Experimental Haematology and Control of Erythropoiesis
The Borg research group contains Malta's most active labs in the fields of Functional Genomics and Transcriptional Regulation. The group focuses on developing new tools for the understanding of fundamental biological processes in erythroid and other cell formation and differentiation as well as a multi -omic analysis of large data sets obtained from both in vivo and in vitro experiments. We are particularly interested in erythropoiesis and the regulation and control of developmental globin gene switching in humans. We collaborate closely with the Cell Biology and Genetics at Erasmus MC and several groups at EMBL (Heidelberg), including the Gene Core and Chemical Core Facilities.
Previous and current research
Our group is using advanced Next Generation Sequencing tools to probe both DNA and RNA profiles from primary erythroid progenitor cells and selected erythroid cell lines under study. These are conducted in order to investigate fundamental and basic mechanisms behind the globin gene switch mechanism in cells in the hope of discovering a cure for beta type haemoglobinopathies. The group is divided into two lab teams each led by a Principal Investigator that focus on molecular gene editing tools, and cellular biology experiments on both established and in-house cell models developed by the group. Extensive bioinformatic analysis are conducted on the data generated and overlaid in an integrative approach that underpins both molecular and cellular based models. New and emerging scripts for data analysis are made accessible via the collaborations with established groups at both EMBL, Heidelberg and Erasmus MC, Rotterdam as shown in the About Us section. Group members design, develop and implement new genetic methods required for blood -omic analysis, and experimental haematology testing to obtain insights into the biological mechanisms of gamma- to beta- globin gene switching building on what has already been achieved thus far (Figure 1)
Borg group has further been among the pioneers in the KLF1 working group to utilise a global network of research laboratories working on the elucidation of a number of patients with unexplained or borderline levels of HbF and/or HbA2. By sequencing the full KLF1 gene in a number of patients, we detected a large number of DNA variants, some being innocuous whilst others were predicted to be damaging and clinically significant. We recently demonstrated that variants in KLF1 are responsible for much more than just HbF control and often are also attributed to high levels of Zinc Protoporphyrin, an increase in total bilirubin count, abnormal red blood cell morphology, cell proliferation defects and many others (Figure 2)
Future projects and goals
Identifying molecular biomarkers and determinants for the induction of Foetal Haemoglobin (HbF) in vitro and in vivo models.
Development of new methodologies to probe and analyze single cell genomic studies of structural variation at the beta globin locus.
Integrating state-of-the-art microscopy methods with single cell sequencing and advanced immuno flowcytometry.
Completion of a full model that depicts the switching mechanism of gamma- to beta- globin gene switching.
Figure 1: The effect of KLF1 on gamma- to beta- globin gene switching (a) in a normal KLF1 sufficient environment (b) in a mutant KLF1 deficient environment
Figure 2: KLF1 target genes and associated clinical phenotypes. KLF1 is a master regulator of ∼700 genes in human erythroid cells involved in a wide variety of molecular processes (blue circles). Deregulated expression of a subset or all of these genes, depending on the KLF1 variant, leads to a diverse array of erythroid phenotypes (gray circles). HbA, adult hemoglobin (α2β2); HbA2, adult hemoglobin 2 (α2δ2); PK, pyruvate kinase; ZnPP, zinc protoporphyrin.
EMBO Lab Management Course
15 to 18 July 2019
EMBO | EMBL Symposium
The non-coding genome