De Novo Chromosome Copy Number Variation in Fanconi Anemia-Associated Hematopoietic Defects
Abstract
The major goal of our research proposal was to determine the role of the FA-BRCA pathway in the suppression of spontaneous and reactive oxygen species (ROS)-induced de novo copy number variation (CNV). A major challenge of our experimental approach has been the isolation of monoclonal populations of hTERT-immortalized FA-A, FA-D2, and FA-G patient-derived cells. This considerable challenge significantly delayed our experimental progress. Nevertheless, through persistent efforts we successfully isolated small numbers of monoclonal populations of FA-D2 and FA-D2 + FANCD2 cells. We performed preliminary studies on these cells and determined that the frequency of spontaneous de novo CNVs is statistically significantly increased in the absence of FANCD2. We also developed an alternative approach to study the role of the FA proteins in the suppression of spontaneous and ROS-inducible CNV using short-interfering RNA (siRNA) in the HCT116 colorectal carcinoma cell line. Using this approach, we have also established that the FANCD2 protein, and not FANCA, is required for the suppression of spontaneous de novo CNV. These findings support a model whereby the FANCD2 protein, possibly independent of the FA core complex proteins, plays a critical role in the prevention of de novo pathogenic CNVs. Future studies will seek to elucidate the molecular mechanism(s) by which FANCD2 performs this key function.
Document Details
- Document Type
- Technical Report
- Publication Date
- Aug 01, 2014
- Accession Number
- ADA611737
Entities
People
- Niall G. Howlett
Organizations
- University of Rhode Island