrane segment secondary structure
against it. It is important to continue the analysis of this noma, either follicular or papillary, is one of the most
topic utilizing experimental systems free of other thyroid curable cancers; the overall survival rate at 10 years for
autoantigens, and to investigate the presence of anti-NIS middle-aged adults with thyroid carcinoma is ⬃80 –90%
autoAb against both linear and conformational epitopes, (97). The reduced I⫺ accumulation detected in the major-
that are no longer present after SDS-PAGE electrophore- ity of thyroid cancers suggests that malignant transforma-
sis. Clearly, a wide range of experimental approaches will tion of these cells may have an effect on the NIS molecule.
be necessary to unequivocally determine the existence, Thus the continued characterization of NIS may shed light
real prevalence, functional effects, and pathological sig- on I⫺ transport changes observed in thyroid cancer.
nificance of autoAb against NIS. Furthermore, another The treatment of differentiated thyroid carcinoma is
interesting aspect to consider is the role of certain cyto- total or near-total thyroidectomy followed by 131I abla-
kines in NIS function, because recent reports indicate that tion. Radioiodide ablation destroys occult microscopic
tumor necrosis factor (TNF)-␣ and, to lesser extent, in- carcinomas and also any remaining normal thyroid tissue,
terleukin (IL)-1␣ inhibit both basal and TSH-induced NIS permitting postablative 131I total body scanning search for
expression (5). High concentrations of IFN-␥ also down- possible persistent carcinoma (119). The tools that allow
regulate TSH-stimulated NIS mRNA expression. Conse- the sensitive and specific detection and subsequent treat-
quently, IL-1␣, TNF-␣, and IFN-␥ all inhibit I⫺ uptake in ment of differentiated thyroid carcinoma are the uptake
FRTL-5 cells. Caturegli et al. (17) have recently generated (mediated by NIS) and organification (mediated by TPO)
Caillou et al. (14) conducted an immunohistochemi- have recently introduced rNIS into melanoma, ovarian,
cal analysis of NIS protein expression in thyroid patho- liver, and colon carcinoma cells. The resulting rNIS trans-
logical tissues, including six benign adenomas, five follic- duced tumor cells exhibited I⫺ uptake activity. In vitro
ular, and nine papillary carcinomas. No immunostaining experiments showed that these transduced cells could be
for hNIS was detected in any of the six benign adenomas destroyed by accumulation of 131I. If positive results were
that appeared to be hypofunctioning on thyroid scintigra- obtained in subsequent in vivo experiments, this gene
phy. hNIS expression was lower or absent in the papillary therapy approach would undoubtedly be one of the most
and follicular well-differentiated carcinomas, but more promising developments concerning the possible uses of
NIS was present in these than in the poorly differentiated the molecular characterization of NIS in the diagnosis and
follicular carcinomas. Consistent with these results, treatment of cancer.
Jhiang et al. (48) detected no hNIS by immunohistochem-
ical staining in any of the three papillary or the single
follicular carcinoma that they studied. In contrast, Saito et VIII. CONGENITAL IODIDE TRANSPORT
al. (92) found increased NIS expression in papillary car- DEFECT DUE TO SODIUM/IODIDE
cinomas. They investigated 31 papillary carcinomas by SYMPORTER MUTATIONS
either Northern blot or immunological analysis. Based on
their observations of higher NIS expression in ⬃50% of Congenital lack of I⫺ transport is a rather uncommon
in nucleotide 1060 in exon 9, resulting in a Pro instead of Subsequently, Levy et al. (65) studied the T354P mu-
Thr at position 354 (T354P) in NIS. The T354P mutation is tation by site-directed mutagenesis and transfection of
located in the putative transmembrane segment IX of the COS cells. They determined that the T354P NIS was not
NIS protein (Figs. 2B, 3, and 7). active, but they observed by immunofluorescence analy-
The patient in the report of Fujiwara et al. (39) was sis that it was properly targeted to the plasma membrane.
homozygous for the T354P mutation and had consanguin- Then various other amino acids substitutions (Ala, Pro,
eous parents; her two siblings were heterozygous for Cys, Tyr, Ser) at position 354 were explored to determine
T354P and were unaffected, suggesting the recessive na- the structural change in putative transmembrane segment
ture of the disease. Extremely high doses of potassium IX, leading to the conclusion that the hydroxyl group at
iodide treatment maintained this patient euthyroid. HEK- the -carbon of the amino acid residue at position 354 is
293 cells (human embryonic kidney cells) transfected essential for NIS function (see sect. II).
