The Effects of Mitochondrial Mt4977 Deletion on Platelet Function in Ischemic Heart Disease Patients

Yıl 2021, , 54 - 58, 01.07.2021

Ahmet Özaydın , Ayla Kerimova İlhan Onaran

https://doi.org/10.38175/phnx.869878

Öz

Objective: It is known that point mutations, duplications and deletions occur in mitochondrial DNAs (mtDNA) of different tissues of individuals. Among the deletions observed, mt4977 mutation, which is located at nucleotide positions 8470-8482 and 13447-13459 and causes the loss of 4977 base pairs, is the most common. mtDNA 4977 deletion leads to the loss of 8 genes encoding subunits of respiratory chain complexes. Consequently, the deletion could be expected to inhibit the oxidative function and reduce ATP production level. It is known that mitochondrial ATP production has an important role on platelet functions. However, there is no information about this in the literature. Since platelet activation in ischemic heart disease (IHD) has been shown to play an important role in the pathophysiology of the disease, we wanted to examine the relationship between platelet function and mtDNA 4977 deletion in ischemic heart disease.
Material and Method: Platelet functions were studied by giving ADP stimulus with the help of lumiaggregometer device to evaluate in terms of secretion and aggregation. ATP measurement was performed with the bioluminescence assay kit. mtDNA 4977 deletion was determined by the modified simultaneous quantitative polymerase chain reaction method.
Results: The frequency of mtDNA 4977 deletion and mtDNA copy number were higher in platelets of the patients compared with the healthy control group (p<0.05). However, no significant differences in platelet ATP content, and in their slope (Ω) and % amplitude values were observed between both groups (p>0.05).
Conclusion: It was observed that increased deletion in patients with IHD did not have a significant effect on
platelet dysfunction compared with healthy control subjects.

Anahtar Kelimeler

mtDNA4977 deletion, ischemic heart disease, platelet, ATP

Destekleyen Kurum

Istanbul University-Cerrahpaşa Scientific Research Projects Coordination Unit

Proje Numarası

770/30498

İskemik Kalp Hastalarında Trombosit Fonksiyonu Üzerine Mitokondri mt4977 Delesyonunun Etkileri

Öz

Amaç: Bireylerin farklı dokularındaki mitokondriyal DNA’larında (mtDNA) nokta mutasyonları, duplikasyonlar ve delesyonlar meydana geldiği bilinmektedir. Gözlenen delesyonlar içerisinde 8470-8482 ile 13447-13459 nükleotid pozisyonlarında yer alan ve 4977 baz çiftinin kaybına yol açan mtDNA 4977 mutasyonu en sık olarak görülmtedir. mtDNA 4977 delesyonu solunum zinciri komplekslerinin alt ünitlerini kodlayan 8 genin kaybolmasına yol açar. Buna bağlı olarak delesyonun oksidatif fonksiyonu engellemesi ve ATP üretim düzeyini azaltması beklenebilir. Mitokondriyal ATP üretiminin trombosit fonksiyonları üzerinde önemli bir rolü olduğu bilinmektedir. Ancak bununla ilişkili olarak literatürde herhangi bir bilgi göze çarpmamaktadır. İskemik kalp hastalığında (İKH) trombosit aktivasyonunun hastalık fizyopatolojisinde önemli bir rol oynadığı gösterildiğinden, bu çalışmamızda trombosit fonksiyonu ile mtDNA 4977 delesyonu arasındaki ilişkiyi ve iskemik kalp hastalığı gelişimindeki olası etkilerini araştırmayı amaçladık .
Gereç ve Yöntem: Trombosit fonksiyonları, sekresyon ve agregasyon açısından değerlendirilmek üzere lumiagregometre cihazı yardımı ile ADP uyaranı verilerek çalışıldı. ATP ölçümü biyolüminesans test kiti ile yapıldı. mt4977 delesyonu, modifiye edilmiş eşzamanlı kantitatif polimeraz zincir reaksiyonu (RT-PCR) yöntemi ile belirlendi.
Bulgular: Hastaların trombositlerinde mtDNA 4977 delesyon sıklığı ve mtDNA kopya sayısı sağlıklı kontrol
grubuna göre daha yüksekti (p <0.05). Ancak her iki grup arasında trombosit ATP içeriği ile bunların eğim (Ω) ve % amplitüd değerlerinde anlamlı farklılık gözlenmedi (p> 0.05).
Sonuç: Sağlıklı kontrol denekleriyle karşılaştırıldığında, İKH hastalarında artmış delesyonun trombosit
disfonksiyonu üzerinde anlamlı bir etkiye sahip olmadığı görülmüştür.

