Project summary
General objectives and the estimated results of the project
Coordinating research tea
Contracting Authority
The activities and responsabilities of the participants
The budget of the project
The results obtained in each stages during the project
Contact details of the project
Final results of the project
Publications
Project summary:
CERVIX-array subscribes to Health area 4.1.3. Investigation and interventional methods based on molecular and cellular medicine, genomics and proteomics.
The clinical need for molecular biomarkers at the transcriptome level in cancer has led to the development of genomics and proteomics-complex tools able to provide complex data to both the researcher and the clinician.
This proposed project is based on a highly complex partnership and seeks to develop the interdisciplinary sciences and high tech fields of genomics and proteomics in Romania in the field of oncology. It would study cervical tumoral pathology and would employ the newest technologies of genomics (microarray) and proteomics (protein-array); to identify new molecular targets that would make possible the creation of personalized therapy and it would have predictive prognostical value for existing and developing treatments in advanced stage IIB-IIIB cervical cancer. In Romania cervical cancer is a very real public health problem as we are in the first place in Europe and 6th place in the world for cervical cancer incidence . Existing treatments of cervical cancer ensures a 5 year survival rate of 66-76% for stage IIB, 45-56% for stage IIIA, and 36-40% for stage IIIB.
The time period of the project is 36 months.
Internationally- the last data (august 2007) provided by the UpToDate data base which collates information from over 375 scientific journals and other sources in the field of epidemiology indicate an inequality in the morbidity and mortality of cervical cancer in the world. The highest rates occur in developing countries; where it is the 2nd leading cause of mortality (50%) and morbidity due to cancer in the female population.
Nationally- according to data provided by CNOASIIDS there is a continuous growth in the number of new cases of cervical cancer. In 2005 cervical cancer represented 12.95% of cancers in the female population (3202 cases out of 24,732) in second place after breast cancer. The data of the Cancer Institute in Cluj Napoca confirms the alarming growth of the number of new cases of cervical cancer diagnosed in an advanced stage (IIB-IIIB) the number almost doubling between 2000 and 2007 from 608 to 1017 new cases. At the national level there are no complex genomics and proteomics studies of advanced stage IIB - IIIB cervical cancer with a concurrent evaluation of the high risk HPV type (13 types), due to the complex nature of analysis, the highly advanced infrastructure required (genomics platform), and the lack of specialists in the domain of functional genomics.
The general objectives of this project consist of: ♦ The identification of the molecular transcriptomic profiles of advanced stage (IIB-IIIB) cervical cancer through functional genomics studies as a predictive factor for the theraputic response to radiotherapy ♦ The correlation of high risk HPV types with the genomic differentiation clusters and the elucidation of the differences in tumor agression and patient survival rates.
These objectives can be accomplished through the creation of an experienced and professional multidisciplinary research team coordinated by IOCN- the cancer institute with renown experience in cell biology , functional genomics, and applied clinical research: P1- by the University of Medicine and Pharmacy Cluj-Napoca- university with European renown in research and the potential to attract FP7 program partners; ; ISP –comparative diagnostic data analysis and the development of screening strategies.
General objectives and the estimated results of the project:
General objectives:
♦ the identification of the transcriptomic molecular profiles of advanced cervical cancer (stage IIB-IIIB) through functional genomic studies as a predictive factor for pacient response to radiation therapy;
♦ the correlation of high risk HPV types with the genomic differentiation clusters in order to define the differences in tumor agression and implicitly, pacient survival.
Specific objectives:
♦ the exact grouping of pacients according to inclusion and exclusion characteristics (age; stage; imagistic evaluation; guided tumor biopsy; hormonal status; tumor serum markers; the lack of anterior chemotherapy);
♦ the analysis of a set of high risk HPV types through qRT-PCR; ♦ the performance of the different stages of the genomic study through whole human genome microarray;
♦ the identification through specific molecular signatures of each different advanced stage for the elucidation of the differential gene expression after radiation therapy;
♦ the correlation of the clinical response to radiation therapy with genomic and proteomic data (molecular signature);
♦ the validation through tissue array of the principal tissue proteins associated with the evaluated genes;
♦ the evaluation of the angiogenic serum markers implicated in the process of invasion and methastasis and immune response (Th1/Th2) through protein-array; ♦ the implication of risk factors in resistance to radiotherapy (the realization of a follow-up protocol for life-style and genetic factors).
Coordinating research team:
„ Prof Dr. Ion Chiricuta” Cancer Institute Cluj-Napoca (IOCN).
