Profiling of microRNAs modulating cytomegalovirus infection in astrocytoma patients
Astrocytoma is recognized as the most common neoplasm of the brain with aggressive progression. The therapeutic regime for glioblastoma, the most aggressive astrocytoma, often consists of aggressive chemo and radiotherapy. The present holistic approaches, however, have failed to influence the quality life of patients. Therefore, it is necessary to understand the underlying mechanisms of its progression for updated therapeutic evaluation. Human cytomegalovirus (HCMV) is reported to be associated with glioblastoma progression. The hypothesis still remains controversial due to the lack of concrete evidences. Here, we report the profile of miRNAs encoded by human host and the cytomegalovirus (CMV) involved in modulation of CMV infection in surgically resected human astrocytoma tissue samples of various malignancy grades (n = 24). Total RNA from the control brain and tumor tissues was extracted by TriZol reagent. The expression levels of the mature form of miRNA were detected by real-time PCR. Primarily, we found the upregulation of miR-210-3p, miR-155-5p, miR-UL-112-3p, miR-183-5p, and miR-223-5p in high-grade astrocytic tumors as compared with low-grade tumor tissues. miR-214-3p is significantly expressed in control brain tissues and its expression decreased with astrocytoma grade progression. This miRNA was reported to be associated with antiviral proprieties. Among CMV-encoded miRNA, miR-UL-112-3p was significantly upregulated in glioblastoma tissue samples and may be involved in providing immune escape to the virus as well as involved in modulating the immune microenvironment of glioblastoma. Taken together, we conclude the possible involvement of miRNAs in modulating the CMV dependent astrocytoma progression.
KeywordsGlioblastoma Astrocytoma CMV miRNA RT PCR
Conception and design: Ravindra Pramod Deshpande
Provision of study material: Manas Panigrahi, Chandrasekhar Y.B.V.K.
Collection and assembly of data: Ravindra Pramod Deshpande
Analysis and interpretation of data: Ravindra Pramod Deshpande, Phanithi Prakash Babu
Manuscript writing: Ravindra Pramod Deshpande, Phanithi Prakash Babu
Final approval: All authors
Authors thank financial assistance from Department of Science and technology (DST-India) (Grant nos. SB/EMEQ-257/2013, SR/CSRI/196/2016), Department of Biotechnology (DBT-India) (Grant nos. BT/PR18168/MED/29/1064/2016, BT/PR13111/MED/29/149/2009), Indian Council of Medical research (ICMR-India) (30/2/2010-01710/SIC/PI/N/162) for lab funding. RDP is thankful to Department of Biotechnology (DBT-India) (Award no: DBT JRF/2011-12/95) for student fellowship.
Compliance with ethical standards
The authors acknowledge KIMS Foundation Research Center (KFRC) and Institutional Ethical Committee (IEC), University of Hyderabad for granting the ethical permissions. Performed studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards.
Informed consent was obtained from all individual participants included in the study.
Conflict of interest
All authors declare that there are no conflicts of interest.
- 1.Louis DN, Perry A, Reifenberger G, von Deimling A, Figarella-Branger D, Cavenee WK, Ohgaki H, Wiestler OD, Kleihues P, Ellison DW (2016) The 2016 World Health Organization classification of tumors of the central nervous system: a summary. Acta Neuropathol 131(6):803–820. https://doi.org/10.1007/s00401-016-1545-1 CrossRefPubMedGoogle Scholar
- 2.Liu B, Pang B, Liu H, Arakawa Y, Zhang R, Feng B, Zhong P, Murata D, Fan H, Xin T, Zhao G, Liu W, Guo H, Luan L, Xu S, Miyamoto S, Pang Q (2015) High mobility group A1 expression shows negative correlation with recurrence time in patients with glioblastoma multiforme. Pathol Res Pract 211(8):596–600CrossRefPubMedGoogle Scholar
