| 1. |
- Barbaro, Michela, et al.
(författare)
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In vitro functional studies of rare CYP21A2 mutations and establishment of an activity gradient for nonclassic mutations improve phenotype predictions in congenital adrenal hyperplasia
- 2015
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Ingår i: Clinical Endocrinology. - : Wiley. - 0300-0664 .- 1365-2265. ; 82:1, s. 37-44
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Tidskriftsartikel (refereegranskat)abstract
- BackgroundA detailed genotype-phenotype evaluation is presented by studying the enzyme activities of five rare amino acid substitutions (Arg233Gly, Ala265Ser, Arg341Trp, Arg366Cys and Met473Ile) identified in the CYP21A2 gene in patients investigated for Congenital adrenal hyperplasia (CAH). ObjectiveTo investigate whether the mutations identified in the CYP21A2 gene are disease causing and to establish a gradient for the degree of enzyme impairment to improve prediction of patient phenotype. Design and patientsThe CYP21A2 genes of seven patients investigated for CAH were sequenced and five mutations were identified. The mutant proteins were expressed in vitro in COS-1 cells, and the enzyme activities towards the two natural substrates were determined to verify the disease-causing state of the mutations. The in vitro activities of these rare mutations were also compared with the activities of four mutations known to cause nonclassic CAH (Pro30Leu, Val281Leu, Pro453Ser and Pro482Ser) in addition to an in silico structural evaluation of the novel mutants. Main outcome measureTo verify the disease-causing state of novel mutations. ResultsFive CYP21A2 mutations were identified (Arg233Gly, Ala265Ser, Arg341Trp, Arg366Cys and Met473Ile). All mutant proteins exhibited enzyme activities above 5%, and four mutations were classified as nonclassic and one as a normal variant. By comparing the investigated protein changes with four common mutations causing nonclassic CAH, a gradient for the degree of enzyme impairment could be established. Studying rare mutations in CAH increases our knowledge regarding the molecular mechanisms that render a mutation pathogenic. It also improves phenotype predictions and genetic counselling for future generations.
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| 2. |
- Hirvikoski, Tatja, et al.
(författare)
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Cognitive functions in children at risk for congenital adrenal hyperplasia treated prenatally with dexamethasone
- 2007
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Ingår i: Journal of Clinical Endocrinology and Metabolism. - : The Endocrine Society. - 0021-972X .- 1945-7197. ; 92:2, s. 542-548
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Tidskriftsartikel (refereegranskat)abstract
- Context and Objective: In Sweden, from 1985 through 1995, 40 fetuses at risk for congenital adrenal hyperplasia (CAH) were treated with dexamethasone (DEX) to prevent virilization of affected females. We report long-term effects on neuropsychological functions and scholastic performance of this controversial treatment. Design and Patients: Prenatally treated children, 7 to 17 yr old, were assessed with standardized neuropsychological tests (A Developmental Neuropsychological Assessment and Wechsler Intelligence Scales for Children) and child-completed questionnaires measuring self-perceived scholastic competence (Self-Perception Profile for Children). A parent-completed questionnaire (Child Behavior Checklist/4-18 School Scale) was used to evaluate whether the treatment had any impact on the children's school performance. In addition, a child-completed questionnaire measuring social anxiety (The Social Anxiety Scale for Children-Revised) was completed by the prenatally treated children aged 8 to 17 yr (n = 21) and age- and sex-matched controls (n = 26). Results: Of 40 DEX-treated children, 26 (median age, 11 yr) participated in the study. Thirty-five sex- and age- matched healthy children were controls. There were no between-group differences concerning psychometric intelligence, measures of cerebral lateralization, memory encoding, and long-term memory. Short-term treated, CAH-unaffected children performed poorer than the control group on a test assessing verbal working memory (P = 0.003), and they rated lower on a questionnaire assessing self-perception of scholastic competence (P = 0.003). This group also showed increased self-rated social anxiety assessed by The Social Anxiety Scale for Children-Revised (P = 0.026). Prenatally treated, CAH-affected children performed poorer than controls on tests measuring verbal processing speed, although this difference disappeared when controlling for the child's full-scale IQ. Conclusions: This study indicates that prenatal DEX treatment is associated with previously not described long-term effects on verbal working memory and on certain aspects of self-perception that could be related to poorer verbal working memory. These findings may thus question future DEX treatment of congenital adrenal hyperplasia. Therefore, we encourage additional retrospective studies of larger cohorts to either confirm or challenge the present findings.
