problendoПроблемы ЭндокринологииProblems of Endocrinology0375-96602308-1430Endocrinology Research Centre10.14341/probl10263problendo-10263Research ArticleКлиническая эндокринологияClinical endocrinologyЭкспрессия циркулирующих микроРНК в плазме у пациентов с акромегалиейExpression of plasma microRNA in patients with acromegalyhttps://orcid.org/0000-0002-9314-7831ЛуценкоАлександр СергеевичLutsenkoAlexander S.

научный сотрудник отделения нейроэндокринологии и остеопатий

research scientist of neuroendocrinology and bone diseases department

some91@mail.ruhttps://orcid.org/0000-0002-6674-6441БелаяЖанна ЕвгеньевнаBelayaZhanna E.

д.м.н., зав. отделения нейроэндокринологии и остеопатий,

профессор кафедры эндокринология, института высшего и дополнительного образования 

MD, PhD, Professor

jannabelaya@gmail.comhttps://orcid.org/0000-0001-9119-2447ПржиялковскаяЕлена ГеоргиевнаPrzhiyalkovskayaElena G.

к.м.н.

PhD

przhiyalkovskaya.elena@gmail.comhttps://orcid.org/0000-0001-9762-3383НикитинАлексей ГеоргиевичNikitinAlexey G.

к.б.н.

PhD

avialn@gmail.comhttps://orcid.org/0000-0001-9512-9277КошкинФилипп АлександровичKoshkinPhilipp A.

к.б.н.

PhD

philipkoshkin@gmail.comhttps://orcid.org/0000-0003-4353-6705ЛапшинаАнастасия МихайловнаLapshinaAnastasiya M.

Кандидат медицинских наук, врач-патологоанатом отдела фундаментальной патоморфологии

MD, PhD

nottoforget@yandex.ruhttps://orcid.org/0000-0002-6993-5096ХандаеваПатимат МагомедовнаKhandaevaPatimat M.

научный сотрудник отделения нейроэндокринологии и остеопатий

Research Scientist of Neuroendocrinological department

pati_khandaeva@mail.ruhttps://orcid.org/0000-0002-5634-7877МельниченкоГалина АфанасьевнаMelnichenkoGalina A.

д.м.н., профессор, академик РАН

MD, PhD, Professor

teofrast2000@mail.ruНациональный медицинский исследовательский центр эндокринологии Минздрава РоссииРоссияEndocrinology Research CentreRussian FederationФГБУ «Национальный медицинский исследовательский центр эндокринологии» Минздрава РоссииРоссияEndocrinology Research CentreRussian FederationНаучно-исследовательский институт пульмонологииРоссияPulmonology Scientific Research Institute under FMBA of RussiaRussian FederationМедико-генетический центр «Геномед»РоссияCenter of medical genetics «Genomed»Russian Federation201923112019655311318Copyright © Луценко А.С., Белая Ж.Е., Пржиялковская Е.Г., Никитин А.Г., Кошкин Ф.А., Лапшина А.М., Хандаева П.М., Мельниченко Г.А., 20192019Луценко А.С., Белая Ж.Е., Пржиялковская Е.Г., Никитин А.Г., Кошкин Ф.А., Лапшина А.М., Хандаева П.М., Мельниченко Г.А.Lutsenko A.S., Belaya Z.E., Przhiyalkovskaya E.G., Nikitin A.G., Koshkin P.A., Lapshina A.M., Khandaeva P.M., Melnichenko G.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.probl-endojournals.ru/jour/article/view/10263Обоснование

Обоснование. МикроРНК – класс малых некодирующих молекул РНК, участвующих в посттранскрипционной регуляции экспрессии генов. Они также обнаруживаются в крови в стабильных концентрациях, что делает их перспективными биомаркерами различных заболеваний.

Цель

Цель. Определить циркулирующие микроРНК, различно экспрессирующиеся у пациентов с акромегалией по сравнению со здоровым контролем.

