Universität zu Köln

Lodrini, Marco, Spruessel, Annika, Astrahantseff, Kathy, Tiburtius, Daniela, Konschak, Robert, Lode, Holger N., Fischer, Matthias, Keilholz, Ulrich, Eggert, Angelika and Deubzer, Hedwig E. (2017). Using droplet digital PCR to analyze MYCN and ALK copy number in plasma from patients with neuroblastoma. Oncotarget, 8 (49). S. 85234 - 85252. ORCHARD PARK: IMPACT JOURNALS LLC. ISSN 1949-2553

Full text not available from this repository.

Abstract

The invasive nature of surgical biopsies deters sequential application, and single biopsies often fail to reflect tumor dynamics, intratumor heterogeneity and drug sensitivities likely to change during tumor evolution and treatment. Implementing molecular characterization of cell-free neuroblastoma-derived DNA isolated from blood plasma could improve disease assessment for treatment selection and monitoring of patients with high-risk neuroblastoma. We established droplet digital PCR (ddPCR) protocols for MYCN and ALK copy number status in plasma from neuroblastoma patients. Our ddPCR protocol accurately discriminated between MYCN and ALK amplification, gain and normal diploid status in a large panel of neuroblastoma cell lines, and discrepancies with reported MYCN and ALK status were detected, including a high-level MYCN amplification in NB-1, a MYCN gain in SH-SY5Y, a high-level ALK amplification in IMR-32 and ALK gains in BE(2)-C, Kelly, SH-SY5Y and LAN-6. MYCN and ALK status were also reliably determined from cell-free DNA derived from medium conditioned by the cell lines. MYCN and ALK copy numbers of subcutaneous neuroblastoma xenograft tumors were accurately determined from cell-free DNA in the mouse blood plasma. In a final validation step, we accurately distinguished MYCN and ALK copy numbers of the corresponding primary tumors using retrospectively collected blood plasma samples from 10 neuroblastoma patients. Our data justify the further development of molecular disease characterization using cell-free DNA in blood plasma from patients with neuroblastoma. This expanded molecular diagnostic palette may improve monitoring of disease progression including relapse and metastatic events as well as therapy success or failure in high-risk neuroblastoma patients.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Lodrini, Marco UNSPECIFIED UNSPECIFIED UNSPECIFIED Spruessel, Annika UNSPECIFIED UNSPECIFIED UNSPECIFIED Astrahantseff, Kathy UNSPECIFIED UNSPECIFIED UNSPECIFIED Tiburtius, Daniela UNSPECIFIED UNSPECIFIED UNSPECIFIED Konschak, Robert UNSPECIFIED UNSPECIFIED UNSPECIFIED Lode, Holger N. UNSPECIFIED UNSPECIFIED UNSPECIFIED Fischer, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Keilholz, Ulrich UNSPECIFIED UNSPECIFIED UNSPECIFIED Eggert, Angelika UNSPECIFIED UNSPECIFIED UNSPECIFIED Deubzer, Hedwig E. UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-214153
DOI:	10.18632/oncotarget.19076
Journal or Publication Title:	Oncotarget
Volume:	8
Number:	49
Page Range:	S. 85234 - 85252
Date:	2017
Publisher:	IMPACT JOURNALS LLC
Place of Publication:	ORCHARD PARK
ISSN:	1949-2553
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language ANAPLASTIC LYMPHOMA KINASE; PARAFFIN-EMBEDDED BREAST; FREE TUMOR DNA; N-MYC; ACTIVATING MUTATIONS; ONCOGENIC MUTATIONS; GENE AMPLIFICATION; CELLULAR ONCOGENE; TARGETING MYCN; HETEROGENEITY Multiple languages Oncology; Cell Biology Multiple languages
Refereed:	Yes
URI:	http://kups.ub.uni-koeln.de/id/eprint/21415