Feasibility of Circulating 5-hydroxymethylcytosine profiling in Osteosarcoma

Aims: To examine the utility and performance of 5-hydroxymethycytosine (5-hmC) profiling of cell-free DNA (cfDNA) as a biomarker of disease in patients with osteosarcoma. Population: A Discovery cohort consisting of 5 patients with primary, localized osteosarcoma and an independent Validation cohort consisting of 17 patients with osteosarcoma. Serial cfDNA samples were collected for the majority of patients.Molecular Technologies Employed: Nano-hmC-Seal libraries were constructed from 1-10 ng of cfDNA. Briefly, cfDNA 5-hmC marks were subjected to enzymatic modification with UDP-N3-glucose, followed by chemical modification with biotin. Biotin-labeled cfDNA fragments were then pulled down using streptavidin beads, and PCR amplified to construct sequencing libraries. Findings: Using genes... (for more see dbGaP study page.)