Abstract 436: Whole body imaging of genetically labeled hematopoietic stem cells in human subjects
Authors: Nowicki Theodore S. , Berent-Maoz Beata , Puig-Saus Cristina , Kaplan-Lefko Paula , Carretero Ignacio Baselga , Champhekar Ameya , Carlucci Giuseppe , Macabali Mignonette , Garcilazo Ivan Perez , Vega-Crespo Agustin , Rodriguez Jonathan , Chmielowski Bartosz , Singh Arun , Allen-Auerbach Martin , Zhu Shaojun , Slavik Roger , Comin-Anduix Begonya , Williams John , Ribas Antoni 
Published: July 2021

Abstract
Background: Despite decades of experience with bone marrow transplantation, there has not been prior whole body visualization of the engraftment of hematopoietic stem cell (HSC) in bone marrow in humans. The herpes simplex virus type 1 sr39 thymidine kinase (sr39tk) gene is a positron emission tomography (PET) reporter/suicide gene that can be used to genetically label cells and track them in vivo based on the ability to retain the PET tracer [18F]FHBG, which is a penciclovir analogue.
Methods: Three patients with metastatic sarcoma were treated with gene-modified autologous CD34+ peripheral blood stem cells (PBSCs) transduced with a lentiviral vector encoding both a transgenic T-cell receptor against the tumor antigen NY-ESO-1 and the sr39tk reporter/suicide gene after a reduced intensity conditioning regimen of busulfan and fludarabine. These gene-modified stem cells were co-administered with gene-modified autologous T-cells also encoding the NY-ESO-1 T-cell receptor. The primary endpoint was safety and feasibility, while an exploratory endpoint was standardized uptake value of [18F]FHBG PET between post-transplant day +25 and day +120 scans to study the biodistribution of the gene-modified PBSCs and progeny T cells.
Results: Two patients (NYSCT-01 and NYSCT-03) demonstrated robust engraftment the gene-modified PBSCs in bone marrow niches by day +25-30, as demonstrated by whole body [18F]FHBG PET. PET signal was evident throughout the body in the bone marrow in the scalp, jaw, vertebrae, ribs, pelvis, femur and other bones with significant areas of bone marrow. The third patient (NYSCT-05), whose product had much lower transduction efficiency, did not display any engraftment signal at day +28. Patient NYSCT-01 demonstrated loss of [18F]FHBG signal at day +120, with no detectable T-cell progeny in peripheral circulation. Patient NYSCT-03 developed a post-transplant de novo multidrug-resistant cytomegalovirus (CMV) viremia at day +58 which ultimately required treatment with systemic ganciclovir, and passed away prior to her day +120 PET scan. Patient NYSCT-05 developed a post-transplant CMV reactivation viremia on day +45, which responded to oral valganciclovir treatment.
Conclusions: PET imaging allowed visualization of the successful engraftment of sr39tk reporter gene-labeled HSCs in bone marrow niches. Further studies are needed to determine mechanisms of loss of engraftment due to immunogenicity of the sr39tk gene, or insufficient transduction efficiency.
Citation Format: Theodore S. Nowicki, Beata Berent-Maoz, Cristina Puig-Saus, Paula Kaplan-Lefko, Ignacio Baselga Carretero, Ameya Champhekar, Giuseppe Carlucci, Mignonette Macabali, Ivan Perez Garcilazo, Agustin Vega-Crespo, Jonathan Rodriguez, Bartosz Chmielowski, Arun Singh, Martin Allen-Auerbach, Shaojun Zhu, Roger Slavik, Begonya Comin-Anduix, John Williams, Antoni Ribas. Whole body imaging of genetically labeled hematopoietic stem cells in human subjects [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 436.