Proiect nucleu PN19.29 (contract 1N/2019) SISPATH
Abordari sistemice in identificarea si/sau validarea de tinte moleculare pentru strategii terapeutice in patologii cu impact major pentru sanatate
Coordinating Institution: Institutul National de Cercetare-Dezvoltare in Domeniul Patologiei si Stiintelor Biomedicale “Victor Babes” (IVB), Director Proiect: Prof. Mihail Eugen Hinescu
Title of project PN19.29.01.03: Technological innovation regarding the detection of genetic fingerprints – applications in the monitoring of residual tumor cells as well as of the hematopoietic regeneration post-transplant
Budget of PN19.29.01.03: 4420000 RON
Principal investigator of PN19.29.01.03:
Dr. Valeriu Cismasiu (2019-2020)
Dr. Laura Cristina Ceafalan (2021-2022)
Dr. Laura Cristina Ceafalan (2021-2022) Team members of PN19.29.01.03: Sevinci Pop, Gisela Gaina, Cristina Niculite, Stefania Rogozea/Alexandra Popa (Gruianu), Ioana Lambrescu, Victor Ionescu, Aurora Arghir, Sorina Papuc, Alina Erbescu
Abstract
Molecular tests are essential for a complete diagnosis and a targeted treatment of the patient. The response to treatment is difficult to predict; therefore, patients should be monitored by molecular testing to determine whether treatment is effective or needs to be adjusted. The large-scale application of modern sequencing methods has led to major advances in molecular diagnostics. Thus, new mutations associated with different types of oncological pathologies were discovered, allowing a better diagnosis and a more efficient prognosis. The field is constantly evolving, with established mutation sets being reclassified annually in terms of their use in diagnosis and prognosis. The aim of the project is the generation of molecular kits suitable for modern PCR methods, for the identification of cellular genetic fingerprints. The study is designed so that (1) the obtained results are useful in laboratory medicine, (2) the selected genes are specific of a diverse spectrum of tumors, and (3) the optimized technique is applicable for the quantification of several types of genetic mutations.
Objectives
The general objective of the project is to produce and optimize kits for the evaluation of genetic fingerprints (mutations) by modern PCR techniques, namely TaqMan, ddPCR and rhPCR. The specific objectives: to set up methods for analysis of genetic anomalies for which there are no such procedures in clinical laboratory; new methods with increased performance compared to those currently used in the clinic.
Patents (pending)
RO134998: Cismasiu VB et al. 2019; Set of two primers and one probe for detection and quantification of the mutant gene NPM1
RO135856: Cismasiu VB et al. 2020; Human Genetic Identification through the detection of mutations like insertions and deletions
Publications
Budisteanu, M., Papuc, S. M., Erbescu, A., Iliescu, C., Dobre, M., Barca, D., Tarta-Arsene, O., Motoescu, C., Dica, A., Sandu, C., Anghelescu, C., Craiu, D., & Arghir, A. (2022). Clinical and genomic findings in brain heterotopia: Report of a pediatric patient cohort from Romania. Experimental and therapeutic medicine, 23(1), 101. https://doi.org/10.3892/etm.2021.11024
Papuc, S. M., Erbescu, A., Cisleanu, D., Ozunu, D., Enache, C., Dumitru, I., Lupoaia Andrus, E., Gaman, M., Popov, V. M., Dobre, M., Stanca, O., Angelescu, S., Berbec, N., Colita, A., Vladareanu, A. M., Bumbea, H., & Arghir, A. (2021). Delineation of Molecular Lesions in Acute Myeloid Leukemia Patients at Diagnosis: Integrated Next Generation Sequencing and Cytogenomic Studies. Genes, 12(6), 846. https://doi.org/10.3390/genes12060846
Lambrescu, I., Popa, A., Manole, E., Ceafalan, L. C., & Gaina, G. (2022). Application of Droplet Digital PCR Technology in Muscular Dystrophies Research. International journal of molecular sciences, 23(9), 4802. https://doi.org/10.3390/ijms23094802
Gaina, G., & Popa Gruianu, A. (2021). Muscular dystrophy: Experimental animal models and therapeutic approaches (Review). Experimental and therapeutic medicine, 21(6), 610. https://doi.org/10.3892/etm.2021.10042
Gaina, G., Vossen, R. H. A. M., Manole, E., Plesca, D. A., & Ionica, E. (2021). Combining Protein Expression and Molecular Data Improves Mutation Characterization of Dystrophinopathies. Frontiers in neurology, 12, 718396. https://doi.org/10.3389/fneur.2021.718396
Lambrescu, I. M., Lambrescu, I. M., Martin, S., Martin, S., Cima, L. N., Cima, L. N., Fica, S., & Fica, S. (2022). Assessment of peripheral blood cells parameters as a valuable tool in patients with neuroendocrine neoplasms. Neuro endocrinology letters, 43(1), 45–54.
Fertig, T. E., Chitoiu, L., Marta, D. S., Ionescu, V. S., Cismasiu, V. B., Radu, E., Angheluta, G., Dobre, M., Serbanescu, A., Hinescu, M. E., & Gherghiceanu, M. (2022). Vaccine mRNA Can Be Detected in Blood at 15 Days Post-Vaccination. Biomedicines, 10(7), 1538. https://doi.org/10.3390/biomedicines10071538
Ionescu, V. S., Popa, A., Alexandru, A., Manole, E., Neagu, M., & Pop, S. (2021). Dietary Phytoestrogens and Their Metabolites as Epigenetic Modulators with Impact on Human Health. Antioxidants (Basel, Switzerland), 10(12), 1893. https://doi.org/10.3390/antiox10121893
Comunicari
1) Improved Chimerism Detection through RNA-based rhAmp Genotyping. Ionescu V et al., Modern Biotechnical Advances for Human Health (BAHH2) – Bucharest, 28-31.05.2019
2) Unlimited Gene Expression Meta-analysis through Microarray Data Mining. Ionescu V et al., the 12th National Pathology Symposium, Bucharest, 21-23.11.2019, Abstract Book page 86, ISSN 2601-0770;
3) Blending the precision of RNase h-dependent PCR with the accuracy of digital droplet PCR – the good, the bad and the “rain”. Lambrescu I et al., the 12th National Pathology Symposium, Bucharest, 21-23.11.2019, Abstract Book pag 92, ISSN 2601-0770;
4) Novel PCR based tools in molecular diagnostics. Cismasiu V., the 12th National Pathology Symposium, Bucharest, 21-23.11.2019, Abstract Book p36, ISSN 2601-0770;
5) Molecular characterization of acute myeloid leukemia by next generation sequencing and microarray technologies. Arghir A et al., European Conference of Cytogenetics, Salzburg, Austria, 6-9.07.2019, Molecular Cytogenetics, Volume 12, Supplement: 1, Meeting Abstract: 2. pag 12;
6) Sanger sequencing and fragment analysis for FLT3-ITD molecular characterization in acute myeloid leukemia patients at diagnostic. Erbescu A et al., Conference of Medical genetics, Timisoara, 18-20.09.2019, Romanian Journal of Rare Diseases, Supplement 2019, pag 47, ISSN 2068-5882
7) Molecular characterization of flt3-itd in acute myeloid leukemia patients at diagnostic, Erbescu A et al., Conference of the Romanian Association of Laboratory Medicine, Bucharest 2020
8) Molecular characterization of acute myeloid leukemia by next generation sequencing, Papuc SM et al., Conference of the Romanian Association of Laboratory Medicine, Bucharest 2020
English 
Romanian