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New and recurrent gain-of-function STAT1 mutations in patients with chronic mucocutaneous candidiasis from Eastern and Central Europe.

Soltész B, Tóth B, Shabashova N, Bondarenko A, Okada S, Cypowyj S, Abhyankar A, Csorba G, Taskó S, Sarkadi AK, Méhes L, Rozsíval P, Neumann D, Chernyshova L, Tulassay Z, Puel A, Casanova JL, Sediva A, Litzman J, Maródi L - J. Med. Genet. (2013)

Bottom Line: The c.1154C>T (T385M) mutation affecting the DNA-binding domain (DBD) resulted in a gain of STAT1 phosphorylation in a Ukrainian patient.Impaired Candida-induced IL-17A and IL-22 secretion by leucocytes and lower levels of intracellular IL-17 and IL-22 production by T cells were found in several patients.Haplotype studies indicated that the c.820C>T (R274W) mutation was recurrent due to a hotspot rather than a founder effect.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious and Pediatric Immunology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary, EU.

ABSTRACT

Background: Chronic mucocutaneous candidiasis disease (CMCD) may result from various inborn errors of interleukin (IL)-17-mediated immunity. Twelve of the 13 causal mutations described to date affect the coiled-coil domain (CCD) of STAT1. Several mutations, including R274W in particular, are recurrent, but the underlying mechanism is unclear.

Objective: To investigate and describe nine patients with CMCD in Eastern and Central Europe, to assess the biochemical impact of STAT1 mutations, to determine cytokines in supernatants of Candida-exposed blood cells, to determine IL-17-producing T cell subsets and to determine STAT1 haplotypes in a family with the c.820C>T (R274W) mutation.

Results: The novel c.537C>A (N179K) STAT1 mutation was gain-of-function (GOF) for γ-activated factor (GAF)-dependent cellular responses. In a Russian patient, the cause of CMCD was the newly identified c.854 A>G (Q285R) STAT1 mutation, which was also GOF for GAF-dependent responses. The c.1154C>T (T385M) mutation affecting the DNA-binding domain (DBD) resulted in a gain of STAT1 phosphorylation in a Ukrainian patient. Impaired Candida-induced IL-17A and IL-22 secretion by leucocytes and lower levels of intracellular IL-17 and IL-22 production by T cells were found in several patients. Haplotype studies indicated that the c.820C>T (R274W) mutation was recurrent due to a hotspot rather than a founder effect. Severe clinical phenotypes, including intracranial aneurysm, are presented.

Conclusions: The c.537C>A and c.854A>G mutations affecting the CCD and the c.1154C>T mutation affecting the DBD of STAT1 are GOF. The c.820C>T mutation of STAT1 in patients with CMCD is recurrent due to a hotspot. Patients carrying GOF mutations of STAT1 may develop multiple intracranial aneurysms by hitherto unknown mechanisms.

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Related in: MedlinePlus

Induction of GAS-dependent reporter gene transcription activity in U3C cells stimulated with IFN-γ. The response to IFN-γ stimulation was measured by determining the activity of a luciferase reporter gene under the control of the GAS promoter. GAS transcriptional activity is shown as fold induction after 16 h of stimulation with IFN-γ (10 U/ml, left columns; 1000 U/ml, right columns), with respect to unstimulated cells. Mock and MSMD-causing Y701C allele-transfected cells served as negative controls. Upon stimulation with IFN-γ, cells transfected with either the N179K or the Q285R allele responded two to three times more strongly than those transfected with the WT or Y701C alleles. Similar results were obtained when cells were transfected with the GOF R274Q allele known to cause CMCD. CMCD, chronic mucocutaneous candidiasis disease; GAS, γ-activated sequences; GOF, gain of function; IFN, interferon; MSMD, Mendelian susceptibility to mycobacterial disease; WT, wild-type.
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JMEDGENET2013101570F1: Induction of GAS-dependent reporter gene transcription activity in U3C cells stimulated with IFN-γ. The response to IFN-γ stimulation was measured by determining the activity of a luciferase reporter gene under the control of the GAS promoter. GAS transcriptional activity is shown as fold induction after 16 h of stimulation with IFN-γ (10 U/ml, left columns; 1000 U/ml, right columns), with respect to unstimulated cells. Mock and MSMD-causing Y701C allele-transfected cells served as negative controls. Upon stimulation with IFN-γ, cells transfected with either the N179K or the Q285R allele responded two to three times more strongly than those transfected with the WT or Y701C alleles. Similar results were obtained when cells were transfected with the GOF R274Q allele known to cause CMCD. CMCD, chronic mucocutaneous candidiasis disease; GAS, γ-activated sequences; GOF, gain of function; IFN, interferon; MSMD, Mendelian susceptibility to mycobacterial disease; WT, wild-type.

