Mesenchymal Neuroblastoma Cells Are Undetected by Current mRNA Marker Panels: The Development of a Specific Neuroblastoma Mesenchymal Minimal Residual Disease Panel

Esther M van Wezel; Lieke M J van Zogchel; Jalenka van Wijk; Ilse Timmerman; Ngoc-Kim Vo; Lily Zappeij-Kannegieter; Boris deCarolis; Thorsten Simon; Max M van Noesel; Jan J Molenaar; Tim van Groningen; Rogier Versteeg; Huib N Caron; C Ellen van der Schoot; Jan Koster; Johan van Nes; Godelieve A M Tytgat

doi:10.1200/PO.18.00413

Mesenchymal Neuroblastoma Cells Are Undetected by Current mRNA Marker Panels: The Development of a Specific Neuroblastoma Mesenchymal Minimal Residual Disease Panel

JCO Precis Oncol. 2019 Oct 3:3:PO.18.00413. doi: 10.1200/PO.18.00413. eCollection 2019.

Authors

Esther M van Wezel^{1

2}, Lieke M J van Zogchel^{1

3}, Jalenka van Wijk^{1

2}, Ilse Timmerman^{1

3}, Ngoc-Kim Vo¹, Lily Zappeij-Kannegieter¹, Boris deCarolis⁴, Thorsten Simon⁴, Max M van Noesel³, Jan J Molenaar³, Tim van Groningen², Rogier Versteeg², Huib N Caron², C Ellen van der Schoot¹, Jan Koster², Johan van Nes², Godelieve A M Tytgat^{2

3}

Affiliations

¹ Sanquin Research Amsterdam, the Netherlands.
² Amsterdam University Medical Center, Amsterdam, the Netherlands.
³ Princess Máxima Center for Pediatric Oncology, Utrecht, the Netherlands.
⁴ Children's Hospital University of Cologne, Cologne, Germany.

Abstract

Patients with neuroblastoma in molecular remission remain at considerable risk for disease recurrence. Studies have found that neuroblastoma tissue contains adrenergic (ADRN) and mesenchymal (MES) cells; the latter express low levels of commonly used markers for minimal residual disease (MRD). We identified MES-specific MRD markers and studied the dynamics of these markers during treatment.

Patients and methods: Microarray data were used to identify genes differentially expressed between ADRN and MES cell lines. Candidate genes were then studied using real-time quantitative polymerase chain reaction in cell lines and control bone marrow and peripheral blood samples. After selecting a panel of markers, serial bone marrow, peripheral blood, and peripheral blood stem cell samples were obtained from patients with high-risk neuroblastoma and tested for marker expression; survival analyses were also performed.

Results: PRRX1, POSTN, and FMO3 mRNAs were used as a panel for specifically detecting MES mRNA in patient samples. MES mRNA was detected only rarely in peripheral blood; moreover, the presence of MES mRNA in peripheral blood stem cell samples was associated with low event-free survival and overall survival. Of note, during treatment, serial bone marrow samples obtained from 29 patients revealed a difference in dynamics between MES mRNA markers and ADRN mRNA markers. Furthermore, MES mRNA was detected in a higher percentage of patients with recurrent disease than in those who remained disease free (53% v 32%, respectively; P = .03).

Conclusion: We propose that the markers POSTN and PRRX1, in combination with FMO3, be used for real-time quantitative polymerase chain reaction-based detection of MES neuroblastoma mRNA in patient samples because these markers have a unique pattern during treatment and are more prevalent in patients with poor outcome. Together with existing markers of MRD, these new markers should be investigated further in large prospective studies.