Cytochrome P450 family 17 subfamily A member 1 mutation causes severe pseudohermaphroditism: A case report

BACKGROUND 17α-Hydroxylase deficiency(17-OHD)is a rare form of congenital adrenal hyperplasia,characterized by hypertension,hypokalemia,and gonadal dysplasia.However,due to the lack of a comprehensive understanding of this disease,it is prone to misdiagnosis and missed diagnosis,and there is no complete cure.CASE SUMMARY We report a female patient with 17-OHD.The patient was admitted to the Department of Neurology of our hospital due to limb weakness.During treatment,it was found that the patient’s condition was difficult to correct except for hypokalemia,and her blood pressure was difficult to control with various antihypertensive drugs.She was then transferred to our department for further treatment.On physical examination,the patient's gonadal development was found to be abnormal,and chromosome analysis demonstrated karyotype 46,XY.Considering the possibility of 17-OHD,the cytochrome P450 family 17 subfamily A member 1(CYP17A1)test was performed to confirm the diagnosis.CONCLUSION The clinical manifestations of 17-OHD are complex.Hormone determination,imaging examination,chromosome determination and CYP17A1 gene test are helpful for early diagnosis.

17α-hydroxylase/17,20 carbon chain lyase deficiency caused by p.Tyr329fs homozygous mutation:Three case reports

p.Tyr329fs is a cytochrome P450c17 mutation among Chinese individuals.However,data on 17-α-hydroxylase deficiency caused by cytochrome P450c17 p.Tyr329fs homozygous mutation are lacking.This paper is a case report of three patients homozygous for p.Tyr329fs who were diagnosed with 17-α-hydroxylase deficiency between 2005 and 2019.CASE SUMMARY Case 1 presented with hypertension,hypokalemia,sexual infantilism and delayed bone age.The patient had a 46,XY karyotype,was homozygous for p.Tyr329fs and was recently treated with dexamethasone 0.375 mg qn.Case 2 presented with hypokalemia,sexual infantilism,osteoporosis and delayed bone age.The patient had a 46,XY karyotype,was homozygous for p.Tyr329fs and was treated with dexamethasone 0.75 mg qn at the last follow-up.Serum potassium and blood pressure could be maintained within normal range for cases 1 and 2.Case 3 presented with amenorrhea,sexual infantilism,osteopenia and delayed bone age.The patient had a 46,XX karyotype,was homozygous for p.Tyr329fs and was treated with dexamethasone 0.75 mg qn and progynova 1 mg qd.Outpatient follow-up revealed an adrenocorticotropic hormone(8 AM)of<5.00 pg/mL.CONCLUSION The homozygous p.Tyr329fs mutation usually manifests as a combined deficiency,and definitive diagnosis depends primarily on genetic testing.

一个含有3例P450c17α缺陷患者的家系分析

目的对一个含有3例P450c17α缺陷患者的家系进行临床和生化分析。方法收集临床资料,用放免、酶免等方法测定患者及家系成员的ACTH及类固醇激素水平,并对患者家系中的杂合子进行了1h ACTH兴奋试验。结果3例患者具有典型的高血压、低血钾和性腺功能低下表现,以先证者临床表现更为突出。患者血ACTH异常升高,而血皮质醇、性激素水平极低,血清17-羟孕酮降低。家系成员的1h ACTH兴奋试验发现,患者父亲在1h ACTH兴奋试验时,表现为高水平的基础17-羟孕酮,ACTH刺激后反而下降。结论3名患者为典型的P450c17缺陷,17α-羟化酶和17,20-裂解酶活性均低下,ACTH兴奋试验发现杂合子具有一定程度的代谢障碍。

Development of MEMS directed evolution strategy for multiplied throughput and convergent evolution of cytochrome P450 enzymes

Directed evolution(DE)inspired by natural evolution(NE)has been achieving tremendous successes in protein/enzyme engineering.However,the conventional"one-protein-for-one-task"DE cannot match the"multi-proteins-for-multi-tasks"NE in terms of screening throughput and efficiency,thus often failing to meet the fast-growing demands for biocatalysts with desired properties.In this study,we design a novel"multi-enzymes-for-multi-substrates"(MEMS)DE model and establish the proof-ofconcept by running a NE-mimicking and higher-throughput screening on the basis of"two-P450 s-against-seven-substrates"(2P×7S)in one pot.With the multiplied throughput and improved hit rate,we witness a series of convergent evolution events of the two archetypal cytochrome P450 enzymes(P450 BM3 and P450 cam)in laboratory.It is anticipated that the new strategy of MEMS DE will find broader application for a larger repertoire of enzymes in the future.Furthermore,structural and substrate docking analysis of the two functionally convergent P450 variants provide important insights into how distinct P450 active-sites can reach a common catalytic goal.

一个三姐妹同患P450c 17α缺陷家系的分子遗传学研究

目的 研究 1个三姐妹同患P45 0c 17α缺陷家系的分子遗传学机制。方法 采用聚合酶链反应 单链构象多态性 (PCR SSCP)、限制性内切酶酶切及自动测序等方法检测患者家系中CYP17基因的突变情况。结果 3例患者CYP 17基因中一等位基因的第 6号外显子有杂合点突变His3 73 →Leu ,用等位基因特异性扩增的方法证明此点突变来自父方 ;另一等位基因第 8号外显子另有9个碱基缺失的突变 ,使位于羧基端 487 489位的Asp Ser Phe缺失 ,并用限制性内切酶酶切的方法证实患者的母亲和一个胞弟是该缺失突变的携带者。结论 1个三姐妹同时患病的中国人P45 0c 17α缺陷的家系是由于CYP 17基因的复合杂合突变所致。