with the mutant T354P NIS cDNA did not exhibit any I⫺ Fujiwara et al. (40) have reported four new cases of
uptake activity. Based on these results, the authors pro- patients with the T354P mutation from two families (Ta-
posed that T354P probably disrupts (due to the presence ble 1, cases 2–5). Case 2 was diagnosed at age 5, treated
of proline), through a structural change, the putative with thyroid powder, and thus maintained euthyroid;
transmembrane segment IX of NIS, where the mutation is however, this patient’s thyroid gland enlarged gradually
located. and developed multinodular goiter at the age of 14. The
The second case (70) was diagnosed 23 years ago patient’s parents were heterozygous for the T354P muta-
with an I⫺ transport defect (44) (Table 1, case 6). The tion without thyroid disorders. The other three patients
patient was euthyroid apparently on account of a very reported by Fujiwara et al. (40) are siblings (Table 1, cases
high iodide dietary intake. However, diffuse goiter with 3–5). These patients and their mother were heterozygous
slightly elevated TSH was noted at 18 yr of age. The for T354P, but their father was negative for this mutation.
patient exhibited greatly increased NIS mRNA levels The patients presented a small goiter that developed late
(⬎100-fold) in his thyroid. Hence, these investigators pro- in life with only slight symptoms of hypothyroidism. Even
posed that the observed increase in NIS transcription may though the patients were heterozygous for the T354 mu-
reflect a compensating mechanism for low NIS activity, tation, they were previously diagnosed as having ITD but
possibly related to transcriptional regulation of NIS by I⫺ were not screened for other NIS mutations.
itself. This patient was born from a consanguineous mar- Kosugi et al. (55) have reported more cases of T354P
riage, and his daughter was heterozygous for the mutation mutations, including three new and four reevaluated
but had no abnormal phenotype, as would be expected in cases with known ITD from five families (Table 1, cases
a recessive condition. 6 –12). All these cases were homozygous for the T354P
July 2000
TABLE 1. Summary of clinical and laboratory parameters of ITD patients and correlation to the NIS molecular defect
1 1986 F Japan 3 mo 8 yr (FA) Normal ⫹ 39, 40 2 0.40 730 4.5 1.61 L-T4 9 A1060C Homozygous T354P TM IX
2 1958 M Japan 5 yr 13 yr (FA) Retarded ⫹ 40 25 1.0 0.94 Thyroid powder 9 A1060C Homozygous T354P TM IX
3 1968 M Japan 3 yr 8 yr Retarded ⫺ 40 27 0.41 71 1.2 1.1 Thyroid powder 9 A1060C/? Heterozygous T354P/? TM IX
4 1966 F Japan 10 yr 10 yr Retarded ⫺ 40 36 1.42 110 4.8 0.93 Thyroid powder 9 A1060C/? Heterozygous T354P/? TM IX
5 1963 F Japan 13 yr 13 yr Normal ⫺ 40 12 0.46 300 1.2 1.2 9 A1060C/? Heterozygous T354P/? TM IX
6 1937 M Japan Euthyroid 18 yr Normal ⫹ 44, 55, 70 69 1.06 3.3 2.5 1.5 L-T4 9 A1060C Homozygous T354P TM IX
7 1942 F Japan Euthyroid 20 yr Normal ⫹ 55 54 1.4 1.3 2.0 1.1 L-T4 9 A1060C Homozygous T354P TM IX
8 1975 F Japan 6 yr 6 yr Retarded ⫹ 46, 55 91 2.35 13.3 2.1 1.0 12.6 mg I/day 9 A1060C Homozygous T354P TM IX
9 1978 M Japan 1 yr None Retarded ⫹ 46, 55 ⬍10 0.61 148 1.3 1.0 12.6 mg I/day 9 A1060C Homozygous T354P TM IX
10 1971 M Japan 8 mo 21 yr Retarded ⫺ 55 11 1.06 37.5 3.0 1.5 1 mg I/day 9 A1060C Homozygous T354P TM IX
11 1989 M Japan 1.5 yr 1.5 yr Retarded ⫹ 55 1.2 1.25 ⬎100 1.4 2.6 L-T4 9 A1060C Homozygous T354P TM IX
12 1974 F Japan 4 mo None Retarded ⫺ 55, 71 5 0.46 430 2.4 1.8 L-T4 9 A1060C Homozygous T354P TM IX
13 1947 M Japan 22 yr 16 yr Normal ⫺ 54, 91, 126 10 0.39 217 0.9 0.7 189 mg I/day 1 G277C Heterozygous G93R TM III
9 A1060C Heterozygous T354P TM IX
14 1984 F Japan 3 yr 10 yr Retarded ⫺ 54 9 0.8 1,044 4.0 1.0 L-T4 13 G1628A Homozygous G543E TM XIII
15 1985 F Japan 2 yr 9 yr Retarded ⫺ 54 4 0.31 484 3.6 1.0 L-T4 13 G1628A Homozygous G543E TM XIII
16 1958 M Brazil 3 mo 3 mo Normal ⫺ 15, 85 37 104 1.0 3.0 12 mg I/day 6 C1163A Homozygous C272X Loop ÷ TM
VII–VIII
6 C1146G Heterozygous Q267E Loop ÷ TM
VII–VIII
17 1958 F USA Neonatal 12 yr (FA) Normal ⫺ 86 67 142 3.0 3 L-T4 13 C1940G Heterozygous Y531X TM XIII
and/or Alt. Spliced 515X Loop ÷ TM
XII–XIII
MOLECULAR ANALYSIS OF THE SODIUM/IODIDE SYMPORTER
S/P, salivary to plasma ratio of I; FA, follicular adenoma; empty box, test not done or unknown; Cons, parental consanguinity; TRIU, thyroid radioiodide uptake (%); T4, thyroxine; T3, triiodothyronine; TSH,
thyrotropin; TM, transmembrane segment. Patients 3–5 only have been screened for the T354P mutation.
