Single cell tracking of malignant transformation in GATA2 deficiency

Projektbeschreibung

Erkenntnisse zur Komplexität der GATA2-Defizienz

GATA2 ist ein Transkriptionsfaktor, der bei der Hämatopoese eine zentrale Rolle spielt. Mutationen des GATA2-Gens wirken sich auf hämatopoetische Vorläuferzellen aus und können zu Knochenmarkversagen und einem erhöhten Risiko für Leukämie führen. Das EU-finanzierte Projekt scGATA2track untersucht die phänotypische Variabilität in Familien mit GATA2-Defizienz. Hauptziel ist es, die zugrunde liegenden genetischen und epigenetischen Mechanismen auf Einzelzellebene zu entschlüsseln, um die verantwortlichen Faktoren der Krankheitsprogression zu bestimmen. Die Ergebnisse werden zur Früherkennung von Leukämie beitragen und eine Grundlage für die Entwicklung neuer personalisierter Therapiestrategien bieten.

Ziel

Precision medicine has improved overall survival of cancer patients; unfortunately, not all diseases can benefit from this practice due to a scarce understanding of their molecular mechanisms. This is the case of GATA2 deficiency, a complex multi-system disorder characterized by bone marrow failure, immunodeficiency and high risk to develop myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML). Chemotherapy and allogenic hematopoietic stem cell (HSC) transplantation are the only available treatments, underlining the lack of predictive tools as a clear medical need.
Penetrance and expressivity within GATA2 families is often variable, suggesting that cooperating somatic mutations and epigenetic events are required to trigger the disease. Here I propose to unravel the molecular mechanisms of malignant progression of GATA2 deficiency by combining single cell multi-OMICs approaches with groundbreaking functional assays in human induced pluripotent stem cells (hiPSCs).
First, to identify pathogenic single nucleotide variations and small insertion-deletions, I will perform an integrative analysis combining whole exome sequencing with whole-genome bisulfite sequencing to analyze the methylation status genome wide on 15 well-annotated GATA2 carriers.
Then, I will model the stepwise progression of normal cells to MDS through the sequential introduction of recurrent GATA2 mutations and known second driver mutations in hiPSCs by CRISPR/Cas9 genome editing. The hiPSC-derived hematopoietic progenitors will be used for simultaneous profiling of transcriptome and chromatin accessibility at single cell resolution to decipher the underlying gene regulatory networks among the different genetic backgrounds.
A thorough understanding of the genetic and epigenetic features of GATA2 carriers and the transcriptional dynamics during cancer initiation/progression will provide valuable insight into early detection and the design of novel personalized therapeutic strategies.