Identification of therapeutic immunometabolic targets in blood cells of sickle cell anemia patients using mass cytometry (CyTOF)

• Grant number: 25/21532-8
• Support Opportunities: Scholarships abroad - Research
• Start date: May 01, 2026
• End date: April 30, 2027
• Field of knowledge: Health Sciences - Medicine
• Principal Investigator: Nicola Amanda Conran Zorzetto
• Grantee: Nicola Amanda Conran Zorzetto
• Host Investigator: John Strouboulis
• Host Institution: Centro de Hematologia e Hemoterapia (HEMOCENTRO). Universidade Estadual de Campinas (UNICAMP). Campinas , SP, Brazil
• Institution abroad: King's College London, England

Abstract

Sickle cell anaemia (SCA) is characterised by recurrent vaso-occlusive crises (VOC) and progressive organ injury, processes that are tightly coupled to inflammatory signalling. Although therapies such as hydroxycarbamide reduce VOC frequency, the cellular mechanisms translating inflammatory stimuli into vaso-occlusion remain poorly defined. Preliminary data from our group show that neutrophils display metabolic alterations in SCA, including increased glucose uptake and mitochondrial stress, where these effects are intensified by hemin. These data highlight the need for high-resolution tools to dissect how inflammatory perturbations reprogram cell metabolism and signalling in the SCA vasculature. This project will apply single-cell mass cytometry (CyTOF) to generate a comprehensive immunometabolic and signalling map of vaso-occlusive effector cells. Peripheral blood from healthy controls, untreated HbSS patients, and HbSS patients on hydroxycarbamide will be stimulated ex vivo with PBS (control) or hemin. A 27-antibody panel will quantify surface, metabolic, and signalling markers in neutrophils, platelets, and neutrophil-platelet aggregates (NPAs). Computational pipelines (CATALYST, difficyt) will cluster cells and compare phenotypes, protein expression and signalling across conditions. Dimensionality reduction (UMAP) and heatmaps will reveal shifts in cellular profiles, while principal component modelling will connect metabolic pathways (e.g. glycolysis, TCA cycle) and signalling networks (e.g. HIF-1, NF-¿B, MAPK) to phenotypic changes such as adhesion, aggregation, and inflammatory mediator release. By integrating these signatures across cell types, CyTOF will provide a systems-level view of metabolic reprogramming and signalling in cells that are crucial to the vaso-occlusive process. The project is unique in applying CyTOF to identify molecular checkpoints where metabolic reprograming modulates signalling cascades to drive pathogenic cell behaviour. This mechanistic atlas will provide the basis for future therapeutic strategies aimed at reprogramming neutrophil, platelet, and NPA function in SCD. King's College London is the ideal environment for this work, with access to one of the largest SCD patient populations in the UK and fully supported CyTOF facilities. This is critical, as Brazil currently lacks mass cytometry infrastructure. Conducting the project at King's ensures feasibility, technical excellence, and impact at a level that cannot be achieved elsewhere. (AU)