- 16.Lau SK, Chen YY, Chen WG, Diamond DJ, Mamelak AN, Zaia JA, Weiss LM (2005) Lack of association of cytomegalovirus with human brain tumors. Modern pathology : an official journal of the United States and Canadian Academy of Pathology, Inc 18(6):838–843. https://doi.org/10.1038/modpathol.3800352 CrossRefGoogle Scholar
- 23.Cobbs C, Khan S, Matlaf L, McAllister S, Zider A, Yount G, Rahlin K, Harkins L, Bezrookove V, Singer E, Soroceanu L (2014) HCMV glycoprotein B is expressed in primary glioblastomas and enhances growth and invasiveness via PDGFR-alpha activation. Oncotarget 5(4):1091–1100CrossRefPubMedPubMedCentralGoogle Scholar
- 27.Stern-Ginossar N, Elefant N, Zimmermann A, Wolf DG, Saleh N, Biton M, Horwitz E, Prokocimer Z, Prichard M, Hahn G, Goldman-Wohl D, Greenfield C, Yagel S, Hengel H, Altuvia Y, Margalit H, Mandelboim O (2007) Host immune system gene targeting by a viral miRNA. Science 317(5836):376–381. https://doi.org/10.1126/science.1140956 CrossRefPubMedPubMedCentralGoogle Scholar
- 29.Li S, Zhu J, Zhang W, Chen Y, Zhang K, Popescu LM, Ma X, Bond Lau W, Rong R, Yu X, Wang B, Li Y, Xiao C, Zhang M, Wang S, Yu L, Chen AF, Yang X, Cai J (2011) Signature microRNA expression profile of essential hypertension and its novel link to human cytomegalovirus infection. Circulation 124(2):175–184. https://doi.org/10.1161/CIRCULATIONAHA.110.012237 CrossRefPubMedGoogle Scholar
- 31.Gyongyosi A, Docs O, Czimmerer Z, Orosz L, Horvath A, Torok O et al (2014) Measuring expression levels of small regulatory RNA molecules from body fluids and formalin-fixed, paraffin-embedded samples. Methods Mol Biol 1182:105–119. https://doi.org/10.1007/978-1-4939-1062-5_10 CrossRefPubMedGoogle Scholar
- 38.Mitchell DA, Xie W, Schmittling R, Learn C, Friedman A, McLendon RE et al (2008) Sensitive detection of human cytomegalovirus in tumors and peripheral blood of patients diagnosed with glioblastoma. Neuro-Oncology 10(1):10–18. https://doi.org/10.1215/15228517-2007-035 CrossRefPubMedPubMedCentralGoogle Scholar
- 39.Goerig NL, Frey B, Korn K, Fleckenstein B, Uberla K, Schmidt MA et al (2016) Frequent occurrence of therapeutically reversible CMV-associated encephalopathy during radiotherapy of the brain. Neuro-Oncology 18(12):1664–1672. https://doi.org/10.1093/neuonc/now120 CrossRefPubMedPubMedCentralGoogle Scholar
- 41.Kawano Y, Kawada J, Kamiya Y, Suzuki M, Torii Y, Kimura H, Ito Y (2016) Analysis of circulating human and viral microRNAs in patients with congenital cytomegalovirus infection. J Perinatol: official journal of the California Perinatal Association 36(12):1101–1105. https://doi.org/10.1038/jp.2016.157 CrossRefGoogle Scholar
- 42.Nallamshetty S, Chan SY, Loscalzo J (2013) Hypoxia: a master regulator of microRNA biogenesis and activity. Free Radic Biol Med 64:20–30. https://doi.org/10.1016/j.freeradbiomed.2013.05.022 CrossRefPubMedPubMedCentralGoogle Scholar
- 46.Zawislak CL, Beaulieu AM, Loeb GB, Karo J, Canner D, Bezman NA, Lanier LL, Rudensky AY, Sun JC (2013) Stage-specific regulation of natural killer cell homeostasis and response against viral infection by microRNA-155. Proc Natl Acad Sci U S A 110(17):6967–6972. https://doi.org/10.1073/pnas.1304410110 CrossRefPubMedPubMedCentralGoogle Scholar
- 48.Santhakumar D, Forster T, Laqtom NN, Fragkoudis R, Dickinson P, Abreu-Goodger C, Manakov SA, Choudhury NR, Griffiths SJ, Vermeulen A, Enright AJ, Dutia B, Kohl A, Ghazal P, Buck AH (2010) Combined agonist-antagonist genome-wide functional screening identifies broadly active antiviral microRNAs. Proc Natl Acad Sci U S A 107(31):13830–13835. https://doi.org/10.1073/pnas.1008861107 CrossRefPubMedPubMedCentralGoogle Scholar