Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment

Chimeric antigen receptor (CAR)-T cells are increasingly used for the treatment of hematologic malignancies. Treatment success relies highly upon sufficient expansion of CAR-T effector cells. Accordingly, longitudinal quantification of CAR-T cells during therapy is clinically important. Techniques t...

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Autores principales:	Thomas Mika, Julia Thomson, Verena Nilius-Eliliwi, Deepak Vangala, Alexander Baraniskin, Gerald Wulf, Susanne Klein-Scory, Roland Schroers
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	CD19-directed chimeric antigen receptor T cells CAR-T cells digital-droplet PCR (ddPCR) liquid biopsy cell-free DNA lymphoma Genetics QH426-470 Cytology QH573-671
Acceso en línea:	https://doaj.org/article/05e6edc5484e40d886ac9ef87c30d4ee
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spelling	oai:doaj.org-article:05e6edc5484e40d886ac9ef87c30d4ee2021-11-20T05:06:44ZQuantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment2329-050110.1016/j.omtm.2021.10.009https://doaj.org/article/05e6edc5484e40d886ac9ef87c30d4ee2021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2329050121001662https://doaj.org/toc/2329-0501Chimeric antigen receptor (CAR)-T cells are increasingly used for the treatment of hematologic malignancies. Treatment success relies highly upon sufficient expansion of CAR-T effector cells. Accordingly, longitudinal quantification of CAR-T cells during therapy is clinically important. Techniques to quantify CAR-T cells in patient blood samples are based on flow cytometry and PCR. However, cellular kinetics of CAR-T cells are very complex and under current investigation. In this study, feasibility of CAR-T cell quantification by cell-free DNA (cfDNA) was analyzed. cfDNA isolated from 74 blood samples of 12 patients during lymphoma treatment with the anti-CD19 CAR-T cell product axicabtagene ciloleucel (axi-cel) were analyzed. Concentrations of cfDNA specific for the CAR-T gene construct (cfCAR-DNA) and a reference gene were quantified by a newly designed digital-droplet PCR (ddPCR) assay. Detection and quantification of cfCAR-DNA was feasible and reliable for all patients included. Relative quantification of cfCAR-DNA compared to a reference gene, suitable for genomic DNA analysis, was heterogeneous in treatment responders and non-responders. In contrast, parallel analyses of cfCAR-DNA and reference cfDNA in a patient-specific approach gave insight into active lymphoma killing and treatment responses. In summary, plasma cfDNA determination in lymphoma patients is a promising tool for future clinical decision making.Thomas MikaJulia ThomsonVerena Nilius-EliliwiDeepak VangalaAlexander BaraniskinGerald WulfSusanne Klein-ScoryRoland SchroersElsevierarticleCD19-directed chimeric antigen receptor T cellsCAR-T cellsdigital-droplet PCR (ddPCR)liquid biopsycell-free DNAlymphomaGeneticsQH426-470CytologyQH573-671ENMolecular Therapy: Methods & Clinical Development, Vol 23, Iss , Pp 539-550 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	CD19-directed chimeric antigen receptor T cells CAR-T cells digital-droplet PCR (ddPCR) liquid biopsy cell-free DNA lymphoma Genetics QH426-470 Cytology QH573-671
spellingShingle	CD19-directed chimeric antigen receptor T cells CAR-T cells digital-droplet PCR (ddPCR) liquid biopsy cell-free DNA lymphoma Genetics QH426-470 Cytology QH573-671 Thomas Mika Julia Thomson Verena Nilius-Eliliwi Deepak Vangala Alexander Baraniskin Gerald Wulf Susanne Klein-Scory Roland Schroers Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment
description	Chimeric antigen receptor (CAR)-T cells are increasingly used for the treatment of hematologic malignancies. Treatment success relies highly upon sufficient expansion of CAR-T effector cells. Accordingly, longitudinal quantification of CAR-T cells during therapy is clinically important. Techniques to quantify CAR-T cells in patient blood samples are based on flow cytometry and PCR. However, cellular kinetics of CAR-T cells are very complex and under current investigation. In this study, feasibility of CAR-T cell quantification by cell-free DNA (cfDNA) was analyzed. cfDNA isolated from 74 blood samples of 12 patients during lymphoma treatment with the anti-CD19 CAR-T cell product axicabtagene ciloleucel (axi-cel) were analyzed. Concentrations of cfDNA specific for the CAR-T gene construct (cfCAR-DNA) and a reference gene were quantified by a newly designed digital-droplet PCR (ddPCR) assay. Detection and quantification of cfCAR-DNA was feasible and reliable for all patients included. Relative quantification of cfCAR-DNA compared to a reference gene, suitable for genomic DNA analysis, was heterogeneous in treatment responders and non-responders. In contrast, parallel analyses of cfCAR-DNA and reference cfDNA in a patient-specific approach gave insight into active lymphoma killing and treatment responses. In summary, plasma cfDNA determination in lymphoma patients is a promising tool for future clinical decision making.
format	article
author	Thomas Mika Julia Thomson Verena Nilius-Eliliwi Deepak Vangala Alexander Baraniskin Gerald Wulf Susanne Klein-Scory Roland Schroers
author_facet	Thomas Mika Julia Thomson Verena Nilius-Eliliwi Deepak Vangala Alexander Baraniskin Gerald Wulf Susanne Klein-Scory Roland Schroers
author_sort	Thomas Mika
title	Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment
title_short	Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment
title_full	Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment
title_fullStr	Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment
title_full_unstemmed	Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment
title_sort	quantification of cell-free dnafor the analysis of cd19-car-t cells during lymphoma treatment
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/05e6edc5484e40d886ac9ef87c30d4ee
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Quantification of cell-free DNAfor the analysis of CD19-CAR-T cells during lymphoma treatment

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