This study was conducted retrospectively on a cohort of 68 patients with steroid 5α-reductase 2(SRD5A2)deficiency and 46,XY disorders of sex development(DSD).Whole-exon sequencing revealed 28 variants of SRD5A2,and f...This study was conducted retrospectively on a cohort of 68 patients with steroid 5α-reductase 2(SRD5A2)deficiency and 46,XY disorders of sex development(DSD).Whole-exon sequencing revealed 28 variants of SRD5A2,and further analysis identified seven novel mutants.The preponderance of variants was observed in exon 1 and exon 4,specifically within the nicotinamide adenine dinucleotide phosphate(NADPH)-binding region.Among the entire cohort,53 patients underwent initial surgery at Sichuan Provincial People’s Hospital(Chengdu,China).The external genitalia scores(EGS)of these participants varied from 2.0 to 11.0,with a mean of 6.8(standard deviation[s.d.]:2.5).Thirty patients consented to hormone testing.Their average testosterone-todihydrotestosterone(T/DHT)ratio was 49.3(s.d.:23.4).Genetic testing identified four patients with EGS scores between 6 and 9 as having this syndrome;and their T/DHT ratios were below the diagnostic threshold.Furthermore,assessments conducted using the crystal structure of human SRD5A2 have provided insights into the potential pathogenic mechanisms of these novel variants.These mechanisms include interference with NADPH binding(c.356G>C,c.365A>G,c.492C>G,and c.662T>G)and destabilization of the protein structure(c.727C>T).The c.446-1G>T and c.380delG variants were verified to result in large alterations in the transcripts.Seven novel variations were identified,and the variant database for the SRD5A2 gene was expanded.These findings contribute to the progress of diagnostic and therapeutic approaches for individuals with SRD5A2 deficiency.展开更多
性类固醇激素与生物生殖、配子排放以及性腺发育密切相关,Srd5a1基因在性类固醇激素合成过程中扮演重要角色。为了探究性类固醇激素合成相关酶基因Srd5a1对雌性三角帆蚌卵巢发育的影响,应用RACE克隆得到了该基因全长cDNA序列,通过实时...性类固醇激素与生物生殖、配子排放以及性腺发育密切相关,Srd5a1基因在性类固醇激素合成过程中扮演重要角色。为了探究性类固醇激素合成相关酶基因Srd5a1对雌性三角帆蚌卵巢发育的影响,应用RACE克隆得到了该基因全长cDNA序列,通过实时荧光定量(qRT-PCR)、原位杂交技术以及不同浓度17β-雌二醇(E2)和17α-甲基睾酮(MT)激素处理来探究基因的表达特性。结果显示,Srd5a1 cDNA全长2224 bp,包括446 bp 5′-UTR、953 bp 3′-UTR和825 bp ORF,编码274个氨基酸。Srd5a1基因在性腺中的表达呈二态性,在卵巢中表达量更高;且在卵巢排放期的表达量最高,极显著高于其他各时期。原位杂交结果显示,Srd5a1基因在卵巢卵母细胞、卵泡以及精巢精母细胞中有表达。不同浓度E2和MT处理24 d后,Srd5a1基因的表达发生了变化。综上,Srd5a1基因可能在雌性三角帆蚌卵巢卵母细胞发育成熟和配子排放中发挥一定作用,对探索三角帆蚌性腺生长发育具有重要意义,同时为实现三角帆蚌单性化养殖提供理论参考价值。展开更多
基金supported by the Sichuan Science and Technology Program(No.2022JDZH0029 to JYY)the Special Fund for Clinical Research and Translational Medicine from Chinese Academy of Medical Sciences(No.2022-I2M-C&T-B-117 to JYY)the Sichuan Key Research and Development Project from the Department of Science and Technology of Sichuan Province(No.2022YFS0237 to YMT).
文摘This study was conducted retrospectively on a cohort of 68 patients with steroid 5α-reductase 2(SRD5A2)deficiency and 46,XY disorders of sex development(DSD).Whole-exon sequencing revealed 28 variants of SRD5A2,and further analysis identified seven novel mutants.The preponderance of variants was observed in exon 1 and exon 4,specifically within the nicotinamide adenine dinucleotide phosphate(NADPH)-binding region.Among the entire cohort,53 patients underwent initial surgery at Sichuan Provincial People’s Hospital(Chengdu,China).The external genitalia scores(EGS)of these participants varied from 2.0 to 11.0,with a mean of 6.8(standard deviation[s.d.]:2.5).Thirty patients consented to hormone testing.Their average testosterone-todihydrotestosterone(T/DHT)ratio was 49.3(s.d.:23.4).Genetic testing identified four patients with EGS scores between 6 and 9 as having this syndrome;and their T/DHT ratios were below the diagnostic threshold.Furthermore,assessments conducted using the crystal structure of human SRD5A2 have provided insights into the potential pathogenic mechanisms of these novel variants.These mechanisms include interference with NADPH binding(c.356G>C,c.365A>G,c.492C>G,and c.662T>G)and destabilization of the protein structure(c.727C>T).The c.446-1G>T and c.380delG variants were verified to result in large alterations in the transcripts.Seven novel variations were identified,and the variant database for the SRD5A2 gene was expanded.These findings contribute to the progress of diagnostic and therapeutic approaches for individuals with SRD5A2 deficiency.
文摘性类固醇激素与生物生殖、配子排放以及性腺发育密切相关,Srd5a1基因在性类固醇激素合成过程中扮演重要角色。为了探究性类固醇激素合成相关酶基因Srd5a1对雌性三角帆蚌卵巢发育的影响,应用RACE克隆得到了该基因全长cDNA序列,通过实时荧光定量(qRT-PCR)、原位杂交技术以及不同浓度17β-雌二醇(E2)和17α-甲基睾酮(MT)激素处理来探究基因的表达特性。结果显示,Srd5a1 cDNA全长2224 bp,包括446 bp 5′-UTR、953 bp 3′-UTR和825 bp ORF,编码274个氨基酸。Srd5a1基因在性腺中的表达呈二态性,在卵巢中表达量更高;且在卵巢排放期的表达量最高,极显著高于其他各时期。原位杂交结果显示,Srd5a1基因在卵巢卵母细胞、卵泡以及精巢精母细胞中有表达。不同浓度E2和MT处理24 d后,Srd5a1基因的表达发生了变化。综上,Srd5a1基因可能在雌性三角帆蚌卵巢卵母细胞发育成熟和配子排放中发挥一定作用,对探索三角帆蚌性腺生长发育具有重要意义,同时为实现三角帆蚌单性化养殖提供理论参考价值。