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| 3. |
- Hirvikoski, Tatja, et al.
(författare)
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Deficient cardiovascular stress reactivity predicts poor executive functions in adults with attention-deficit/hyperactivity disorder
- 2011
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Ingår i: Journal of Clinical and Experimental Neuropsychology. - : Informa UK Limited. - 1380-3395 .- 1744-411X. ; 33:1, s. 63-73
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Tidskriftsartikel (refereegranskat)abstract
- Associations between cardiovascular stress markers, subjective stress reactivity, and executive functions were studied in 60 adults (30 with attention-deficit/hyperactivity disorder, ADHD, and 30 controls) using the Paced Auditory Serial Addition Test (PASAT, a test of executive functions) as a cognitive stressor. Despite higher self-perceived stress, the adults with ADHD showed lower or atypical cardiovascular stress reactivity, which was associated with poorer performance on PASAT. Using cardiovascular stress markers, subjective stress, and results on PASAT as predictors in a logistic regression, 83.3% of the ADHD group and 86.9% of the controls could be classified correctly.
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| 4. |
- Hirvikoski, Tatja, et al.
(författare)
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Gender role behaviour in prenatally dexamethasone-treated children at risk for congenital adrenal hyperplasia - a pilot study
- 2011
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Ingår i: Acta Paediatrica. - : Wiley. - 0803-5253 .- 1651-2227. ; 100:9, s. e112-E119
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Tidskriftsartikel (refereegranskat)abstract
- Aim: To investigate the effects of prenatal dexamethasone (DEX) exposure on gender role behaviour. Methods: The participants were 25 of the 40 children (62%, mean age 11 years) at risk for CAH treated with DEX prenatally during the years 1985-1995 in Sweden. The control group consisted of 35 sex- and age-matched healthy children. A new inventory, the Karolinska Inventory of Gender Role Behaviour (KI-GRB), was developed to assess directly school-age children's behaviour, and was evaluated using a separate sample of 160 school-age children. Results: DEX-treated CAH-unaffected boys showed more neutral behaviours than the controls (p = 0.04), while the DEX-treated CAH-unaffected girls did not differ from the controls after adjusting for the site of residence. There was a larger variation in the behaviour of the DEX-treated boys (p < 0.05) and a tendency for less-masculine behaviours in the DEX-treated CAH-unaffected children (p = 0.13). There were no between-group differences in the feminine behaviours. Recalculation of the analyses including the CAH-affected children showed analogous results. Conclusions: This pilot study indicates that the gender role behaviour may be affected in boys as an effect of DEX exposure in early pregnancy. Larger retrospective studies are needed for more conclusive results.
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| 5. |
- Hirvikoski, Tatja, et al.
(författare)
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High self-perceived stress and many stressors, but normal diurnal cortisol rhythm, in adults with ADHD (attention-deficit/hyperactivity disorder).
