Disordered fetal adrenal steroidogenesis can cause marked clinical effects including virilization of female fetuses. In postnatal life, adrenal disorders can be life-threatening due to the risk of adrenal crisis and must be carefully managed. However, testing explicit adrenal steroidogenic inhibitory effects of therapeutic drugs is challenging due to species-specific characteristics, and particularly the impact of adrenocorticotropic hormone (ACTH) stimulation on drugs targeting steroidogenesis has not previously been examined in human adrenal tissue. Therefore, this study aimed to examine the effects of selected steroidogenic inhibitors on human fetal adrenal (HFA) steroid hormone production under basal and ACTH-stimulated conditions. This study used an established HFA ex vivo culture model to examine treatment effects in 78 adrenals from 50 human fetuses (gestational weeks 8–12). Inhibitors were selected to affect enzymes critical for different steps in classic adrenal steroidogenic pathways, including CYP17A1 (Abiraterone acetate), CYP11B1/2 (Osilodrostat), and a suggested CYP21A2 inhibitor (Efavirenz). Treatment effects were examined under basal and ACTH-stimulated conditions in tissue from the same fetus and determined by quantifying the secretion of adrenal steroids in the culture media using liquid chromatography-tandem mass spectrometry. Statistical analysis was performed on ln-transformed data using one-way ANOVA for repeated measures followed by Tukey’s multiple comparisons test. Treatment with Abiraterone acetate and Osilodrostat resulted in potent inhibition of CYP17A1 and CYP11B1/2, respectively, while treatment with Efavirenz reduced testosterone secretion under basal conditions. ACTH-stimulation affected the inhibitory effects of all investigated drugs. Thus, treatment effects of Abiraterone acetate were more pronounced under stimulated conditions, while Efavirenz treatment caused a non-specific inhibition on steroidogenesis. ACTH-stimulation prevented the Osilodrostat-mediated CYP11B1 inhibition observed under basal conditions. Our results show that the effects of steroidogenic inhibitors differ under basal and ACTH-stimulated conditions in the HFA ex vivo culture model. This could suggest that in vivo effects of therapeutic drugs targeting steroidogenesis may vary in conditions where patients have suppressed or high ACTH levels, respectively. This study further demonstrates that ex vivo cultured HFAs can be used to evaluate steroidogenic inhibitors and thereby provide novel information about the local effects of existing and emerging drugs that targets steroidogenesis.

中文翻译：

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所选抑制剂对人胎儿肾上腺类固醇生成的影响在基础和 ACTH 刺激条件下不同

胎儿肾上腺类固醇生成障碍可引起显着的临床影响，包括女性胎儿男性化。在产后生活中，由于肾上腺危象的风险，肾上腺疾病可能会危及生命，必须小心处理。然而，由于物种特异性特征，测试治疗药物的显式肾上腺类固醇生成抑制作用具有挑战性，特别是促肾上腺皮质激素 (ACTH) 刺激对靶向类固醇生成药物的影响之前尚未在人类肾上腺组织中进行过检查。因此，本研究旨在检查选定的类固醇生成抑制剂在基础和 ACTH 刺激条件下对人胎儿肾上腺 (HFA) 类固醇激素产生的影响。本研究使用已建立的 HFA 离体培养模型来检查来自 50 个人类胎儿（孕周 8-12）的 78 个肾上腺的治疗效果。选择抑制剂来影响对经典肾上腺类固醇生成途径的不同步骤至关重要的酶，包括 CYP17A1（醋酸阿比特龙）、CYP11B1/2（Osilodrostat）和建议的 CYP21A2 抑制剂（Efavirenz）。在来自同一胎儿的组织中在基础和 ACTH 刺激条件下检查治疗效果，并通过使用液相色谱-串联质谱法量化培养基中肾上腺类固醇的分泌来确定治疗效果。使用单向方差分析对 ln 转换的数据进行统计分析，以进行重复测量，然后进行 Tukey 的多重比较检验。用醋酸阿比特龙和奥西洛他治疗分别导致对 CYP17A1 和 CYP11B1/2 的有效抑制，而用依法韦仑治疗减少基础条件下的睾酮分泌。ACTH 刺激影响所有研究药物的抑制作用。因此，醋酸阿比特龙的治疗效果在刺激条件下更为显着，而依法韦仑治疗对类固醇生成造成非特异性抑制。ACTH 刺激阻止了在基础条件下观察到的 Osilodrostat 介导的 CYP11B1 抑制。我们的研究结果表明，在 HFA 离体培养模型中，类固醇生成抑制剂的作用在基础和 ACTH 刺激条件下不同。这可能表明，针对类固醇生成的治疗药物的体内作用可能会在患者分别抑制或高 ACTH 水平的情况下发生变化。该研究进一步表明，离体培养的 HFA 可用于评估类固醇生成抑制剂，从而提供有关针对类固醇生成的现有和新兴药物的局部作用的新信息。