0013-7227/05/$15.

00/0 Endocrinology 146(7):28612863

Printed in U.S.A. Copyright 2005 by The Endocrine Society
doi: 10.1210/en.2005-0394

Thyroid Hormones: Rapid Reply by Surface

Delivery Only
T4 is usually thought to function as a prohormone, yielding call these actions rapid in the experimental paradigms we
T3 by deiodination at target tissues. T3, in turn, gains access design, the ambient levels of thyroid hormone in intact or-
to the cell interior and the cell nucleus, binding to a nuclear ganisms are constant, and thus we assume that these non-
receptor protein that acts as heterodimeric or homodimeric genomic actions contribute to basal levels of activities of ion
transactivator for thyroid hormone-regulated genes. This pumps and channels or to rates of transcription.
takes time, which is quite all right for a hormone traditionally Bergh et al. (1) demonstrates that isolated V3 integrin
considered to be a regulator of protein synthesis, cell me- binds T4 with very high affinity. The bound ligand can be
tabolism, and the slow business of differentiation and de- displaced by the thyroid hormone metabolite tetraiodothy-
velopment. Actually, it is also true that T3 and T4 sometimes, roacetic acid, which previously has been shown to be able to
when the experimental design permits, induce very fast re- inhibit nongenomic effects of T4, and binding is also inhibited
sponses in cells, called nongenomic or extranuclear effects by specific anti-V3 antibodies and a peptide ligand that
because they appear within minutes or even seconds; al- binds to the integrin Arg-Gly-Asp (RGD) recognition site (1).
though such effects have been known for many years, it is not Using a fibroblast cell line expressing plasma membrane
clear how they are brought about or what their physiological V3 but lacking the nuclear thyroid hormone receptor, they
roles are. As so often happens when how and why of actions show that exposure to T4 leads to rapid activation of MAPK
are left unanswered, these nongenomic aspects of thyroid and induction of angiogenesis; these effects were prevented
hormone action have been tacitly ignored. by tetraiodothyroacetic acid, the anti-V3 antibodies, and
That may be about to change now. In this issue of Endo- the inhibitory peptide. In addition, cells devoid of the V- or
crinology, Bergh et al. (1) identified the cell surface T4 receptor, the 3-protein did not respond to the hormone treatment,
and surprisingly it turns out that the long-searched-for leaving little doubt that the integrin is the receptor.
plasma membrane receptor is a well-known membrane pro- Integrins are a family of transmembrane glycoproteins
tein, V3 integrin. It is likely that this protein, or similar that form noncovalent heterodimers. The extracellular do-
membrane receptors, will turn out to be the trigger for most mains of the integrins interact with a variety of ligands in-
of the nongenomic actions of thyroid hormone reported in cluding matrix glycoproteins (14), whereas the intracellular
the literature, which include alterations in solute transport domains are linked to the cytoskeleton (15). Integrin V3
(Ca2, Na, H, glucose), changes in activity of signal trans- has a large number of extracellular protein ligands, including
ducing kinases like protein kinase A, protein kinase C, pyru- growth factors, and in most cases ligand binding can activate
vate kinase M2, phosphatidylinositol 3-kinase and the the MAPK pathway (16, 17). Many integrins contain the RGD
MAPK pathway (2 4). We can see these actions to be rapid recognition site that is important for the interaction with
onset when we make cells thyroprival and reexpose them to peptide ligands containing an Arg-Gly-Asp sequence (14).
thyroid hormone. Some of these effects rapidly lead to post- Recently Hoffman et al. (18) have shown that blocking the
translational modifications of nucleoproteins, e.g. serine V3 integrin RGD site prevented T4-induced bone resorp-
phosphorylation of p53, the estrogen receptor, and even the tion. Another novelty of the paper by Bergh et al. (1) is that
nuclear thyroid hormone receptor TR1 (57). The effects on a small molecule like T4 can bind to an integrin; usually
nucleoproteins are mediated by MAPK (ERK1/2), and pro- integrins bind polypeptides. The location of the ligand bind-
tein kinase C and the phosphatidylinositol pathway are ing site for T4 is not known, but the results obtained with
probably activated upstream of MAPK (8). Recently an as- inhibitory peptides show it must be located at or near the
sociation between thyroid hormone receptor-1 and phos- RGD binding groove (19, 20). It is curious that the crystal
phatidylinositol 3-kinase-Akt/protein kinase B cascade has structure of the membrane T4 receptor actually was known
also been reported (9). In any case, the mechanisms by which before the protein was identified as a receptor.
thyroid hormones nongenomically affect the activity of The finding that thyroid hormone uses an integrin as a
plasma membrane ion channels and ion pumps are not well surface receptor may explain several of the nongenomic and
understood (3, 10). Membrane binding sites for thyroid hor- genomic effects reported in the literature. Angiogenesis, the
mones were identified many years ago in cell membranes growth of new blood vessels, plays a key role in develop-
from erythrocytes and hepatocytes (11, 12; for a review, see ment, wound repair, inflammation, and tumor growth and
Ref. 13), but the linkage between binding sites and hormone is a process known to be mediated by thyroid hormones in
actions has not been established until now. Although we can vivo (21, 22). Recently the role of nongenomic effects of T4 in
angiogenesis has also been shown in vitro by the group of
Abbreviations: IFN, Interferon; RGD, Arg-Gly-Asp. Davis (23) with the classical chick chorioallantoic membrane
Endocrinology is published monthly by The Endocrine Society (http://
assay, a system that has a high level of expression of the
www.endo-society.org), the foremost professional society serving the V3-integrin (24). The mechanism by which thyroid hor-
endocrine community. mone causes angiogenesis in the chorioallantoic membrane

