MGUS'ta azalmış GADD45A gen ekspresyonu seviyesi

Yıl 2023, Cilt: 16 Sayı: 4, 594 - 601, 01.10.2023

Ilknur Suer , Aynur Dağlar Aday , Gözde Öztan

https://doi.org/10.31362/patd.1291199

Öz

Amaç: Belirsiz öneme sahip monoklonal gamopati (MGUS), bir plazma hücre diskrazisidir. MGUS'un Multipl miyelom’a (MM) ilerleme riskini artırdığı, ayrıca Waldenstrom makroglobulinemi (WM), non-Hodgkin lenfoma ve kronik lenfositik lösemi (KLL) gibi hastalıklara zemin hazırladığı bilinmektedir. Çalışmamızda bazı önemli p53 yolağı genlerinin MGUS ve sağlıklı bireylerde ekspresyon açısından farklılık gösterip göstermediğini değerlendirmeyi amaçladık.
Gereç ve yöntem: Çalışma için 8 sağlıklı ve 8 MGUS tanılı bireyden kemik iliği toplandı ve RNA örnekleri izole edildi. p53 yolağında yer alan çeşitli genlerin (BAX, CDKN2A, APAF1, ATM, ATR, CASP9, CDK4, CDKN1A, CHEK2, E2F1, E2F3, MCL1, MDM2, MDM4, PTEN, RB1, P53, BCL2, CHEK1, GADD45A, PCNA, PTX3) ekspresyon seviyeleri, RT2-profiler PCR array yöntemiyle karşılaştırıldı. Normalizasyon için β-Aktin gen ekspresyon seviyesi kullanıldı. Pearson Korelasyon ve Receiver Operating Characteristic (ROC) analizleri yapıldı.
Bulgular: Bu çalışmada ekspresyon seviyeleri incelenen genlerden sadece GADD45A geninin ekspresyon seviyesinin kontrol grubuna göre MGUS'ta anlamlı olarak azaldığı belirlendi (p=0,027). Pearson korelasyon verileri, GADD45A gen ekspresyonunun, p53 yolağında incelenen diğer 12 gen (APAF1, CDK4, PCNA, BAX, CDKN2A, CASP9, CHEK2, MDM2, RB1, P53, BCL2, CHEK1) ile yüksek oranda ilişkili olduğunu gösterdi (r>0,7). Ayrıca ROC analizine göre GADD45A'nın MGUS ile sağlıklı bireyler arasında güçlü bir ayrım gücüne sahip olduğu saptandı (AUC=0,797 ve p=0,015).
Sonuç: GADD45A geninin MGUS grubunda kontrol grubuna göre azalmış ekspresyonu, MGUS gelişimini saptamak için yeni bir biyobelirteç olarak faydalı olabilir.

Anahtar Kelimeler

MGUS, p53 yolağı, GADD45A

Decreased GADD45A gene expression level in MGUS

Öz

Purpose: Monoclonal gammopathy of undetermined significance (MGUS) is a plasma cell dyscrasia. It is known that MGUS has an increased risk of progression to multiple myeloma (MM), and prepares the ground for diseases such as Waldenstrom macroglobulinemia (WM), non-Hodgkin lymphoma, and chronic lymphocytic leukemia (CLL). Our study aimed to evaluate whether some important p53 pathway genes differ in terms of expression between MGUS and healthy individuals.
Materials and methods: Bone marrow was collected from eight healthy individuals and eight individuals diagnosed with MGUS, and RNA samples were isolated. The expression levels of various genes involved in the p53 pathway were compared using an RT2-profiler PCR array. β-Actin housekeeping gene expression level was used for normalization. Pearson’s Correlation and Receiver Operating Characteristic (ROC) analyses were conducted.
Results: Among the genes whose expression levels were examined in this study, it was determined that the expression level of only the GADD45A gene decreased significantly in the MGUS group compared to the control group (p=0.027). Pearson’s correlation data showed that GADD45A gene expression was highly correlated with 12 of the other genes (APAF1, CDK4, PCNA, BAX, CDKN2A, CASP9, CHEK2, MDM2, RB1, P53, BCL2, CHEK1) examined in the p53 pathway (r>0.7). In addition, according to the ROC analysis, GADD45A was detected to have strong discrimination power between MGUS and healthy individuals (AUC=0.797 and p=0.015).
Conclusion: The decreased expression of the GADD45A gene in the MGUS group compared to the control group may be useful as a new biomarker to detect the development of MGUS.

