LEUKEMOGENESISAPL

An in-vivo Screen for the Identification of Leukemia-Promoting Factors

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 Obiettivo del progetto (Objective)

'The proposed research aims to identify and characterize factors conferring leukemic properties to hematopoietic cells. This screen will be based upon a murine leukemia model I have developed in my time at the Salk Institute (La Jolla, CA) these mice express a fusion of RARalpha with three FKBP12 Rapamycin-regulated dimerization domains” FKBP12 F36M (F3-RARa). These F3-RARa TG mice, though healthy, exhibit a “preleukemic” state that is more accessible to transformation by cellular factors, such as activated cytokine receptors. (Sternsdorf et al, 2006). At low frequency, these mice spontaneously develop myeloid leukemia with APL features. From these leukemias I want to identify leukemogenic factors. Freshly expanded Leukemia cells will be used to generate a retroviral cDNA library. Initially, I will use this library to transduce non-transplantable murine cell lines and test, which factors convey transplantability. In the following, I will transduce /transplant pre-leukemic bone marrow from younger F3-RARa mice. This should lead to leukemias, which will be tested, isolated by cell sorting, according to presence of viral integrate (GFP). Leukemias will be FACS-isolated by phenotype (GFP, Surface markers). The integrated retroviral vectors will be isolated by PCR. The goal of this analysis is to identify and characterize novel cellular factors, involved in the process of leukemia development. This will enable me to identify novel potential targets for therapy. I am an experienced scientist in the earlier part of my career: after spending nine years in the US, I am returning to the European Union. A Heisenberg Fellowship (Germany) made this possible. This fellowship, though prestigious, covers only a small amount of funding besides my salary. Therefore I consider the IRG a critical component of funding in this critical time of my career and the successful transfer of my knowledge to the hosting lab in Hamburg.'

Introduzione (Teaser)

Understanding the molecular determinants of cancer is the first step for designing new targeted therapies. Researchers worked on understanding acute promyelocytic leukaemia (APL) at the molecular level.

Descrizione progetto (Article)

Myeloid leukaemia is a type of blood cancer, which very often arises from chromosomal rearrangements in the blood-forming cells in the bone marrow. These rearrangements lead to fusion proteins and in the case of APL this protein is called PML-RAR.

PML-RAR is believed to disrupt an important anti-tumour mechanism known as cellular senescence. Senescence is the gradual deterioration of function, characteristic of biological ageing and is thought to represent a first-line of defence against cancerous changes.

In APL cells, cellular senescence involves the assembly of a protein complex that regulates the organisation of cellular DNA. The EU-funded (LEUKEMOGENESISAPL) (An in-vivo screen for the identification of leukemia-promoting factors) project worked on further delineating the mechanism by which chromatin organisation is altered in APL.

Researchers developed a novel ex vivo method for studying and quantifying the activity of the PML-RAR protein and other oncogenic factors. This assay significantly reduced the required number of performed animal experiments to unveil the role of this fusion product.

Scientists identified that in cellular senescence, the protein PAX-complex regulates the chromatin landscape and implicates PML. The PML-RAR fusion protein disrupts PAX assembly causing a failure to enter senescence, and cells are thus more sensitive to leukemogenicity.

Given the enzymatic activity of the PAX-complex, the consortium proposed that it could serve as a target for therapeutic purposes. Therefore, drugs which work through induction of senescence could be developed as an anti-cancer strategy without the need of genotoxic drugs.