2861

The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 05 July 2016. at 23:00 For personal use only. No other uses without permission. . All rights reserved.
2862 Endocrinology, July 2005, 146(7):28612863 Incerpi News & Views

model is complex but is clearly initiated at the plasma mem- another integrin receptor in the plasma membrane? The as-
brane by the hormone and involves transduction of the hor- sociation of integrins with thyroid hormones appears even
mone signal via the MAPK pathway into a fibroblast growth more appealing in the light of recent findings concerning the
factor-2-dependent angiogenic response (23). Thyroid hor- structure-activity relationship of these proteins: through con-
mones have been shown to increase intracellular pH through formation changes they can transmit both outside-in and
nongenomic activation of the Na/H exchanger, a plasma inside-out signaling (34). Either intracellular (cytoplasmic/
membrane protein that exchanges extracellular Na with nuclear) or extracellular interaction can give rise to active or
internal protons, and this is also a proliferative and proan- inactive states of the integrin, i.e. the integrin itself or the
giogenic factor (2527). In fact, the higher intracellular pH is integrin-ligand complex can shuttle between different con-
an important requisite for cell spreading and appears to be formations. This could enable cells to expose either high- or
regulated by several integrins (28, 29). It is known that both low-affinity binding sites on the surface and perhaps provide
human and chick blood vessels involved in angiogenesis an explanation for the interaction between nongenomic and
have enhanced expression of V3, and consistently the genomic effects of thyroid hormones. In fact, thyroid hor-
expression of V3 in cultured endothelial cells can be in- mones display such a wide variety of effects that the integrin
duced by various cytokines in vitro (24). Thyroid hormones mechanism appears to be a sort of deus ex machina able to
enhance the actions of several cytokines and growth factors, account for all the effects and mechanisms that cannot be
such as interferon (IFN)- and epidermal growth factor. explained at present. The delivery is going to take some time,
Davis and colleagues have shown that there are two mech- though.
anisms by which thyroid hormone can potentiate the IFN
Sandra Incerpi
effect (30): the first is a protein synthesis-dependent mech- Department of Biology
anism evidenced by enhancement of IFN antiviral action University Roma Tre
upon incubation with T3, T4, or the analog rT3 and inhibition 00146 Rome, Italy
of this enhancement by tetraiodothyroacetic acid or cyclo-
heximide; the second is a protein synthesis-independent Acknowledgments
(posttranslational) mechanism induced by incubation of T4
or T3, but not reverse T3, with IFN and is not inhibited by Received April 4, 2005. Accepted April 4, 2005.
tetraiodothyroacetic acid or cycloheximide (30). Address all correspondence and requests for reprints to: Dr. Sandra
Incerpi, Department of Biology, University of Rome, Roma Tre, Viale
Thyroid hormones are required for the normal develop- Marconi, 446, 00146 Rome, Italy. E-mail: incerpi@uniroma3.it.
ment and differentiation of the cells of the central nervous
system, in particular the oligodendrocyte precursor cells, the References
most important source of axons remyelination in the adult.
1. Bergh JJ, Lin H-Y, Lansing L, Mohamed SN, Mousa S, Davis FB, Mousa S,
Furthermore the cell-cycle blocking mechanism, terminal dif- Davis PJ 2005 Integrin V3 contains a cell surface receptor site for thyroid
ferentiation, and myelin production all depend on thyroid hormone that is linked to activation of mitogen-activated protein kinase and