- 2009
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Ingår i: Hormones and Behavior. - : Elsevier B. V.. - 0018-506X .- 1095-6867. ; 55:3, s. 418-424
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Tidskriftsartikel (refereegranskat)abstract
- Attention-deficit/hyperactivity disorder (ADHD) in adults is associated with significant impairment in many life activities and may thus increase the risk of chronic stress in everyday life. We compared adults with a DSM-IV ADHD diagnosis (n=28) with healthy controls (n=28) regarding subjective stress and amounts of stressors in everyday life, diurnal salivary cortisol in the everyday environment and salivary cortisol before and after cognitive stress in a laboratory setting. The association between cortisol concentrations and impulsivity was also investigated. Consistent with assumptions, individuals with ADHD reported significantly more self-perceived stress than controls, and subjective stress correlated with the amount of stressors in everyday life. The two groups were comparable with respect to overall diurnal cortisol levels and rhythm, as well as in pre- and post-stress cortisol concentrations. Post-stress cortisol (but not baseline cortisol) concentration was positively correlated with impulsivity. The group with high post-stress cortisol also reported more symptoms of depression and anxiety, as well as self-perceived stress and stressors in everyday life. The diagnosis of ADHD significantly increased the risk of belonging to the group with high post-stress cortisol levels. The results in this study warrant a focus not only on the primary diagnosis of ADHD, but also calls for a broader assessment of stressors and subjective stress in everyday life, as well as support comprising stress management and coping skills.
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| 6. |
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| 7. |
- Hirvikoski, Tatja, et al.
(författare)
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Long-term follow-up of prenatally treated children at risk for congenital adrenal hyperplasia : Does dexamethasone cause behavioural problems?
- 2008
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Ingår i: European Journal of Endocrinology. - 0804-4643 .- 1479-683X. ; 159:3, s. 309-316
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Tidskriftsartikel (refereegranskat)abstract
- Objectives: To investigate the long-term effects of prenatal treatment of congenital adrenal hyperplasia (CAH) with emphasis on behavioural problems and temperament. Design: A population-based long-term follow-up study of Swedish children at risk for virilising CAH, who had received treatment prenatally with dexamethasone (DEX). The questionnaire-based follow-up was performed when the children had reached school age. Methods: Standardised parent-completed questionnaires were used to evaluate adaptive functioning, behavioural/emotional problems and psychopathology, social anxiety and temperament in DEX-exposed school-aged children (n=26) and matched controls (n=35). In addition, the association between parental questionnaires and children's self-ratings was investigated. Results: There were no statistically significant differences between DEX-exposed children and controls in measures of psychopathology, behavioural problems and adaptive functioning. In a questionnaire on temperamental traits, DEX-exposed children were described by their parents as being more sociable than controls (P=0.042). The correlation analysis showed only modest parent–child agreement on social anxiety, i.e. the increased social anxiety in children's self-ratings was not confirmed by their parents. Conclusions: DEX-treated children showed good overall adjustment. The parent–child agreement with respect to social anxiety was modest, highlighting the importance of multiple information sources and assessment methods. The clinical significance of the observed difference in sociability cannot be determined within the frameworks of this study. Additional studies of larger cohorts are essential to make more decisive conclusions on the safety of the treatment. Until then, it is important that parents are thoroughly informed about the benefits and potential risks and uncertainties of this controversial treatment.
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| 8. |
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| 9. |
- Hirvikoski, Tatja, et al.
(författare)
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Prenatal Dexametasone treatment of children at risk for congenital adrenal hyperplasia affects cognitive functions.
- 2007
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Ingår i: Journal of Clinical Endocrinology and Metabolism. - : The Endocrine Society. ; 92
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Tidskriftsartikel (refereegranskat)abstract
- In Sweden, during 1985-1995, 40 foetuses at risk for congenital adrenal hyperplasia (CAH) were treated with dexamethasone (DEX) in order to prevent virilisation of affected females. We report long-term effects on neuropsychological functions and scholastic performance of this controversial treatment.Prenatally treated children, 7-17 years, were assessed with standardized neuropsychological tests (NEPSY and WISC-III) and child-completed questionnaires measuring self-perceived scholastic competence (SPPC). A parent-completed questionnaire (CBCL/4-18 School Scale) was used to evaluate whether the treatment had any impact on the children’s school performance.Of 40 DEX treated children, 26 (median age 11 years) participated in the study. Thirty-five sex- and age matched healthy children were controls. There were no between-group differences concerning psychometric intelligence, measures of cerebral lateralization, memory encoding, and long term memory. Short term treated, CAH unaffected children performed worse than the control group on a test assessing verbal working memory (p=0.003), and on self-perception of scholastic competence (p=0.003). Prenatally treated, CAH affected children performed poorer than controls on tests measuring verbal processing speed, although this difference disappeared when controlling for the child’s Full-Scale IQ (FSIQ).This study indicates that prenatal DEX treatment is associated with previously not described long-term effects on verbal working memory and certain aspects of self-perception that could be related to poorer verbal working memory. These findings may thus question future DEX treatment of congenital adrenal hyperplasia. Therefore, we encourage additional retrospective studies of larger cohorts to either confirm or challenge the present findings.
