Phenotype-genotype correlation in aldosterone-producing adenomas characterized by intracellular cholesterol metabolism
Phenotype-genotype correlation in aldosterone-producing adenomas characterized by intracellular cholesterol metabolism
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DOI：
10.1016/j.jsbmb.2022.106116
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发表时间：
2022-05-10
影响因子：
4.1
通讯作者：
Sasano, Hironobu
中科院分区：
文献类型：
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作者：
Harashima, Shogo;Yamazaki, Yuto;Sasano, Hironobu
醛固酮生成腺瘤（APA）在组织学上是由透明致密的肿瘤细胞组成。据报道，与非kcnj5突变的APAs相比，kcnj5突变的APAs与更高的血浆醛固酮浓度和更丰富的含脂滴的透明肿瘤细胞相关。然而，胆固醇摄取和/或合成、细胞形态和基因型之间的关系仍然未知。因此，为了探讨这些差异，52例APA病例(KCNJ5 mt: n = 33，非KCNJ5 mt: n = 19；ATP1A1: n = 3, ATP2B3: n = 3, CACNA1D: n = 5, CTNNB1: n = 1，以上无任何突变的肿瘤：n = 7)、APA附近的肾小球带（ZG）组织和10个非病理性肾上腺（NAs）进行肿瘤形态定量组织病理学分析和胆固醇受体（SRB1、LDL-R）、胆固醇代谢酶（ACAT1、ACAT2、HSL、DHCR24、StAR）和类固醇合成所需酶（CYP11A1、CYP17A、3 β HSD、CYP11B1、CYP11B2）的免疫组织化学分析。采用气相色谱-质谱（GC-MS）分析了21例APA患者（KCNJ5 mt: n = 16，非KCNJ5 mt: n = 5）和14例肾上腺皮质邻近肾上腺组织的胆固醇前体和代谢物。结果表明，SR-B1和DHCR24在ZG中的表达明显低于NAs的束状组织和网状组织，而LDL-R在免疫组化分析中无显著差异。SR-B1和DHCR24在APA中的表达均显著高于APA附近的ZG组织。在GC-MS分析中，除羊毛甾醇外，大多数胆固醇前体和代谢物及其代谢比（=胆固醇/前体浓度）在APAs中高于邻近的肾上腺皮质组织。LDL-R、ACAT1/2、HSL、DHCR24均明显低于APA致密性肿瘤细胞。KCNJ5突变APAs的LDL-R显著低于非KCNJ5突变APAs，胆固醇/毛甾醇比值显著高于非KCNJ5突变APAs。我们证明SR-B1介导的胆固醇酯的选择性摄取和新生胆固醇合成都在APAs中增强。此外，透明肿瘤细胞和致密肿瘤细胞的胆固醇摄取和代谢也不同。KCNJ5突变的APAs主要由含有丰富胆固醇酯的透明肿瘤细胞组成，但激活的LDL-R介导的摄取较少，新生合成增加。上述结果表明，在甾体生成和细胞内胆固醇代谢方面，它们与正常ZG细胞的功能偏差更为明显。
Aldosterone-producing adenoma (APA) is histologically composed of clear and compact tumor cells. KCNJ5mutated APAs were reported to be associated with higher plasma aldosterone concentration and more abundant clear tumor cells containing lipid droplets than non-KCNJ5- mutated APAs. However, the association among cholesterol uptake and/or synthesis, cellular morphology and genotypes has remained unknown. Therefore, in order to explore these differences, 52 APA cases (KCNJ5 mt: n = 33, non-KCNJ5 mt: n = 19; ATP1A1: n = 3, ATP2B3: n = 3, CACNA1D: n = 5, CTNNB1: n = 1, tumors without any mutation above: n = 7), zona glomerulosa (ZG) tissue adjacent to APA and 10 non-pathological adrenal glands (NAs) were examined for quantitative histopathological analysis of tumor morphology and immunohistochemical analysis of cholesterol receptors (SRB1, LDL-R), cholesterol metabolic enzymes (ACAT1, ACAT2, HSL, DHCR24, StAR), and the enzymes required for steroid synthesis (CYP11A1, CYP17A, 3 beta HSD, CYP11B1, CYP11B2). Gas chromatography-mass spectrometry (GC-MS) analysis was further performed to profile cholesterol precursors and metabolites in 21 APA cases (KCNJ5 mt: n = 16, non-KCNJ5 mt: n = 5) and 14 adrenal cortex of adjacent adrenal tissues. Results demonstrated that both SR-B1 and DHCR24 were significantly lower in the ZG than in fasciculata or reticularis of NAs but LDL-R was not significantly different among them in immunohistochemical analysis. SR-B1 and DHCR24 were both significantly higher in APAs than in ZG tissue adjacent to APA. In GC-MS analysis, most cholesterol precursors and metabolites, except for lanosterol, and their metabolic ratios (= concentration of cholesterol/ precursor) were higher in APAs than in the adjacent adrenal cortex tissue. LDL-R, ACAT1/2, HSL, DHCR24 were all significantly lower in clear than in compact tumor cells of APA. LDL-R was significantly lower and cholesterol/lanosterol ratio was significantly higher in KCNJ5- mutated than non-KCNJ5- mutated APAs. We demonstrated SR-B1 mediated selective uptake of cholesterol ester and de novo cholesterol synthesis were both enhanced in APAs. In addition, cholesterol uptake and metabolism were different between clear and compact tumor cells. KCNJ5- mutated APAs were predominantly composed of clear tumor cells containing abundant cholesteryl ester but less activated LDL-R mediated uptake and increased de novo synthesis. Those findings above indicated their more pronounced functional deviation from the normal ZG cells in terms of their steroidogenic and intracellular cholesterol metabolism.