urmjУральский медицинский журналUral Medical Journal2949-4389Ural State Medical University10.52420/2071-5943-2023-22-2-93-101urmj-1232Research ArticleОригинальные статьи | Original articlesOriginal articlesИммуноморфологическая характеристика экстрацеллюлярного матрикса костного мозга при ph-негативных миелопролиферативных заболеванияхImmunohistochemistry of bone marrow extracellular matrix in ph-negative myeloproliferative diseaseshttps://orcid.org/0000-0001-5425-6472ГоголеваД. В.GogolevaD. V.

Дарья Витальевна Гоголева − аспирант

Челябинск

Darya V. Gogoleva − Post-graduated student

Chelyabinsk

pathology249@mail.ruhttps://orcid.org/0000-0003-3251-6944СычуговГ. В.SychugovG. V.

Глеб Вячеславович Сычугов − кандидат медицинских наук

Челябинск

Gleb V. Sychugov − Ph.D. in medicine

Chelyabinsk

docsgv@gmail.comЮжно-Уральский государственный медицинский университетРоссияSouth Ural State Medical UniversityRussian Federation20233004202322293101Copyright © Гоголева Д.В., Сычугов Г.В., 20232023Гоголева Д.В., Сычугов Г.В.Gogoleva D.V., Sychugov G.V.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.umjusmu.ru/jour/article/view/1232Введение

Введение. Оценка иммуногистохимической экспрессии матриксных металлопротеиназ (MMP-2, MMP9), их ингибиторов (TIMP-1, TIMP-2), фибробластического фактора роста 2 (FGF2), трансформирующего фактора роста b1 (TGFB1) и коллагена 3 типа в трепанобиопсиях костного мозга пациентов с Ph-негативными миелопролиферативными заболеваниями (МПЗ) имеет большое значение.

Цель работы оценка экспрессии компонентов экстрацеллюлярного матрикса (MMP-2, MMP-9, TIMP1, TIMP-2, FGF2, TFGB1, Collagen III), участвующих в прогрессировании миелофиброза в трепанобиопсиях костного мозга в зависимости от мутационного статуса пациентов с ХМПЗ.

Материалы и методы

Материалы и методы. Было проанализировано 108 трепанобиопсий костного мозга пациентов с МПЗ, которые были разделены на 3 группы: JAK2-позитивные (n=62), CALR-позитивные (n=25) и трижды негативные (n=21). Все гистопрепараты были окрашены иммуногистохимическим методом с использованием антител к MMP-2, MMP-9, TIMP-1, TIMP-2, FGF2, TGFB1 и коллагену III и были оценены в программе ImageJ. Для выявления статистически значимых различий использовались критерии Краскелла-Уоллиса и Манна-Уитни. Также был подсчитан коэффициент корреляции Спирмена. Статистически достоверными считались различия p <0,05. Результаты. Экспрессия металлопротеиназы MMP2 определялась в цитоплазме мегакариоцитов. Экспрессия MMP9 определялась в цитоплазме нейтрофилов, макрофагов, а также в экстрацеллюлярном матриксе (ЭМ) микроокружения кроветворного костного мозга. Ингибитор металлопротеиназ TIMP1 экспрессировался только в ЭМ. Экспрессия TIMP2, Collagen III, FGF2 и TGFB1 определялась в цитоплазме мегакариоцитов и ЭМ. Выявлены различия между всеми тремя группами, наибольшие различия между группами JAK2 и CALR. Выявлена слабая и умеренная положительная корреляция между мутацией JAK2 и иммуногистохимической экспрессией компонентов ЭМ и слабая негативная корреляция между мутацией CALR и иммуногистохимической экспрессией компонентов ЭМ.

Обсуждение

Обсуждение. Наибольшее количество компонентов ЭМ выявлено в JAK2-позитивной группе, наименьшее – в CALR-позитивной группе. Так как экспрессия оценивалась во всех миелоидных клетках и компонентах костномозговых ниш, можно говорить, что данный вид оценки более полный, чем в ранее опубликованных исследованиях, где оценка производилась только в сыворотке крови, по культуре клеток и др.

Заключение

Заключение. Иммуногистохимическая экспрессия MMP-2, MMP-9, TIMP-1, TIMP-2, FGF2, TGFB1, collagen type III зависит от драйверных мутаций. Это может быть полезно для понимания патогенеза фиброза и оценки прогноза Ph-негативных МПЗ. 

Introduction Evaluation of immunohistochemical expression of matrix metalloproteinases 2 and 9 (MMP2, MMP9), their inhibitors (TIMP1, TIMP2), fibroblast growth factor (FGF2), transforming growth factor beta (TGFB1) and collagen type III in the bone marrow of patients with Ph-negative myeloproliferative neoplasms (MPN) is of great importance.

The aim of the study was the evaluation of expression of extracellular matrix components (MMP-2, MMP-9, TIMP1, TIMP-2, FGF2, TFGB1, Collagen III) involved in myelofibrosis progression in bone marrow trepan biopsies depending on mutational status of patients with CMPD.

Materials and methods We analyzed 108 bone marrow biopsies of patients with MPN, which were divided into 3 groups: JAK2-positive (n=62), CARL-positive (n=25) and triple-negative (n=21). Whole-slide sections were immunostained using antibodies against MMP-2, MMP-9, TIMP-1, TIMP-2, FGF2, TGFB1, collagen type III and scored by ImageJ plugin software. We used Kruskal- Wallis test and Mann-Whitney U-test for comparisons of differences in medians. Spearman’s rank order correlation was calculated. Statistical significance was set at p<0,05.

Results and Discussion MMP2 expression was observed in megakaryocytes. MMP9 expression was observed in neutrophils, macrophages and the bone marrow extracellular matrix (EM). TIMP1 expression was observed in the EM. TIMP-2, FGF2, TGFB1 and collagen type III expression was observed in megakaryocytes and the EM. Kruskal-Wallis test determined the differences between all 3 groups (MMP- 2 p< 0,001, MMP-9 p=0,023, TIMP-1 p< 0,001, TIMP-2 p< 0,001, FGF2 p< 0,001, TGFB1 p< 0,001, collagen type III p< 0,001). Mann-Whitney U-test determined the most differences between JAK2- and CALR-groups (MMP-2 p=0,001, MMP-9 p=0,001, TIMP-1 p=0,001, FGF2 p=0,001, TGFB1 p=0,001, collagen type III p=0,001), except TIMP-2. There was the weak and moderate positive correlation between JAK2-mutation and the immunohistochemistry expression of EM components, also the weak negative correlation between CALR-mutation and the immunohistochemistry expression of EM components.

Conclusion The bone marrow immunohistochemistry expression of MMP-2, MMP-9, TIMP-1, TIMP-2, FGF2, TGFB1, collagen type III depends on driver mutations. It may be useful for understanding of fibrosis pathogenesis and prognosis estimate of Ph-negative MPN.

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