CLIC-01: Manufacture and distribution of non-cryopreserved CAR-T cells for patients with CD19 positive hematologic malignancies

Abstract

Access to commercial CD19 CAR-T cells remains limited even in wealthy countries like Canada due to clinical, logistical, and financial barriers related to centrally manufactured products. We created a non-commercial academic platform for end-to-end manufacturing of CAR-T cells within Canada’s publicly funded healthcare system. We report initial results from a single-arm, open-label study to determine the safety and efficacy of in-house manufactured CD19 CAR-T cells (entitled CLIC-1901) in participants with relapsed/refractory CD19 positive hematologic malignancies. Using a GMP compliant semi-automated, closed process on the Miltenyi Prodigy, T cells were transduced with lentiviral vector bearing a 4-1BB anti-CD19 CAR transgene and expanded. Participants underwent lymphodepletion with fludarabine and cyclophosphamide, followed by infusion of non-cryopreserved CAR-T cells. Thirty participants with non-Hodgkin’s lymphoma (n=25) or acute lymphoblastic leukemia (n=5) were infused with CLIC-1901: 21 males (70%), median age 66 (range 18-75). Time from enrollment to CLIC-1901 infusion was a median of 20 days (range 15-48). The median CLIC-1901 dose infused was 2.3 × 106 CAR-T cells/kg (range 0.13-3.6 × 106/kg). Toxicity included ≥ grade 3 cytokine release syndrome (n=2) and neurotoxicity (n=1). Median follow-up was 6.5 months. Overall response rate at day 28 was 76.7%. Median progression-free and overall survival was 6 months (95%CI 3-not estimable) and 11 months (95% 6.6-not estimable), respectively. This is the first trial of in-house manufactured CAR-T cells in Canada and demonstrates that administering fresh CLIC-1901 product is fast, safe, and efficacious. Our experience may provide helpful guidance for other jurisdictions seeking to create feasible and sustainable CAR-T cell programs in research-oriented yet resource-constrained settings.Clinical trial registrationhttps://clinicaltrials.gov/ct2/show/NCT03765177, identifier NCT03765177.

Discovery and preclinical development of a therapeutically active nanobody-based chimeric antigen receptor targeting human CD22

“…Based on the results of the preliminary assessments above, we selected 4 CD22sdCARs (1ug36, 1ug74, 1ug13, and Pas33) for testing in a CAR backbone that has previously been used for CAR delivery in an ongoing CD19 CAR-T clinical trial. 22 , 23 There were several differences between the initial CAR screening plasmid and the clinical trial such as plasmid used, the most significant being a change in the CAR structure from a CD28TM domain to a CD8TM domain (see Figure 3 A). In the CAR-Jurkat assay, CD28TM and CD8TM CAR constructs showed similar responses, although baseline CD69 expression and nonspecific response to Ramos-CD22KO cells were consistently higher with CD8TM CAR-T constructs compared to CD28TM CAR-T constructs ( Figure 3 B).…”

“…To identify a lead CD22sdCAR construct, we next performed a functional comparison of the 4 CD8TM-CD22sdCAR candidate molecules implementing full-sample randomization and blinding and using a CAR-T production process mimicking the cell manufacturing approach for an ongoing CD19 CAR-T clinical trial. 22 CAR-T products were generated and characterized (see Table S2 ) before being cryopreserved, anonymized, and transferred to a separate site for in vitro and in vivo testing. Consistent with the results above, in vitro functional testing showed highly similar CD22-specific repression of target cell growth across all constructs ( Figures S7 A–S7F).…”

Analyzing functional heterogeneity of effector cells for enhanced adoptive cell therapy applications

“…D) Autologous/allogeneic TILs vs. autologous wildtype (WT) or MHCI-deficient ( B2M - ) tumour. E) CD19-specific CAR-T 27 or non-transduced T-cells vs. CD19+ Daudi target cells.…”

“…All procedures were approved by the Ethics Committee of the Capital Region of Denmark and national regulations for biomedical research (Ethical approval reference: H-20070020; Data Protection approval P-2021-303). The CD19-specific CAR-T cell construct 27 was generously provided by the Holt group at the BC Cancer Research Institute (Vancouver, Canada). Biorender.com, GraphPad Prism v10 (GraphPad Software, Boston, Massachusetts, USA), and FlowJo™ v10.0 Software (BD Life Sciences, Ashland, Oregon, USA) were used for figure generation.…”

CAR-T Cells in Canada; Perspective on How to Ensure We Get Our Value’s Worth

“…This has been done in Spain, for example [ 27 , 28 ]. In Canada, the ExCELLirate Canada Platform and the CLIC (Canadian Led Immunotherapies in Cancer) program are also working toward Canadian-made CAR-T products within the academic setting, which will hopefully prove to be cheaper than but equally effective as existing commercial products [ 29 ]. This could be a game-changer for the economic impact of CAR-T cells, assuming the incidence and severity related to adverse events with this manufacturing model are unchanged.…”