Use of ligands for treatment of diseases responsive to...

Multicellular living organisms and unmodified parts thereof and – Method of using a transgenic nonhuman animal in an in vivo...

Reexamination Certificate

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C800S008000, C800S018000, C435S006120, C435S007100, C435S007200, C435S325000

Reexamination Certificate

active

06521814

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
The invention relates to methods for treatment of neurological disease by administering an agent which interacts with a retinoid receptor associated with the neurological disease. The invention also relates to a method of modulating dopamine receptor synthesis by introducing an agent that interacts with a retinoid receptor associated with the dopamine receptor synthesis. The invention further relates to a transgenic mouse which is deficient in the normal synthesis of one or more receptors of RAR&agr;, &bgr;, &ggr; and RXR, and cell line thereof.
2. Related Art
Retinoids
A number of studies have demonstrated that retinoids (vitamin A derivatives) are essential for normal growth, vision, tissue homeostasis, reproduction and overall survival (for reviews and references, see Sporn et al.,
The Retinoids,
Vols. 1 and 2, Spom et al., eds., Academic Press, Orlando, Fla. (1984)). Retinoids are also apparently crucial during embryogenesis, since offspring of dams with vitamin A deficiency (VAD) exhibit a number of developmental defects (Wilson, J. G., et al.,
Am. J. Anat.
92:189-217 (1953)). With the exceptions of vision (Wald, G., et al.,
Science
162:230-239 (1968)) and spermatogenesis in mammals (van Pelt, H. M. M., and De Rooij, D. G.,
Endocrinology
128:697-704 (1991)), most of the effects generated by VAD in animals and their fetuses can be prevented and/or reversed by retinoic acid (RA) administration (Wilson, J. G., et al.,
Am. J. Anat.
92:189-217 (1953); Thompson et al.,
Proc. Royal Soc.
159:510-535 (1964)). The dramatic teratogenic effects of maternal RA administration on mammalian embryos (Shenefelt, R. E.,
Teratology
5, 103-108 (1972); Kessel, M.,
Development
115:487-501 (1992); Creech Kraft, J., In
Retinoids in Normal Development and Teratogenesis,
G. M. Morriss-Kay, ed., Oxford University Press, Oxford, UK, pp. 267-280 (1992)), and the marked effects of topical administration of retinoids on embryonic development of vertebrates and limb regeneration in amphibians (Mohanty-Hejmadi et al.,
Nature
355:352-353 (1992); Tabin, C. J.,
Cell
66:199-217 (1991)), have contributed to the notion that RA may have critical roles in morphogenesis and organogenesis.
Retinoid Receptors
Except for those involved in visual perception (Wald, G. et al.,
Science
162:230-239 (1968)), the molecular mechanisms underlying the highly diverse effects of retinoids have until recently remained obscure. The discovery of nuclear receptors for RA (Petkovich et al.,
Nature
330:444-450 (1987); Giguère et al.,
Nature
330:624-629 (1987)) has greatly advanced the understanding of how the retinoids may exert their pleiotropic effects (Leid, M., et al.,
TIBS
17:427-433 (1992); Linney, E.,
Current Topics in Dev. Biol.
27:309-350 (1992)). It is thought that the effects of the RA signal are mediated through two families of receptors—the RAR family and the RXR family—which belong to the superfamily of ligand-inducible transcriptional regulatory factors that include steroid/thyroid hormone and vitamin D3 receptors (for reviews, see Leid, M., et al.,
TIBS
17:427-433 (1992); Chambon, P., Semin.
Cell Biol.
5:115-125 (1994); Chambon, P.,
FASEB J.
10:940-954 (1996); Giguere, V.,
Endocrinol. Rev.
15:61-79 (1994); Mangelsdorf, D. J., and Evans, R. M.,
Cell
83:841-850 (1995); Gronemeyer, H., and Laudet, V.,
Protein Profile
2:1173-1236 (1995)).
RAR Receptors
Receptors belonging to the RAR family (RAR&agr;, &bgr; and &ggr; and their isoforms) are activated by both all-trans- and 9-cis-RA (Leid, M., et al.,
TIBS
17:427-433 (1992); Chambon, P., Semin.
Cell Biol.
5:115-125 (1994); Dollé, P., et al.,
Mech. Dev.