1099
mutation. Whenever the patients’ parents were analyzed, segments VII and VIII (Fig. 6), and no I⫺ uptake activity
they were heterozygous for the T354P mutation and had was detected after transfecting mutant NIS cDNA into
no obvious thyroid disorders. The first case (case 6) was COS-7 cells. The patient was homozygous for the muta-
previously described by the same group (see above) (70). tion. His parents were not consanguineous; his mother,
Cases 6 and 7, siblings, were euthyroid due to an ex- son, and a paternal aunt were heterozygous for C272X,
tremely high I⫺ diet. They presented large diffuse goiters and his father was not investigated. All of the carriers
at ages 18 and 20, respectively. In cases 9 and 12, hypo- were euthyroid with normal or slightly enlarged thyroid
thyroidism was diagnosed early so that L-T4 treatment was glands. The patient’s goiter was noted at 3 mo of age and
started and goiter development was prevented. The rest treated with L-T4, but examination after several years
of the cases had different degrees of diffuse goiter and revealed low T4 and high TSH levels. A nodule developed
hypothyroidism. NIS protein determinations were carried in each thyroid lobe. Subsequent treatment with high I⫺
out in cases 6 and 10 and were found to be increased restored the patient to the euthyroid state.
⬃10-fold with respect to normal tissue. Immunohisto- Case 17 in Table 1 was reported by Pohlenz et al.
chemical staining of NIS in these two patients indicated (86). This was a patient of Hispanic ancestry who carried
that NIS was overexpressed and properly localized in the a compound heterozygous mutation. From the paternal
basolateral plasma membrane of the thyroid cells (55). allele she presented a substitution of cytosine for guani-
The same group has subsequently reported three pa- dine in nucleotide 1146 in exon 6 that resulted in a Gln for
nonspecific channels, as suggested by Wolff (126). More- cases (cases 6 and 7) remained euthyroid. Most probably,
over, Cl⫺ channels have been shown to be able to trans- this variability reflects the roles played by multiple factors
port other halides with different affinities, and in some in the onset and evolution of ITD due to NIS mutations,
cases, these channels display a considerable affinity for I⫺ including the type of mutation, time of detection of the
(26, 27, 52). Administration of high doses of potassium condition, time and type of administered therapy, and I⫺
iodide (KI) (1–200 mg/day) has been reported to maintain content in the diet. In any case, the considerable strides
these patients euthyroid, without apparent long-term side made in this area of research in recent years illustrate the
effects. powerful impact that detailed function/structure studies
The inheritance pattern of ITD is autosomal recessive of this key molecule can have on health and disease.
in all cases. In the investigated families, the heterozygous
carriers were always clinically unaffected, and only a few
of them presented slight goiter. Nevertheless, cases 3–5 in IX. CONCLUDING REMARKS
Table 1, reported by Fujiwara et al. (40), were heterozy-
gous for T354P. However, other mutations could have The landscape of NIS research has been dramatically
been present in these patients and missed in the study transformed in a very short time. Just six years ago, in
because only T354P was screened. 1993, the following was written in a review on “Iodide
Most of the patients suffering from this condition Transport in the Thyroid Gland” (16): “The chemical na-
man cells, which then exhibited active iodide transport 14. CAILLOU B, TROALEN F, BAUDIN E, TALBOT M, FILETTI S,
SCHLUMBERGER M, AND BIDART JM. Na⫹/I⫺ symporter distribu-
and became susceptible to destruction with radioiodide. tion in human thyroid tissues: an immunohistochemical study.
Hence, the impact of the continued molecular analysis of J Clin Endocrinol Metab 83: 4102– 4106, 1998.
NIS has already reached farther than predicted a few 15. CAMARGO RYA, GROSS JL, SILVEIRO SP, KNOBEL M, AND ME-
DEIROS-NETO G. Pathological findings in dyshormonogenetic goi-
years ago, given that it extends not only to structure/ ter with defective iodide transport. Endocr Pathol 9: 225–233, 1998.
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