FMF Kliniğine Alternatif Bir Bakış Açısı: MCP-1 (A-2518G) ve CCR2 (G190A) Polimorfizmleri ve MCP1 Ekspresyonu

Yıl 2022, , 804 - 810, 30.09.2022

Şenol Çitli , Nadir Koçak

https://doi.org/10.16899/jcm.1164970

Öz

Amaç: Ailevi Akdeniz Ateşi (AAA) otoinflamatuar bir hastalıktır ve çeşitli klinik bulgular olarak kendini gösterebilir. Kemokinler, inflamatuar sürecin önemli unsurlarıdır. MCP-1 ve onun reseptörü CCR2, FMF'de kritik roller oynayabilen monositler/makrofajlar için ana kemokinlerdir. Bundan dolayı MEFV gen fonksiyonunu etkileyebilecek MCP-1 (A-2518G) ve CCR2 (G190A) polimorfizmlerinin ve MCP-1 ekspresyon düzeyinin araştırılması amaçlanmıştır.
Gereç ve Yöntem: FMF'li hastalar Tel-Hashomer kriterlerine göre belirlendi. Elde edilen kan örneklerinden DNA ve RNA izole edildi. Genotipleme analizi, PCR-RFLP tekniği ile yapıldı. Ayrıca Real-time PCR yöntemi ile ekspresyon analizleri yapıldı. Elde edilen sonuçlar istatistiksel olarak değerlendirildi.
Bulgular: Çalışmaya toplam 229 birey (125 erkek ve 104 kadın) dahil edildi. Bunlardan 120 kişide FMF kliniği bulunurken, 107 kişide yoktu. Kalan iki kişi şüpheli klinik duruma sahipti. Çalışmaya alınan bireyler MEFV genotiplemesine göre değerlendirildiğinde ise 75 birey homozigot mutant, 77 birey Heterozigot saptanırken 77 birey ise MEFV geninde mutasyon taşımıyordu. Yapılan analizde Hem FMF kliniği hem de MEFV genotipleri ile MCP-1 (A-2518G) ve CCR2 (G190A) genotipleri arasında anlamlı bir ilişki bulunmadı. Ekspresyon analizinde, FMF kliniği olan hastalarda olmayanlara göre MCP-1 ekspresyonu artmış olarak saptandı. Ayrıca heterozigot MEFV grubunda mutasyonu olmayanlara göre MCP-1 ekspresyonu artmış olarak saptandı, Dahası homozigot MEFV grubunda MCP-1 ekspresyonu en yüksek düzeydeydi. Ek olarak, MCP-1 (A-2518G) genotiplendirmesine göre, MCP-1 ekspresyonu, Wild type gruba kıyasla hem homozigot hem de heterozigot gruplarda yükselmiştir.
Sonuç: FMF hastalığında MCP-1 ekspresyonu artmış olup, bu durum FMF hastaları arasındaki klinik farklılıkları açıklayabilir. MEFV mutasyonları, MCP-1 transkripsiyonunu artırarak inflamasyonu şiddetlendirebilir. MCP-1(A-2518G) mutasyonlu hastalarda MCP-1 ekspresyonu artar, bu da FMF kliniğini ağırlaştırır.
Anahtar Kelimeler: Ailevi Akdeniz Ateşi, MCP-1, CCR2, Expresyon analizi

Anahtar Kelimeler

Ailevi Akdeniz Ateşi, MCP-1, CCR2, Expresyon analizi

Destekleyen Kurum

cübap

Proje Numarası

t-511

Teşekkür

Yazarlar, çalışmanın planlanmasındaki desteğinden dolayı emekli Prof. Dr. İlhan SEZGİN'e teşekkürlerini sunar.

An Alternative Perspective to the FMF Clinic: MCP-1 (A-2518G) and CCR2 (G190A) Polymorphisms and MCP1 Expression

Öz

Background: Familial Mediterranean Fever (FMF) is an autoinflammatory disease and may express as various clinical findings. Chemokines are crucial elements of the inflammatory process. MCP-1 and its’ receptor CCR2 are the main chemokines for monocytes/macrophages that may play critical roles in FMF. Thus, it was aimed to investigate the MCP-1 (A-2518G) and CCR2 (G190A) polymorphisms and MCP-1 expression level, which may affect MEFV gene function.
Material and Method: Patients with FMF were identified according to the Tel-Hashomer criteria. DNA and RNA were isolated from the obtained blood samples. Genotyping analysis was performed by PCR-RFLP technique. In addition, expression analyzes were performed by Real-time PCR method. The obtained results were evaluated statistically.
Results: A total of 229 individuals (125 male and 104 female) were included in the study. While 120 individuals had FMF clinic, and 107 individuals did not have. The remaining two individuals had suspicious clinical status. In addition, while 75 individuals were homozygous mutants, 77 individuals were heterozygous mutants, and 77 individuals did not carry mutation in the MEFV gene. No significant relationship was found in between both FMF clinic and MEFV genotypes, and MCP-1 (A-2518G) and CCR2 (G190A) genotypes. In the expression analysis, MCP-1 expression increased in patients with FMF clinic compared to those without. In addition, MCP-1 expression was increased in the heterozygous MEFV group compared to those without mutation, moreover, the expression level was highest in homozygous MEFV group. In addition, according to the MCP-1 (A-2518G) genotyping, MCP-1 expression elevated in the homozygous as well as the heterozygous groups, compared to the Wild type group.
Conclusion: MCP-1 expression is increased in FMF disease, which may explain the clinical differences between FMF patients. MEFV mutations may exacerbate inflammation by increasing MCP-1 transcription. MCP-1 expression is increased in patients with MCP-1(A-2518G) mutations, which aggravates FMF clinic. MCP-1 expression may be assessed as a marker in suspicious cases.
Keywords: Familial Mediterranean Fever, MCP-1, CCR2, expression