45:91-104 (1994)). Within a given species, the DNA binding (C) and the ligand binding (E) domains of the three RAR subtypes are highly similar, whereas the C-terminal domain F and the middle domain D exhibit no or little similarity. The amino acid sequences of the three RAR subtypes are also notably different in their B regions, and their main isoforms (&agr;1 and &agr;2, &bgr;1 to &bgr;4, and &ggr;1 and &ggr;2) further differ in their N-terminal A regions (Leid, M., et al,
TIBS
17:427-433 (1992)). Amino acid sequence comparisons have revealed that the interspecies conservation of a given RAR subtype is greater than the similarity found between the three RAR subtypes within a given species (Leid, M., et al.,
TIBS
17:427-433 (1992)). This interspecies conservation is particularly striking in the N-terminal A regions of the various RAR&agr;, &bgr; and &ggr; isoforms, whose A region amino acid sequences are quite divergent. Taken together with the distinct spatio-temporal expression patterns observed for the transcripts of each RAR and RXR subtype in the developing embryo and in various adult mouse tissues (Zelent, A., et al.,
Nature
339:714-717 (1989); Dollé, P., et al., Nature 342:702-705 (1989); Dollé et al.,
Development
110:1133-1151 (1990); Ruberte et al.,
Development
108:213-222 (1990); Ruberte et al.,
Development
111:45-60 (1991); Mangelsdorf et al.,
Genes
&
Dev.
6:329-344 (1992)), this interspecies conservation has suggested that each RAR subtype (and isoform) may perform unique functions. This hypothesis is further supported by the finding that the various RAR isoforms contain two transcriptional activation functions (AFs) located in the N-terminal A/B region (AF-1) and in the C-terminal E region (AF-2), which can synergistically, and to some extent differentially, activate various RA-responsive promoters (Leid, M., et al.,
TIBS
1 7:427-433 (1992); Nagpal, S., et al.,
Cell
70:1007-1019 (1992); Nagpal, S., et al.,
EMBO J.
12:2349-2360 (1993)). Knock-outs of RAR&agr;, &bgr; and &ggr; have also provided some insight into the physiological functions of these receptors (see, WO 94/26100; Ghyselinck et al.,
Intl. J. Dev. Biol.
41:425-447 (1997)).
RXR Receptors
Unlike the RARs, members of the retinoid X receptor family (RXR&agr;, &bgr; and &ggr;) are activated exclusively by 9-cis-RA (Chambon, P.,
Semin. Cell Biol.
5:115-125 (1994); Dollé, P., et al.,
Mech. Dev.
45:91-104 (1994); Linney, E.,
Current Topics in Dev. Biol.
27:309-350 (1992); Leid, M., et al.,
TIBS
1 7:427-433 (1992); Kastner et al., in
Vitamin A in Health and Disease,
R. Blomhoff, ed., Marcel Dekker, New York (1993)). However, the RXRs characterized to date are similar to the RARs in that the different RXR subtypes also differ markedly in their N-terminal A/B regions (Leid, M., et al.,
TIBS
17:427-433 (1992); Leid, M., et al.,
Cell
68:377-395 (1992); Mangelsdorf et al.,
Genes
&
Dev.
6:329-344 (1992)), and contain the same transcriptional activation functions in their N-terminal A/B region and C-terminal E region (Leid, M., et al.,
TIBS
1 7:427-433 (1992); Nagpal, S., et al.,
Cell
70:1007-1019 (1992); Nagpal, S., et al.,
EMBO J.
12:2349-2360 (1993)).
It is currently unclear whether all the molecular properties of RXRs characterized in vitro are relevant for their physiological functions in vivo. In particular, it is unknown under what conditions these receptors act as 9-cis-RA-dependent transcriptional regulators (Chambon, P.,
Semin. Cell Biol.
5:115-125 (1994)). The knock-outs of RXR&agr; and RXR&bgr; in the mouse have provided some insight into the physiological functions of these receptors. For example, the ocular and cardiac malformations observed in RXR&agr;
−/−
fetuses (Kastner, P., et al.,
Cell
78:987-1003 (1994); Sucov, H. M., et al.,
Genes
&
Dev.
8:1007-1018 (1994)) are similar to those found in the fetal VAD syndrome, thus suggesting an important function of RXR&agr; in the transduction of a retinoid signal during development. The involvement of RXRs in retinoid signaling is further supported by studies of compound RXR&agr;/RAR mutants, which reveal defects that are either absent or less severe in the single mutants (Kastner, P., et al.,
Cell
78:987-1003 (1994); Kastner, P., et al.,
Cell
83:859-869 (1995); Chiba, H., et al.,
J. Cell Biol.
139:735-747 (1997)). Moreover, it has be

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