Consortium components:
Coordinator of the multidisciplinary consortium:
The“ Ion Chiricuta” Cancer Institute – Comprehensive Cancer Center (IOCN),
Project partners:
the University of Medicine and Pharmacy Cluj-Napoca- (UMFCN
The Institute of Public Health “Iuliu Moldovan„ Cluj-Napoca (ISPCN),
Contracting Authority:
the National Program of RDI.—PN II-partnership.
Project code: 42-160/2008
Project time span:: (36 month) 01.10.2008— 30.09.2011
The activities and responsabilities of the participants:
A1. Basic Research
Literature databases search and research of the data in accordance with proposed topic
1.1 Literature data analysis in accordance with proposed topic (A1.1)
1.2 Selection criteria definition (A 1.2)
1.3 Web page design (A 1.2)
Responsabilities
CO-Update in genomics and proteomics studies of cervical cancer
P1-Oncological updates in cervical cancer; Work procedures harmonized to the EU legislation and international regulations regarding human tissues processing as support for biomedical research.
P2-Literature data review and analysis regarding the risk factors related to lifestyle and environment involved in cervical cancer ethiopathogenesis, progression and prognosis
A.2 Industrial Research.
Starting of CERVIX-array study.
Patients selection in conformity with the Project’s Execution Plan
2.1 Study design
2.2 Set-up patients groups in accordance with the approved study criteria (A 2.2)
2.3 Establishment of patients’ databases. Biobank establishment; Collection of biological samples (tissue,
blood) (A 2.4; A2.6)s
2.4 Evaluation of high risk viral HPV load (A 2.1)
2.5 Processing of collected biological samples and completion of the data base (A 2.5; A 2.6)
B. Support activities
2.5 Partial results dissemination (B2; B3)
2.6 Participation in specializing and training courses (B 4)
Responsabilities
CO - Experimental Design and Functional Genomics and Proteomics Protocols; Setup of the patients selection files clearly defining selection and exclusion criteria; Collection of biological samples; Standardization of selection methods for biological samples into Biobank and management of collected specimens.
P1 – Defines the activities necessary for the genomics and proteomics study
-establishes the ethical conditions and standardization of working protocols for biological samples according to European and international standards
P2 - Design of working outputs; Consent Forms, Risk Factors Questionnaires about lifestyle and environment, Questionnaire Validation;
CO, P1, P2 -assurance of support activities
Industrial Research.
Biobank enhancement; Setting-up Genomics and Proteomics Study
3.1 Biobank Enhancement. Prelevation of new biological samples (A 2.1)
3.2 Biological samples processing and completion of the data base (A 2.5; A 2.6)
3.3 Microarray probes synthesis (Cy-cRNA) and quantitative and qualitative evaluation of microarray probes (A 2.1)
Support activities
3.4 Partial results dissemination (B2; B3)
3.5 Participation in specializing and training courses (B 4)
Responsabilities
CO – completion of IT databases ; collection of new biological samples; Biobank enhancement with new biological samples; tumor RNA extraction; Microarray probes synthesis (Cy-cDNA);
P1 – Nanotechnology analysis of microarray probes; processing of biological samples;
CO, P1-assurance of support activities
A.2 Industrial Research.
Genomics and Proteomics Study Implementation
4.1 Setting-up microarray study on WHG (Whole Human Genome) glass slides (A 2.1)
4.2 Bioinformatics analysis of microarray data (A 2.6)
4.3 Protein-array study (Th1/Th2 and angiogenesis) (A 2.1)
4.4 Correlation of microarray and protein-array data with post-treatment clinical evaluation (A 2.6)
B. Support activities.
4.5 Partial results dissemination (B 2; B 3)
4.6 Participation in specializing and training courses (B 4)
CO-Microarray analysis on WHG (Whole Human Genome) glass slides and bioinformatics analysis of microarray data
P1 - evaluation of serum proteins involved in angiogenesis and immune response through Fast Quant technology; processing and realization of protein array study (Fast Quant)
CO, P1 - Correlation of microarray and protein-array data with post-treatment clinical evaluation
A.2 Industrial Research.
Data Validation.
Results dissemination and revaluation.
