JAK2V617F-Positive Endothelial Cells Induce Apoptosis and
Release JAK2V617F-Positive Microparticles

TOPRAK, SELIN FULYA; Sozer Tokdemir, Selcuk; Hekimoğlu, Hilal

doi:10.4274/tjh.galenos.2021.2021.0607

JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles

Hilal HEKİMOĞLU, (İstanbul Üniversitesi, Aziz Sancar Deneysel Tıp Enstitüsü, Genetik Anabilim Dalı, İstanbul, Türkiye)

Selin Fulya TOPRAK, (İstanbul Üniversitesi, Aziz Sancar Deneysel Tıp Enstitüsü, Genetik Anabilim Dalı, İstanbul, Türkiye)

Selçuk Sözer TOKDEMİR (İstanbul Üniversitesi, Aziz Sancar Deneysel Tıp Enstitüsü, Genetik Anabilim Dalı, İstanbul, Türkiye)

Turkish Journal of Hematology

9 0

Yıl: 2022 Cilt: 39 Sayı: 1 Sayfa Aralığı: 13 - 21 Metin Dili: İngilizce DOI: 10.4274/tjh.galenos.2021.2021.0607 İndeks Tarihi: 21-06-2022

JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles

Öz:

Objective: Philadelphia chromosome-negative myeloproliferative neoplasms (MPNs) have a high propensity for thrombosis, which has been attributed to increased blood counts, endothelial cell (EC) dysfunction, and inflammation. The presence of the JAK2V617F mutation in the ECs of MPN patients has been confirmed, but the consequences of EC involvement by JAK2V617F in the pathogenesis of thrombosis are unclear. Endothelial microparticles (EMPs) released from ECs play an important role in endothelial dysfunction and also in the intercellular exchange of biological signals and information. Several studies have revealed that patients with JAK2V617F and a thrombosis history have increased numbers of MPs in their circulation. Materials and Methods: The current study utilized a lentiviral transduction model of JAK2 wild type (JAK2wt) or JAK2V617F encoding green fluorescent protein (GFP) into human umbilical vein ECs to determine the effect of JAK2V617F on ECs. EC infected with JAK2V617F, JAK2WT, and only-GFP were tested after two days of culture. Results: The proteins of ECs that potentially play a role in the development of thrombosis, including endothelial protein C receptor, thrombomodulin, and tissue factor, were detected by flow cytometry analysis with no statistical significance. Increased annexin-V uptake of JAK2V617F and JAK2wt ECs compared to GFP-alone ECs was detected. The EMP production in the supernatants of the EC culture was investigated. Genotyping of the EMPs revealed the presence of genomic DNA and RNA fragments in EMP cargos. JAK2V617F-positive DNA was detected in EMPs released from JAK2V617F-infected ECs and EMPs were shown to carry the genotype of the cell of origin. Conclusion: JAK2V617F-positive EMPs were shown for the first time in the literature. This novel research provides the first evidence that EMPs might regulate neighboring and distant cells via their cargo materials. Thus, the direct effect of JAK2V617F on ECs and their functions suggests that different mechanisms might play a role in the pathogenesis of thrombosis in MPNs.

Anahtar Kelime:

JAK2V617F-Pozitif Endotel Hücreleri Apoptozu İndükler ve JAK2V617F-Pozitif Mikropartikül Salınımı Yapar

Öz:

