ometendoОжирение и метаболизмObesity and metabolism2071-87132306-5524Endocrinology Research Centre10.14341/omet12741ometendo-12741Research ArticleНаучные исследованияOriginal studiesРезультаты иммуногистохимического исследования антител к CYP11B2 при первичном гиперальдостеронизмеThe results of immunohistochemical study of antibodies to CYP11B2 in primary hyperaldosteronismhttps://orcid.org/0000-0002-9117-4908РомановаН. Ю.RomanovaN. Yu.

Романова Наталья Юрьевна; eLibrary SPIN: 5288-2968

ул. Дмитрия Ульянова, д. 11, 117036 Москва, Россия

Natalia Yu. Romanova, MD

11 Dm. Ulyanova street, 117036 Moscow, Russia

natasharomanova1991@gmail.comhttps://orcid.org/0000-0001-6891-0009СеливановаЛ. С.SelivanovaL. S.

Селиванова Лилия Сергеевна, к.м.н.; eLibrary SPIN: 5151-3675

Москва

Liliya S. Selivanova, MD, PhD

Moscow

liselivanova89@yandex.ruhttps://orcid.org/0000-0001-6388-1544ПлатоноваН. М.PlatonovaN. M.

Платонова Надежда Михайловна, д.м.н.; eLibrary SPIN: 4053-3033

Москва

Nadezhda M. Platonova, MD, PhD

Moscow

doc-platonova@inbox.ruhttps://orcid.org/0000-0001-7098-4584БельцевичД. Г.BeltsevichD. G.

Бельцевич Дмитрий Германович, д.м.н., профессор; eLibrary SPIN: 4475-6327

Москва

Dmitriy G. Beltsevich, MD, PhD, professor

Moscow

belts67@gmail.comhttps://orcid.org/0000-0002-8520-8702ТрошинаЕ. А.TroshinaE. A.

Трошина Екатерина Анатольевна, д.м.н., профессор; eLibrary SPIN: 8821-8990

Москва

Ekaterina A. Troshina, MD, PhD, professor

Moscow

troshina@inbox.ruНациональный медицинский исследовательский центр эндокринологииРоссияEndocrinology Research CentreRussian Federation202117062021183245253Copyright © Романова Н.Ю., Селиванова Л.С., Платонова Н.М., Бельцевич Д.Г., Трошина Е.А., 20212021Романова Н.Ю., Селиванова Л.С., Платонова Н.М., Бельцевич Д.Г., Трошина Е.А.Romanova N.Y., Selivanova L.S., Platonova N.M., Beltsevich D.G., Troshina E.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.omet-endojournals.ru/jour/article/view/12741Обоснование

Обоснование. Альдостерон-продуцирующая аденома (АПА) — наиболее частая причина диагностированного первичного гиперальдостеронизма (ПГА). Известно, что ключевым ферментом синтеза альдостерона, участвующим в патогенезе ПГА, является альдостеронсинтаза (CYP11B2). Недавние исследования показали, что различные факторы влияют на экспрессию и функцию CYP11B2 при АПА. В частности, соматические мутации, приводящие к повышенной и сниженной функции CYP11B2, были идентифицированы в нескольких генах, каждый из которых кодирует ключевой белок, влияющий на кальциевый сигнальный путь, экспрессию CYP11B2 и продукцию альдостерона. С другой стороны, CYP11B2 также катализирует превращение кортизола в 18-гидроксикортизол и впоследствии превращает 18-­гидроксикортизол в 18-оксокортизол. Также в статье обсуждается клиническое значение 18-оксокортизола — важного биомаркера для диагностики ПГА. Соматические мутации в генах-драйверах альдостерона тесно связаны с экспрессией CYP11B2 и были обнаружены только в CYP11B2-положительной области опухоли, что указывает на неоднородную экспрессию CYP11B2 в опухолевых клетках.

Цель

Цель. Оценить экспрессию антител к CYP11B2 в новообразованиях коркового слоя надпочечников при первичном гиперальдостеронизме.

