The Effect of Anti-Inflammatory Drugs on MEFV, PSTPIP1, Siva, and ASC Gene Expression Levels

Year 2023, Volume: 5 Issue: Supplement (1) - Innovations in Medicine and Healthcare in the 100th Year of the Republic, 144 - 9, 19.10.2023

Yeliz Z. Akkaya-ulum

https://doi.org/10.37990/medr.1348540

Abstract

Aim: Familial Mediterranean Fever (FMF) is the one of the most common autoinflammatory diseases. FMF is characterized by fever attacks and inflammation and colchicine treatment reduces the frequency and severity of FMF attacks. The FMF gene, MEditerranean FeVer (MEFV), encodes a protein called Pyrin, which regulates inflammation through its interactions with several proteins. These proteins are; Apoptosis-associated speck like protein with a CARD (ASC), Proline serine threonine phosphatase interacting protein 1 (PSTPIP1), 14.3.3 proteins and Siva proteins. In this study, we aimed to study the effect of anti-inflammatory drugs with different mechanisms of action on MEFV, PSTPIP1, Siva, and ASC gene expression levels.
Material and Methods: We used differentiated monocytic cell line called THP-1 cells. Cells treated with colchicine, naproxen, prednol-L, acetylsalicylic acid, or azathioprine w and w/o lipopolysaccharide (LPS). After incubation, quantitative RT-PCR (qRT-PCR) was performed to measure MEFV, PSTPIP1, Siva, and ASC gene expression levels.
Results: MEFV gene expression level was down regulated in colchicine, naproxen, and azathioprine treated cells whereas PSTPIP1 gene expression level was down regulated in naproxen and azathioprine treated cells with LPS. Siva gene expression level was up regulated in all treatments although ASC gene expression level was up regulated in only prednol-L treated cells with LPS.
Conclusion: These anti-inflammatory drugs are known to have different mechanisms of action however they are all used to treat pain or inflammation. Since Pyrin, PSTPIP1, Siva, and ASC have pro and anti-inflammatory roles, the results showing an alteration in gene expression levels with specific drugs may indicate the possible mechanisms of therapeutic action.

Keywords

Familial Mediterranean Fever (FMF), Anti-inflammatory drugs, Inflammation, Inflammation-related genes

Anti-inflamatuvar ilaçların MEFV, PSTPIP1, Siva ve ASC gen ifade düzeylerine olan etkisi

Abstract

Amaç: Ailevi Akdeniz Ateşi (AAA) en sık görülen ve tekrarlayan ateş atakları ve ağrılı inflamasyon ile karakterize olan otoinflamatuvar bir hastalıktır. Kolşisin tedavisi, AAA ataklarının sıklığını ve şiddetini azaltır. Pyrin adlı bir proteini kodlayan AAA geni, MEditerranean FeVer (MEFV) geni tarafından kodlanmakta ve ilişkili olduğu proteinler aracılığıyla inflamasyonu düzenlemektedir. İlişkili olduğu bu proteinler; Apoptozla ilişkili benek benzeri protein CARD(ASC), Prolin serin treonin fosfataz etkileşimli protein 1(PSTPIP1), 14.3.3 proteini ve Siva proteinleridir. Bu çalışmada, farklı etki mekanizmalarına sahip anti-inflamatuvar ilaçların MEFV, PSTPIP1, Siva ve ASC gen ifade seviyeleri üzerindeki etkilerinin araştırılması amaçlanmıştır.
Gereç ve Yöntem: Hücre hattı olarak farklılaşmış monositik hücreleri olan THP-1 hücreleri kullanılmıştır. Kolşisin, naproksen, prednol-L, asetilsalisilik asit veya azatiyoprin ilaçları, lipopolisakkarit (LPS) varlığında ve yokluğunda hücrelere uygulanmıştır. MEFV, PSTPIP1, Siva ve ASC gen ifade seviyelerini ölçmek için kantitatif RT-PCR(qRT-PCR) yapılmıştır.
Bulgular: LPS varlığında, MEFV geni ifade seviyesi, kolşisin, naproksen ve azatioprin uygulanan hücrelerde azalırken, PSTPIP1 gen ifade seviyesi, naproksen ve azatioprin uygulanan hücrelerde azalmıştır. LPS varlığında, Siva gen ifade seviyesi tüm ilaç uygulamalarında artarken, ASC gen ifadesi sadece LPS ve prednol-L uygulamalarında artmıştır.
Sonuç: Tüm bu anti-inflamatuvar ilaçların farklı etki mekanizmaları olduğu bilinmektedir, ancak hepsi ağrı veya iltihabı tedavi etmek için kullanılmaktadır. Pyrin, PSTPIP1, Siva ve ASC proteinleri pro ve anti-inflamatuvar rollere sahip olduğundan, genlerin ifade düzeylerindeki değişiklik ilaçların terapötik etkilerinin olası mekanizmalarını aydınlatmada yol gösterici olabilecektir.

