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Multi-user equipment approved in grant 19/18886-1: PeriCam PSI NR


The proposed Thematic Project "Pathophysiological Mechanisms and Treatment of Red Blood Cell Abnormalities" will be developed by the Red Blood Cell Research Group at the Hematology Center, the Department of Clinical Pathology and the Center for Molecular Biology and Genetic Engineering (CBMEG), UNICAMP, in collaboration with various Brazilian and international researchers. The project aims to expand and implement new therapeutic approaches in red blood cell diseases, elucidate mechanisms of these diseases with potential for the development of treatments and continue our structural and functional analyzes of variant hemoglobins. We present a consistent set of well-focused investigations: The identification of as yet elucidated pathophysiological mechanisms responsible for the major red cell diseases and the testing of innovative therapeutic strategies that may contribute significantly to the cure of these diseases. The proposal is based on results obtained in recent years by the group of researchers involved and on the major methodological advances that have occurred in this area, with special emphasis on the use of new gene editing techniques, modern methods for studying gene expression, imaging cytometry, the use of animal models of diseases and the use of in vitro mechanisms for the study of cellular function. The infrastructure available is remarkable; we can highlight: Cytometry equipment, such as the Amnis Image Stream, for visualization of interactions and morphological characteristics of cells, and the CytoFlex for detection of microvesicles; the Cellix Venaflux microfluidic equipment for in vitro erythrocyte studies; and ECIS for monitoring the endothelial cell barrier. Our strategy is to identify three major areas of study related to red cell diseases that include a total of 14 subprojects. The first area concerns investigations related to the functional study of abnormal hemoglobins. The second area of study comprises subprojects that relate to various aspects of the pathophysiological mechanisms of red blood cell disease. Important findings previously obtained in this field of investigation may be translated into therapeutic options. Among the subprojects included in this area are studies of the association of macrophages and monocytes with erythropoiesis and iron metabolism; and the use of animal models of sickle cell disease to analyze cellular functions, new models of clinical changes such as retinopathy and the production of an animal with sickle cell disease associated with PHHF. The third area of study encopasses projects related to the treatment of red blood cell diseases, including strategies to increase fetal hemoglobin (HbF) production, identification of genes related to this event, and the evaluation of the efficacy of this increase in HbF in patients treated with hydroxyurea; projects that aim to development approaches to reduce the deleterious effects of hemolysis; projects aimed at preventing or reversing the effects of vascular occlusion and the analysis of new molecules with iron-chelating properties. The goal of this area is to investigate new therapeutic possibilities, in particular, the use of CRISPR-Cas9 gene editing to induce HbF production; in this case, our preliminary results are highly promising. (AU)

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