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| 10. |
- Karlsson, Leif, et al.
(författare)
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Novel non-classic CYP21A2 variants, including combined alleles, identified in patients with congenital adrenal hyperplasia
- 2019
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Ingår i: Clinical Biochemistry. - : Elsevier BV. - 0009-9120 .- 1873-2933. ; 73, s. 50-56
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Tidskriftsartikel (refereegranskat)abstract
- Objective: Congenital adrenal hyperplasia (CAH) is an inborn error of metabolism and a common disorder of sex development where >90% of all cases are due to 21-hydroxylase deficiency. Novel and rare pathogenic variants account for 5% of all clinical cases. Here, we sought to investigate the functional and structural effects of four novel (p.Val358Ile, p.Arg369Gln, p.Asp377Tyr, and p.Leu461Pro) and three combinations of CYP21A2 variants (i.e. one allele containing two variants p.[Ile172Asn;Val358Ile], p.[Val281Leu;Arg369Gln], or p.[Asp377Tyr;Leu461Pro]) identified in patients with CAH.Methods: All variants were reconstructed by in vitro site-directed mutagenesis, the proteins were transiently expressed in COS-1 cells and enzyme activities directed toward the two natural substrates (17-hydroxyprogesterone and progesterone) were determined. In parallel, in silico prediction of the pathogenicity of the variants based on the human CYP21 X-ray structure was performed.Results: The novel variants, p.Val358Ile, p.Arg369Gln, p.Asp377Tyr, and p.Leu461Pro exhibited residual enzymatic activities within the range of non-classic (NC) CAH variants (40–82%). An additive effect on the reduction of enzymatic activity (1–17%) was observed when two variants were expressed together, as identified in several patients, resulting in either NC or more severe phenotypes. In silico predictions were in line with the in vitro data except for p.Leu461Pro.Conclusions: Altogether, the combination of clinical data, in silico prediction, and data from in vitro studies are important for establishing a correct genotype and phenotype correlation in patients with CAH.
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| 11. |
- Lajic, Svetlana
(författare)
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Molecular analysis of mutated P450c21 in congenital adrenal hyperplasia
- 1998
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Defects in cytochrome P450c21 (21-hydroxylase, 210H) are the most common causes of congenital adrenal hyperplasia (CAH). This recessively inherited disorder demonstrates a wide spectrum of severity as a consequence of impaired production of cortisol and aldosterone and excessive secretion of adrenal androgens. A limited number of mutations accounts for the majority (95%) of mutated alleles and genetic diagnostics based on detection of these common mutations is well established. However, additional rare mutations, usually specific for a single family or population, are responsible for this disorder in certain patients. Severe forms of 210H deficiency result in prenatal virilizing malformations of external genitalia in affected girls. In an attempt to prevent these malformations and avoid reconstructive surgery, prenatal treatment with dexamethasone (DEX) has been employed. Because eukaryotic cytochromes P450 are integral membrane proteins, it has been difficult to purify and crystallize these proteins in order to determine their three dimensional structures. Useful predictions regarding structure-function relationships for P450c21 can be obtained by studying bacterial members of the cytochrome P450 family, in combination with functional analyses of different P450c21 mutants affecting structurally and/or functionally important residues of the protein. Using the approaches of site-directed mutagenesis and transient expression in COS-I cells, the functional consequences of eight novel missense mutations in CYP21 have been characterized. All of these mutant proteins demonstrated impaired enzyme function and were thus responsible for the disease in the patients in whom they were identified. The degree of functional impairment was generally in agreement with the severity of the symptoms displayed by the patient. By