Pharmacological chaperone action on G-protein-coupled receptors

doi:10.1016/j.coph.2004.08.001

Current Opinion in Pharmacology

Volume 4, Issue 5, October 2004, Pages 528-533

https://doi.org/10.1016/j.coph.2004.08.001 Get rights and content

An increasing number of genetic diseases are found to result from mutations that lead to retention of the affected proteins in the endoplasmic reticulum, where they are recognized as misfolded by the quality control system. Several of these conformational diseases involve mutations in G-protein-coupled receptors. Recent studies demonstrated that pharmacologically selective compounds, termed pharmacological chaperones, can stabilize the misfolded receptors, facilitating their export from the endoplasmic reticulum to the plasma membrane, where they can be active. Such functional rescue suggests that pharmacological chaperones could represent novel therapeutic agents for the treatment of conformational diseases. Although only a few examples are currently available, the observation that pharmacological chaperones can also favour the folding of wild-type G-protein-coupled receptors indicates that these compounds could have wide applications.

Introduction

Conformational diseases often result from mutations in proteins that are recognized as misfolded by quality control systems 1., 2.. Such recognition can lead to two different phenotypes: some misfolded proteins can be efficiently ubiquitinated and degraded by the proteasome, leading to a loss of function [3], whereas others accumulate in cells, forming aggregates that can have toxic consequences and are often referred to as gain of functions [4]. Studies carried out in the past decade have linked these two types of quality control outcomes to the aetiology of a growing list of congenital and acquired conformational diseases. In parallel, efforts to overcome these defects have led to the development of various interventions that successfully rescue proteins from both aggregation and degradation pathways. In particular, treatments with chemical compounds known as either chemical or pharmacological chaperones have been found to stabilize some conformational mutants, promoting their proper transport to their site of action where, in many cases, they can be functional 5., 6., 7.. Identifying compounds that can bind to the mutant proteins has been easier for proteins such as channels and receptors for which selective ligands have already been characterized. Because of their involvement in many pathophysiological conditions and the rich pharmacological diversity generated through various drug screening campaigns, G-protein-coupled receptors (GPCRs) have attracted considerable attention for the identification of pharmacological chaperones. At least ten congenital diseases have been linked to mutations in GPCRs that lead to their retention in the endoplasmic reticulum (ER) (Table 1), and pharmacological chaperones have been identified for three of these (Table 2). Here, we review the studies that led to the discovery of these potential therapeutic agents, with a special emphasis on their proposed mechanisms of action.

Section snippets

The discovery of pharmacological chaperones acting on GPCRs

The first demonstration that pharmacologically selective agents could rescue cell surface expression and function of GPCR mutants, which were otherwise retained in the ER, came from work carried out on V₂ vasopressin receptor (V₂R) mutants responsible for nephrogenic diabetes insipidus (NDI). NDI is a rare X-linked disease characterized by a loss of antidiuretic response to the hormone arginine-vasopressin (AVP) that results in the inability of the affected patients to concentrate their urine,

Generalization of the pharmacological chaperone action on GPCRs

Pharmacological chaperones were soon identified for another GPCR, the gonadotropin-releasing hormone receptor (GnRHR). Indeed, ER-retained mutant forms of GnRHR, which are responsible for hypogonadotropic hypogonadism, were rescued by treatment with selective non-peptidic GnRHR antagonists [20]. As with the V₂R, treatment with an antagonist (an indole known as IN3) was first shown to restore both cell surface expression and signaling activity of 11 of the 13 disease-linked mutant GnRHRs tested

Pharmacological chaperones can act on wild-type receptors

Although pharmacological chaperones have been described mainly in the context of mutant proteins, their action has also been reported for a few wild-type GPCRs. For example, the wild-type δ-opioid receptor is inefficiently processed, with less than 40% of the synthesized receptors reaching the mature form and being targeted to the plasma membrane under basal conditions [31]. Treatment with non-peptidic selective opioid ligands (Table 2) significantly increased the maturation efficacy, leading

