Increased frequency of TIGIT+CD73-CD8+ T cells with a TOX+ TCF-1low profile in patients with newly diagnosed and relapsed AML

Abstract

The inhibitory receptor TIGIT, as well as theectonucleotidases CD39 and CD73 constitute potential exhaustion markers for T cells. Detailed analysis of these markers can shed light into dysregulation of the T-cell response in acute myeloid leukemia (AML) and will help to identify potential therapeutic targets. The phenotype and expression of transcription factors was assessed on different T-cell populations derived from peripheral blood (PB, n = 38) and bone marrow (BM, n = 43). PB and BM from patients with AML diagnosis, in remission and at relapse were compared with PB from healthy volunteers (HD) (n = 12) using multiparameter flow cytometry. An increased frequency of terminally differentiated (CD45R^-CCR7^-)CD8⁺ T cells was detected in PB and BM regardless of the disease state. Moreover, we detected an increased frequency of two distinct T-cell populations characterized by the co-expression of PD-1 or CD39 on TIGIT⁺CD73^-CD8⁺ T cells in newly diagnosed and relapsed AML in comparison to HDs. In contrast to the PD-1⁺TIGIT⁺CD73^-CD8⁺ T-cell population, the frequency of CD39⁺TIGIT⁺CD73^-CD8⁺ T cells was normalized in remission. PD-1⁺- and CD39⁺TIGIT⁺CD73^-CD8⁺ T cells exhibited additional features of exhaustion by decreased expression of CD127 and TCF-1 and increased intracellular expression of the transcription factor TOX. CD8⁺ T cells in AML exhibit a key signature of two subpopulations, PD-1⁺TOX⁺TIGIT⁺CD73^-CD8⁺- and CD39⁺TOX⁺TIGIT⁺CD73^-CD8⁺ T cells that were increased at different stages of the disease. These results provide a rationale to analyze TIGIT blockade in combination with inhibition of the purinergic signaling and depletion of TOX to improve T-cell mediated cytotoxicity in AML. Abbreviations: AML: Acute myeloid leukemia; pAML: newly diagnosed AML; rAML: relapse AML; lrAML: AML in remission; HD: healthy donor; PB: peripheral blood; BM: bone marrow; TIGIT: T-cell immunoreceptor with Ig and ITIM domains; PD-1: Programmed cell death protein 1; CD73: ecto-5'-nucleotidase; CD39: ectonucleoside triphosphate diphosphohydrolase 1; ATP: adenosine triphosphate; ADO: adenosine; CD127: interleukin-7 receptor; CAR-T cell: chimeric antigen receptor T cell; TCF-1: transcription factor T-cell factor 1; TOX: Thymocyte selection-associated high mobility group box protein; NFAT: nuclear factor of activated T cells; NA: Naïve; CM: Central Memory; EM Effector Memory; EMRA: Terminal Effector Memory cells; FMO: Fluorescence minus one; PVR: poliovirus receptor; PVRL2: poliovirus receptor-related 2; IFN-γ: Interferon-γ; IL-2: interleukin-2; MCF: multiparametric flow cytometry; TNFα: Tumornekrosefaktor α; RT: room temperature.

Keywords: Acute myeloid leukemia (AML); CD127; CD39; CD73; PD-1; T cells; TCF-1; TIGIT; TOX.

Figure 1.

Expression of PD-1, TIGIT, CD73 and CD39 on CD8⁺ and conCD4⁺ and regCD4⁺ T cells in pAML and HDs

Figure 2.

Increased expression of PD-1 and CD39 on TIGIT⁺CD73⁻CD8⁺ T cells in pAML

Figure 3.

Comparison of the expression of TIGIT, PD-1, CD73 and CD39 on CD8⁺ T cells in pAML, rAML and lrAML

Figure 4.

Reduced frequencies of TCF-1^+- and CD127⁺ CD8⁺ T cells in pAML and rAML vs. HDs

Figure 5.

Increased TOX expression in pAML and rAML in comparison to HDs