Adoptive transfer of murine T cells expressing a chimeric‐PD1‐Dap10 receptor as an immunotherapy for lymphoma

Abstract

Adoptive transfer of T cells is a promising cancer therapy and expression of chimeric antigen receptors can enhance tumour recognition and T-cell effector functions. The programmed death protein 1 (PD1) receptor is a prospective target for a chimeric antigen receptor because PD1 ligands are expressed on many cancer types, including lymphoma. Therefore, we developed a murine chimeric PD1 receptor (chPD1) consisting of the PD1 extracellular domain fused to the cytoplasmic domain of CD3ζ. Additionally, chimeric antigen receptor therapies use various co-stimulatory domains to enhance efficacy. Hence, the inclusion of a Dap10 or CD28 co-stimulatory domain in the chPD1 receptor was compared to determine which domain induced optimal anti-tumour immunity in a mouse model of lymphoma. The chPD1 T cells secreted pro-inflammatory cytokines and lysed RMA lymphoma cells. Adoptive transfer of chPD1 T cells significantly reduced established tumours and led to tumour-free survival in lymphoma-bearing mice. When comparing chPD1 receptors containing a Dap10 or CD28 domain, both receptors induced secretion of pro-inflammatory cytokines; however, chPD1-CD28 T cells also secreted anti-inflammatory cytokines whereas chPD1-Dap10 T cells did not. Additionally, chPD1-Dap10 induced a central memory T-cell phenotype compared with chPD1-CD28, which induced an effector memory phenotype. The chPD1-Dap10 T cells also had enhanced in vivo persistence and anti-tumour efficacy compared with chPD1-CD28 T cells. Therefore, adoptive transfer of chPD1 T cells could be a novel therapy for lymphoma and inclusion of the Dap10 co-stimulatory domain in chimeric antigen receptors may induce a preferential cytokine profile and T-cell differentiation phenotype for anti-tumour therapies.

A Jurkat 76 based triple parameter reporter system to evaluate TCR functions and adoptive T cell strategies

“…CD28 can be regarded as the primary costimulatory receptor and lack of CD28 signals might limit the success of adoptive therapy with TCR-tg T cells. To overcome this problem, Ankri et al and others have designed so-called switch receptors that comprise for example the extracellular domain of PD-1 fused to intracellular CD28 sequences and sustain CD28 signaling in the immune-suppressive tumor microenvironment [ 25 , 47 , 48 ]. We have adopted this strategy and devised chimeric receptors where CD28-signaling domains were fused to the extracellular binding domains of either CD2, CD226 or TIGIT and thus can be engaged by CD58 or CD155 expressed on the tumor targets.…”

Toll‐like receptor‐2/7‐mediated T cell activation: An innate potential to augment CD8+ T cell cytokine production

“…To generate effective T cell products, costimulatory signals can also be provided by chimeric costimulatory receptors. For instance, fusing the extracellular domains of PD‐1 or CTLA‐4 with costimulatory intracellular domains (eg CD28) showed promising results in several murine cancer models, including human xenograft melanoma, mesothelioma and prostate cancer models 149‐153 . Cancer treatment via chimeric receptors could be another opportunity where TLR signalling could be exploited, either through PD‐1‐/TLR2‐TIR chimeric receptors or in combination with a CD28 costimulatory domain.…”

CARs: Beyond T Cells and T Cell-Derived Signaling Domains

“…PD1-DAP10/CD3ζ CAR-T cells employed against syngeneic murine models of a variety of solid cancers (renal, pancreatic, liver, colon, breast, prostate, and bladder cancer, and melanoma) were also capable of inducing remission [193]. Both groups accredited the good performance to the preferential (IL-10-free) cytokine profile and memory phenotype differentiation induced by the DAP10 domain [192,193].…”

“…Comparing the cytokine profile of different CAR cells, secreted cytokines usually include pro-inflammatory IFNγ, TNF, IL-2, GM-CSF, IL-17, and IL-21, as well as anti-inflammatory IL-10 [152]. Unlike most other signaling domains, DAP10 was shown not to induce IL-10 secretion, but to strongly enhance T cell effector functions via pro-inflammatory cytokines [152,182,[185][186][187][188][189][190][191][192][193]. Similar to CD28-bearing receptors, DAP10 promoted differentiation towards an effector memory phenotype in T cells, which was associated with enhanced persistence in vivo [27,121,[147][148][149][150][151][152]182,[185][186][187][188][189][190][191][192].…”

“…CARs based on the extracellular binding domain of PD-1 can be employed against a broad variety of cancer types that upregulate PD-1 ligands on their surface. When testing PD1-CD3ζ/DAP10 CAR-T cells in a mouse model of lymphoma, Lynch et al saw significantly enhanced persistence compared to a CD28 co-stimulated CAR, leading to long-term tumor-free survival [192]. PD1-DAP10/CD3ζ CAR-T cells employed against syngeneic murine models of a variety of solid cancers (renal, pancreatic, liver, colon, breast, prostate, and bladder cancer, and melanoma) were also capable of inducing remission [193].…”