Functional analysis of rare anti-Müllerian hormone protein-altering variants identified in women with PCOS.
Functional analysis of rare anti-Müllerian hormone protein-altering variants identified in women with PCOS.
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在多囊卵巢综合征患者中发现的罕见的抗苗勒氏激素蛋白改变变体的功能分析。
DOI：
10.1093/molehr/gaad011
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发表时间：
2023-04-29
影响因子：
4
通讯作者：
Visser, J. A.
中科院分区：
文献类型：
--
作者：
Meng, L.;McLuskey, A.;Dunaif, A.;Visser, J. A.
最近，在多囊卵巢综合征（PCOS）女性中发现了罕见的杂合AMH蛋白改变变体，导致抗苗勒管激素（AMH）信号转导减少。然而，确切的功能机制仍然未知。在这里，我们分析了这些AMH变体的加工，分泌和信号传导。在小鼠颗粒细胞系KK-1中对六种PCOS特异性AMH变体（V12 G、P151 S、P270 S、P352 S、P362 S、H506 Q）和一种对照特异性变体（A519 V）进行功能分析。与野生型（wt）AMH信号传导相比，人（h）AMH-151 S和hAMH-506 Q的AMH信号传导降低了约90%。hAMH-151 S或hAMH-506 Q与wt-hAMH的共表达剂量依赖性地抑制wt-hAMH信号传导。Western blotting显示hAMH-151 S和hAMH-506 Q蛋白在细胞裂解物中被检测到，但在上清液中未被检测到。共聚焦显微镜显示，与表达wt-hAMH的细胞相比，表达hAMH-151 S和hAMH-506 Q的HEK 293细胞具有更高的细胞AMH蛋白水平和内质网（ER）滞留。使用两种AMH ELISA试剂盒，在细胞裂解物中检测到hAMH-151 S，而在上清液中仅检测到非常低的水平。使用自动AMH ELISA可检测到hAMH-362 S和hAMH-519 V，但在手动ELISA中显示出严重降低的免疫活性。令人惊讶的是，使用任一ELISA在细胞裂解物和上清液中都检测不到hAMH-506 Q。然而，在PCOS病例中，P151 S和H506 Q变异体的杂合携带者在两种检测中仍有可检测到的AMH。因此，P151 S和H506 Q破坏AMH的正常加工和分泌，导致ER滞留。此外，AMH变体可损害AMH免疫活性。当PCOS患者血清AMH水平相对较低时，可考虑AMH变异。
Recently, rare heterozygous AMH protein-altering variants were identified in women with polycystic ovary syndrome (PCOS), causing reduced anti-Müllerian hormone (AMH) signaling. However, the exact functional mechanism remains unknown. Here, we analyzed the processing, secretion, and signaling of these AMH variants. Functional analysis of six PCOS-specific AMH variants (V12G, P151S, P270S, P352S, P362S, H506Q) and one control-specific variant (A519V) was performed in the mouse granulosa cell-line KK-1. Human (h) AMH-151S and hAMH-506Q have ∼90% decreased AMH signaling compared to wild-type (wt) AMH signaling. Coexpression of hAMH-151S or hAMH-506Q with wt-hAMH dose-dependently inhibited wt-hAMH signaling. Western blotting revealed that hAMH-151S and hAMH-506Q proteins were detected in the cell lysate but not in the supernatant. Confocal microscopy showed that HEK293 cells expressing hAMH-151S and hAMH-506Q had higher cellular AMH protein levels with endoplasmic reticulum (ER) retention compared to cells expressing wt-hAMH. Using two AMH ELISA kits, hAMH-151S was detected in the cell lysate, while only very low levels were detected in the supernatant. Both hAMH-362S and hAMH-519V were detectable using the automated AMH ELISA but showed severely reduced immunoactivity in the manual ELISA. Surprisingly, hAMH-506Q was undetectable in both the cell lysate and supernatant using either ELISA. However, in PCOS cases, heterozygous carriers of the P151S and H506Q variants still had detectable AMH in both assays. Thus, P151S and H506Q disrupt normal processing and secretion of AMH, causing ER retention. Additionally, AMH variants can impair the AMH immunoactivity. An AMH variant may be considered when serum AMH levels are relatively low in PCOS cases.
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影响因子：
11
作者：
de Vrij FM;Bouwkamp CG;Gunhanlar N;Shpak G;Lendemeijer B;Baghdadi M;Gopalakrishna S;Ghazvini M;Li TM;Quadri M;Olgiati S;Breedveld GJ;Coesmans M;Mientjes E;de Wit T;Verheijen FW;Beverloo HB;Cohen D;Kok RM;Bakker PR;Nijburg A;Spijker AT;Haffmans PMJ;Hoencamp E;Bergink V;GROUP Study Consortium;Vorstman JA;Wu T;Olde Loohuis LM;Amin N;Langen CD;Hofman A;Hoogendijk WJ;van Duijn CM;Ikram MA;Vernooij MW;Tiemeier H;Uitterlinden AG;Elgersma Y;Distel B;Gribnau J;White T;Bonifati V;Kushner SA
通讯作者：
Kushner SA
DOI：
10.1073/pnas.2104809118
发表时间：
2021-06-29
影响因子：
11.1
作者：
Hart KN;Stocker WA;Nagykery NG;Walton KL;Harrison CA;Donahoe PK;Pépin D;Thompson TB
通讯作者：
Thompson TB
影响因子：
5.8
作者：
Gorsic, Lidija K.;Kosova, Gulum;Urbanek, Margrit
通讯作者：
Urbanek, Margrit
影响因子：
6.7
作者：
Chang, Hsun-Ming;Klausen, Christian;Leung, Peter C. K.
通讯作者：
Leung, Peter C. K.
影响因子：
6.1
作者：
Fauser, BCJM;Chang, J;Lobo, R
通讯作者：
Lobo, R