Cell-free DNA profiling in retinoblastoma patients with advanced intraocular disease: An MSKCC experience

Cancer Med. 2020 Sep;9(17):6093-6101. doi: 10.1002/cam4.3144. Epub 2020 Jul 7.


Purpose: The enucleation rate for retinoblastoma has dropped from over 95% to under 10% in the past 10 years as a result of improvements in therapy. This reduces access to tumor tissue for molecular profiling, especially in unilateral retinoblastoma, and hinders the confirmation of somatic RB1 mutations necessary for genetic counseling. Plasma cell-free DNA (cfDNA) has provided a platform for noninvasive molecular profiling in cancer, but its applicability in low tumor burden retinoblastoma has not been shown. We analyzed cfDNA collected from 10 patients with available tumor tissue to determine whether sufficient tumorderived cfDNA is shed in plasma from retinoblastoma tumors to enable noninvasive RB1 mutation detection.

Methods: Tumor tissue was collected from eye enucleations in 10 patients diagnosed with advanced intra-ocular unilateral retinoblastoma, three of which went on to develop metastatic disease. Tumor RB1 mutation status was determined using an FDA-cleared tumor sequencing assay, MSK-IMPACT. Plasma samples were collected before eye enucleation and analyzed with a customized panel targeting all exons of RB1.

Results: Tumor-guided genotyping detected 10 of the 13 expected somatic RB1 mutations in plasma cfDNA in 8 of 10 patients (average variant allele frequency 3.78%). Without referring to RB1 status in the tumor, de novo mutation calling identified 7 of the 13 expected RB1 mutations (in 6 of 10 patients) with high confidence.

Conclusion: Plasma cfDNA can detect somatic RB1 mutations in patients with unilateral retinoblastoma. Since intraocular biopsies are avoided in these patients because of concern about spreading tumor, cfDNA can potentially offer a noninvasive platform to guide clinical decisions about treatment, follow-up schemes, and risk of metastasis.

Keywords: RB1 mutation; liquid biopsy; molecular profiling in retinoblastoma; plasma cell-free DNA; retinoblastoma.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cancer Care Facilities
  • Child, Preschool
  • Circulating Tumor DNA / blood
  • Circulating Tumor DNA / genetics*
  • DNA Mutational Analysis / methods
  • Exons / genetics
  • Eye Enucleation
  • Feasibility Studies
  • Genes, Retinoblastoma / genetics*
  • Genotyping Techniques
  • Humans
  • Infant
  • Infant, Newborn
  • New York City
  • Retinal Neoplasms / blood
  • Retinal Neoplasms / genetics*
  • Retinal Neoplasms / therapy
  • Retinoblastoma / blood
  • Retinoblastoma / genetics*
  • Retinoblastoma / therapy


  • Circulating Tumor DNA