Azacitidine Response in Myelodysplastic Syndromes is Marked by NK-like CD8 T-Cell Expansion and CXCL12+ Reticular Cell Remodeling

Hampton, H. R.; Pan, A.; Carnell, M.; Wang, B.; Shinko, D.; Kasherman, M.; Slapetova, I.; Joshi, S.; Nguyen, M. N. T.; Yan, F.; Davidson, S.; Choi, N. F. Y.; Wong, J. W. H.; Tedla, N.; Hiwase, D. K.; Tobiasson, M.; Polizzotto, M. N.; McGuire, H. M.; Abbas, H. A.; Javed, A.; Olivier, J.; Thoms, J. A. I.; Jolly, C. J.; Pimanda, J. E.

doi:10.64898/2026.07.07.736922

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bioRxiv · 10.64898/2026.07.07.736922

Azacitidine Response in Myelodysplastic Syndromes is Marked by NK-like CD8 T-Cell Expansion and CXCL12+ Reticular Cell Remodeling

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Abstract

Myelodysplastic syndromes (MDS) are driven by somatic mutations in hematopoietic stem and progenitor cells (HSPCs), leading to clonal expansion and ineffective hematopoiesis. Hypomethylating agents (HMAs; azacitidine or decitabine) are the standard of care for higher-risk MDS. However, their effects on the bone marrow (BM) microenvironment, and the extent to which these changes correlate with clinical response, remain poorly understood. We performed longitudinal analyses of BM aspirates, trephine biopsies, and peripheral blood samples from MDS patients treated with azacitidine in a clinical trial (NCT03493646), integrating CyTOF, 5' single-cell RNA and TCR sequencing, plasma proteomics, and multiplex immunofluorescence microscopy to characterize changes associated with azacitidine response. Clinical responders showed expansion of GzmBCD56CD8 T cells together with increased type I and type II interferon signaling within the T-cell compartment. Responders also exhibited marked alterations in circulating platelet- and myeloid-derived factors with the potential to remodel the BM niche. Spatial analyses revealed expansion of "neighborhoods" enriched for CXCL12-abundant reticular cells and CD8 T cells in responders, whereas HSPC-enriched neighborhoods were largely unchanged. In contrast, several HSPC-enriched neighborhoods expanded in non-responders. These microenvironmental changes were accompanied by evidence of enhanced myelopoiesis in clinical responders. Our findings support a model in which azacitidine response extends beyond direct effects on malignant hematopoietic cells to involve coordinated remodeling of the BM microenvironment which may be reinforced by platelet- and myeloid-derived signals that establish a feed-forward circuit promoting productive hematopoiesis. One Sentence SummaryLongitudinal bone marrow profiling reveals stromal and immune correlates of azacitidine response in myelodysplastic syndrome.

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Hampton, H. R., Pan, A., Carnell, M., Wang, B., Shinko, D., Kasherman, M., Slapetova, I., Joshi, S., Nguyen, M. N. T., Yan, F., Davidson, S., Choi, N. F. Y., Wong, J. W. H., Tedla, N., Hiwase, D. K., Tobiasson, M., Polizzotto, M. N., McGuire, H. M., Abbas, H. A., Javed, A., Olivier, J., Thoms, J. A. I., Jolly, C. J., Pimanda, J. E.. 2026-07-08. Azacitidine Response in Myelodysplastic Syndromes is Marked by NK-like CD8 T-Cell Expansion and CXCL12+ Reticular Cell Remodeling. https://doi.org/10.64898/2026.07.07.736922

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