Mentions: The two new mutations affecting the STAT1 CCD, N179K and Q285R, were found in a Czech patient (patient P1) and a Russian patient, respectively (P7). These patients were not available for biochemical studies. We therefore analysed the possible pathological consequences of these mutations by site-directed mutagenesis and studies of γ-activated factor (GAF)-dependent cellular responses to IFN-γ. We transfected STAT1-deficient U3C cells with wild-type (WT) STAT1 and c.537C>A (N179K) and c.854A>G (Q285R) STAT1 mutant constructs, and with the previously characterised CMCD-causing c. 821G>A (R274Q)9 and autosomal-dominant MSMD-causing Y701C STAT1 alleles Hirato O et al (manuscript submitted). Responses to cytokine stimulation were investigated by measuring the luciferase activity of the reporter gene under the control of the γ-activated sequence (GAS) promoter. Stimulation with various concentrations (10 and 1000 U/ml) of IFN-γ resulted in responses that were two to three times stronger in cells transfected with the two new mutant alleles or with the CMCD-causing R274Q allele than in those transfected with WT or MSMD-causing Y701C alleles (figure 1). These data suggest that the new heterozygous alleles reported in this study are GOF for GAF-dependent cellular responses to IFN-γ, one of the key STAT1-activating cytokines.


New and recurrent gain-of-function STAT1 mutations in patients with chronic mucocutaneous candidiasis from Eastern and Central Europe.

Soltész B, Tóth B, Shabashova N, Bondarenko A, Okada S, Cypowyj S, Abhyankar A, Csorba G, Taskó S, Sarkadi AK, Méhes L, Rozsíval P, Neumann D, Chernyshova L, Tulassay Z, Puel A, Casanova JL, Sediva A, Litzman J, Maródi L - J. Med. Genet. (2013)

Induction of GAS-dependent reporter gene transcription activity in U3C cells stimulated with IFN-γ. The response to IFN-γ stimulation was measured by determining the activity of a luciferase reporter gene under the control of the GAS promoter. GAS transcriptional activity is shown as fold induction after 16 h of stimulation with IFN-γ (10 U/ml, left columns; 1000 U/ml, right columns), with respect to unstimulated cells. Mock and MSMD-causing Y701C allele-transfected cells served as negative controls. Upon stimulation with IFN-γ, cells transfected with either the N179K or the Q285R allele responded two to three times more strongly than those transfected with the WT or Y701C alleles. Similar results were obtained when cells were transfected with the GOF R274Q allele known to cause CMCD. CMCD, chronic mucocutaneous candidiasis disease; GAS, γ-activated sequences; GOF, gain of function; IFN, interferon; MSMD, Mendelian susceptibility to mycobacterial disease; WT, wild-type.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3756505&req=5

