CD3xCD19 DART molecule treatment induces non-apoptotic killing and is efficient against high-risk chemotherapy and venetoclax-resistant chronic lymphocytic leukemia cells
Open Access
- 1 January 2020
- journal article
- research article
- Published by BMJ in Journal for ImmunoTherapy of Cancer
- Vol. 8 (1), e000218
- https://doi.org/10.1136/jitc-2019-000218
Abstract
Background Bispecific antibodies are promising new therapeutics in B cell malignancies. Whether they lead to potent T cell activation despite described T cell dysfunction in chronic lymphocytic leukemia (CLL), and are able to effectively target high-risk or venetoclax-resistant samples, is currently unknown. Methods CD19(+)cell lines or primary (high-risk) CLL were cocultured in vitro with healthy donor (HD) or CLL-derived T cells in the presence of a CD3xCD19 dual affinity retargeting molecule (CD3xCD19 DART). Cell cytotoxicity, T cell activation, proliferation and effector molecule production were analyzed using flow cytometry. Results Here, we report that a bispecific CD3xCD19 DART mediates efficient killing by HD T cells of CD19(+)cell-lines and primary CLL cells, regardless of immunoglobulin heavy chain variable region (IGHV) mutational status TP53 status or chemotherapy, ibrutinib or venetoclax sensitivity. Whereas TCR stimulation of CLL-derived T cells resulted in dysfunctional T cell activation and proliferation, treatment with CD3xCD19 DART led to a similar activation profile in CLL-derived and HD-derived T cells. Consistently, co-culture of CLL derived T cells with JeKo-1 or CLL cells in the presence of CD3xCD19 DART resulted in significant cytotoxicity by both CD4(+)and CD8(+)T cells. On stimulation of CLL cells with CD40L, CLL cells become resistant to the specific inhibitor of anti-apoptotic Bcl-2 protein venetoclax, due to upregulation of Bcl-2 family members such as Bcl-XL. Nevertheless, CD40L stimulated CLL cells were as efficiently lysed on CD3xCD19 DART treatment as unstimulated CLL cells. Further examination of the mechanism of CD3xCD19 DART mediated killing showed that lysis was dependent on granules, but was independent of BAX/BAK or caspase activity, indicating non-apoptotic cell death. Conclusions These data show that CD3xCD19 DART in CLL leads to robust T cell activation and lysis of high-risk venetoclax resistant CLL cells through a non-apoptotic mechanism.Funding Information
- Janssen Pharmaceuticals
- ZonMw
This publication has 61 references indexed in Scilit:
- Long-term follow-up of hematologic relapse-free survival in a phase 2 study of blinatumomab in patients with MRD in B-lineage ALLBlood, 2012
- Safety and persistence of adoptively transferred autologous CD19-targeted T cells in patients with relapsed or chemotherapy refractory B-cell leukemiasBlood, 2011
- Tumor-reactive CD4+ T cells develop cytotoxic activity and eradicate large established melanoma after transfer into lymphopenic hostsThe Journal of Experimental Medicine, 2010
- Immunogenic and tolerogenic cell deathNature Reviews Immunology, 2009
- Chronic lymphocytic leukemia T cells show impaired immunological synapse formation that can be reversed with an immunomodulating drugJCI Insight, 2008
- Cytotoxic T lymphocytes overcome Bcl-2 inhibition: target cells contribute to their own demiseBlood, 2008
- Granzyme B-induced cell death exerted by ex vivo CTL: discriminating requirements for cell death and some of its signsCell Death & Differentiation, 2007
- High expression of Bfl‐1 contributes to the apoptosis resistant phenotype in B‐cell chronic lymphocytic leukemiaInternational Journal of Cancer, 2004
- Granzyme B activates procaspase-3 which signals a mitochondrial amplification loop for maximal apoptosisThe Journal of cell biology, 2003
- T lymphocyte subpopulations in chronic lymphocytic leukemia of B cell type in relation to immunoglobulin isotype(s) on the leukemic clone and to clinical featuresEuropean Journal of Haematology, 1987