direct sequencing of CYP21, genetic diagnostics was provided for families who do not segregate the common mutations. The P105L and P453S mutations were found on the same allele in two siblings with late-onset disease. Functional studies of each of these mutant proteins individually revealed an enzymatic activity which was 60-70% of the wild-type, while these two mutations in combination acted in a synergistic manner to reduce the enzymatic activity to 10%. The exact mechanism underlying the functional impairment for these mutant enzymes is not fully understood, since only the Vmax and not the KM value was affected. Loss of proline residues might disrupt the three-dimensional structures of the proteins, resulting in reduced stability. If only one of these mutations is present in a patient with CAH, the phenotype will most likely be subtle, not including prenatal virilization. Two other mutations (R356P and R356Q) were identified at the same position of P450c21. R356P (0.15% of wild-type activity) was found in a boy with the most severe form of the disease and R356Q (0.65% activity) was present in a girl with prenatal virilization but no salt loss. The region where these mutations are located is likely to be involved in interaction with the NADPH-cytochrome P450 reductase. With this addition of R356P and R356Q, there is a clear clustering of missense mutations in arginine residues in this portion of P450c21, supporting the concept that the region is involved in interactions with redox partners, since such interactions apparently involve charged amino acid residues. Other structurally and/or functionally important portions of P450c21 include the regions where the mutations delE196, G291S and R483P are located. The delE196 and R483P mutant proteins retained some enzymatic activity and were found in patients with more moderate disease manifestations. These mutant proteins were more rapidly degraded in COS-I cells, as assessed by pulse-chase experiments followed by immunoprecipitation. Thus, the decreased catalytic capacity may be explained by altered protein structure and consequently reduced levels of these mutant forms of P450c21. In the bacterial enzymes P450BM-3 and P450cam, the residue corresponding to G291 of P450c21 has been proposed to participate in the electron transfer required for catalysis. Our data on the function of the G291S mutant suggest that this residue is indeed essential for maintenance of enzyme activity. One fundamental difference between prokaryotic and eukaryotic P450s is the membrane localization of the latter. Membrane-anchoring of P450c21 was determined using cell-free in vitro transcription-translation in the presence of microsomal membranes. Our data indicate that P450c21 binds to the endoplasmic reticulum via only one trans-membrane domain in the most proximal amino-terminal portion of the protein, in agreement with previous reports. We have also demonstrated that the polymorphism (-L10) and the severe P30Q mutation, found in the first and second hydrophobic segments of P450c21 respectively, do not impair the membrane-binding capacity of the protein. Scandinavian experience (44 cases) with prenatal treatment of CAH indicates that such treatment is effective in preventing virilization in affected females. The majority of treated children are born normal in size and they develop and grow normally, compared to untreated controls. However, several adverse events were observed in treated children, such as failure to thrive and late psychomotor development. Women receiving DEX report more side-effects and gain more weight during early pregnancy when treatment is initiated. Therefore, until more experience is gained this treatment must still be considered to be experimental and prospective studies should be designed to exclude possible negative effects on the mother and child.
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| 12. |
- Michelatto, Debora de Paula, et al.
(författare)
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Functional and Structural Consequences of Nine CYP21A2 Mutations Ranging from Very Mild to Severe Effects
- 2016
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Ingår i: International Journal of Endocrinology. - : Hindawi Limited. - 1687-8337 .- 1687-8345.
