Heart Failure Impairs Bone Marrow Hematopoietic Stem Cell Function and Responses to Injury

Figures

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  Figure 1. Heart failure (HF) reduces engraftment potential in mice. A, Experimental design for the bone marrow transplant (BMT) study. EGFP+ (Enhanced green fluorescent protein) mice were subject to sham or left anterior descending coronary artery (LAD) ligation (HF), and 8 weeks (8w) later, bone marrow (BM) was collected. 5×106 BM mononuclear cells were transplanted into wild‐type C57BL/6J mice, and BM populations assessed 3 months later. B, BM cellularity 3 months after reconstitution with sham or HF BM. C, Engrafted EGFP+ lineage negative, Sca‐1+, c‐Kit+ (LSK) cells in sham‐BMT or HF‐BMT mice 3 months after reconstitution. D, Representative flow plots of EGFP+ LSK cells. E, EGFP+ hematopoietic stem/progenitor cell (HSPC) populations in BMT‐sham and BMT‐HF mice. F, Relative abundance of EGFP+ blood B cells (B220+), T cells (CD3+), and myeloid cells (CD11b+), as well as (G) myeloid cell subpopulations in mice transplanted with sham or HF BM. H, Representative flow cytometry plots of blood T‐cell and myeloid lineages (n=3–7 per group). *P≤0.05 by Mann‐Whitney test. Values are mean±SEM. LAD indicates left anterior descending coronary artery; LT‐HSC indicates long‐term repopulating…

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  Figure 2. Patient selection flowchart. Patient enrollment for the current study. A total of 90 patients were recruited, and 71 met the study inclusion criteria listed in Table S1. HF indicates heart failure.

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  Figure 3. Complete blood count analysis of control patients and patients with heart failure (HF). When compared with the control group, patients with HF have significantly lower levels of erythrocyte (A) and hemoglobin (B). Patients with HF also show significantly increased numbers of circulating leukocytes (C), which is driven specifically by increased neutrophils (D) and monocytes (E). F, No differences in total lymphocytes were observed (n=20–51 per group). *P<0.05 control vs HF by unpaired 2‐tailed Student's t test. Values are mean±SEM. RBC indicates red blood cell; and WBC, white blood cell.

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  Figure 4. Analysis of patient bone marrow (BM). Total mononuclear cellularity (MNC) (A) and CD34+ cell frequency (B) in BM from control and patients with heart failure (HF). *P<0.05 by unpaired two‐tailed Student's t test. C, Colony formation ability of BM CD34+ cells from control patients and patients with HF. **P<0.01 by unpaired 2‐tailed Student's t test. D, Frequency of CD34+/CD38− cells was increased in BM of patients with HF. P=0.055 by unpaired t test with Welch correction. E, This increase appears to be driven by a greater number of multipotent progenitors (MPPs), whereas hematopoietic stem cells (HSCs) and multipotent lymphoid progenitors (MLPs) were not different (n=5–6 per group). *P≤0.05 by unpaired t test with Welch correction. F, Myeloid progenitor relative abundance was not different between BM of control patients and patients with HF (n=5–10 per group). Representative gating for identification of BM neutrophils and monocytes (G) and quantification of the populations in the BM (H) (n=8–12 per group). Values are mean±SEM.

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  Figure 5. Reduced engraftment of bone marrow (BM) CD34+ cells from patients with heart failure (HF) in NOD‐SCID‐IL2rγnull (NSG) mice. A, Human CD45 (hCD45) and (B) human CD34+ (hCD34) cells in the BM of NSG mice 3 months after reconstitution (n=7–25 per group). *P<0.05 by Mann‐Whitney test. C, Frequency of BM progenitor populations 3 months after reconstitution (n=6–9 per group). *P≤0.05 by Mann‐Whitney test. D, hCD45 frequency in the blood and spleen in reconstituted animals (n=6–15 per group). B‐cell and myeloid‐cell frequency in the spleen (E) and blood (F) (n=6–15 per group). *P≤0.05 by unpaired t test with Welch correction. Values are mean±SEM. HSC indicates hematopoietic stem cell; MLP, multilymphoid progenitor; MPP, multipotent progenitor; and MNC, mononuclear cell.

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  Figure 6. Altered cardiac immune responses after myocardial infarction (MI) in mice reconstituted with bone marrow (BM) from patients with heart failure (HF). A, Schematic illustrating experimental design. B, Akinetic wall length/left ventricular (LV) endocardial length at 4 weeks after MI in reconstituted mice (n=5–11 per group). Unpaired 2‐tailed Student's t test. Echocardiographic analysis of LV dimensions at diastole (LVIDd) (C), at systole (LVIDs) (D), and fractional shortening (FS) (E) at baseline (0 weeks), 1 week, 2 weeks, and 4 weeks after MI (n=5–11 per group). Flow cytometry quantification of human (h) CD45+ (F), T cell (CD3+) (G), and myeloid (CD33+) (H) cell infiltration in the heart at baseline, 3 days, and 7 days after MI (n=3–14 per group). *P≤0.05 by 2‐way ANOVA followed by Sidak multiple comparison test. Flow cytometry quantification of mouse (m) CD45 (I), monocytes (J), neutrophils (K), and macrophages (L) in the heart at 3 days after MI (n=5–6 per group). *P≤0.05 by unpaired 2‐tailed Student's t test. Values are mean±SEM. LAD indicates left anterior descending coronary artery; and NSG, NOD‐SCID‐IL2rγnull.