JMEDGENET2013101570F1: Induction of GAS-dependent reporter gene transcription activity in U3C cells stimulated with IFN-γ. The response to IFN-γ stimulation was measured by determining the activity of a luciferase reporter gene under the control of the GAS promoter. GAS transcriptional activity is shown as fold induction after 16 h of stimulation with IFN-γ (10 U/ml, left columns; 1000 U/ml, right columns), with respect to unstimulated cells. Mock and MSMD-causing Y701C allele-transfected cells served as negative controls. Upon stimulation with IFN-γ, cells transfected with either the N179K or the Q285R allele responded two to three times more strongly than those transfected with the WT or Y701C alleles. Similar results were obtained when cells were transfected with the GOF R274Q allele known to cause CMCD. CMCD, chronic mucocutaneous candidiasis disease; GAS, γ-activated sequences; GOF, gain of function; IFN, interferon; MSMD, Mendelian susceptibility to mycobacterial disease; WT, wild-type.
Mentions: The two new mutations affecting the STAT1 CCD, N179K and Q285R, were found in a Czech patient (patient P1) and a Russian patient, respectively (P7). These patients were not available for biochemical studies. We therefore analysed the possible pathological consequences of these mutations by site-directed mutagenesis and studies of γ-activated factor (GAF)-dependent cellular responses to IFN-γ. We transfected STAT1-deficient U3C cells with wild-type (WT) STAT1 and c.537C>A (N179K) and c.854A>G (Q285R) STAT1 mutant constructs, and with the previously characterised CMCD-causing c. 821G>A (R274Q)9 and autosomal-dominant MSMD-causing Y701C STAT1 alleles Hirato O et al (manuscript submitted). Responses to cytokine stimulation were investigated by measuring the luciferase activity of the reporter gene under the control of the γ-activated sequence (GAS) promoter. Stimulation with various concentrations (10 and 1000 U/ml) of IFN-γ resulted in responses that were two to three times stronger in cells transfected with the two new mutant alleles or with the CMCD-causing R274Q allele than in those transfected with WT or MSMD-causing Y701C alleles (figure 1). These data suggest that the new heterozygous alleles reported in this study are GOF for GAF-dependent cellular responses to IFN-γ, one of the key STAT1-activating cytokines.

Bottom Line: The c.1154C>T (T385M) mutation affecting the DNA-binding domain (DBD) resulted in a gain of STAT1 phosphorylation in a Ukrainian patient.Impaired Candida-induced IL-17A and IL-22 secretion by leucocytes and lower levels of intracellular IL-17 and IL-22 production by T cells were found in several patients.Haplotype studies indicated that the c.820C>T (R274W) mutation was recurrent due to a hotspot rather than a founder effect.

View Article: PubMed Central - PubMed

Affiliation: Department of Infectious and Pediatric Immunology, Medical and Health Science Center, University of Debrecen, Debrecen, Hungary, EU.

ABSTRACT

Background: Chronic mucocutaneous candidiasis disease (CMCD) may result from various inborn errors of interleukin (IL)-17-mediated immunity. Twelve of the 13 causal mutations described to date affect the coiled-coil domain (CCD) of STAT1. Several mutations, including R274W in particular, are recurrent, but the underlying mechanism is unclear.

Objective: To investigate and describe nine patients with CMCD in Eastern and Central Europe, to assess the biochemical impact of STAT1 mutations, to determine cytokines in supernatants of Candida-exposed blood cells, to determine IL-17-producing T cell subsets and to determine STAT1 haplotypes in a family with the c.820C>T (R274W) mutation.

Results: The novel c.537C>A (N179K) STAT1 mutation was gain-of-function (GOF) for γ-activated factor (GAF)-dependent cellular responses. In a Russian patient, the cause of CMCD was the newly identified c.854 A>G (Q285R) STAT1 mutation, which was also GOF for GAF-dependent responses. The c.1154C>T (T385M) mutation affecting the DNA-binding domain (DBD) resulted in a gain of STAT1 phosphorylation in a Ukrainian patient. Impaired Candida-induced IL-17A and IL-22 secretion by leucocytes and lower levels of intracellular IL-17 and IL-22 production by T cells were found in several patients. Haplotype studies indicated that the c.820C>T (R274W) mutation was recurrent due to a hotspot rather than a founder effect. Severe clinical phenotypes, including intracranial aneurysm, are presented.

Conclusions: The c.537C>A and c.854A>G mutations affecting the CCD and the c.1154C>T mutation affecting the DBD of STAT1 are GOF. The c.820C>T mutation of STAT1 in patients with CMCD is recurrent due to a hotspot. Patients carrying GOF mutations of STAT1 may develop multiple intracranial aneurysms by hitherto unknown mechanisms.

Show MeSH
Related in: MedlinePlus