Oral Presentation Australasian Cytometry Society 44th Annual Conference and Workshop

Phase 1 Clinical Trial of CD19 CAR-T Cells Manufactured at the Point-of-Care (24581)

C J Hutchins 1 , A J Henderson 1 , C W Leung 1 , E C Lynam 1 , K J Harvey 1 , M E Abaca-Cleopas 1 , M Acworth 1 , E C Barnes 1 , A M McLean 2 , K L Mudie 2 , M O'Donnell 2 , A S Henden 2 3 , G A Kennedy 2 , S K Tey 2 3
  1. Cellular Therapy Laboratory, Royal Brisbane and Women's Hospital, Brisbane, QLD, Australia
  2. Bone Marrow Transplant / Clinical Haematology, Royal Brisbane and Women's Hospital, Brisbane, QLD, Australia
  3. QIMR Berghofer Medical Research Institute, Herston, QLD, Australia

Point-of-care (POC) manufacturing of CAR-T cells has the potential to expand the availability of CAR-T cell treatment to patients who are ineligible for government funded CAR-T cells, or enrolment onto a sponsored clinical trial protocol. 

We initiated a phase I clinical trial of POC manufactured CD19 CAR-T cells for the treatment of adults with relapsed and / or refractory CD19-positive haematological malignancy (Trial ID: ACTRN12621000762853). To manufacture CAR-T cells, mononuclear cells (MNC) were collected by apheresis and 1 - 3 x109 CD3+ T cells were loaded onto the Miltenyi Biotec CliniMACS Prodigy® for selection of CD4+ and CD8+ T cells. Following selection, 1 x108 T cells were activated with Transact® and transduced 24h later with the CD19 4-1BB-based CAR lentiviral vector, LTG1563 supplied by Lentigen. Cells were expanded for a further 11 days in TexMACS GMP medium supplemented with IL-7 and IL-15. Quality assurance testing was performed on the MNC, CD4+/CD8+ selected MNC, in-process samples on days 5, 9 and 12, and on the formulated CAR-T cells. CAR-T cells were released and infused into 8 of 9 patients without cryopreservation on day 12 of manufacture. One patient required overnight cryopreservation of the formulated CAR-T cells prior to infusion. The target cell doses were 0.5 x106 viable CD3+CAR+ cells / kg for patients at high risk for cytokine release syndrome (CRS) and 2.0 x106 viable CD3+CAR+ cells per kg for those at standard risk. 

CAR-T cells manufactured at the POC met specification for viability, transduction efficiency, microbial contamination, mycoplasma, endotoxin, and qPCR for replication competent lentivirus (RCL) and lentiviral copy number. Eight of the nine patients received target cell doses of CAR-T cells ranging from 32 to 236 x 106. One patient with a target cell dose of 38 x 106 received only 6.7 x106 CAR T cells. CAR-T cells were readily detectable by flow cytometry in the peripheral blood of patients from day 7 post-infusion. Ongoing CAR-T cell persistence has been monitored by flow cytometry and qPCR in the patients 87 to 457 days following infusion.

POC manufacture in a clinical hospital laboratory is feasible and enables the infusion of a CAR-T product with a short vein-to-vein time of 12-13 days. The development of POC manufacture provides a platform for future clinical translation of novel CAR-T cell products.