5.1 Validation of the genomics data through qRT-PCR and identification of predictive therapy biomarkers
B. Support activities
5.2 Large scale data dissemination through communication and publication (B 2; B 3)
5.3 Connection to national/European networks (B 6)
5.4 Working visits/ Training exchange (B 7)
5.5 Identification and assignation of intellectual property rights over the results (B 1)
Responsabilities
CO - Genomics data validation by RT-PCR
P1-In silico study for identification of optimal conditions for qRT-PCR (structure of primers and fluorescent probes-UPL)
CO, P1, P2 –dissemination and revaluation of the results
The budget of the project
Currency project:
2.000.0000 RON state budget; 0 RON co
The results obtained in each stages during the project :
Stage I (01.10.2008-15.02.2009)
– Documentations (Report of state-of-the art) on studies involving genomics and proteomics in cervical cancer
– Documentations (Report of state-of the art) on the methods of genomic analysis
– Diagrams (Questionnaires) concerning the definition of lots of patients on the basis of inclusion and exclusion criterions for advanced cervical cancer
– Methods (Questionnaires) assesment of risk factors related to lifestyle and environment of etiopatogenia,evolution and prognosis of cervical.
– Methods of analysis, biological samples; working protocols for collection and processing of human biological samples that support biomedical research
Stage I I : 16.02.2009-15.12 2009
– Documentations (Report of state-of-the art) on the optimal conditions and the types of evaluations of genomics and proteomics.
– The conceptual model (data base) – to include patients in the data base (clinical data and molecular biology). Storing samples in biobanks
– Methods of analysis, biological samples collection and processing of biological samples (tissue, blood, vaginal citology)
Stage III : 16.12.2009-10.12 2010
– The conceptual model (data base) – to include patients in the data base (clinical data and molecular biology). Storing samples in biobanks
– Methods of analysis, biological samples collection and processing of biological samples (tissue, blood, vaginal citology);
– Methods of analysis, biological samples collection, HPV genotyping
– Analysis of risk factors related to lifestyle and environment of etiopatogenia, evolution and prognosis of cervical cancer.
Contact details of the project:
Contact:
Dr. Ovidiu Balacescu
„Prof Dr. Ion Chiricuta” Cancer Institute Cluj-Napoca
Department of Functional Genomics, Proteomics and Experimental Pathology
Republicii str, no 34-36, Cluj Napoca, Cod 400015
Tel: 0264-590638, Fax: 0264-590638
Email: obalacescu@yahoo.com; ovidiubalacescu@iocn.ro
Final results of the project :
A modern method of investigation in medicine that may be a predictor of effective treatment with radio-chemotheraphy in cervical cancer with a known load viral HPV.
P. Publications
1. Balacescu O, Balacescu L, Tudoran O, Todor N, Rus M, Buiga R, Susman S, Fetica B, Pop L, Maja L, Visan S, Ordeanu C, Berindan-Neagoe I, Nagy V. Gene expression profiling reveals activation of the FA/BRCA pathway in advanced squamous cervical cancer with intrinsic resistance and therapy failure. BMC Cancer. 2014 Apr 8;14:246. doi: 10.1186/1471-2407-14-246.
2. Oana Tudoran, Olga Soritau, Ovidiu Balacescu, Loredana Balacescu, Cornelia Braicu, Meda Rus, Claudia Gherman, Florin Irimie, Ioana Berindan Neagoe. Early transcriptional pattern of angiogenesis induced by EGCG treatment in cervical tumor cells - J Cell Mol Med. 2012 Mar;16(3):520-30. doi: 10.1111/j.1582-4934.2011.01346.x.
3. Berindan-Neagoe I, Braicu C, Tudoran O, Balacescu O, Irimie A, Early apoptosis signals induced by a low dose of epigallocatechin 3-gallate interfere with apoptotic and cell death pathways, Journal of Nanoscience and Nanotechnology, manuscript accepted for publication 21.09.2011.
4. Sorina Irimie, Mariana Vlad, Ileana Maria Mirestean, Ovidiu Balacescu, Rus Meda, #Loredana Balacescu, Ioana Berindan-Neagoe, Rares Buiga, Oana Tudoran, Viorica Nagy, Alexandru Irimie. Risk profile in a sample of patients with advanced cervical cancer. Applied Medical Informatics, volumul 29(4), 2011; pag 1-10 (BDI).
5. Meda Rus, Loredana Balacescu, Oana Tudoran, Cornelia Braicu, Ioana Berindan-Neagoe, R.Buiga, Viorica Nagy, N.Todor, Manuela Banciu, N.Dragos, O.Balacescu. Could HPV hight risk genotypes predict the response to the therapy in cervical cancer? Annals of RSCB, (2011),vol XVI, nr 2, 121-127 (BDI).
National Patent (OSIM): No 128780 from 26.02.2016, (BOPI nr 2/2016) entitled Method of prediction of response to chemoradiotherapy in cervical cancer through functional genomics studies. (Dr. Ovidiu Balacescu, Dr. Loredana Balacescu, Prof. Dr. Ioana Neagoe, Prof Dr. Alexandru Irimie)