Molecular landscapes of the hematopoietic niche: single-cell insights and inflammatory signatures in sickle cell anemia

Abstract

Sickle Cell Anemia (SCA) is a complex hematological disorder encompassing dysregulated hematopoiesis and chronic inflammation within the bone marrow. This interdisciplinary project investigates the molecular intricacies of bone marrow aspirates from three distinct subject groups: healthy donors (control group), chronically transfused SCA patients, and hydroxyurea-treated SCA patients (study groups). Leveraging single-cell RNA sequencing on bone marrow-derived mononuclear cells, this study aims to unveil unique gene expression profiles at the single-cell level, employing the Chromium Fixed RNA Profiling technology, which enables comprehensive gene expression measurements in fixed samples, enabling up to 16 uniquely barcoded samples pooled within a single library construction reaction. Additionally, fluorescence-activated cell sorting will dissect the cell constitution of the hematopoietic stem cell and immunological compartments. In addition, the proposed methodology extends beyond transcriptomics to quantifying cytokines and chemokines related to hematopoiesis and inflammation in bone marrow plasma. This integrative approach delineates the dynamic cellular and molecular interactions governing hematopoiesis and inflammation in SCA. Expected results include the identification of novel cell subpopulations, dysregulated molecular pathways, and inflammatory signatures specific to SCA. Furthermore, the project seeks to establish correlations between transcriptomic alterations and changes in the hematopoietic niche. This research holds significant relevance for advancing our understanding of SCA pathophysiology, providing insights into potential therapeutic targets, and improving the way for personalized treatment strategies such as gene therapies. By bridging the gap between single-cell transcriptomics and inflammation profiling, this project contributes to a holistic comprehension of the bone marrow microenvironment in SCA, fostering new avenues for innovative clinical interventions. (AU)