Adipocyte enhancer binding protein 1 (AEBP1) inhibition mediates cardiac reverse remodeling by regulating MRTFB and RUNX2 signaling

Figures

• 

  Figure 1:. Increased expression of AEBP1 is an indicator of pathological remodeling in the heart. a, Masson trichrome analysis of human non-failing donor and advanced HF myocardium (n=14 donor, n=61 HF). b, Differential expression analysis between donor and advanced HF myocardium (n=9 donor, n=41 HF). c, Representative immunohistochemistry image of non-failing myocardium and infarct and peri-infarct regions of human HF myocardium (n=3). d, UMAP of fibroblast cluster showing the topology of ACTA2, POSTN and AEBP1 expression in HF compared to donor fibroblasts respectively (n=14 donor and n=13 HF). e, Mouse myocardial infarction (MI) model depiction. f, Serum ELISA quantifying ACLP levels (n=4 each). g, Representative immunohistochemistry of mouse infarct/peri-infarct and distal myocardium at 2-, 4- and 8-days post MI (n=4 each). p-value: unpaired t-test (panel a), One-way ANOVA (panel f).

• 

  Figure 2:. Aebp1 inhibition improves cardiac function in mice with MI. a, Schematic of fibroblast-specific Aebp1 KO along with MI induction (MI+inj). b-c, Left ventricular ejection fraction (EF%) and fractional shortening (FS%) data in MI+inj group (n=5 each) d, Heart weight to body weight ratio in MI+inj group (n=5 each). e, Representative immunohistochemistry data in WT and fibroblast-specific Aebp1 KO (n=5 each). f, Schematic of fibroblast-specific Aebp1 KO 2 weeks post tamoxifen injection (2wpi-MI). g-h, EF% and FS% data in 2wpi-MI (n=5 each) i, Heart weight to body weight ratio in WT and KO mice (n=5 each). j, Representative immunohistochemistry data in WT and fibroblast-specific Aebp1 KO 2wpi (n=5 each). k, Schematic of global Aebp1 KD using AAV9 specifically in the heart. l-m, EF% and FS% data following Aebp1 KD (n=5 each). n, Representative immunohistochemistry data in WT GFP and Aebp1 KD mice (n=5 each). p-value: One way ANOVA (panels b-c, g-h and l-m), unpaired t-test (panels d&i).

• 

  Figure 3:. Aebp1 inhibition suppresses interstitial fibrosis progression and cardiac hypertrophy in mice with pressure overload. a, Schematic of AngII/PE infusion and fibroblast-specific Aebp1 KO. b-c, Left ventricular ejection fraction (EF%) and fractional shortening (FS%) data (n=5 each). d, Heart weight to body weight ratio (n=5 each). e, Representative immunohistochemistry of WT and KO myocardium following AngII/PE infusion (n=5 each). f, Schematic of AngII/PE infusion and global Aebp1 KD. g-h, EF% and FS% data (n=5 each). i, Heart weight to body weight ratio (n=4 saline, n=5 each of GFP and AEBP1 KD). j, Representative immunohistochemistry of mouse myocardium (n=4 saline, n=5 each GFP and AEBP1 KD). k-m, Quantification of periostin signal, ECM percentage measured from WGA signal intensity and cell surface area. p-value: Two-way ANOVA (panel b-c, g-h); unpaired t-test (panel d); One-way ANOVA (panel i, k, l and m).

• 

  Figure 4:. AEBP1 plays a crucial role in cardiac fibrosis and cardiomyocyte remodeling in advanced human HF. a, Human myocardial tissue culture methodology. b, Representative immunohistochemistry data of cultured non-failing human myocardium (n=3). c, Quantification of cell surface area. d, ECM percentage measured from WGA signal intensity (n=3 donors, multiple images/donor). e-f, cell length to width ratio (LWR) and width to length ratio (WLR) from immunohistochemistry (n=3 donors, multiple images/donor). g, Representative immunohistochemistry data of cultured human HF myocardium (n=6). h, Quantification of cell surface area, i, ECM percentage measured from WGA signal intensity. j-k, LWR and WLR from immunohistochemistry (n=6 patients, multiple images/patient). p-value: unpaired t-test.

• 

  Figure 5:. Increased ACLP drives cardiac fibrosis by regulating multiple pro-fibrotic targets. a, Schematic of primary human cardiac fibroblast (HCF) culture and ACLP overexpression (OE). b, Heatmap of GFP US and ACLP OE HCF (n=4 each). c-f, Western blot image and quantification of MRTFB, COL1A1, SM22 and RUNX2 in GFP US and ACLP OE (n=6 each for all targets, RUNX2 – n=5 GFP US and n=6 ACLP OE). g, Schematic of primary human cardiac fibroblast culture and AEBP1 knockdown (KD). h, Heatmap of GFP S and AEBP1 KD HCF (n=3 each). i-l, Western blot image and quantification of MRTFB, COL1A1, SM22 and RUNX2 in GFP S and AEBP1 KD (n=6 each for all targets, RUNX2 – n=5 GFP US and n=6 ACLP OE). p-value: unpaired t-test.

• 

  Figure 6:. ACLP regulates cardiac fibroblast activation via the RUNX2 signaling pathway. a, Schematic of cell culture using isolated mouse cardiac fibroblasts (MCF). b, CUT&RUN data on mouse cardiac fibroblasts (MCF) using H3K27Ac. c-e, Western blot image and quantification of SM22 and RUNX2 in WT vs fibroblast-specific Aebp1 knockout (KO) MCF (n=6). f-n, Western blot image and quantification of RUNX2, ACLP, MRTFB, COL1A1 and SM22 in WT GFP, RUNX2 KD following and prior to ACLP OE respectively (n=3 each). p-value: Two-tailed unpaired t-test (d&e), One way ANOVA (g-k).