Objective
The project will be applied to the study of common event which occurs in chronic myeloid leukemia (CML) cells, namely the expression of a group of tumor associated antigens (TAA) such as the PRAME protein and other MAGE type antigens, which usually are not expressed in normal tissues, and are recognizable by autologous cytotoxic lymphocytes (CTL) in the contex of specific HLA-I allospecifical restrictions.
Aims of the project include:
- Defining the expression patterns both at mRNA and protein level, of PRAME genes and the other TAA genes in hemopoietic CML precursors and in cell lines.
- Clarifying the molecular mechanisms associated to the expression of the genes in CML cells.
- Elucidating the contribution of PRAME and other TAA gene expression in CML progression.
- Establishing the clinical significance of detection of PRAME and other TAA gene expression in monitoring and prognosis of CML.
- Investigating the possibility to use PRAME gene expression for development of active immunotherapy of CML through the generation of autologous leukemia-derived dendridic cells (LD-DCs).
Co-ordinator of the project is F. Pane, Professor, MD, head of Molecular Hematology Unit Department of Biochemistry and Biotechnology Medicine University Napoli Federico II. In the realization of the project 4 other teams take part: Italian team (leader - G. Saglio, Professor, MD), Latvian (leader - Petuchov V, Professor, MD), Russian (leader - Misyurin A, Dr), and Ukrainian (leader - Abramenko I., MD).
It is planned to develop PRAME-containing construct using the sheep metallothionine promoter, plasmid pMT-CB6, retrovirus and adenovirus vectors; to perform transfection of this vector into Ph+ and Ph- human hematopoietic cell lines to induce the expression of PRAME gene; to study cell cycle parameters and sensitivity to drug-induced apoptosis of the PRAME-expressing cell lines.
Significance part of investigation will be devoted to evaluation of PRAME gene expression in conditions of down-regulation of BCR/ABL gene expression (by the use of specific antisense oligonucleotides complementary to the breakpoint between BCR and ABL sequences and IFN-?) and under inhibition of BCR/ABL kinase activity (STI571).
The influence of PI3K-dependent pathway on the expression of PRAME gene will be investigated under the drugs wortmannin and Ly 294002, specific inhibitors of PI3K. For study Jak/STAT pathway will plan to use AG490 as an inhibitor of the Janus kinase 2 (JAK2).
During all period of duration of the project joint efforts will be move to elucidate the clinical significance of detection of the PRAME gene expression in CML and to develop of new therapeutic strategies of CML based on CTL immune response.
So, we will plan to receive new data about oncogenic role of PRAME expression, signal transduction pathways for induction and regulation of PRAME gene expression during CML progression; to develop of scheme accurate monitoring CML patients under treatment (INF-?, STI571) or after BMT, to create of innovative approach to generate and purify PRAME-specific CTL.
The project has significance relevance to human health problems and should answer some fundamental questions about PRAME gene expression in CML progression and should provide valuable information that may improve treatment for this disease.
Topic(s)
Call for proposal
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80131 Napoli
Italy