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Tidskriftsartikel (refereegranskat)abstract
- We present the functional and structural effects of seven novel (p.Leu12Met, p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, p.Gln389 Ala391del, and p.Thr450Met) and two previously reported but not studied (p.Ser113Phe and p.Thr450Pro) CYP21A2 mutations. Functional analyses were complemented with in silico prediction of mutation pathogenicity based on the recently crystallized human CYP21A2 structure. Mutated proteins were transiently expressed in COS-1 cells and enzyme activities towards 17-hydroxyprogesterone and progesterone were determined. Residual enzyme activities between 43% and 97% were obtained for p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, and p.Thr450Met, similar to the activities of the well-known nonclassic mutations p.Pro453Ser and p.Pro482Ser, whereas the p.Leu12Met variant showed an activity of 100%. Conversely, the novel p.Ser113Phe, p. Gln389 Ala391del, and p.Thr450Promutations drastically reduced the enzyme function below 4%. The K-m values for all novel variants were in the same order of magnitude as for the wild-type protein except for p.The450Met. The maximum velocity was decreased for all mutants except for p.Leu12Met. We conclude that p. Leu12Met is a normal variant; the mutations p.Arg16Cys, p.Ser101Asn, p.Ser202Gly, p.Pro267Leu, and p.Thr450Met could be associated with very mild nonclassic CAH, and the mutations p.Ser113Phe, p.Gln389 Ala391del, and p.Thr450Pro are associated with classic CAH. The obtained residual activities indicated a good genotype-phenotype correlation.
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| 13. |
- Van't Westeinde, A., et al.
(författare)
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Increased Resting-State Functional Connectivity in Patients With Autoimmune Addison Disease
- 2024
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Ingår i: Journal of Clinical Endocrinology & Metabolism. - 0021-972X .- 1945-7197. ; 109:3, s. 701-710
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Tidskriftsartikel (refereegranskat)abstract
- Context Individuals with autoimmune Addison disease (AAD) take replacement medication for the lack of adrenal-derived glucocorticoid (GC) and mineralocorticoid hormones from diagnosis. The brain is highly sensitive to these hormones, but the consequence of having AAD for brain health has not been widely addressed.Objective The present study compared resting-state functional connectivity (rs-fc) of the brain between individuals with AAD and healthy controls.Methods Fifty-seven patients with AAD (33 female) and 69 healthy controls (39 female), aged 19 to 43 years were scanned with 3-T magnetic resonance imaging (MRI).Results Independent component and subsequent dual regression analyses revealed that individuals with AAD had stronger rs-fc compared to controls in 3 networks: the bilateral orbitofrontal cortex (OFC), the left medial visual and left posterior default mode network. A higher GC replacement dose was associated with stronger rs-fc in a small part of the left OFC in patients. We did not find any clear associations between rs-fc and executive functions or mental fatigue.Conclusion Our results suggest that having AAD affects the baseline functional organization of the brain and that current treatment strategies of AAD may be one risk factor.
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| 14. |
- Van't Westeinde, Annelies, et al.
(författare)
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Young adult Swedish patients with autoimmune Addison's disease report difficulties with executive functions in daily life despite overall good cognitive performance
- 2022
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Ingår i: Psychoneuroendocrinology. - : Elsevier. - 0306-4530 .- 1873-3360. ; 140
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Tidskriftsartikel (refereegranskat)abstract
- OBJECTIVES: Sub-optimal replacement of glucocorticoids (GC) in autoimmune Addison's disease (AAD) may affect cognitive functioning. The present study therefore sought to investigate cognitive performance and self-reported problems with executive functions in a cohort of young adult patients with AAD.DESIGN AND METHODS: 67 patients with AAD (39 females), mean age 32 yrs. (range 19-41), and 80 control participants (43 females), mean age 29 yrs. (range 19-43), completed neuropsychological tests estimating verbal and non-verbal intellectual ability, learning, memory and executive functioning, in addition to self-report scales assessing problems with executive functions, fatigue and symptoms of anxiety and depression.RESULTS: Patients performed within the average range on all cognitive tests compared to population norms. However, female AAD patients reported more problems than controls with both hot (emotion regulation) and cold (cognitive regulation) executive functions in daily life. Moreover, experienced problems with executive functions in both male and female patients were associated with increased mental fatigue and lower GC replacement doses.CONCLUSIONS: Despite average performance in neuropsychological tests by both sexes, young adult female patients with AAD experience problems with executive functions in daily life. Coping with mental fatigue and optimization of pharmacotherapy may be important factors to be addressed in order to provide timely support for patients. Future research is needed to further determine other risk factors for experiencing executive function impairments in AAD.
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