Simone Krebs1, Afruja Ahad1, Lukas M Carter2, Justin Eyquem3, Christian Brand4, Meghan Bell1, Vladimir Ponomarev4, Thomas Reiner4, Claude F Meares5, Stephen Gottschalk6, Michel Sadelain3, Steven M Larson1, Wolfgang A Weber1,7. 1. Molecular Imaging and Therapy Service, Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, New York. 2. Radiochemistry and Molecular Imaging Sciences Service, Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, New York krebss@mskcc.org. 3. Center for Cell Engineering and Immunology Program, Sloan Kettering Institute, New York, New York. 4. Radiochemistry and Molecular Imaging Sciences Service, Department of Radiology, Memorial Sloan Kettering Cancer Center, New York, New York. 5. Chemistry Department, University of California, Davis, California. 6. Department of Bone Marrow Transplant and Cellular Therapy, St. Jude Children's Research Hospital, Memphis, Tennessee; and. 7. Department of Nuclear Medicine, Technical University of Munich, Munich, Germany.
Abstract
There remains an urgent need for the noninvasive tracking of transfused chimeric antigen receptor (CAR) T cells to determine their biodistribution, viability, expansion, and antitumor functionality. DOTA antibody reporter 1 (DAbR1) comprises a single-chain fragment of the antilanthanoid-DOTA antibody 2D12.5/G54C fused to the human CD4-transmembrane domain and binds irreversibly to lanthanoid (S)-2-(4-acrylamidobenzyl)-DOTA (AABD). The aim of this study was to investigate whether DAbR1 can be expressed on lymphocytes and used as a reporter gene as well as a suicide gene for therapy of immune-related adverse effects. Methods: DAbR1 was subcloned together with green fluorescent protein into an SFG-retroviral vector and used to transduce CD3/CD28-activated primary human T cells and second-generation 1928z (CAR) T cells. Cell surface expression of DAbR1 was confirmed by cell uptake studies with radiolabeled AABD. In addition, the feasibility of imaging of DAbR1-positive T cells in vivo after intravenous injection of 86Y/177Lu-AABD was studied and radiation doses determined. Results: A panel of DAbR1-expressing T cells and CAR T cells exhibited greater than 8-fold increased uptake of 86Y-AABD in vitro when compared with nontransduced cells. Imaging studies showed 86Y-AABD was retained by DAbR1-positive T cells while it continuously cleared from normal tissues, allowing for in vivo tracking of intravenously administered CAR T cells. Normal-organ dose estimates were favorable for repeated PET/CT studies. Selective T cell ablation in vivo with 177Lu-AABD seems feasible for clustered T-cell populations. Conclusion: We have demonstrated for the first time that T cells can be modified with DAbR1, enabling their in vivo tracking via PET and SPECT. The favorable biodistribution and high image contrast observed warrant further studies of this new reporter gene.
There remains an urgent need for the noninvasive tracking of transfused chimeric antigen receptor (CAR) T cells to determine their biodistribution, viability, expansion, and antitumor functionality. DOTA antibody reporter 1 (DAbR1) comprises a single-chain fragment of the antilanthanoid-DOTA antibody 2D12.5/G54C fused to the human CD4-transmembrane domain and binds irreversibly to lanthanoid(S)-2-(4-acrylamidobenzyl)-DOTA (AABD). The aim of this study was to investigate whether DAbR1 can be expressed on lymphocytes and used as a reporter gene as well as a suicide gene for therapy of immune-related adverse effects. Methods: DAbR1 was subcloned together with green fluorescent protein into an SFG-retroviral vector and used to transduce CD3/CD28-activated primary human T cells and second-generation 1928z (CAR) T cells. Cell surface expression of DAbR1 was confirmed by cell uptake studies with radiolabeled AABD. In addition, the feasibility of imaging of DAbR1-positive T cells in vivo after intravenous injection of 86Y/177Lu-AABD was studied and radiation doses determined. Results: A panel of DAbR1-expressing T cells and CAR T cells exhibited greater than 8-fold increased uptake of 86Y-AABD in vitro when compared with nontransduced cells. Imaging studies showed 86Y-AABD was retained by DAbR1-positive T cells while it continuously cleared from normal tissues, allowing for in vivo tracking of intravenously administered CAR T cells. Normal-organ dose estimates were favorable for repeated PET/CT studies. Selective T cell ablation in vivo with 177Lu-AABD seems feasible for clustered T-cell populations. Conclusion: We have demonstrated for the first time that T cells can be modified with DAbR1, enabling their in vivo tracking via PET and SPECT. The favorable biodistribution and high image contrast observed warrant further studies of this new reporter gene.
