通过测定内源性雄激素受体（AR）活性鉴定一类AR突变阴性的雄激素不敏感症
Identification of an AR Mutation-Negative Class of Androgen Insensitivity by Determining Endogenous AR Activity.
作者信息
Hornig N C, Ukat M, Schweikert H U, Hiort O, Werner R, Drop S L S, Cools M, Hughes I A, Audi L, Ahmed S F, Demiri J, Rodens P, Worch L, Wehner G, Kulle A E, Dunstheimer D, Müller-Roßberg E, Reinehr T, Hadidi A T, Eckstein A K, van der Horst C, Seif C, Siebert R, Ammerpohl O, Holterhus P-M
机构信息
Department of Pediatrics (N.C.H., M.U., J.D., P.R., A.E.K., P.-M.H.), Division of Pediatric Endocrinology and Diabetes, and Institute of Human Genetics (L.W., R.S., O.A.), Christian-Albrechts-University Kiel and University Hospital Schleswig-Holstein, Campus Kiel, Schwanenweg 20, 24105 Kiel, Germany; Rheinische Friedrich-Wilhelms-Universität Bonn, Department of Medicine III (H.U.S., G.W.), Institute for Biochemistry and Molecular Biology, Nussallee 11, 53115 Bonn, Germany; Department of Pediatrics (O.H., R.W.), Division of Experimental Pediatric Endocrinology, University of Luebeck, 23538 Luebeck, Germany; Department of Pediatrics (S.L.S.D.), Division of Pediatric Endocrinology, Sophia Childreńs Hospital, Erasmus Medical Center, 's-Gravendijkwal 230, 3015 CE Rotterdam, The Netherlands; Department of Pediatric Endocrinology (Medical Center), Ghent University Hospital, Ghent University, 9000 Ghent, Belgium; Department of Pediatrics (I.A.H.), University of Cambridge, Cambridge CB2 0QQ, United Kingdom; Pediatric Endocrinology Research Unit (L.A.), Vall d'Hebron Institut de Recerca, Hospital Universitari Vall d'Hebron, Centro de Investigación Biomédica en Red Enfermedades Raras, Instituto de Salud Carlos III, Passeig Vall d'Hebron 119, 08035 Barcelona, Spain; Developmental Endocrinology Research Group (S.F.A.), School of Medicine, University of Glasgow, Yorkhill Glasgow G3 8SJ, United Kingdom; Kinderklinik (D.D.), Klinikum Augsburg, 86156 Augsburg, Germany; Klinikum Esslingen (E.M.-R.), 73730 Esslingen, Germany; Department of Pediatrics (T.R.), Division of Pediatric Endocrinology, Diabetes, and Nutrition, University Witten/Herdecke, 45711 Datteln, Germany; Hypospadiezentrum (A.T.H.), 63500 Seligenstadt, Germany; Gemeinschaftspraxis für Kinderchirurgie (A.K.E.), 24119 Kronshagen, Germany; Urologische Gemeinschaftspraxis (C.v.d.H), and UROLOGIE Zentrum Kiel (C.S.), 24103 Kiel, Germany; and Institute of Human Genetics (R.S.), University of Ulm and University Hospital of Ulm, 89081 Ulm, Germany.
出版信息
J Clin Endocrinol Metab. 2016 Nov;101(11):4468-4477. doi: 10.1210/jc.2016-1990. Epub 2016 Sep 1.
CONTEXT
Only approximately 85% of patients with a clinical diagnosis complete androgen insensitivity syndrome and less than 30% with partial androgen insensitivity syndrome can be explained by inactivating mutations in the androgen receptor (AR) gene.
OBJECTIVE
The objective of the study was to clarify this discrepancy by in vitro determination of AR transcriptional activity in individuals with disorders of sex development (DSD) and male controls.
DESIGN
Quantification of DHT-dependent transcriptional induction of the AR target gene apolipoprotein D (APOD) in cultured genital fibroblasts (GFs) (APOD assay) and next-generation sequencing of the complete coding and noncoding AR locus.
SETTING
The study was conducted at a university hospital endocrine research laboratory.
PATIENTS
GFs from 169 individuals were studied encompassing control males (n = 68), molecular defined DSD other than androgen insensitivity syndrome (AIS; n = 18), AR mutation-positive AIS (n = 37), and previously undiagnosed DSD including patients with a clinical suspicion of AIS (n = 46).
INTERVENTION(S): There were no interventions.
MAIN OUTCOME MEASURE(S): DHT-dependent APOD expression in cultured GF and AR mutation status in 169 individuals was measured.
RESULTS
The APOD assay clearly separated control individuals (healthy males and molecular defined DSD patients other than AIS) from genetically proven AIS (cutoff < 2.3-fold APOD-induction; 100% sensitivity, 93.3% specificity, P < .0001). Of 46 DSD individuals with no AR mutation, 17 (37%) fell below the cutoff, indicating disrupted androgen signaling.
CONCLUSIONS
AR mutation-positive AIS can be reliably identified by the APOD assay. Its combination with next-generation sequencing of the AR locus uncovered an AR mutation-negative, new class of androgen resistance, which we propose to name AIS type II. Our data support the existence of cellular components outside the AR affecting androgen signaling during sexual differentiation with high clinical relevance.
背景
临床诊断为完全性雄激素不敏感综合征的患者中只有约85%，而部分性雄激素不敏感综合征患者中不到30%可通过雄激素受体（AR）基因的失活突变来解释。
目的
本研究的目的是通过体外测定性发育障碍（DSD）个体和男性对照者的AR转录活性来阐明这一差异。
设计
在培养的生殖器成纤维细胞（GFs）中对AR靶基因载脂蛋白D（APOD）的双氢睾酮（DHT）依赖性转录诱导进行定量（APOD测定），并对完整的AR编码和非编码基因座进行二代测序。
地点
本研究在一家大学医院的内分泌研究实验室进行。
患者
研究了169名个体的GFs，包括对照男性（n = 68）、除雄激素不敏感综合征（AIS）外分子明确的DSD（n = 18）、AR突变阳性的AIS（n = 37）以及包括临床怀疑为AIS的患者在内的既往未确诊的DSD（n = 46）。
干预措施
无干预措施。
主要观察指标
测定培养的GF中DHT依赖性APOD表达以及169名个体的AR突变状态。
结果
APOD测定能明确区分对照个体（健康男性和除AIS外分子明确的DSD患者）与基因证实的AIS（临界值<2.3倍APOD诱导；敏感性100%；特异性93.3%；P <.0001）。在46名无AR突变的DSD个体中，17名（37%）低于临界值，表明雄激素信号传导中断。
结论
APOD测定可可靠地识别AR突变阳性的AIS。其与AR基因座的二代测序相结合，发现了一类新的AR突变阴性的雄激素抵抗，我们建议将其命名为II型AIS。我们的数据支持在性分化过程中存在AR之外影响雄激素信号传导的细胞成分，具有高度临床相关性。
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