Abstract
Objective
To investigate genetic subtypes of inherited bone marrow failure syndrome Fanconi anemia (FA) in Sebia. FA-D2 subtype was found to be the most frequent genetic subtype among investigated FA patients; specific observations of FA-D2 phenotype are pointed out.
Methods
Several biological endpoints of FA cells in vitro such as radiation-induced level of lymphocyte micronuclei (radiosensitivity), base line and radiation induced level of the DNA double strand breaks (DSBs), leukocyte apoptosis, and telomere capping function were assessed.
Results
The results indicate that all FA-D2 patients display radioresistant in vitro response, which is seen as significantly reduced yield of radiation-induced micronuclei. On the contrary, FA-A patients display radiosensitive in vitro response seen as increased number of radiation-induced micronuclei (MN). A massive elimination of irradiated cells via apoptosis is found in both FA-A and FA-D2 subtypes. In FA-A subtype apoptosis positively relates with the yield of radiation-induced MN, whereas in FA-D2 subtype apoptosis relates with a high percentage of cells carrying dysfunctional telomeres. The present results unequivocally demonstrate that cytokinesis-block micronucleus (CBMN) assay and analyses of telomere capping function can be used to distinguish FA-D2 and FA-A complementation groups.
Conclusions
Considering all biological endpoints were analyzed, it can be concluded that all FA patients are radiosensitive, regardless of their complementation group. Thus, using CBMN test and telomere capping function analysis can discriminate FA-A from FA-D2 complementation groups, which could be important for assessment the conditioning regimens prior to bone marrow transplantation.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Dokal I, Vulliamy T. Inherited bone marrow failure syndromes. Haematologica. 2010;95:1236–40.
Alter BP. Diagnosis, genetics, and management of inherited bone marrow failure syndromes. Hematology Am Soc Hematol Educ Program. 2007:29–39.
Al-Haggar M, Al-Morsy Z, Yahia S, Chalaby N, Ragab A, Mesbah A. Correlation of thyroid and growth hormones to chromosomal instability in Egyptian Fanconi anemia patients. Indian J Pediatr. 2008;75:679–84.
D'Souza F, Usha MK, Subba Rao SD. Fanconi's anemia in monozygotic twins. Indian J Pediatr. 2007;74:859–61.
Buchwald M. Complementation groups: One or more per gene? Nat Genet. 1995;11:228–30.
Sasaki MS, Tonomura A. A high susceptibility of Fanconi's anemia to chromosome breakage by DNA cross-linking agents. Cancer Res. 1973;33:1829–36.
Auerbach AD. Diagnosis of fanconi anemia by diepoxybutane analysis. Curr Protoc Hum Genet. 2003;Chapter 8:Unit 8.7.
Pinto FO, Leblanc T, Chamousset D, Le Roux G, Brethon B, Cassinat B, et al. Diagnosis of Fanconi anemia in patients with bone marrow failure. Haematologica. 2009;94:487–95.
Fenech M. The cytokinesis-block micronucleus technique: A detailed description of the method and its application to genotoxicity studies in human populations. Mutat Res. 1993;285:35–44.
Crompton NE, Ozsahin M. A versatile and rapid assay of radiosensitivity of peripheral blood leukocytes based on DNA and surface-marker assessment of cytotoxicity. Radiat Res. 1997;147:55–60.
Joksic G, Petrovic-Novak A, Stankovic M, Kovacevic M. Radiosensitivity of human lymphocytes in vitro correlates more with proliferative ability of cells than with the incidence of radiation-induced damages of the genome. Neoplasma. 1999;46:40–9.
Dickey JS, Redon CE, Nakamura AJ, Baird BJ, Sedelnikova OA, Bonner WM. H2AX: Functional roles and potential applications. Chromosoma. 2009;118:683–92.
Fenech M. The cytokinesis-block micronucleus technique and its application to genotoxicity studies in human populations. Environ Health Perspect. 1993;101:S101–7.
Pinar B, Henriquez-Hernandez LA, Lara PC, Bordon E, Rodriguez-Gallego C, Lloret M, et al. Radiation induced apoptosis and initial DNA damage are inversely related in locally advanced breast cancer patients. Radiat Oncol. 2010;5:85.
Fenech M. Cytokinesis-block micronucleus cytome assay. Nat Protoc. 2007;2:1084–104.
Djuzenova C, Flentje M, Plowman PN. Radiation response in vitro of fibroblasts from a fanconi anemia patient with marked clinical radiosensitivity. Strahlenther Onkol. 2004;180:789–97.
Hucl T, Gallmeier E. DNA repair: Exploiting the Fanconi anemia pathway as a potential therapeutic target. Physiol Res. 2011;60:453–65.
Acknowledgments
The authors are thankful to Prof. Dr. J. Surralles (Center for Biomedical Network Research on Rare Diseases (CIBERER), Bellaterra, Barcelona, Spain) for FA complementation group analysis. This research was supported by Ministry of Health and Ministry of Education, Science and Technological Development of the Republic of Serbia (Grant No. 173046).
Contribution
Joksić Gordana will act as guarantor for this paper.
Conflict of Interest
None.
Role of Funding Source
None.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Dragana, V., Sandra, P., Emilija, L. et al. Prevalence of FA-D2 Rare Complementation Group of Fanconi Anemia in Serbia. Indian J Pediatr 81, 260–265 (2014). https://doi.org/10.1007/s12098-013-1284-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12098-013-1284-4