Authors: Amer M Najjar; Pallavi R Manuri; Simon Olivares; Leo Flores; Tiejuan Mi; Helen Huls; Elizabeth J Shpall; Richard E Champlin; Nashaat Turkman; Vincenzo Paolillo; Jason Roszik; Brian Rabinovich; Dean A Lee; Mian Alauddin; Juri Gelovani; Laurence J N Cooper Journal: Mol Imaging Biol Date: 2016-12 Impact factor: 3.488
Authors: Khun Visith Keu; Timothy H Witney; Shahriar Yaghoubi; Jarrett Rosenberg; Anita Kurien; Rachel Magnusson; John Williams; Frezghi Habte; Jamie R Wagner; Stephen Forman; Christine Brown; Martin Allen-Auerbach; Johannes Czernin; Winson Tang; Michael C Jensen; Behnam Badie; Sanjiv S Gambhir Journal: Sci Transl Med Date: 2017-01-18 Impact factor: 17.956
Authors: Pat Zanzonico; Guenther Koehne; Humilidad F Gallardo; Mikhail Doubrovin; Ekaterina Doubrovina; Ronald Finn; Ronald G Blasberg; Isabelle Riviere; Richard J O'Reilly; Michel Sadelain; Steven M Larson Journal: Eur J Nucl Med Mol Imaging Date: 2006-04-11 Impact factor: 9.236
Authors: Liu H Wei; Tove Olafsen; Caius Radu; Isabel J Hildebrandt; Mark R McCoy; Michael E Phelps; Claude Meares; Anna M Wu; Johannes Czernin; Wolfgang A Weber Journal: J Nucl Med Date: 2008-10-16 Impact factor: 10.057
Authors: Mikhail M Doubrovin; Ekaterina S Doubrovina; Pat Zanzonico; Michel Sadelain; Steven M Larson; Richard J O'Reilly Journal: Cancer Res Date: 2007-12-15 Impact factor: 12.701
Authors: Lucia Baratto; K Elizabeth Hawk; Lisa States; Jing Qi; Sergios Gatidis; Louise Kiru; Heike E Daldrup-Link Journal: J Nucl Med Date: 2021-10 Impact factor: 10.057
Authors: Mark A Sellmyer; Sarah A Richman; Katheryn Lohith; Catherine Hou; Chi-Chang Weng; Robert H Mach; Roddy S O'Connor; Michael C Milone; Michael D Farwell Journal: Mol Ther Date: 2019-10-15 Impact factor: 11.454
Authors: Megan M Dacek; Darren R Veach; Sarah M Cheal; Lukas M Carter; Michael R McDevitt; Blesida Punzalan; Daniela Burnes Vargas; Thomas Z Kubik; Sebastien Monette; Brian H Santich; Guangbin Yang; Ouathek Ouerfelli; Adam L Kesner; Nai-Kong V Cheung; David A Scheinberg; Steven M Larson; Simone Krebs Journal: Bioconjug Chem Date: 2021-04-05 Impact factor: 4.774
Authors: David Akhavan; Darya Alizadeh; Dongrui Wang; Michael R Weist; Jennifer K Shepphird; Christine E Brown Journal: Immunol Rev Date: 2019-07 Impact factor: 12.988
Authors: Federico Simonetta; Israt S Alam; Sanjiv S Gambhir; Robert S Negrin; Juliane K Lohmeyer; Bita Sahaf; Zinaida Good; Weiyu Chen; Zunyu Xiao; Toshihito Hirai; Lukas Scheller; Pujan Engels; Ophir Vermesh; Elise Robinson; Tom Haywood; Ataya Sathirachinda; Jeanette Baker; Meena B Malipatlolla; Liora M Schultz; Jay Y Spiegel; Jason T Lee; David B Miklos; Crystal L Mackall Journal: Clin Cancer Res Date: 2020-10-21 Impact factor: 13.801