Cercetător Senior Constantin Carolina

Cercetător Senior Constantin Carolina

Institutul Naţional de Cercetare-Dezvoltare în Domeniul Patologiei şi Ştiinţelor Biomedicale “Victor Babeş” Bucureşti
Serviciul de Anatomie Patologică, Spitalul Clinic Colentina, Bucureşti

Perspective epigenomice ca sursă de biomarkeri în tumorile cutanate
Autori: Carolina Constantin1,2, Mihaela Surcel1, Adriana Narcisa Munteanu1,3, Ana Căruntu4, C. Căruntu5, Sabina Andrada Zurac2, 6, Monica Neagu1,2,3
1Secția Imunologie, Institutul Naţional de Cercetare-Dezvoltare în Domeniul Patologiei şi Ştiinţelor Biomedicale “Victor Babeş” Bucureşti
2Serviciul de Anatomie Patologică, Spitalul Clinic Colentina, Bucureşti
3Școala Doctorală, Facultatea de Biologie, Universitatea Bucureşti
4Departmentul de Chirurgie Orală și Maxilofacială, Spitalul de Urgență Militar Central “Dr. Carol Davila”, Bucureşti
5Departamentul de Fiziologie, Universitatea de Medicină şi Farmacie “Carol Davila”, Bucureşti
6Departamentul de Patologie, Facultatea de Medicină Dentară, Universitatea de Medicină şi Farmacie “Carol Davila”, Bucureşti

Cuvinte cheie: biomarker, epigenom, metilare, tumori piele

Introducere: Epigenomul include o componenta relativ recentă – epitranscriptomul, ce definește modificările suferite de ARN, și care se alatură epigeneticii în lărgirea arsenalului de markeri în contextul tumoral. Majoritatea studiilor legate de epigenetica proceselor tumorale se focusează pe căile oncogene tumorale la nivelul tumorii primare, astfel încât efectele și rolul modificării de tipul N6-metiladenozina (m6A) în ARN-ul din celulele sângelui periferic, sunt extrem de puțin abordate, fiind în mare parte necunoscute. În tumorile cutanate, studiile privind valorile potențiale de diagnostic și prognostic ale leucocitelor periferice prin prisma modificărilor epitranscriptomice m6A pot furniza noi markeri pentru managementul bolii.
Material și metodă: Celule mononucleare (PBMC) au fost izolate din sange periferic de la subiecti cu carcinom bazocelular (BCC), spinocelular (SCC) sau melanom cutanat. Din suspensiile celulare s-a extras ARN total la care s-a evaluat gradul de metilare m6A printr-o platformă de tip ELISA bazată pe anticorpi specifici anti-m6A.
Rezultate: Indicele cât și procentul de metilare m6A la nivelul ARN din leucocite PBMC este crescut la tumorile de piele comparativ cu normal. De asemenea, există diferențe în nivelul de metilare la tumori non-melanom versus melanom, în cazul melanomului cutanat indicele m6A fiind dublu față de SCC și de cel putin 10x mai ridicat comparativ cu BCC.
Concluzii: Nivelul m6A poate conferi identitate transcriptomică în leucocitele periferie și poate constitui un nou biomarker de graniță epi-imun, neinvaziv, cu utilitate potențială atât în discriminarea subtipurilor de tumori cutanate dar și în monitorizarea clinică a pacientului.
Studiu finanțat prin proiectele: PN-III-P1-1.2-PCCDI-2017-0341/2018, PN 19.29.01.01/2019, EPITRAN COST ACTION 16120/2018.

Epigenomic perspectives as biomarkers source in skin cancers
Autori: Carolina Constantin1,2, Mihaela Surcel1, Adriana Narcisa Munteanu1,3, Ana Căruntu4, C. Căruntu5, Sabina Andrada Zurac 2, 6, Monica Neagu 1, 2, 3
1Immunology Department, „Victor Babes” National Institute of Pathology, Bucharest
2Pathology Department, „Colentina” University Hospital, Bucharest
3Doctoral School, Faculty of Biology, University of Bucharest, Bucharest
4Department of Oral and Maxillofacial Surgery, “Dr. Carol Davila” Central Military Emergency Hospital, Bucharest
5Department of Physiology, “Carol Davila” University of Medicine and Pharmacy, Bucharest
6Pathology Department, Faculty of Dental Medicine, “Carol Davila” University of Medicine and Pharmacy, Bucharest

Keywords: biomarker, epigenom, methylation, skin tumors

Introduction. The epigenome includes a relatively recent component – the epitranscriptome, which defines the changes in RNA, and joins epigenetics in expanding the arsenal of tumor markers. Most studies related to the epigenetics of tumor processes focus on tumor oncogenic pathways in the primary tumor, so the effects and role of N6-methyladenosine (m6A) modification in RNA in peripheral blood cells are extremely poorly addressed. In skin tumors, studies on the potential diagnostic and prognostic values of peripheral leukocytes in the light of m6A epitranscriptomic changes may provide new markers for disease management.
Material and method. Mononuclear cells (PBMC) were isolated from peripheral blood from subjects with basal cell carcinoma (BCC), squamous cell carcinoma (SCC) or cutaneous melanoma. From the cell suspensions, total RNA was extracted and the m6A methylation level was assessed by an ELISA-type platform based on specific anti-m6A antibodies.
Results. The index and the percentage of m6A methylation in RNA from PBMC leukocytes is increased in skin tumors compared to normal. There are also differences in the level of methylation in non-melanoma versus melanoma, in the case of cutaneous melanoma the m6A index is double that of SCC and at least ten fold higher compared to BCC.
Conclusions. The m6A level can confer transcriptomic identity in the peripheral leukocytes and can be a new biomarker of epi-immune border, non-invasive, with potential utility both in discriminating skin tumor subtypes and in clinical monitoring of the patient.
Study financed by the following projects: PN-III-P1-1.2-PCCDI-2017-0341/2018, PN 19.29.01.01/2019, EPITRAN COST ACTION 16120/2018.

Scurt CV

Senior researcher I, PhD, biochemist, “Victor Babeș” National Institute of Pathology, Immunology Laboratory & Colentina Clinical Hospital, Research Core of Pathology Department, Bucharest.
Research topics: immune parameters analysis and biomarker discovery in melanoma and non-melanoma skin cancer; co-inventor for 4 patents related to photodynamic therapy PDT in skin pathologies. Dr. Constantin conducted national research projects related to novel photosensitizers for PDT in cancer, and drug-delivery nanosystems for skin regeneration; partner responsible for MNT_ERANET project 7030-1/2010 related to novel porphyrins in theranostics. Specialization in molecular biology at Ludwig Institute for Cancer Research, Signal Transduction Unit (Bruxelles); cellular biology specialization at University of Athens, Biochemistry and Physiology Department, in the frame of NATO SfP 982838/2007 project. Technical expert in 5 EU projects for biomedical assays; teaching experience in protein microarray technology; volunteer reviewer for several ISI journals.
Currently involved as MC member for COST Action CA18127 – International Nucleome Consortium (INC); MC substitute of the management committee for COST action CA16120 – European Epitranscriptomics Network (EPITRAN); alternate member of the management committee for CA18125 – Advanced Engineering and Research of Aerogels for Environment and Life Sciences (AEROGELS).