Toxicity Analysis of Recombinant L-asparaginase I and II in Zebrafish
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Toxicity Analysis of Recombinant L-asparaginase I and II in Zebrafish Susan Aishwarya Suresh1 • Selvarajan Ethiraj1 • K. N. Rajnish1
Received: 11 February 2020 / Accepted: 23 May 2020 Ó Association of Microbiologists of India 2020
Abstract L-asparaginases are extensively applied in the treatment of Acute Lymphoblastic Leukemia (ALL). The treatment regime of ALL consists of asparaginase from E. coli or Erwinia. The survival rate post-chemotherapy has increased to \ 90% in recent years. Asparaginase therapy has also resulted in numerous toxicities to patients receiving therapy. This study demonstrates the reaction of normal cells of Danio rerio to asparaginase therapy. L-asparaginase I and II used in the present study are from two probiotic Lactobacillus species in comparison with a commercial L-asparaginase of E. coli origin. Zebrafish adults were injected with 2500 U/kg body weight of L-asparaginase treatments. The expression of SOD 2, CAT, GST, GTP BP3, FADS2 were analyzed with EF1a as house-keeping gene. The p value obtained proves that the data are significant. The histology of the L-asparaginase I treated fishes showed dilated sinusoids in the liver and pseudocyst in the pancreas. The L-asparaginase II and commercial asparaginase showed no pathology. Keywords L-asparaginase Zebrafish RT-PCR Lactobacillus Histology
Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12088-020-00890-7) contains supplementary material, which is available to authorized users. & Selvarajan Ethiraj [email protected] & K. N. Rajnish [email protected] 1
Department of Genetic Engineering, School of Bioengineering, SRM Institute of Science and Technology, Kattankulathur, Tamil Nadu, India
Introduction Acute Lymphoblastic Leukemia is characterized by the synthesis of immature lymphocytes by the bone marrow and is the second most common non-Hodgkin lymphoma occurring in children [1]. The treatment regimen includes the induction phase (4–6 weeks) with a combination of L-asparaginase and few drugs. Consolidation therapy is carried out for 4-8 weeks [2]. Reinduction phase is a repetition of the induction phase with additional drugs for a 3 to 6 months time period. The maintenance phase with antimetabolite and few drugs are administered for 2–3 years [3]. Asparaginases are found in diverse organisms wherein bacterial asparaginases are extensively studied [4]. A study on asparaginases from halophilic bacteria revealed the asparaginase activity of moderate halophilic bacteria [5]. Marine actinobacteria have shown to express asparaginases along with other compounds of interest [6]. One international unit of L-asparaginase is the amount of enzyme required to release 1 lM of ammonia per minute at 37 °C and pH 8.6. In the L-asparaginase therapy, L-asparagine is depleted in the bloodstream and the cancer cells are eradicated due to starvation of L-asparagine [7]. L-asparaginases from Escherichia coli and Erwinia chrysanthemi are employed in the treatment of ALL [8]. Adverse
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