Amaç: Philadelphia kromozom negatif miyeloproliferatif neoplazi’ler (MPN), yüksek tromboz riski taşıyarak inflamasyon, endotel hücre (EH) disfonksiyonu ve artan kan sayıları ile karakterize edilirler. Önceki çalışmalarda, MPN hastalarının EH’de JAK2V617F mutasyonunun varlığı tespit edilmiştir. Ancak tromboz patogenezinde JAK2V617F mutasyonu taşıyan EH’in rolü belirsizdir. EH’den salınan endotel mikropartikülleri (EMP’ler) biyolojik sinyallerin ve hücreler arası bilginin değişiminde ve endotel disfonksiyonunda önemli rol oynarlar. Birçok çalışmada, JAK2V617F mutasyonu taşıyan ve tromboz öyküsü olan hastaların dolaşımlarında artan sayıda mikropartikül olduğunu ortaya konulmuştur. Gereç ve Yöntemler: Çalışmamızda, JAK2V617F’nin EH üzerindeki etkisini belirlemek için yeşil fluoresan proteini (GFP) ile işaretli JAK2 yabanıl tip (JAK2WT) veya JAK2V617F’li lentiviral transdüksiyon modeli kullanılarak insan göbek kordonundan elde edilmiş endotel hücre hattı (HUVEC) enfekte edildi. JAK2V617F, JAK2WT ve sadece- GFP ile enfekte EH’ler iki gün süren kültür sonrasında test edildi. Bulgular: Akım ölçer analizinde, Endotelyal protein C reseptörü, trombomodulin ve doku faktörü dahil olmak üzere tromboz gelişiminde potansiyel rol oynayan EH proteinlerinde istatistiksel anlamlılık olmadığı tespit edildi. JAK2V617F ve JAK2WT enfekte EH’de annexin-V alımının sadece-GFP enfekte EH’ye kıyasla arttığı tespit edildi. EH kültüründen elde edilen süpernatanlarda EMP üretimi araştırıldı. EMP genotiplemesinde EMP kargolarında genomik DNA ve RNA fragmanları varlığı tespit edildi. JAK2V617F enfekte EH’den salınan EMP’lerde JAK2V617F pozitif DNA tespit edildi ve EMP’lerin köken aldıkları hücre genotipini taşıdığı gösterildi. Sonuç: Bu çalışma ile literatürde ilk defa, JAK2V617F pozitif EMP’ler gösterilmiştir. Bu yeni araştırma, EMP’lerin taşıdıkları kargolar aracılığıyla komşu ve uzaktaki hücreleri düzenleme potansiyeli olduğunu göstermektedir. Bu sayede JAK2V617F’nin EH ve işlevleri üzerindeki direkt etkisi ile MPN’lerde tromboz patogenezinde farklı mekanizmaların rolü olduğunu ileri sürer.

Anahtar Kelime:

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	Hekimoğlu H, TOPRAK S, Sozer Tokdemir S (2022). JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. , 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
Chicago	Hekimoğlu Hilal,TOPRAK SELIN FULYA,Sozer Tokdemir Selcuk JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. (2022): 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
MLA	Hekimoğlu Hilal,TOPRAK SELIN FULYA,Sozer Tokdemir Selcuk JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. , 2022, ss.13 - 21. 10.4274/tjh.galenos.2021.2021.0607
AMA	Hekimoğlu H,TOPRAK S,Sozer Tokdemir S JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. . 2022; 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
Vancouver	Hekimoğlu H,TOPRAK S,Sozer Tokdemir S JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. . 2022; 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
IEEE	Hekimoğlu H,TOPRAK S,Sozer Tokdemir S "JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles." , ss.13 - 21, 2022. 10.4274/tjh.galenos.2021.2021.0607
ISNAD	Hekimoğlu, Hilal vd. "JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles". (2022), 13-21. https://doi.org/10.4274/tjh.galenos.2021.2021.0607

APA	Hekimoğlu H, TOPRAK S, Sozer Tokdemir S (2022). JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. Turkish Journal of Hematology, 39(1), 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
Chicago	Hekimoğlu Hilal,TOPRAK SELIN FULYA,Sozer Tokdemir Selcuk JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. Turkish Journal of Hematology 39, no.1 (2022): 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
MLA	Hekimoğlu Hilal,TOPRAK SELIN FULYA,Sozer Tokdemir Selcuk JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. Turkish Journal of Hematology, vol.39, no.1, 2022, ss.13 - 21. 10.4274/tjh.galenos.2021.2021.0607
AMA	Hekimoğlu H,TOPRAK S,Sozer Tokdemir S JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. Turkish Journal of Hematology. 2022; 39(1): 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
Vancouver	Hekimoğlu H,TOPRAK S,Sozer Tokdemir S JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles. Turkish Journal of Hematology. 2022; 39(1): 13 - 21. 10.4274/tjh.galenos.2021.2021.0607
IEEE	Hekimoğlu H,TOPRAK S,Sozer Tokdemir S "JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles." Turkish Journal of Hematology, 39, ss.13 - 21, 2022. 10.4274/tjh.galenos.2021.2021.0607
ISNAD	Hekimoğlu, Hilal vd. "JAK2V617F-Positive Endothelial Cells Induce Apoptosis and Release JAK2V617F-Positive Microparticles". Turkish Journal of Hematology 39/1 (2022), 13-21. https://doi.org/10.4274/tjh.galenos.2021.2021.0607