Anahtar Kelimeler

mtDNA4977 delesyonu, iskemik kalp hastalığı, trombosit, ATP

Proje Numarası

770/30498

Kaynakça

• Sherratt EJ, Thomas AW, Alcolado JC. Mitochondrial DNA defects: a widening clinical spectrum of disorders. Clin Sci (Lond). 1997;92(3):225-235. DOI:10.1042/cs0920225
• Mohamed SA, Wesch D, Blumenthal A, Bruse P, Windler K, Ernst M, et al. Detection of the 4977 bp deletion of mitochondrial DNA in different human blood cells. Exp Gerontol. 2004;39:181-8. DOI: 10.1016/j.exger.2003.10.011.
• Hu H, Lin Y, Xu X, Lin S, Chen X, Wang S. The alterations of mitochondrial DNA in coronary heart disease. Exp Mol Pathol. 2020;114:104412. DOI: 10.1016/j.yexmp.2020.104412.
• Chen T, He J, Shen L, Fang H, Nie H, Jin T, Wei X, Xin Y, Jiang Y, Li H, Chen G, Lu J, Bai Y. The mitochondrial DNA 4,977-bp deletion and its implication in copy number alteration in colorectal cancer. BMC Med Genet. 2011 Jan 13;12:8. DOI: 10.1186/1471-2350-12-8.
• Yusoff AAM, Abdullah WSW, Khair SZNM, Radzak SMA. A comprehensive overview of mitochondrial DNA 4977-bp deletion in cancer studies. Oncol Rev. 2019;13(1):409. DOI:10.4081/oncol.2019.409
• Kakimoto M, Inoguchi T, Sonta T, et al. Accumulation of 8-hydroxy-2’-deoxyguanosine and mitochondrial DNA deletion in kidney of diabetic rats. Diabetes. 2002;51(5):1588-1595. doi:10.2337/diabetes.51.5.1588
• Wei YH, Wu SB, Ma YS, Lee HC. Respiratory function decline and DNA mutation in mitochondria, oxidative stress and altered gene expression during aging. Chang Gung Med J. 2009;32:113-32.
• Zabihi Diba L, Mohaddes Ardebili SM, Gharesouran J, Houshmand M. Age-related decrease in mtDNA content as a consequence of mtDNA 4977 bp deletion. Mitochondrial DNA A DNA Mapp Seq Anal. 2016 Jul;27(4):3008-3012. DOI: 10.3109/19401736.2015.1063046.
• Malik AN, Czajka A. Is mitochondrial DNA content a potential biomarker of mitochondrial dysfunction? Mitochondrion. 2013 Sep;13(5):481-92. DOI: 10.1016/j.mito.2012.10.011.
• Huang YH, Chen CM, Lee YS, Chang KH, Chen HW, Chen YC. Detection of mitochondrial DNA with 4977 bp deletion in leukocytes of patients with ischemic stroke. PLoS One. 2018 Feb 23;13(2):e0193175. DOI: 10.1371/journal.pone.0193175.
• Talebi E, Karimian M, Nikzad H. Association of sperm mitochondrial DNA deletions with male infertility in an Iranian population. Mitochondrial DNA A DNA Mapp Seq Anal. 2018 May;29(4):615-623. DOI: 10.1080/24701394.2017.1331347.
• Bacalhau M, Simões M, Rocha MC, Hardy SA, Vincent AE, Durães J, et al. Disclosing the functional changes of two genetic alterations in a patient with Chronic Progressive External Ophthalmoplegia: Report of the novel mtDNA m.7486G>A variant. Neuromuscul Disord. 2018 Apr;28(4):350-360. DOI: 10.1016/j.nmd.2017.11.006.
• Daniel JL, Dangelmaier C, Jin J, Ashby B, Smith JB, Kunapuli SP. Molecular basis for ADP-induced platelet activation. I. Evidence for three distinct ADP receptors on human platelets. J Biol Chem. 1998 Jan 23;273(4):2024-9. doi: 10.1074/jbc.273.4.2024.
• Collins AS. Pivotal role of platelets in critical illness: evidence behind clinical interventions. Dimens Crit Care Nurs. 2004;23(5):217-221. DOI:10.1097/00003465-200409000-00006.
• Vecoli C, Borghini A, Pulignani S, Mercuri A, Turchi S, Carpeggiani C, et al. Prognostic value of mitochondrial DNA4977 deletion and mitochondrial DNA copy number in patients with stable coronary artery disease. Atherosclerosis. 2018 Sep;276:91-97. DOI: 10.1016/j.atherosclerosis.2018.07.015.
• Lott MT, Leipzig JN, Derbeneva O, Xie HM, Chalkia D, Sarmady M, et al. mtDNA Variation and Analysis Using Mitomap and Mitomaster. Curr Protoc Bioinformatics. 2013 Dec;44(123):1.23.1-26. DOI: 10.1002/0471250953.bi0123s44.
• Ye C, Shu XO, Wen W, Pierce L, Courtney R, Gao YT, et al. Quantitative analysis of mitochondrial DNA 4977-bp deletion in sporadic breast cancer and benign breast diseases. Breast Cancer Res Treat. 2008 Apr;108(3):427-34. DOI: 10.1007/s10549-007-9613-9.
• Meissner C, Bruse P, Mohamed SA, Schulz A, Warnk H, Storm T, et al. The 4977 bp deletion of mitochondrial DNA in human skeletal muscle, heart and different areas of the brain: a useful biomarker or more? Exp Gerontol. 2008 Jul;43(7):645-52. DOI: 10.1016/j.exger.2008.03.004.
• Zhang Y, Ma Y, Bu D, Liu H, Xia C, Zhang Y,et al. Deletion of a 4977-bp Fragment in the Mitochondrial Genome Is Associated with Mitochondrial Disease Severity. PLoS One. 2015 May 29;10(5):e0128624. DOI: 10.1371/journal.pone.0128624.
• von Wurmb N, Schwark T, Caliebe A, Drenske C, Nikolaus S, Screiber S, et al. Low level of the mtDNA4977 deletion in blood of exceptionally old individuals. Mech Ageing Dev. 2010; 131: 179–184. DOI: 10.1016/j.mad.2010.01.005.
• Shao JY, Gao HY, Li YH, Zhang Y, Lu YY, Zeng YX. Quantitative detection of common deletion of mitochondrial DNA in hepatocellular carcinoma and hepatocellular nodular hyperplasia. World J Gastroenterol. 2004; 10: 1560–1564.
• Nie HZ, Shu HY, Vartak R, Milstein AC, Mo Y, Hu X, et al. Mitochondrial common deletion, a potential biomarker for cancer occurrence, is selected against in cancer background: a meta-analysis of 38 studies. PLOS One. 2013; 8: e67953. DOI: 10.1371/journal.pone.0067953.