Single-cell genetic analysis validates cytopathological identification of circulating cancer cells in patients with clear cell renal cell carcinoma

Lucile Broncy; Basma Ben Njima; Arnaud Méjean; Christophe Béroud; Khaled Ben Romdhane; Marius Ilie; Veronique Hofman; Jane Muret; Paul Hofman; Habiba Chaabouni Bouhamed; And Patrizia Paterlini-Bréchot

doi:10.18632/oncotarget.25102

Single-cell genetic analysis validates cytopathological identification of circulating cancer cells in patients with clear cell renal cell carcinoma

Oncotarget. 2018 Apr 13;9(28):20058-20074. doi: 10.18632/oncotarget.25102.

Authors

Affiliations

¹ INSERM Unit 1151, Faculté de Médecine Paris Descartes, Paris, France.
² Genetics and Pathology Departments, University of Tunis, Tunis, Tunisia.
³ Service d'Urologie, Hôpital Européen Georges Pompidou, Paris, France.
⁴ Aix Marseille University, INSERM, MMG, Marseille, France.
⁵ APHM, Hôpital TIMONE Enfants, Laboratoire de Génétique Moléculaire, Marseille, France.
⁶ Laboratoire de pathologie clinique et Biobank BB-0033-00025, Centre Hospitalo-Universitaire de Nice, Nice, France.
⁷ Institut Curie, PSL Research University, Département d'Anesthésie Réanimation Douleur, Paris, France.
⁸ Laboratoire de Biochimie A, Hôpital Necker-Enfants Malades, Paris, France.

Abstract

Context: Circulating Rare Cells (CRC) are non-haematological cells circulating in blood. They include Circulating Cancer Cells (CCC) and cells with uncertain malignant features (CRC-UMF) according to cytomorphology. Clear cell renal cell carcinomas frequently bear a mutated Von Hippel-Lindau (VHL) gene.

Aim: To match blind genetic analysis of CRC and tumor samples with CRC cytopathological diagnosis.

Results: 29/30 patients harboured CRC (20 harboured CCC, 29 CRC-UMF) and 25/29 patients carried VHL mutations in their tumour. 205 single CRC (64 CCC, 141 CRC-UMF) provided genetic data. 57/57 CCC and 104/125 CRC-UMF from the 25 patients with VHL-mutated tumor carried the same VHL mutation detected in the tumor. Seven CCC and 16 CRC-UMF did not carry VHL mutations but were found in patients with wild-type VHL tumor tissue.

Conclusions: All the CCC and 83,2% (104/125) of the CRC-UMF were found to carry the same VHL mutation identified in the corresponding tumorous tissue, validating cytopathological identification of CCC in patients with clear cell renal cell carcinoma.

Methods: The blood of 30 patients with clear cell renal cell carcinoma was treated by ISET^® for CRC isolation, cytopathology and single-cell VHL mutations analysis, performed blindly and compared to VHL mutations of corresponding tumor tissues and leukocytes.

Keywords: ISET® (Isolation by Size of Tumour/Trophoblastic Cells) technology; VHL mutation; circulating cancer cells; clear cell renal cell carcinoma; liquid biopsy.