Research Projects

Novel therapeutic targets for the treatment of leukemia

BRIC (UCPH) — Biotech Research and Innovation Centre

University of Copenhagen

Denmark

4/13

Supervision

Kristian Helin
BRIC, 1st Supervisor

Saverio Minucci
IEO, 2nd Supervisor

Objectives

To identify potential novel therapeutic targets for the treatment of leukemia, we have performed CRISPR-Cas9 based screens using libraries targeting chromatin-associated proteins in mouse models for leukemia and in human cell lines. These screens have led to the identification of several interesting hits, which should be validated in vivo before they will be further characterized.

The objective of the project is to understand the role of the validated hits in normal hematopoiesis and in leukemia with the goal of translating the findings into potential new treatments for leukemia patients.

The specific objectives include: i) validating the in vitro hits in transplantation models for mixed-lineage and Npm1-mutated leukemia, ii) perform structure-function analysis of the prioritized hit(s), including the identification of associated proteins; iii) performing studies to understand the role of the prioritized hits in transcriptional regulation and in regulating genome structures during hematopoietic differentiation and in leukemia; iv) testing (or developing) specific inhibitors for the prioritized hits in mouse and xenograft models for leukemia.

Methodology

Mouse transplantation experiments, CRISPR-Cas9 KO tiling screens, epitope tagging of endogenous genes using CRISPR-Cas9 technology, purification of proteins, mass spectrometry, chromatin-immunoprecipitations experiments, transcriptional studies, Hi-C studies, bioinformatics analyses, chemical inhibitor studies.

Expected Results

Validation of one-two new targets for the treatment of leukemia, including providing an understanding of the role in regulating transcription, genomic structures, haematopoiesis and leukemia.

Planned Secondments

CNAG-CRG, Spain (2 weeks):
Bioinformatics analysis
EMBL, Germany (1 month):
Hi-C experiments
IEO, Italy (1 month):
Drug testing

Enrolment in doctoral programs

PhD from the Graduate School of Health and Medical Sciences, University of Copenhagen

References

Agger, K. et al. Jmjd2/Kdm4 demethylases are required for expression of Il3ra and survival of acute myeloid leukemia cells. Genes Dev 30, 1278-1288, doi:10.1101/gad.280495.116 (2016).

Mohammad, F. et al. EZH2 is a potential therapeutic target for H3K27M-mutant pediatric gliomas. Nat Med 23, 483-492, doi:10.1038/nm.4293 (2017).

Rasmussen, K. D. et al. Loss of TET2 in hematopoietic cells leads to DNA hypermethylation of active enhancers and induction of leukemogenesis. Genes Dev 29, 910-922, doi:10.1101/gad.260174.115 (2015).