hormones (31). Deficiency of thyroid hormones during the induction of angiogenesis. Endocrinology 146:2864 2871
2. Davis PJ, Davis FB 1996 Nongenomic actions of thyroid hormones. Thyroid
perinatal period results in severe mental and physical retar- 6:497504
dation, known in humans as cretinism. Beside the known 3. Incerpi S 2002 Actions of thyroid hormone on ion transport. Curr Opin En-
genomic action of thyroid hormones mediated by classical docrinol Diabetes 9:381386
4. Davis PJ, Davis FB 2003 Nongenomic actions of thyroid hormone. In: Braver-
nuclear receptors, also nongenomic actions of thyroid hor- man LE, ed. Diseases of the thyroid. 2nd ed. Totowa, NJ: Humana Press; 19 37
mone, such as actin polymerization and extracellular orga- 5. Davis PJ, Shih A, Lin H-Y, Martino LJ, Davis FB 2000 Thyroxine promotes
association of mitogen-activated protein kinase and nuclear thyroid hormone
nization of laminin, play a role in brain development and in receptor (TR) and causes serine phosphorylation of TR. J Biol Chem 275:38032
particular in neuronal migration (31). T4 is converted to T3 by 38039
a 5-deiodinase in astrocytes, and T3 is then transferred to 6. Tang HY, Lin H-Y, Zhang S, Davis FB, Davis PJ 2004 Thyroid hormone causes
mitogen-activated protein kinase-dependent phosphorylation of the nuclear
neurons where it binds to nuclear receptors to regulate gene estrogen receptor. Endocrinology 145:32653272
expression. Probably T3 in these cells is directly regulating 7. Shih A, Lin H-Y, Davis FB, Davis PJ 2001 Thyroid hormone promotes serine
gene expression of proteins such as myelin basic protein, phosphorylation of p53 by mitogen-activated protein kinase. Biochemistry
40:2870 2878
neurotropins, and reelin (32). Interestingly, Farwell and 8. Lin H-Y, Davis FB, Gordinier JK, Martino LJ, Davis PJ 1999 Thyroid hormone
Dubord-Tomasetti (33) have shown that thyroid hormones induces activation of mitogen-activated protein kinase in cultured cells. Am J
Physiol 276:C1014 C1024
can modulate the arrangement of laminin, an extracellular 9. Cao X, Kambe F, Moeller LC, Refetoff S, Seo H 2005 Thyroid hormone
matrix protein that plays a pivotal role in the nerve cell induces rapid activation of Akt/protein kinase B-mammalian target of rapa-
migration during central nervous system morphogenesis mycin-p70S6K cascade through phosphatidylinositol 3-kinase in human fi-
broblasts. Mol Endocrinol 19:102112
through the interaction between integrins and components 10. Lei J, Mariash CN, Ingbar DH 2004 3,3,5-Triiodo-l-thyronine up-regulation
of the cytoskeleton. of Na,K-ATPase activity and cell surface expression in alveolar epithelial cells
One particular property of the V3 integrin is that T4 in Src kinase- and phosphoinositide 3-kinase-dependent. J Biol Chem 279:
47589 47600
binds with a very high affinity, whereas T3 is a weak ligand, 11. Pliam MB, Goldfine ID 1977 High affinity thyroid hormone binding sites on
whereas exactly the opposite is true for the nuclear receptors. purified rat liver plasma membranes. Biochem Biophys Res Commun 79:166
172
This demonstrates that T4 is not just a precursor of T3 but has 12. Botta JA, de Mendoza D, Morero RD, Farias RN 1983 High affinity l-tri-
a life of its own. However, also T3 gives rise to nongenomic iodothyronine binding sites on washed rat erythrocyte membranes. J Biol
effects after binding to a receptor on the cell surface, as shown Chem 258:6690 6692
13. Hennemann G, Docter RE, Friesema CH, de Jong M, Krenning EP, Visser TJ
by the use of membrane-impermeant T3 derivatives (S. In- 2001 Plasma membrane transport of thyroid hormones and its role in thyroid
cerpi, unpublished results). Does this mean that there is hormone metabolism and bioavailability. Endocr Rev 22:451 476

