MIUR: FIRB “Major Strategic Projects "2007-2011 - RBIP0695BB_001" Research and development of new agents for cancer immunotherapy through the study of genomics, proteomics and immunoproteomics # 27394B5
Project Objectives
The Research Unit aims at developing DNA vaccines and mono/multi epitope protein vaccine coding adjuvant and costimulatory molecules. The objective is to stimulate an antitumor immune response using single antigen epitopes, selected on their binding affinity for MHC I. The adjuvant function of CD4+ T helper cells will be ensured by the inclusion of T helper epitopes and molecules such as IL-2, IL-12, able to stimulate lymphocyte proliferation and activation.
Regarding immunotherapy against Ph+ leukemia, the first objective is to demonstrate the immunogenicity of out-of-frame peptides generated by alternative splicing of the BCR/ABL fusion gene through the study of the native protein sequence derived from the splicing and the identification, in the same sequence, of immunogenic epitopes to be optimized for increasing their binding ability to MHC molecules.
Epitope optimization and prediction of their binding ability to specific MHC molecules I and II will be performed by using the SYFPEITHI database and Bimas peptide ligands. Constructs for DNA and protein vaccination will be designed to also include promiscuous T helper epitopes, identified within the out-of-frame protein, thus increasing vaccine efficacy through the ability to activate numerous clones of T helper cells within the cellular population.
Regarding the study of human colorectal cancer, a tumor-associated antigen (Frizzled-like antigen) to be used for vaccine production has been detected. The effects of DNA vaccination encoding the Frizzled-like tumor-associated antigen and those of the corresponding peptide are evaluated to allow for the comparison of the efficacy of DNA vaccine versus the peptide vaccine. To this aim, the DNA vaccine is associated with chemo immunotherapy to increase expression and presentation of the Frizzled-like antigen.
Indeed, data from literature confirms that vaccination and chemotherapy-combined treatments are more effective. In order to design clinical trials on patients, the vaccines produced in this research project will be used in experimental protocols performed on specific animal models. For this purpose, transgenic animal models for human HLA antigens are selected because they have a antigen mechanism presentation that is more similar to that occurring in patients, thus allowing for study of immune response.
In particular, for the study of chronic myeloid leukemia, the murine strain C57/BL6, expressing the transgenic molecule HLA-A2.1 homozygous H-2b in the K region, is used. This is a suitable experimental model for the study of leukemia vaccination, since it expresses the human transgene HLA-A2.1.
The study of colorectal cancer involves the inoculation of DHD/K12 cells into a syngeneic strain of BDIX rats, a suitable model for the study of colorectal cancer vaccination because it naturally expresses the Frizzled-like human antigen.
Following experiments on animal models, parameters and criteria for submission to phase 1-2 clinical trials of human oriented vaccination will be evaluated. This Research Unit also includes the study of diagnostic imaging systems to be carried out within the project.
Start/End Date
July 18, 2007 - July 18, 2011
Principal Investigator
Prof. Vito Michele Fazio - Coordinator
Host Institution
Campus Bio-Medico University of Rome
Other Institutions involved
- Consiglio Nazionale delle Ricerche - Rome
- Consiglio Nazionale delle Ricerche - Milano
- Università degli Studi di Bari Aldo Moro
- Università degli Studi di Torino
- Università degli Studi di Cagliari
Source of funding
Italian Ministry of Education