Keywords

Ailevi Akdeniz Ateşi (AAA), Anti-inflamatuvar ilaçlar, İnflamasyon, İnflamasyonla ilişkili genler

References

• Livneh A, Langevitz P, Zemer D, et al. Criteria for the diagnosis of familial Mediterranean fever. Arthritis Rheum. 1997;40:1879-85.
• Ancient missense mutations in a new member of the RoRet gene family are likely to cause familial Mediterranean fever. The International FMF Consortium. Cell. 1997;90:797-807.
• Diaz A, Hu C, Kastner DL, et al. Lipopolysaccharide-induced expression of multiple alternatively spliced MEFV transcripts in human synovial fibroblasts: a prominent splice isoform lacks the C-terminal domain that is highly mutated in familial mediterranean fever. Arthritis Rheum. 2004;50:3679-89.
• Chae JJ, Aksentijevich I, Kastner DL. Advances in the understanding of familial Mediterranean fever and possibilities for targeted therapy. Br J Haematol. 2009;146:467-78.
• Xu H, Yang J, Gao W, et al. Innate immune sensing of bacterial modifications of Rho GTPases by the Pyrin inflammasome. Nature. 2014;513:237-41.
• Park YH, Wood G, Kastner DL, Chae JJ. Pyrin inflammasome activation and RhoA signaling in the autoinflammatory diseases FMF and HIDS. Nat Immunol. 2016;17:914-21.
• Shoham NG, Centola M, Mansfield E, et al. Pyrin binds the PSTPIP1/CD2BP1 protein, defining familial Mediterranean fever and PAPA syndrome as disorders in the same pathway. Proc Natl Acad Sci U S A. 2003;100:13501-6.
• Chitu V, Stanley ER. Pombe Cdc15 homology (PCH) proteins: coordinators of membrane-cytoskeletal interactions. Trends Cell Biol. 2007;17:145-56.
• Wise CA, Gillum JD, Seidman CE, et al. Mutations in CD2BP1 disrupt binding to PTP PEST and are responsible for PAPA syndrome, an autoinflammatory disorder. Hum Mol Genet. 2002;11:961-9.
• Cortis E, De Benedetti F, Insalaco A, et al. Abnormal production of tumor necrosis factor (TNF) -- alpha and clinical efficacy of the TNF inhibitor etanercept in a patient with PAPA syndrome [corrected]. J Pediatr. 2004;145:851-5.
• Stichweh DS, Punaro M, PASCual V. Dramatic improvement of pyoderma gangrenosum with infliximab in a patient with PAPA syndrome. Pediatr Dermatol. 2005;22:262-5.
• Church LD, Cook GP, McDermott MF. Primer: inflammasomes and interleukin 1beta in inflammatory disorders. Nat Clin Pract Rheumatol. 2008;4:34-42.
• Yu JW, Fernandes-Alnemri T, Datta P, et al. Pyrin activates the ASC pyroptosome in response to engagement by autoinflammatory PSTPIP1 mutants. Mol Cell. 2007;28:214-27.
• Waite AL, Schaner P, Richards N, et al. Pyrin Modulates the Intracellular Distribution of PSTPIP1. PLoS One. 2009;4:e6147.
• Taskiran EZ, Cetinkaya A, Balci-Peynircioglu B, et al The effect of colchicine on pyrin and pyrin interacting proteins. J Cell Biochem. 2012;113:3536-46.
• Cooper KM, Bennin DA, Huttenlocher A. The PCH family member proline-serine-threonine phosphatase-interacting protein 1 targets to the leukocyte uropod and regulates directed cell migration. Mol Biol Cell. 2008;19:3180-91.
• Akkaya-Ulum YZ, Balci-Peynircioglu B, Purali N, Yilmaz E. Pyrin-PSTPIP1 colocalises at the leading edge during cell migration. Cell Biol Int. 2015;39:1384-94.
• Masumoto J, Taniguchi S, Ayukawa K, et al. ASC, a novel 22-kDa protein, aggregates during apoptosis of human promyelocytic leukemia HL-60 cells. J Biol Chem. 1999;274:33835-8.
• Richards N, Schaner P, Diaz A, et al. Interaction between pyrin and the apoptotic speck protein (ASC) modulates ASC-induced apoptosis. J Biol Chem. 2001;276:39320-9.
• Py B, Slomianny C, Auberger P, et al. Siva-1 and an alternative splice form lacking the death domain, Siva-2, similarly induce apoptosis in T lymphocytes via a caspase-dependent mitochondrial pathway. J Immunol. 2004;172:4008-17.
• Balci-Peynircioglu B, Waite AL, Hu C, et al. Pyrin, product of the MEFV locus, interacts with the proapoptotic protein, Siva. J Cell Physiol. 2008;216:595-602.
• Goulielmos GN, Petraki E, Vassou D, et al. The role of the pro-apoptotic protein Siva in the pathogenesis of Familial Mediterranean fever: a structural and functional analysis. Biochem Biophys Res Commun. 2010;402:141-6.
• Zemer D, Pras M, Sohar E, et al. Colchicine in the prevention and treatment of the amyloidosis of familial Mediterranean fever. N Engl J Med. 1986;314:1001-5.
• Daigneault M, Preston JA, Marriott HM, et al. The identification of markers of macrophage differentiation in PMA-stimulated THP-1 cells and monocyte-derived macrophages. PLoS One. 2010;5:e8668.
• Hentgen V, Vinit C, Fayand A, Georgin-Lavialle S. The use of interleukine-1 inhibitors in familial mediterranean fever patients: a narrative review. Front Immunol. 2020;11:971.
• Liu F, Ma R, Riordan SM, et al. Azathioprine, mercaptopurine, and 5-aminosalicylic acid affect the growth of IBD-associated campylobacter species and other enteric microbes. Front Microbiol. 2017;8:527.
• Sayarlioglu H, Erkoc R, Sayarlioglu M, et al. Successful treatment of nephrotic syndrome due to FMF amyloidosis with azathioprine: report of three Turkish cases. Rheumatol Int. 2006;27:197-9.
• Schellevis MA, Stoffels M, Hoppenreijs EP, et al. Variable expression and treatment of PAPA syndrome. Ann Rheum Dis. 2011;70:1168-70.