PS-BPB06-10: DISTRIBUTION OF CACNA1D SOMATIC MUTATIONS AMONG CYP11B2 IMMUNOHISTOCHEMISTRY-GUIDED SAMPLES OF ALDOSTERONE-PRODUCING ADENOMA (APA)
PS-BPB06-10: DISTRIBUTION OF CACNA1D SOMATIC MUTATIONS AMONG CYP11B2 IMMUNOHISTOCHEMISTRY-GUIDED SAMPLES OF ALDOSTERONE-PRODUCING ADENOMA (APA)
复制标题
PS-BPB06-10：CACNA1D 体细胞突变在 CYP11B2 免疫组织化学引导的醛固酮腺瘤 (APA) 样本中的分布
DOI：
10.1097/01.hjh.0000915608.20633.e6
复制
发表时间：
2023
影响因子：
4.9
通讯作者：
Pauzi F
中科院分区：
文献类型：
--
作者：
Pauzi F
目的：过去对非洲-加勒比地区原发性醛固酮增多症（PA）患者的研究发现，CACNA1D突变是其队列中最常见的醛固酮驱动突变；这与KCNJ5突变最常见的高加索人和东方人人群存在显著差异。然而，采用醛固酮合酶（CYP11B2）免疫组化（IHC）引导下的全CACNA1D基因测序方法的研究并不多，而是采用非引导的腺瘤取样和热点测序。因此，我们对来自东方队列（马来西亚）和高加索队列（捷克共和国）的APAs进行了CYP11B2 ihc引导的全基因测序以进行比较。设计与方法：对100例马来西亚和捷克单侧PA患者手术切除肾上腺进行研究。从CYP11B2 IHC染色阳性的腺瘤FFPE切片中分离基因组DNA，并通过已知醛固酮驱动基因的下一代测序（NGS）进行体细胞突变分析。预测的致病突变通过Sanger测序进行验证。结果：在100份送交NGS的FFPE样本中，有36份因数据质量问题被排除。64例APAs中58例（87.5%）存在致病性醛固酮驱动体细胞突变。KCNJ5突变最为常见（n= 37, 57.8%），其次是CACNA1D （n= 11, 20.3%）、ATP1A1 （n= 6, 9.4%）、ATP2B3 （n= 3, 4.7%）和CTNNB1 （n= 2, 3.1%）突变。特别有趣的是，一个APA样本有两个致病体细胞突变，CACNA1D p. Val1373Met和CTNNB1: p. Thr41Ala。SIFT和Polyphen2评分预测这两种突变都是有害的。这两种突变先前分别被报道引起APAs。CACNA1D/CTNNB1突变体APA是孤立的，CYP11B2表达均匀，CYP17A1表达阴性。对一名53岁捷克男性患者进行肾上腺切除术。大多数携带CACNA1D突变的APAs患者为男性（n= 8, 72.7%），来自捷克队列（n= 7, 64%）。CACNA1D突变APA患者（45±3）与非CACNA1D突变APA患者（48±2）的肾上腺切除术年龄无显著差异。结论：孤立的ZG APAs可以同时携带CACNA1D和CTNNB1突变。在这项CYP11B2-IHC引导的研究中，CACNA1D的患病率与先前在东方和高加索人群中报道的结果一致。尽管在这些人群中，CACNA1D并不是导致APAs的首要基因改变，但CACNA1D仍可能是特发性醛固酮增多症和醛固酮生成微分子的潜在关键因素。因此，需要进一步研究CACNA1D致病突变在双侧PA中的分布特征。
Objectives:Past studies in Afro-Caribbean primary aldosteronism (PA) patients have found CACNA1D mutations to be the most common aldosterone-driving mutation among their cohort; a striking difference to Caucasian and Oriental cohorts where KCNJ5 mutations are reported to be the most common. However not many studies had used aldosterone synthase (CYP11B2) immunohistochemistry (IHC)-guided sequencing approach of the whole CACNA1D gene, performing instead unguided adenoma sampling and hotspot sequencing. We thus performed CYP11B2 IHC-guided whole gene sequencing on APAs from an Oriental cohort (Malaysia) and a Caucasian cohort (Czech Republic) for comparison.Design and method:Surgically removed adrenals from 100 Malaysian and Czech PA patients with unilateral disease were studied. Genomic DNA was isolated from adenoma FFPE sections that had a positive CYP11B2 IHC staining and somatic mutations analysis were performed by next generation sequencing (NGS) of known aldosterone-driver genes. Predicted pathogenic mutations were verified by Sanger Sequencing.Results:Out of 100 FFPE samples sent for NGS, 36 were excluded due to the quality of the data. In total 58 of 64 APAs (87.5%) had a pathogenic aldosterone-driver somatic mutation. KCNJ5 mutations were the most common (n= 37, 57.8%), followed by CACNA1D (n= 11, 20.3%), ATP1A1 (n= 6, 9.4%), ATP2B3 (n= 3, 4.7%) and CTNNB1 (n= 2, 3.1%) mutations. Of particular interest, one APA sample had two pathogenic somatic mutations, CACNA1D p. Val1373Met and CTNNB1: p. Thr41Ala. SIFT and Polyphen2 scores predicted both mutations to be deleterious. Both mutations have previously been reported separately to cause APAs. The CACNA1D/CTNNB1 mutant APA, was solitary, with homogeneous CYP11B2 expression while negative for CYP17A1 expression. The APA was adrenalectomized from a 53-year-old male Czech patient. The majority of patients with APAs harboring CACNA1D mutations were male (n= 8, 72.7%) and from the Czech cohort (n= 7, 64%). There were no significant differences in age at adrenalectomy between patients harboring a CACNA1D mutant APA (45±3) and those with a non-CACNA1D mutant APA (48±2).Conclusions:Solitary ZG APAs can harbor both a CACNA1D and a CTNNB1 mutation. The prevalence of CACNA1D in this CYP11B2-IHC guided study is consistent with previous results reported in Oriental and Caucasian cohorts. Although CACNA1D is not the top genetic alteration responsible for APAs in these cohorts, CACNA1D may still be a potential key player in idiopathic hyperaldosteronism and aldosterone-producing micronodules. Thus further study is required to characterize the distribution of CACNA1D pathogenic mutations in bilateral PA.