Exploration of neuroblastoma xenograft models for tumor extracellular RNA profiling in murine blood plasma

Jill Deleu; Hanne Van Droogenbroeck; Jasper Anckaert; Anneleen Decock; Jilke De Wilde; Kaat Durinck; Liselot M. Mus; Justine Nuytens; Muhammad Rishfi; Kathleen Schoofs; Frank Speleman; Maaike Van Trimpont; Kimberly Verniers; Nurten Yigit; Jo Vandesompele; Bram De Wilde; Tom Van Maerken

doi:10.55092/exrna20230007

Article

Open Access

Exploration of neuroblastoma xenograft models for tumor extracellular RNA profiling in murine blood plasma

Jill Deleu^1,²^,†Hanne Van Droogenbroeck^1,²^,†Jasper Anckaert^1,²Anneleen Decock^1,²Jilke De Wilde^2,³Kaat Durinck^2,⁴Liselot M. Mus^2,^4,⁵Justine Nuytens^1,²Muhammad Rishfi^2,⁴Kathleen Schoofs^1,^2,⁶Frank Speleman^2,⁴Maaike Van Trimpont^2,^7,⁸Kimberly Verniers^1,²Nurten Yigit^1,²Jo Vandesompele^1,²^,∗Bram De Wilde^1,^2,^4,^5,⁹^,‡Tom Van Maerken^1,^2,¹⁰^,‡

¹ OncoRNALab, Cancer Research Institute Ghent (CRIG), Ghent, Belgium

² Department of Biomolecular Medicine, Ghent University, Ghent, Belgium

³ Department of Pathology, Ghent University Hospital, Ghent, Belgium

⁴ Pediatric Precision Oncology Lab (PPOL), Cancer Research Institute Ghent (CRIG), Ghent, Belgium

⁵ Department of Paediatric Haematology Oncology and Stem Cell Transplantation, Ghent University Hospital, Ghent, Belgium

⁶ TOBI lab, VIB – Ghent University, Zwijnaarde, Belgium

⁷ Department of Diagnostic Sciences, Ghent University, Ghent, Belgium

⁸ Lab Normal and Malignant Hematopoiesis, Cancer Research Institute Ghent (CRIG), Ghent, Belgium

⁹ Department of Internal Medicine and Pediatrics, Ghent University, Ghent, Belgium

¹⁰ Department of Laboratory Medicine, AZ Groeninge, Kortrijk, Belgium

†They are joint first authors and contributed equally to this work.

‡They are joint last authors and contributed equally to this work.

* jo.vandesompele@ugent.be

Volume
Volume 5 Issue 2, 2023
Citation
Deleu J, Van Droogenbroeck H, Anckaert J, Decock A, De Wilde J, et al. Exploration of neuroblastoma xenograft models for tumor extracellular RNA profiling in murine blood plasma. ExRNA 2023(2):0007, https://doi.org/10.55092/exrna20230007.
DOI
10.55092/exrna20230007
Copyright
Copyright2023 by the authors. Published by ELSP.

Abstract

Background: Minimally invasive liquid biopsies are becoming increasingly important in the diagnosis and treatment follow-up of cancer patients, including children with neuroblastoma. Such biopsies contain various biomarker analytes, including extracellular RNA (exRNA) with the potential to reflect dynamic changes in the tumor. However, it is challenging to distinguish tumor-derived exRNA from normal RNA. To overcome this limitation, xenograft models serve as a practical tool. In a mouse engrafted with human tumor cells, human exRNA is by definition originating from the tumor, whereas murine exRNA is host-derived. To study treatment response by monitoring tumor-derived exRNA, xenograft models with a high release of tumor exRNA into the circulation are desirable. Methods: The aim of this study was to evaluate whether and to what extent the cell line, its engraftment site, or the tumor size influence the amount of tumoral exRNA detected in blood plasma. To that end, four different neuroblastoma cell lines were engrafted in nude mice, either subcutaneously in the flank or orthotopically in the adrenal gland. Tumor sizes were monitored by caliper measurements (subcutaneous grafts) or MRI scans (orthotopic grafts) and blood was collected via terminal cardiac puncture to evaluate the tumoral exRNA fraction. Results: We demonstrate that the tumoral exRNA levels are correlated with the size of the subcutaneous tumor grafts. These levels are also highly dependent on the engrafted cell line. Furthermore, orthotopic engraftment potentially results in superior levels of tumoral exRNA, likely because of higher vascularity of the tumor tissue. Conclusions: Factors as cell line, tumor size and injections site should carefully be considered when performing experiments to study circulating RNA biomarkers.

Keywords

liquid biopsies; extracellular RNA; cell-free RNA; murine xenografts; neuroblastoma

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