A coding variant in NLRP1 is associated with autoimmune Addison's disease

doi:10.1016/j.humimm.2010.02.004

Human Immunology

Volume 71, Issue 5, May 2010, Pages 530-534

https://doi.org/10.1016/j.humimm.2010.02.004 Get rights and content

Abstract

Autoimmune Addison's disease (AAD) is a complex disorder with several susceptibility loci. Variations in the NLRP1 (previously, NALP1) gene have recently been reported to confer risk for vitiligo and associated autoimmune conditions. We hypothesized that polymorphisms in this gene may affect susceptibility to AAD. The aim of this study was to analyze the associations of six NLRP1 single-nucleotide polymorphisms (SNPs) with AAD within a Polish cohort. The study comprised 101 AAD patients and 254 healthy control individuals. Genotyping was performed by polymerase chain reaction followed by restriction fragment length polymorphism and single strand conformation polymorphism methods. The minor allele of the coding SNP rs12150220 appeared significantly more frequently in AAD compared with healthy individuals (OR = 1.5, 95% CI, 1.08–2.08, p = 0.015). The distribution of genotypes also demonstrated significant differences. The frequency of high-risk genotype AA of rs12150220 SNP was significantly increased among AAD subjects versus controls (p = 0.006 and p = 0.036, respectively; significant after Bonferroni correction), yielding an OR of 2.96 (95% CI, 1.34–6.55). Likewise, the heterozygous genotype TA was observed more frequently in the patient group [OR = 3.09 (95% CI, 1.53–6.24), p = 0.001 and p = 0.006 after Bonferroni correction]. In conclusion, this study confirms an association between the coding polymorphism in NLRP1 and AAD.

Introduction

Autoimmune Addison's disease (AAD), also known as primary adrenocortical insufficiency, is an endocrine disease, targeting the adrenal glands. Progressive destruction of adrenal cortex results in deficits in mineralocorticoids, glucocorticoids, and adrenal androgens. Inappropriate hormonal therapy or delayed diagnosis may lead to life-threatening adrenal crisis [1]. AAD is a rare condition, which affects 6–14 individuals per 100,000, with relatively higher prevalence among women [2]. Notably, primary adrenocortical insufficiency tends to be associated with other autoimmune disorders, giving rise to autoimmune polyendocrine syndromes (APSs in nearly 60% of AAD cases [3]. Thus, patients with AAD are at substantial risk for immune deregulation.

The pathogenesis of autoimmune endocrine diseases is a complex process, influenced by both genetic and environmental factors. Monogenetic inheritance is limited to a few rare polyendocrine syndromes. Mutations in AIRE gene are associated with autoimmune polyendocrine syndrome type 1 (APS1). Nonetheless, inherited factors for most autoimmune disorders remain largely unidentified [4]. Numerous genetic variants that contribute to autoimmune endocrinopathy have been identified using candidate gene approaches and familial linkage studies [5]. The major histocompatibility complex, the cytotoxic T-lymphocyte antigen-4 (CTLA4) gene, and the gene encoding the lymphoid tyrosine phosphatase (PTPN22) are significant immune-response loci [6], [7], [8].