The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 05 July 2016. at 23:00 For personal use only. No other uses without permission. . All rights reserved.
Incerpi News & Views
14. Calderwood DA, Shattil SJ, Ginsberg MH 2000 Integrins and actin filaments:
reciprocal regulation of cell adhesion and signaling. J Biol Chem 275:22607
22610
15. Plow EF, Haas TA, Zhang L, Loftus J, Smith JW 2000 Ligand binding to
integrins. J Biol Chem 275:2178521788
16. Hood JD, Frausto R, Kiosses WB, Schwartz MA, Cheresh DA 2003 Differ-
ential V integrin-mediated Ras-ERK signaling during two pathways of an-
giogenesis. J Cell Biol 162:933943
17. Pereira JJ, Meyer T, Docherty SE, Reid HH, Marshall J, Thompson EW,
Rossjohn J, Price JT 2004 Bimolecular interaction of insulin-like growth factor
(IGF) binding protein-2 with V3 negatively modulates IGF-1-mediated mi-
gration of tumor growth. Cancer Res 64:977984
18. Hoffman SJ, Vasko-Moser J, Miller WH, Lark MW, Gowen M, Stroup G 2002
Rapid inhibition of thyroxine-induced bone resorption in the rat by an orally
active vitronectin receptor antagonist. J Pharmacol Exp Ther 302:205211
19. Xiong JP, Stehle T, Goodman SL, Arnaout MA 2003 New insights into the
structural basis of integrin activation. Blood 102:11551159
20. Xiong JP, Stehle T, Zhang R, Joachimiak A, Frech M, Goodman SL, Arnaout
MA 2002 Crystal structure of the extracellular segment of integrin V3 in
complex with an Arg-Gly-Asp ligand. Science 296:151155
21. Chilian WM, Wangler RD, Peters KG, Tomanek RJ, Marcus ML 1985 Thy-
roxine-induced left ventricular hypertrophy in the rat. Anatomical and phys-
iological evidence for angiogenesis. Circ Res 57:591598
22. Tomanek RJ, Busch TL 1998 Coordinated capillary and myocardial growth in
response to thyroxine treatment. Anat Rec 251:44 49
23. Davis FB, Mousa SA, OConnor L, Mohamed S, Lin H-Y, Cao HJ, Davis PJ
2004 Proangiogenic action of thyroid hormone is fibroblast growth factor-
dependent and is initiated at the cell surface. Circ Res 94:1500 1506
24. Brooks PC, Clark RA, Cheresh DA 1994 Requirement of vascular integrin
V3 for angiogenesis. Science 264:569 571
Endocrinology is published monthly by The Endocrine Society (http://www.endo-society.org), the foremost professional society serving the
endocrine community.
The Endocrine Society. Downloaded from press.endocrine.org by [${individualUser.displayName}] on 05 July 2016. at 23:00 For personal use only. No other uses without permission. . All rights reserved.
Endocrinology, July 2005, 146(7):28612863 2863
25. Incerpi S, Luly P, De Vito P, Farias RN 1999 Short-term effects of thyroid
hormones on the Na/H antiport in L-6 myoblasts: high molecular specificity
for the 3,3,5-triiodo-l-thyronine. Endocrinology 140:683 689
26. Incerpi S, De Vito P, Luly P, Spagnuolo S, Leoni S 2002 Short-term effects
of thyroid hormones and 3,5-diiodothyronine on membrane transport systems
in chick embryo hepatocytes. Endocrinology 143:1660 1668
27. DArezzo S, Incerpi S, Davis FB, Acconcia F, Marino M, Farias RN, Davis PJ
2004 Rapid nongenomic effects of 3,5,3-triiodo-l-thyronine on the intracel-
lular pH of L-6 myoblasts are mediated by intracellular calcium mobilization
and kinase pathways. Endocrinology 145:5694 5703
28. Schwartz MA, Ingber DE 1994 Integrating with integrins. Mol Biol Cell 5:389
393
29. Barillari G, Albonici L, Incerpi S, Bogetto L, Pistritto G, Volpi A, Ensoli B,
Manzari V 2001 Inflammatory cytokines stimulate vascular smooth muscle
cells locomotion and growth by enhancing 51 integrin expression and func-
tion. Atherosclerosis 154:377385
30. Lin H-Y, Thacore HR, Davis FB, Davis PJ 1996 Thyroid hormone analogues
potentiate the antiviral action of interferon- by two mechanisms. J Cell
Physiol 167:269 276
31. Koibuchi N, Chin WW 2000 Thyroid hormone action and brain development.
Trends Endocrinol Metab 11:123128
32. Siegrist-Kaiser CA, Juge-Aubry C, Tranter Mp, Ekenbarger DM, Leonard JL
1990 Thyroxine-dependent modulation of actin polimerization in cultured
astrocytes. A novel, extranuclear action of thyroid hormone. J Biol Chem
265:5296 5302
33. Farwell AP, Dubord-Tomasetti SA 1999 Thyroid hormone regulates the ex-
tracellular organization of laminin on astrocytes. Endocrinology 140:5014
5021
34. Hynes RO 2002 Integrins: bidirectional, allosteric signaling machines. Cell
110:673 687