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Single-cell polyfunctional proteomics of CD4 cells from patients with AML predicts responses to anti-PD-1-based
Hussein A Abbas1,2, Zoe Alaniz1, Sean Mackay3
1Department of Leukemia.
Abstract:
Acute myeloid leukemia (AML) remains a difficult disease to treat disease. In a phase 2 clinical trial in patients with relapsed/refractory AML, combining the hypomethylating agent, azacitidine, with the PD-1 checkpoint inhibitor, nivolumab, demonstrated encouraging response rates (33%), median event-free, and overall survival, compared with a historical cohort of contemporary patients treated with azacitidine-based therapies, with an acceptable safety profile. Biomarkers of response are yet to be determined. In this study, we leveraged a multiplexed immune assay to assess the functional states of CD4+ and CD8+ cells at a single-cell level in pretherapy bone marrows in 16 patients with relapsed/refractory AML treated with azacitidine/nivolumab. Effector CD4+ but not CD8+ cells had distinct polyfunctional groups and were associated with responses and better outcomes. Further evaluation of the polyfunctional strength index composition across cell types revealed that interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were the major drivers of enhanced polyfunctionality index of pretherapy CD4+ subset, whereas Granzyme B, IFN-γ, MIP-1b, and TNF-α drove the nonsignificantly enhanced pretreatment Polyfunctional Strength Index of CD8+ subset in the responders. Single-cell polyfunctional assays were predictive of response in AML and may have a potential role as a biomarker in the wider sphere of immunotherapy.
Insights
Single-cell immune cell analysis in relapsed/refractory acute myeloid leukemia (AML) identified specific CD4+ T cell functions as predictive biomarkers. These findings offer potential for guiding immunotherapy in AML treatment.
Area of Science:
- Immunology
- Hematology
- Oncology
Background:
- Acute myeloid leukemia (AML) presents significant treatment challenges, particularly in relapsed/refractory cases.
- Combining azacitidine with nivolumab shows promise in clinical trials for AML, but response biomarkers are needed.
Purpose of the Study:
- To investigate single-cell immune cell functional states as potential biomarkers of response to azacitidine/nivolumab therapy in relapsed/refractory AML.
- To identify specific immune cell subsets and their functional characteristics associated with treatment outcomes.
Main Methods:
- Utilized a multiplexed immune assay for single-cell, single-cell level functional analysis of CD4+ and CD8+ T cells.
- Analyzed pretherapy bone marrow samples from 16 patients with relapsed/refractory AML treated with azacitidine and nivolumab.
- Assessed polyfunctional strength index and cytokine production (IFN-γ, TNF-α, Granzyme B, MIP-1b).
Main Results:
- Distinct polyfunctional CD4+ T cell subsets were associated with treatment response and improved outcomes in AML patients.
- Interferon-gamma (IFN-γ) and tumor necrosis factor-alpha (TNF-α) were key drivers of CD4+ T cell polyfunctionality in responders.
- While CD8+ T cells showed some enhanced polyfunctionality, it was less pronounced and associated with different cytokine profiles.
Conclusions:
- Single-cell immune cell polyfunctionality assays can predict treatment response in acute myeloid leukemia.
- Specific functional profiles of CD4+ T cells may serve as valuable biomarkers for immunotherapy in AML.
- Further validation of these biomarkers could enhance patient selection for immunotherapy strategies.
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