Background/Purpose: Autologous anti-CD19 chimeric antigen receptor (CAR) T cells show early clinical evidence of safety and efficacy for treating several autoimmune diseases (Müller F. N Engl J Med. 2024). Allogeneic CAR T cells bring the promise of an off-the-shelf therapy with the scalability to address the growing B-cell driven autoimmune disease population, no need for patient apheresis, and reduced treatment wait times (Van Blarcom T. CAR TCR Summit. 2023). KYV-201 is an allogeneic anti-CD19 CAR T-cell therapy for B-cell driven autoimmune diseases. KYV-201 combines the same fully human anti-CD19 CAR construct as KYV-101 (a first-in-class autologous CAR T-cell therapy being investigated in patients with autoimmune disease) with a differentiated allogeneic platform developed by Intellia Therapeutics involving selected CRISPR/Cas9-mediated gene edits. These edits intend to avoid graft versus host disease by removing the endogenous T-cell receptor and provide immune evasion to prolong CAR T-cell persistence by removing key cell surface proteins involved in host recognition of an allogeneic product. This study aims to generate clinical-scale drug product characterization and preclinical data to support advancement of KYV-201 to first-in-human clinical studies.

Methods: CAR-mediated and target-dependent activity of KYV-201 against CD19⁺ target cell lines and primary B cells was assessed with in vitro cytotoxicity, cytokine production, and proliferation assays, and an in vivo xenograft model. Immune evasion editing functionality was assessed by in vitro assays modeling host T and NK cell-mediated rejection mechanisms. Genotoxicity concerns of the product were assessed with off-target editing, chromosomal translocation, karyotyping, and cytokine independent growth assays.

Results: High efficiency CAR transduction and on-target gene editing was achieved in clinical-scale lots of KYV-201 produced under good manufacturing practices. Preclinically, KYV-201 demonstrated CAR-mediated and CD19-dependent cytotoxicity, cytokine production, and proliferation against human CD19⁺ NALM6 cells. In vivo, KYV-201 controlled CD19⁺ NALM6 xenograft growth in a dose-dependent manner, similar to unedited anti-CD19 CAR T cells not modified with CRISPR/Cas9-mediated gene edits. In co-culture with allogeneic peripheral blood mononuclear cells from either healthy donors or patients with systemic lupus erythematosus, KYV-201 eliminated B cells and proliferated in a CAR-dependent manner (Figure) without evidence of rejection by alloreactive T and NK cells. Genotoxicity assays performed on clinical-scale lots revealed minimal levels of off-target editing, translocations, or chromosomal abnormalities in KYV-201 and lack of transformation by cytokine-independent growth assays.

Conclusion: The preclinical and drug product characterization data generated for KYV-201 demonstrate the functionality and safety necessary to advance to first-in-human studies, bringing the promise of allogeneic anti-CD19 CAR T cells for autoimmune diseases closer to fruition. A phase 1, open-label, multicenter study of the safety, tolerability, and clinical activity of KYV-201 in refractory lupus nephritis is planned.

« Back to ACR Convergence 2024

ACR Meeting Abstracts - https://acrabstracts.org/abstract/preclinical-development-and-manufacturability-of-kyv-201-an-investigational-allogeneic-anti-cd19-chimeric-antigen-receptor-t-cell-for-the-treatment-of-autoimmune-disease/