Recently, the NLRP1 gene, mapped to chromosome 17p13, has been linked to vitiligo and associated autoimmunity [9]. The NLRP1 gene encodes NLRP1, a leucine-rich repeat protein 1, a member of the nucleotide oligomerization domains-like receptors (NLRs) family. The NLRs are a group of cytoplasmic pattern recognition receptors, which nonspecifically recognize microbial products, such as lipopolysaccharide, thereby stimulating innate immunity. NLRP1 contains a protein–protein interaction N-terminal pyrin domain followed by centrally located NACHT (NAIP CIIA HET-E and TP1 family proteins) domain responsible for oligomerization and activation of the NLRs, a C-terminal domain comprised of five tandem leucine-rich repeat (LRR) domains that is speculated to bind microbial ligands, a FIIND domain, and a finally a C-terminal caspase-recruitment (CARD) domain [10]. Activated NLRP1 interacts directly with caspase-1 and caspase-5, forming a multiprotein complex known as the inflammasome, which promotes the processing and maturation of cytokines: IL-1β, IL-18, and IL-33 [11], [12]. Trough its multiple domain structure, NLRP1 plays a crucial role in the assembly of the apoptosome, a complex composed of NLRP1, caspase-2, and caspase-9. High levels of expression in immune cells, particularly T cells and Langerhans' cells, emphasize the role of NLRP1 in regulation of the immune system [13]. Variations in the NLRP1 gene have been reported to confer risk for vitiligo and extended autoimmune disorders in Caucasian patients from the United Kingdom, the United States, and Romania [9], [14]. A recent report demonstrated that in a Norwegian population, polymorphisms in NLRP1 are associated with the organ-specific autoimmune conditions, Addison's disease, and type 1 diabetes [15]. These results prompted us to investigate whether single-nucleotide polymorphisms (SNPs) in the NLRP1 gene are a risk factor in Polish patients with AAD, especially as no data on this association in Eastern European populations have been published. NLRP1 is an intriguing functional candidate for autoimmune disease and further study will contribute considerable insight into genetic bases of autoimmune disorders, particularly in the context of our previous demonstration of the significant association between AAD and a variant of the PTPN22 gene [16].

Section snippets

Subjects

The study comprised 101 unrelated patients with AAD (72 females and 29 males) and 254 healthy control subjects (173 females and 81 males), all issued from Polish population of Caucasian origin. The mean age (±SD) of disease onset was 35.8 (±12.6) years. AAD was diagnosed as primary adrenocortical insufficiency with positive serum autoantibodies to 21-hydroxylase (RIA, RSR, UK), and no evidence of tuberculosis or X-linked adrenoleukodystrophy was found in any of the patients. Patients with APS1

Results

The six SNPs (rs6502867, rs12150220, rs2670660, rs878329, rs8182352, rs4790797) of the NLRP1 region were genotyped for 101 unrelated patients with AAD and 254 healthy control subjects. All SNPs were in Hardy–Weinberg equilibrium (HWE) in both patients and control subjects (p > 0.2), with the exception of marker rs12150220 in AAD patients (p = 0.003).

Discussion

Because of its important role in innate immunity and inflammatory processes, the NLRP1 gene represents an interesting candidate for linkage to autoimmune disease, including AAD.

The present study confirms a significant association of the rs12150220 SNP in the NLRP1 gene, with the AAD within a Polish population. The same polymorphism has been reported to confer risk for vitiligo and vitiligo-associated autoimmune disorders in Caucasian patients from the United Kingdom and the United States [9].

Acknowledgments

The study was supported by The Ministry of Science and Higher Education (MNiSW), Poland, under MNiSW grants NN402 226635, NN402 359738.

