Pyruvate kinase deficiency: optimizing pkrl gene sequencing and analysis of genotypic data in relation to malaria risk

Abstract

Pyruvate kinase (PK) is the glycolytic enzyme responsible for catalyzing the transphosphorylation of phosphoenolpyruvate into adenosine diphosphate in human red blood cells, producing pyruvate and ATP as the final product of the second step of glycolysis. Pyruvate kinase deficiency (PKD), with autosomal recessive inheritance, is the most common defect associated with hereditary non-spherocytic hemolytic anemia, characterized by anemia and chronic hemolysis resulting from loss-of-function mutations in the pklr gene. It leads to ATP depletion and increased concentration of 2,3-bisphosphoglycerate (2,3-BPG) within erythrocytes, reducing their half-life. There is evidence that PKD may contribute to resistance to falciparum malaria, but whether the same applies to vivax malaria remains unknown.The main objective of Dr. Ana Paula Arez's visit is to optimize techniques to be applied in our laboratory in São Paulo and to start data analysis to investigate whether there is an association between PKD and the risk of malaria in Amazonian populations. During her 3-week visit to Brazil, Dr. Ana Paula Arez is expected to participate directly in the training of our laboratory teams in the following activities:1. Optimization of protocols for the complete sequencing of the promoter region and the coding region (13 exons) of the pklr gene in Amazonian populations with previously phenotypically determined PKD;2. Standardization of techniques for analyzing the relationship between polymorphisms in the genomic region that comprises the pklr locus and the risk of malaria in Amazonian populations;3. Seminar open to all members of the Institute of Biomedical Sciences of USP, with wide dissemination throughout the USP campus, on the most recent results of her cellular and molecular studies on the impact of PKD on the development of malaria parasites and its epidemiological consequences, as part of the traditional seminar program of the Department of Parasitology at the Institute of Biomedical Sciences oft USP (graduate discipline BMP 5761, "Seminars of Parasitology II").4. Individualized discussion on research projects, open to all members of the Department of Parasitology of the Institute of Biomedical Sciences of USP (teachers, post-doctoral students, and postgraduate and scientific initiation students).5. Symposium on Malaria, open to all members of the Institute of Biomedical Sciences of USP, with wide dissemination throughout the USP campus. This is a mixed event (in-person and online) bringing together all associated and main researchers of the thematic project 2022/11963-3 and their students and post-doctoral fellows, lasting three days. (AU)