Материалы и методы

Материалы и методы. Ретроспективная оценка опухолей надпочечников, удаленных в ходе адреналэктомии, у пациентов с лабораторно подтвержденным ПГА (n=20). По данным мультиспиральной компьютерной томографии односторонняя аденома была выявлена у 19 пациентов (95% от общего числа), во всех случаях подтверждена односторонняя гиперпродукция альдостерона данными сравнительного селективного забора крови из надпочечниковых вен. На отобранных опухолях проводилось иммуногистохимическое исследование с использованием антител к CYP11B2. В соответствии со стандартными протоколами использовался biotin-free Bond Polymer Refine Detection system (Leica) с автоматизированной системой Bond max (Leica). В каждой опухоли было выбрано окрашенное поле, увеличенное в 200 раз. Эти поля были оцифрованы в виде изображений JPEG с использованием цифровой камеры (Leica, DFC 490). После чего была оценена экспрессия CYP11B2 в аденоме и в соседней коре надпочечников.

Результаты

Результаты. При иммуногистохимическом исследовании опухоли надпочечников у пациентов с ПГА (n=20) получена положительная экспрессия с антителом к CYP11B2 во всех случаях, при этом: в 50% случаев (n=10) — выявлена интенсивная реакция в опухоли и прилежащем корковом слое, в 10% случаев (n=2) — умеренная реакция в опухоли и прилежащем корковом слое, в 15% (n=3) — слабая реакция в опухоли и прилежащем корковом слое, а также в 25% случаев (n=5) — интенсивная реакция в прилежащем корковом слое более выражена, чем в опухоли надпочечника.

Заключение

Заключение. Доказана функциональная гетерогенность опухоли надпочечников при ПГА.

Background

Background: Aldosterone-producing adrenocortical adenoma (APA) is responsible for the majority of cases clinically diagnosed as primary aldosteronism. Aldosterone synthase (CYP11B2) is one of the enzymes that play essential roles in aldosterone synthesis and is involved in the pathogenesis of primary aldosteronism. Recent studies have demonstrated that various factors influence the expression and function of CYP11B2 in APA. In particular, somatic mutations, such as gain-of-function and loss-of-function mutations have been identified in several genes, each of which encodes a pivotal protein that affects the calcium signaling pathway, the expression of CYP11B2, and aldosterone production. On the other hand, CYP11B2 also catalyzes the conversion of cortisol to 18-hydroxycortisol and subsequently converts 18-hydroxycortisol to 18-oxocortisol. The article also discusses the clinical significance of 18-oxocortisol, an important biomarker for the diagnosis of primary aldosteronism. Somatic mutations in aldosterone-driver genes are strongly associated with CYP11B2 expression and have been only detected in the CYP11B2-positive tumor area, that indicating heterogeneous expression of CYP11B2 in tumor.

Aim

Aim: to assess the functional heterogeneity of adrenal tumors in primary aldosteronism

Materials and methods

Materials and methods: Retrospective evaluation adrenal tumors from patients with primary aldosteronism (n=20). According to CT unilateral macrohyperplasia was detected in 19 patients (95% of total), all of them were confirmed to have unilateral hyperproduction of aldosterone according to AVS. Selected tumors werestained with anty-CYP11B2 antibody. We evaluated the CYP11B2 expression in the adenoma and in the adjacent adrenal cortex.

Results

Results: Immunohistochemistry studies of the resected adrenals from 20 patients with PA operated due to unilateral production of aldosterone using CYP11B2 staining showed that 10 of those with an adenoma on CT scanning showed CYP11B2 staining in the adenoma. Furthermore, 5 cases of an unilateral adenoma, showed CYP11B2 staining in the adjacent adrenal cortex and an absence of staining for CYP11B2 in the adenoma. 5 cases showed CYP11B2 expression is heterogeneously immunolocalized throughout the tumor area.

Conclusion

Conclusion: Thus, the functional heterogeneity of adrenal tumors in primary aldosteronism has been proven. It is necessary to compare the data of immunohistochemical studies on the expression of CYP11B2 with the indicators of the level of 18-hydroxycortisol, 18-oxocortisol.

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The authors declare that there are no conflicts of interest present.