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Type 1 diabetes (T1D) is an autoimmune disease characterized by the destruction of pancreatic β cells. We show here that the protein NOD-like receptor family pyrin domain containing 1 (NLRP1) has a key role in the pathogenesis of mouse and human T1D. More specifically, downregulation of NLRP1 expression occurs during T helper 17 (Th17) differentiation, alongside greater expression of several molecules related to Th17 cell differentiation in a signal transducers and activators of transcription 3 (STAT3)-dependent pathway. These changes lead to a consequent increase in interleukin 17 (IL-17) production within the pancreas and higher incidence of diabetes in streptozotocin (STZ)-injected mice. Finally, in patients with T1D and a SNP (rs12150220) in NLRP1, there is a robust decrease in IL-17 levels in serum and in memory Th17 cells from peripheral blood mononuclear cells. Our results demonstrate that NLRP1 acts as a negative regulator of the Th17 cell polarization program, making it an interesting target for intervention during the early stages of T1D.
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In humans, multiple mutations in NLRP1 have been found linked or associated with autoimmune disease phenotypes. For example, the marker rs12150220, a nonsynonymous coding region variant that is highly conserved in primate species, was found to be associated with multiple autoimmune diseases such as Addison's Disease (Alkhateeb et al., 2013; Jin et al., 2007b; Li et al., 2017; Magitta et al., 2009; Pontillo et al., 2012b; Zurawek et al., 2010). Additionally, the marker rs2670660, conserved in both mice and humans, is associated with general autoimmunity, interferes with binding motifs for transcription factors, and functionally impacts proliferation and apoptosis (Alkhateeb et al., 2013; Jin et al., 2007b; Pontillo et al., 2012b).
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Autoimmunity against the adrenal cortex is the leading cause of Addison's disease in industrialized countries, with prevalence estimates ranging from 93–220 per million in Europe. The immune-mediated attack on adrenocortical cells cripples their ability to synthesize vital steroid hormones and necessitates life-long hormone replacement therapy. The autoimmune disease etiology is multifactorial involving variants in immune genes and environmental factors. Recently, we have come to appreciate that the adrenocortical cell itself is an active player in the autoimmune process. Here we summarize the complex interplay between the immune system and the adrenal cortex and highlight unanswered questions and gaps in our current understanding of the disease.
L’auto-immunité contre le cortex surrénalien est la principale cause de la maladie d’Addison dans les pays industrialisés, avec des estimations de prévalence allant de 93 à 220 cas par million d'habitants en Europe. L’attaque immunitaire des cellules corticosurrénales affaiblit leur capacité à synthétiser des hormones stéroïdes vitales et nécessite un traitement hormonal substitutif définitif. L’étiologie de la maladie auto-immune est multifactorielle impliquant des polymorphismes dans les gènes immunitaires et des facteurs environnementaux. Récemment, nous avons appris que la cellule corticosurrénale elle-même joue un rôle actif dans le processus auto-immun. Nous résumons ici l’interaction complexe entre le système immunitaire et le cortex surrénalien et soulignons les questions et les lacunes sans réponse dans notre compréhension actuelle de la maladie d'Addison.
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Citation Excerpt :
Among those, a nonsynonymous L155H substitution (rs12150220) and a variant in the promoter region (rs2670660) were the most significantly associated with disease. This association was further observed not only in vitiligo [64] but also in Addison's disease [65,66], type 1 diabetes [66], systemic lupus erythematosus [18], inflammatory bowel disease [67], and celiac disease [17]. NLRP1 rs8182352 variant was identified in patients with giant cell arteritis [68] and systemic sclerosis-related fibrosing alveolitis [69].
NLRP1 was the first NOD-like receptor described to form an inflammasome, recruiting ASC to activate caspase-1, which processes interleukin-1β and interleukin-18 to their active form. A wealth of new genetic information has now redefined our understanding of this innate immune sensor. Specifically, rare loss-of-function variants in the N-terminal pyrin domain indicate that this part of NLRP1 is autoinhibitory and normally acts to prevent a familial autoinflammatory skin disease associated with cancer. In the absence of a ligand to trigger human NLRP1, these mutations have now confirmed the requirement of NLRP1 autolytic cleavage within the FIIND domain, which had previously been implicated in NLRP1 activation. Autolytic cleavage generates a C-terminal fragment of NLRP1 containing the CARD domain which then forms an ASC-dependent inflammasome. The CARD domain as an inflammasome linker is consistent with the observation that under some conditions, particularly for mouse NLRP1, caspase-1 can be engaged directly, and although it is no longer processed, it is still capable of producing mature IL-1β. Additional rare variants in a linker region between the LRR and FIIND domains of NLRP1 also cause autoinflammatory disease in both humans and mice. This new genetic information is likely to provide for more mechanistic insight in the years to come, contributing to our understanding of how NLRP1 functions as an innate immune sensor of infection and predisposes to autoimmune or autoinflammatory diseases.

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A coding variant in NLRP1 is associated with autoimmune Addison's disease

Abstract

Introduction

Section snippets

Subjects

Results

Discussion

Acknowledgments

Trends Mol Med

Trends Immunol

Mol Cell

J Invest Dermatol

Am J Hum Genet

Am J Hum Genet

Normal overall mortality rate in Addison's disease, but young patients are at risk of premature death

Eur J Endocrinol

Autoimmune adrenal insufficiency and autoimmune polyendocrine syndromes: Autoantibodies, autoantigens, and their applicability in diagnosis and disease prediction

Endocr Rev

Polyglandular autoimmune diseases

Positional cloning of the APECED gene

Nat Genet

Localizationof type 1 diabetes susceptibility to the MHC class I genes HLA-B and HLA-A

Nature

The emerging role of the CTLA4 gene in autoimmune endocrinopathies

Eur J Endocrinol