Методы

Методы: проведено одноцентровое, одномоментное, выборочное исследование случай–контроль: оценка экспрессии микроРНК в плазме крови у пациентов с акромегалией по сравнению со здоровым контролем. Забор крови проводился натощак, в течение 30 мин после забора крови образцы цельной крови однократно центрифугировались при температуре +5 °С на скорости вращения 3000 об/мин в течение 20 мин. Далее образцы плазмы замораживались и хранились при температуре –80 °C. Выделение РНК и подготовка адаптеров и библиотек для секвенирования выполнялись согласно протоколам производителей. Анализ экспрессии произведен на секвенаторе NextSeq. Биоинформатический анализ проводился с помощью программного обеспечения atropos (удаление адаптеров), STAR (выравнивание), FastQC (контроль качества), seqbuster/seqcluster/miRge2 (аннотация микроРНК, поиск isomiR, поиск новых микроРНК, анализ экспрессии). Первичная конечная точка исследования – показатели экспрессии микроРНК плазмы у пациентов с акромегалией в сравнении со здоровым контролем.

Результаты

Результаты. В исследование включены 12 пациентов с акромегалией – возраст 33,1 года [20; 47], ИМТ 29,3 кг/м2 [24,0; 39,6], ИРФ-1 686,10 нг/мл [405,90; 1186,00] и 12 здоровых добровольцев – возраст 36,2 года [26; 44], ИМТ 26,7 кг/м2 [19,5; 42,5], ИРФ-1 210,40 нг/мл [89,76; 281,90]; соотношение по полу в обеих группах – 4 мужчин, 8 женщин. Группы не различались по полу, возрасту и индексу массы тела (p>0,05). У пациентов с акромегалией выявлено снижение экспрессии четырех микроРНК в плазме крови: miR-4446-3p –1,317 (p=0,001), miR-215-5p –3.040 (p=0,005), miR-342-5p –1,875 (p=0,013) и miR-191-5p –0,549 (p=0,039). Однако, после поправки на множественность сравнений ни одно из различий не достигло статистической достоверности (q >0,1).

Заключение

Заключение. В ходе исследования методом высокопроизводительного секвенирования определены четыре микроРНК, экспрессия которых может отличаться у пациентов с активной акромегалией по сравнению со здоровым контролем. Для подтверждения полученных результатов необходима валидизация другим методом оценки экспрессии на большей выборке.

BACKGROUND

BACKGROUND: microRNA is a class of small non-coding RNA molecules involved in posttranscriptional regulation of gene expression. MicroRNAs are detectable in blood in stable concentrations, which makes them promising biomarkers for various diseases.

AIM

AIM: to assess plasma microRNA expression in patients with active acromegaly compared with healthy controls.

MATERIAL AND METHODS

MATERIAL AND METHODS: single-center, case-control study: assessment of plasma microRNA in patients with acromegaly compared with healthy controls. Fasting blood samples were drawn and centrifuged at +5°С temperature and 3000 rpm for 20 minutes, then aliquoted and frozen at –80°C until further analysis. MicroRNA extraction and library preparation was done according to manufacturer’s instructions. Expression analysis was performed on NextSeq sequencer. Bioinformatic analysis using atropos (adapted deletion), STAR (aligning), FastQC (quality control), seqbuster/seqcluster/miRge2 (microRNA annotation, isomiR and new microRNA search, expression analysis). Primary endpoint of the study – differential expression of plasma microRNA in patients with acromegaly compared with healthy controls.

RESULTS

RESULTS: we included 12 patients with acromegaly – age 33.1 [20; 47], BMI 29.3 kg/m2 [24.0; 39.6], IGF-1 686.1 ng/mL [405.9; 1186.0] and 12 healthy subjects – age 36.2 [26; 44], BMI 26.7 kg/m2 [19.5; 42.5], IGF-1 210.4 ng/mL [89.76; 281.90]; gender ratio for both groups – 4 males, 8 females. The groups did not differ in gender (p=0.666), age (p=0.551) and BMI (p=0.378). We found decreased expression of four microRNAs in patients with acromegaly: miR-4446-3p –1.317 (p=0.001), miR-215-5p –3.040 (p=0.005), miR-342-5p –1.875 (p=0.013) and miR-191-5p –0.549 (p=0.039). However, none of these changes were statistically significant after adjustment for multiple comparisons (q >0.1).

CONCLUSION

CONCLUSION: we found four microRNAs, which could potentially be downregulated in plasma of patients with acromegaly. The result need to be validated using different measurement method with larger sample size.

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