MSACL 2019 EU Abstract

Mass Spectrometry Studies of Glutathione S Transferase P1 Protein and Gene and 5-Methylcytosine Level in Acute Lymphoblastic Leukemia

Negin Fasih Ramandi (1), Maryam Hasan Dihree (1), Mojan Esmaeillou (1), Mohammad Faranoush (2), Alireza Ghassempour (1)
(1) Medicinal Plants and Drugs Research Institute, Shahid Beheshti University, Evin, Tehran, Iran (2) Pediatric Growth and Development of Endocrinology, Iran University of Medicine Sciences, Tehran, Iran

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Alireza Ghassempour (Presenter) Shahid Beheshti University >> POSTER (PDF)

Presenter Bio: I am Alireza Ghassempour Professor of Analytical Chemistry in Medicinal Plants and Drugs Research Institute. My team focus on mass spectrometry techniques of natural compounds and clinical samples. Also, we could purify natural compounds and microorganisms by preparative HPLC. The study of the interaction between purified natural compounds with sick tissues and microorganisms by mass spectrometry is our goal.

Relevant Financial Disclosures (within past 24 months) Grant/Research Support Shahid Beheshti University Conflict mitigation NOT REQUIRED.

Abstract
INTRODUCTION: Childhood acute lymphoblastic leukemia (ALL) is considered for more than 30% of childhood cancer per a year throughout the world. The routine ALL diagnosis and prognosis methods are considered of peripheral blood analysis, the bone marrow aspiration and biopsy, flow cytometry and immunophenotyping assays. They rely on well-known compounds and the interpretation of their results are user dependently and in this regard some mistakes can be happened. Meanwhile, there are some evidences that biomarkers based on mass spectrometry (MS) techniques can be considered as well replacement of these clinical diagnosis and prognosis techniques. Some reports confirm the role of Glutathione S Transferase P1 (GST-P1) protein in childhood ALL risk. In addition, previous reports showed the role of epigenetic in ALL. Figuring out epigenetic modifications in their biological context includes several aspects of DNA methylation analysis. MS techniques can determine epigenetic levels and gene region involve to methylation.
METHODS: In the present study, we have monitored the changes in the GST-P1 protein in blood plasma samples of 19 ALL patients at the diagnosis stage and after finishing their first chemotherapy. We used GST-P1 gene-promoter region to show that a ratio between cytosine and 5-methylcytosine DNA content by gas chromatography-mass spectrometry (GC-MS). In addition, the analysis of PCR samples for GST-P1 gene-promoter was done by MALDI-TOF-MS. We performed similar analyses in the children blood sample of same sex and age.
RESULTS: The affinity magnetic sorbent enriched of GST-P1 protein from blood plasma sample without any pretreatment, followed by MALDI-TOF-MS detection. Our results show the significant variation on GST-P1 protein levels compare to normal blood children and diagnosis procedure of ALL due to chemotherapy of ALL patients. The results of GC-MS are sufficient for discovering methylation variation within a selected target region of these patients. Also, a method for DNA methylation analysis that utilizes MALDI-TOF MS analysis of base-specifically cleaved amplification products has been developed to discover the exact sites of methylation.