Mechanistic features of pharmacological chaperones

The emerging hypothesis for the action of pharmacological chaperones suggests that selective lipophilic ligands can penetrate the plasma and ER membranes to bind to the partially folded receptor early during biosynthesis. In this context, ligand binding might alter the thermodynamic equilibrium in favour of the correctly folded protein, increasing the likelihood of the protein escaping the stringent ER quality control, and ultimately leading to an increase in the steady-state level of

Conclusions

Pharmacological chaperones represent a promising avenue for the treatment of conformational diseases, such as familial hypocalciuric hypercalcemia, hirschsprung disease, hypogonadotropic hypogonadism, hypothyroidism, neonatal hyperparathyroidism, NDI, obesity and retinitis pigmentosa (see Table 1), that result from GPCR misfolding. Although proof-of-principle for their action has been obtained for only three of these GPCR-related diseases, the apparent generality of the concept, even for

References and recommended reading

Papers of particular interest, published within the annual period of review, have been highlighted as:

• of special interest
•• of outstanding interest

Acknowledgements

We are grateful to Dr Monique Lagacé for critical reading of the manuscript.

References (31)

R.W. Carrell et al.
Conformational disease
Lancet
(1997)
V. Bernier et al.
Pharmacological chaperones: potential treatment for conformational diseases
Trends Endocrinol Metab
(2004)
S. Sato et al.
Glycerol reverses the misfolding phenotype of the most common cystic fibrosis mutation
J Biol Chem
(1996)
S.M. Noorwez et al.
Pharmacological chaperone-mediated in vivo folding and stabilization of the P23H-opsin mutant associated with autosomal dominant retinitis pigmentosa
J Biol Chem
(2003)
R.R. Kopito et al.
Conformational disease
Nat Cell Biol
(2000)
R. Sitia et al.
Quality control in the endoplasmic reticulum protein factory
Nature
(2003)
D.J. Selkoe
Folding proteins in fatal ways
Nature
(2003)
P.M. Conn et al.
Protein origami: therapeutics rescue of misfolded gene products
Molecular Interventions
(2002)
D.H. Perlmutter
Chemical chaperones: a pharmacological strategy for disorders of protein folding and trafficking
Pediatr Res
(2002)
J.P. Morello et al.
Nephrogenic diabetes insipidus
Annu Rev Physiol
(2001)

Y. Ala et al.

Functional studies of twelve mutant V2 vasopressin receptors related to nephrogenic diabetes insipidus: molecular basis of a mild clinical phenotype

J Am Soc Nephrol

(1998)

H.M. Sadeghi et al.

An X-linked NDI mutation reveals a requirement for cell surface V2R expression

Mol Endocrinol

(1997)

H. Tsukaguchi et al.

Binding-, intracellular transport-, and biosynthesis-defective mutants of vasopressin type 2 receptor in patients with X-linked nephrogenic diabetes insipidus

J Clin Invest

(1995)

T. Schoneberg et al.

Functional rescue of mutant V2 vasopressin receptors causing nephrogenic diabetes insipidus by a co-expressed receptor polypeptide

EMBO J

(1996)

A. Oksche et al.

Vasopressin V2 receptor mutants that cause X-linked nephrogenic diabetes insipidus: analysis of expression, processing, and function

Mol Pharmacol

(1996)

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Pharmacological chaperone action on G-protein-coupled receptors

Introduction

Section snippets

The discovery of pharmacological chaperones acting on GPCRs

Generalization of the pharmacological chaperone action on GPCRs

Pharmacological chaperones can act on wild-type receptors

Mechanistic features of pharmacological chaperones

Conclusions

References and recommended reading

Acknowledgements

Lancet

Trends Endocrinol Metab

J Biol Chem

J Biol Chem

Conformational disease

Nat Cell Biol

Quality control in the endoplasmic reticulum protein factory

Nature

Folding proteins in fatal ways

Nature

Protein origami: therapeutics rescue of misfolded gene products

Molecular Interventions

Chemical chaperones: a pharmacological strategy for disorders of protein folding and trafficking

Pediatr Res

Nephrogenic diabetes insipidus

Annu Rev Physiol