Anahtar Kelimeler

MGUS, p53 pathway, GADD45A

Kaynakça

• 1. Landgren O. Advances in MGUS diagnosis, risk stratification, and management: introducing myeloma-defining genomic events. Hematology Am Soc Hematol Educ Program 2021;2021:662-672. https://doi.org/10.1182/hematology.2021000303
• 2. van de Donk NWCJ, Pawlyn C, Yong KL. Multiple myeloma. Lancet 2021;397:410-427. https://doi.org/10.1016/S0140-6736(21)00135-5
• 3. Akhtar S, Najafzadeh M, Isreb M, Newton L, Gopalan RC, Anderson D. An in vitro investigation into the protective and genotoxic effects of myricetin bulk and nano forms in lymphocytes of MGUS patients and healthy individuals. Toxicol Lett 2020;327:33-40. https://doi.org/10.1016/j.toxlet.2020.03.012
• 4. Xiong W, Wu X, Starnes S, et al. An analysis of the clinical and biologic significance of TP53 loss and the identification of potential novel transcriptional targets of TP53 in multiple myeloma. Blood 2008;112:4235-4246. https://doi.org/10.1182/blood-2007-10-119123
• 5. Awada H, Thapa B, Awada H, et al. A comprehensive review of the genomics of multiple myeloma: evolutionary trajectories, gene expression profiling, and emerging therapeutics. Cells 2021;10:1961. https://doi.org/10.3390/cells10081961
• 6. Mikulasova A, Wardell CP, Murison A, et al. The spectrum of somatic mutations in monoclonal gammopathy of undetermined significance indicates a less complex genomic landscape than that in multiple myeloma. Haematologica 2017;102:1617-1625. https://doi.org/10.3324/haematol.2017.163766
• 7. Geraldes C, Gonçalves AC, Cortesão E, et al. Aberrant p15, p16, p53, and DAPK gene methylation in myelomagenesis: clinical and prognostic ımplications. Clin Lymphoma Myeloma Leuk 2016;16:713-720. https://doi.org/10.1016/j.clml.2016.08.016
• 8. Oka S, Ono K, Nohgawa M. The effect of azacitidine therapy on the M protein of MDS patients with concomitant MGUS. Am J Hematol 2018;93:220-222. https://doi.org/10.1002/ajh.25160
• 9. Wang W, Shim YK, Michalek JE, et al. Serum microRNA profiles among dioxin exposed veterans with monoclonal gammopathy of undetermined significance. J Toxicol Environ Health A 2020;83:269-278. https://doi.org/10.1080/15287394.2020.1749919
• 10. Ho M, Patel A, Goh CY, Moscvin M, Zhang L, Bianchi G. Changing paradigms in diagnosis and treatment of monoclonal gammopathy of undetermined significance (MGUS) and smoldering multiple myeloma (SMM). Leukemia 2020;34:3111-3125. https://doi.org/10.1038/s41375-020-01051-x
• 11. Liebermann DA, Hoffman B. Gadd45 in stress signaling. J Mol Signal 2008;3:15. https://doi.org/10.1186/1750-2187-3-15
• 12. Patel K, Murray MG, Whelan KA. Roles for GADD45 in development and cancer. Adv Exp Med Biol 2022;1360:23-39. https://doi.org/10.1007/978-3-030-94804-7_2
• 13. Higashi H, Vallböhmer D, Warnecke Eberz U, et al. Down-regulation of Gadd45 expression is associated with tumor differentiation in non-small cell lung cancer. Anticancer Res 2006;26:2143-2147.
• 14. Guo W, Dong Z, Guo Y, Chen Z, Kuang G, Yang Z. Methylation-mediated repression of GADD45A and GADD45G expression in gastric cardia adenocarcinoma. Int J Cancer 2013;133:2043-2053. https://doi.org/10.1002/ijc.28223
• 15. Wang W, Huper G, Guo Y, Murphy SK, Olson JA, Marks JR. Analysis of methylation-sensitive transcriptome identifies GADD45a as a frequently methylated gene in breast cancer. Oncogene 2005;14:2705-2714. https://doi.org/10.1038/sj.onc.1208464
• 16. Reddy SP, Britto R, Vinnakota K, et al. Novel glioblastoma markers with diagnostic and prognostic value identified through transcriptome analysis. Clin Cancer Res 2008;15:2978-2987. https://doi.org/10.1158/1078-0432.CCR-07-4821
• 17. Myint KZ, Kongpracha P, Rattanasinganchan P, et al. Gadd45β silencing impaired viability and metastatic phenotypes in cholangiocarcinoma cells by modulating the EMT pathway. Oncol Lett 2018;15:3031-3041. https://doi.org/10.3892/ol.2017.7706
• 18. Borges GA, Guilatco AJ, Hachfeld CM, et al. Abstract PRO12: Pre-malignant plasma cells exhibit a senescence-like phenotype and accumulation of transposable elements. Cancer Research 2023;83. https://doi.org/10.1158/1538-7445.AGCA22-PR012
• 19. López Corral L, Corchete LA, Sarasquete ME, et al. Transcriptome analysis reveals molecular profiles associated with evolving steps of monoclonal gammopathies. Haematologica 2014;99:1365-1372. https://doi.org/10.3324/haematol.2013.087809
• 20. Salvador JM, Brown Clay JD, Fornace AJ Jr. Gadd45 in stress signaling, cell cycle control, and apoptosis. Adv Exp Med Biol 2013;793:1-19. https://doi.org/10.1007/978-1-4614-8289-5_1
• 21. Aljabban J, Chen D, Cottini F, et al. Characterization of monoclonal gammopathy of undetermined significance progression to multiple myeloma through meta-analysis of geo data. 2019;134:4395. https://doi.org/10.1182/blood-2019-129699