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Induced dendritic cells co-expressing GM-CSF/IFN-α/tWT1 priming T and B cells and automated manufacturing to boost GvL

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Julia K. Bialek-Waldmann, Sabine Domning, Ruth Esser, Wolfgang Glienke, Mira Mertens, Krasimira Aleksandrova, Lubomir Arseniev, Suresh Kumar, Andreas Schneider, Johannes Koenig, Sebastian J. Theobald, Hsin Chieh Tsay, Angela D.A. Cornelius, Agnes Bonifacius, Britta Eiz-Vesper, Constanca Figueiredo, Dirk Schaudien, Steven R. Talbot, Andre Bleich, Loukia M. Spineli & 8 more Constantin von Kaisenberg, Caren Clark, Rainer Blasczyk, Michael Heuser, Arnold Ganser, Ulrike Köhl, Farzin Farzaneh, Renata Stripecke

Original languageEnglish
Pages (from-to)621-641
Number of pages21
JournalMolecular Therapy - Methods and Clinical Development
Volume21
DOIs
Published11 Jun 2021

Bibliographical note

Funding Information: The authors thank other members of the Regenerative Immune Therapies Applied Laboratory, in particular, Simon Danisch, for revisions of the animal protocol; Benjamin Ostermann for technical assistance with animals; and Laura Gerasch for technical assistance with cell cultures. We thank Dr. Thea B?hm (Miltenyi Biotec) for valuable technical advice for the development of process protocol for automated cell manufacturing and Dr. Anne Richter and her staff (Miltenyi Biotec) for technical advice for detection of WT1-specific CTL responses. We acknowledge the generous support of this project provided by the Else-Kr?ner Fresenius Stiftung/Forschungstransfer Program (2017_T04 to R.S.) and thank Prof. Dr. Martin Z?rnig for conceptual input toward translation. This work was also partially financed by grants from the German Center for Infections Research (DZIF-TTU07.805 to R.S.), by a research collaboration grant from ?The Jackson Laboratory,? and by the German Research Council (DFG/REBIRTH Unit 6.4 to R.S.; FOR2830 to B.E.-V.). Work in the Molecular Medicine Group at King's College London was supported by CRUK, the Experimental Cancer Medicine Centre, and the NIHR Biomedical Research Centres (BRC) based at King's Health Partners. J.K.B.-W. co-coordinated and managed the project, planned and performed experiments, analyzed data, wrote the first draft of the manuscript, and edited the final manuscript. S.D. and F.F. produced and tested the GL vector. R.E. W.G. M.M. K.A. L.A. and U.K. developed the process for automated cell manufacturing and QC for batch release. S.K. A.S. J.K. S.J.T. H.-C.T. A.D.A.C. and C.F. performed experiments and analyzed data. A.B. and B.E.-V. planned and executed assays to detect WT1-specific responses. D.S. performed the histopathology and immunohistochemistry analysis. S.R.T. and L.M.S. revised or conducted the statistical analyses. C.v.K. provided cord blood, R.B. provided leukapheresis, and C.C. provided technical assistance for adaptation of the manufacturing protocol for the Prodigy. A.B. assisted with the mouse breeding. M.H. assisted with the development of the mouse study protocol. M.H. and A.G. provided clinical input for the discussion with the regulatory agency. R.S. planned and managed the project with the collaborating consortium, provided overall conceptualization and project leadership, obtained funding, supervised the data analyses, and wrote and revised the final manuscript. R.S. and A.G. are co-inventors in the US- and EU-granted patent ?Induced Dendritic Cells and Uses Thereof,? publication number WO/2014/122035; PCT/EP2014/051422 (in national phases in China, Japan, and Canada). R.S. received honoraria and research funding from The Jackson Laboratory, a not-for-profit organization developing and commercializing humanized mouse models. Funding Information: R.S. and A.G. are co-inventors in the US- and EU-granted patent “Induced Dendritic Cells and Uses Thereof,” publication number WO/2014/122035; PCT/EP2014/051422 (in national phases in China, Japan, and Canada). R.S. received honoraria and research funding from The Jackson Laboratory, a not-for-profit organization developing and commercializing humanized mouse models. Funding Information: The authors thank other members of the Regenerative Immune Therapies Applied Laboratory, in particular, Simon Danisch, for revisions of the animal protocol; Benjamin Ostermann for technical assistance with animals; and Laura Gerasch for technical assistance with cell cultures. We thank Dr. Thea Böhm (Miltenyi Biotec) for valuable technical advice for the development of process protocol for automated cell manufacturing and Dr. Anne Richter and her staff (Miltenyi Biotec) for technical advice for detection of WT1-specific CTL responses. We acknowledge the generous support of this project provided by the Else-Kröner Fresenius Stiftung/Forschungstransfer Program ( 2017_T04 to R.S.) and thank Prof. Dr. Martin Zörnig for conceptual input toward translation. This work was also partially financed by grants from the German Center for Infections Research ( DZIF-TTU07.805 to R.S.), by a research collaboration grant from “The Jackson Laboratory,” and by the German Research Council ( DFG/REBIRTH Unit 6.4 to R.S.; FOR2830 to B.E.-V.). Work in the Molecular Medicine Group at King’s College London was supported by CRUK , the Experimental Cancer Medicine Centre , and the NIHR Biomedical Research Centres (BRC) based at King’s Health Partners. Publisher Copyright: © 2021 The Authors Copyright: Copyright 2021 Elsevier B.V., All rights reserved.

King's Authors

Abstract

Acute myeloid leukemia (AML) patients with minimal residual disease and receiving allogeneic hematopoietic stem cell transplantation (HCT) have poor survival. Adoptive administration of dendritic cells (DCs) presenting the Wilms tumor protein 1 (WT1) leukemia-associated antigen can potentially stimulate de novo T and B cell development to harness the graft-versus-leukemia (GvL) effect after HCT. We established a simple and fast genetic modification of monocytes for simultaneous lentiviral expression of a truncated WT1 antigen (tWT1), granulocyte macrophage-colony-stimulating factor (GM-CSF), and interferon (IFN)-α, promoting their self-differentiation into potent “induced DCs” (iDCtWT1). A tricistronic integrase-defective lentiviral vector produced under good manufacturing practice (GMP)-like conditions was validated. Transduction of CD14+ monocytes isolated from peripheral blood, cord blood, and leukapheresis material effectively induced their self-differentiation. CD34+ cell-transplanted Nod.Rag.Gamma (NRG)- and Nod.Scid.Gamma (NSG) mice expressing human leukocyte antigen (HLA)-A∗0201 (NSG-A2)-immunodeficient mice were immunized with autologous iDCtWT1. Both humanized mouse models showed improved development and maturation of human T and B cells in the absence of adverse effects. Toward clinical use, manufacturing of iDCtWT1 was up scaled and streamlined using the automated CliniMACS Prodigy system. Proof-of-concept clinical-scale runs were feasible, and the 38-h process enabled standardized production and high recovery of a cryopreserved cell product with the expected identity characteristics. These results advocate for clinical trials testing iDCtWT1 to boost GvL and eradicate leukemia.

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