Ruxolitinib improves hematopoietic regeneration by restoring mesenchymal stromal cell function in acute graft-versus-host disease
Yan Lin, … , Hui Cheng, Xiaoxia Hu
Yan Lin, … , Hui Cheng, Xiaoxia Hu
Published June 20, 2023
Citation Information: J Clin Invest. 2023;133(15):e162201. https://doi.org/10.1172/JCI162201.
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Research Article

Ruxolitinib improves hematopoietic regeneration by restoring mesenchymal stromal cell function in acute graft-versus-host disease

Abstract

Acute graft-versus-host disease (aGVHD) is a severe complication of allogeneic hematopoietic stem cell transplantation. Hematopoietic dysfunction accompanied by severe aGVHD, which may be caused by niche impairment, is a long-standing clinical problem. However, how the bone marrow (BM) niche is damaged in aGVHD hosts is poorly defined. To comprehensively address this question, we used a haplo-MHC–matched transplantation aGVHD murine model and performed single-cell RNA-Seq of nonhematopoietic BM cells. Transcriptional analysis showed that BM mesenchymal stromal cells (BMSCs) were severely affected, with a reduction in cell ratio, abnormal metabolism, compromised differentiation potential, and defective hematopoiesis-supportive function, all of which were validated by functional assays. We found that ruxolitinib, a selective JAK1/2 inhibitor, ameliorated aGVHD-related hematopoietic dysfunction through a direct effect on recipient BMSCs, resulting in improved proliferation ability, adipogenesis/osteogenesis potential, mitochondria metabolism capacity, and crosstalk with donor-derived hematopoietic stem/progenitor cells. By inhibiting the JAK2/STAT1 pathway, ruxolitinib maintained long-term improvement of aGVHD BMSC function. Additionally, ruxolitinib pretreatment in vitro primed BMSCs to better support donor-derived hematopoiesis in vivo. These observations in the murine model were validated in patient samples. Overall, our findings suggest that ruxolitinib can directly restore BMSC function via the JAK2/STAT1 pathway and, in turn, improve the hematopoietic dysfunction caused by aGVHD.

Authors

Yan Lin, Quan Gu, Shihong Lu, Zengkai Pan, Zining Yang, Yapu Li, Shangda Yang, Yanling Lv, Zhaofeng Zheng, Guohuan Sun, Fanglin Gou, Chang Xu, Xiangnan Zhao, Fengjiao Wang, Chenchen Wang, Shiru Yuan, Xiaobao Xie, Yang Cao, Yue Liu, Weiying Gu, Tao Cheng, Hui Cheng, Xiaoxia Hu

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Figure 3

Impaired BM niche in a murine model of aGVHD.

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Impaired BM niche in a murine model of aGVHD. (A) Overview of murine aGV...
(A) Overview of murine aGVHD model study. (B) Frequency of LepR+ cells; n = 10–15 per group. (C) Quantitation of BMSCs by CFU-F assay; n = independent replicates. (D and E) Osteogenesis (D) and adipogenesis (E) of BMSCs from BMT and aGVHD mice after in vitro induction. Red arrows indicate calcium nodules (D), and yellow arrows indicate fatty droplets (E). Scale bars: 200 μm. (F) Quantitation of Col2.3-GFPhi osteoblasts; n = 10–15 per group. (G) Immunofluorescence images of femurs at day 21; three independent replicates. Scale bars: 500 μm. (H and I) High-magnification views of metaphysis areas (H) and diaphysis area (I). White dotted lines represent the growth plate, and red arrows indicate Col2.3-GFPhi osteoblasts. Scale bars: 200 μm. (J) Quantitative analysis of Col2.3-GFPhi area versus DAPI+ area in BMT and aGVHD mice. Numbers above columns represent the fold change compared with BMT mice; n = 3 independent replicates. (K) Images of calcein-stained femurs; 3 independent replicates. Scale bars: 50 μm. (L) Immunofluorescence images of adipocytes; n = independent replicates. Scale bars: 500 μm. (M and N) High-magnification views of metaphysis (M) and diaphysis (N) areas. Scale bars: 200 μm. (O) Quantitative analysis of perilipin+ area versus DAPI+ area; n = independent replicates. **P < 0.01 and ***P < 0.001, by 1-way ANOVA followed by an unpaired, 2-tailed t test (B and F), and unpaired, 2-tailed t test (C).