Anahtar Kelimeler

Familial Mediterranean Fever, MCP-1, CCR2, expression

Proje Numarası

t-511

Kaynakça

• Ben-Chetrit E, Levy M. Familial mediterranean fever. Lancet 11998;351:659-64.
• Mortensen SB, Hansen A, Byg K-E, et al. Monocyte secretory profiling in a clinical and MEFV genotype-characterized cohort of Danish familial Mediterranean fever patients: diagnostic potential of CCL1 and CXCL1. J Scand J Rheumatol 2022; 1-9.
• Masters SL, Simon A, Aksentijevich I, Kastner DL. Horror autoinflammaticus: the molecular pathophysiology of autoinflammatory disease. J Ann Rev Immunol 2009;27:621.
• Ben‐Chetrit E, Touitou I. Familial Mediterranean fever in the world. J Arthritis Care Res 2009;61:1447-53.
• Ben-Chetrit E, Yazici H. Familial Mediterranean fever: different faces around the world. J Clin Exp Rheumatol 2019;37:S18-22.
• Van Gijn ME, Ceccherini I, Shinar Y, et al. New workflow for classification of genetic variants’ pathogenicity applied to hereditary recurrent fevers by the International Study Group for Systemic Autoinflammatory Diseases (INSAID). J Med Gen 2018;55:530-7.
• Akkaya-Ulum YZ, Akbaba TH, Tavukcuoglu Z, Chae JJ, Yilmaz E, Ozen S, et al. Familial Mediterranean fever-related miR-197-3p targets IL1R1 gene and modulates inflammation in monocytes and synovial fibroblasts. J Scientific Reports 2021;11:1-10.
• Gunpinar S, Alptekin NO, Ucar VB, Acar H. Frequency of MCP-1 (rs1024611) and CCR2 (rs1799864) gene polymorphisms and its effect on gene expression level in patients with AgP. J Arch Oral Biol 2017;80:209-16.
• Sima C, Glogauer M. Macrophage subsets and osteoimmunology: tuning of the immunological recognition and effector systems that maintain alveolar bone. J Periodontol 2013;63:80-101.
• Walczak A, Przybyłowska-Sygut K, Sygut A, et al. An association of the MCP-1 and CCR2 single nucleotide polymorphisms with colorectal cancer prevalence. J Polish J Surg 2017;89:1-5.
• Taghavi Y, Hassanshahi G, Kounis NG, Koniari I, Khorramdelazad H. Monocyte chemoattractant protein-1 (MCP-1/CCL2) in diabetic retinopathy: latest evidence and clinical considerations. J Cell Commun Signal 2019;13:451-62.
• Liehn EA, Piccinini A-M, Koenen RR, et al. A new monocyte chemotactic protein-1/chemokine CC motif ligand-2 competitor limiting neointima formation and myocardial ischemia/reperfusion injury in mice. J Am Coll Cardiol 2010;56:1847-57.
• Leuschner F, Dutta P, Gorbatov R, et al. Therapeutic siRNA silencing in inflammatory monocytes in mice. J Nat Biotechnol 2011;29(11):1005-10.
• Zarebska JM, Chanalaris A, Driscoll C, et al. CCL2 and CCR2 regulate pain-related behaviour and early gene expression in post-traumatic murine osteoarthritis but contribute little to chondropathy. J Osteoarthritis Cartilage 2017;25:406-12.
• Ben-Zvi I, Herskovizh C, Kukuy O, Kassel Y, Grossman C, Livneh A. Familial Mediterranean fever without MEFV mutations: a case–control study. J Orphanet J Rare Dis 2015;10:1-6.
• Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2− ΔΔCT method. Methods 2011;25:402-8.
• Bouomrani S, Masmoudi I, Teber SB. Familial Mediterranean fever: What associations to screen for?. Reumatologia 2020;58:150-4.
• Ozen S. Changing concepts in familial Mediterranean fever: is it possible to have an autosomal-recessive disease with only one mutation?. Arthritis Rheum 2009;60:1575-7.
• Touitou I. Inheritance of autoinflammatory diseases: shifting paradigms and nomenclature. J Med Gen 2013;50:349-59.
• Zhu S, Liu M, Bennett S, Wang Z, Pfleger KD, Xu JJ. The molecular structure and role of CCL2 (MCP‐1) and C‐C chemokine receptor CCR2 in skeletal biology and diseases. J Cell Physiol 2021;236:7211-22.
• Ozturk O, Cakmakoglu B, Ozturk GN, Unur M. An association of the MCP-1 and CCR2 gene polymorphisms with oral lichen planus. J Oral Surg Oral Med Oral Pathol Oral Radiol 2021;132:708-14.
• Turner MD, Nedjai B, Hurst T, Pennington DJ. Cytokines and chemokines: At the crossroads of cell signalling and inflammatory disease. J Biochimica et Biophysica Acta -Molecular Cell Res 2014;1843:2563-82.