Research ArticleHeart Failure

Histone deacetylase activity governs diastolic dysfunction through a nongenomic mechanism

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Science Translational Medicine  07 Feb 2018:
Vol. 10, Issue 427, eaao0144
DOI: 10.1126/scitranslmed.aao0144
  • Fig. 1 HDAC inhibition attenuates diastolic dysfunction in DSS rats.

    (A) Study overview. DSS rats were fed NS diet or a 4% NaCl–containing diet (HS) for 10 weeks. Beginning at the time of HS feeding, animals were treated with ITF2357 (pan-HDAC inhibitor) (3 or 30 mg/kg) or vehicle control by oral gavage (PO) once a day (QD). Serial echocardiography was performed before HS feeding and after 6, 8, and 10 weeks. (B) Doppler measurement of mitral inflow velocity (E/A) in DSS rats treated with vehicle or HDAC inhibitor. (C) Representative E/A Doppler echocardiography images. The ratio of the early filling (E) phase of the LV during diastole and the late filling (A) phase is a common echocardiographic measurement of diastolic function. Measurements of septal mitral annulus velocities (E′/A′) (D) and isovolumic relaxation time (IVRT) (E). (F) EF throughout the experiment. (G) LV end diastolic pressure (LVEDP). (H) Mean systemic pressure (MSP). For (G) and (H), ITF (3) and ITF (30) represent the dose of ITF in mg/kg. For all graphs, mean ± (or +) SEM values are shown and were compared by one-way analysis of variance (ANOVA) with Newman-Keuls post-test. *P < 0.05 versus NS + vehicle, #P < 0.05 versus HS + vehicle. For (B) and (D) to (F), rat numbers for each condition and time are provided in table S7. Each figure includes the number of animals used in the analysis. When such format was not practical, table S7 summarizes the number of animals used.

  • Fig. 2 HDAC inhibition improves diastolic function in DSS rats independently of effects on cardiac hypertrophy or fibrosis.

    Echocardiographic assessment of LV posterior (LVPW) (A) and anterior (LVAW) (B) wall thickness in NS- and HS-fed rats treated with vehicle or ITF2357. LV-to-tibia length (C) and images and quantification of myocyte cross-sectional area (D and E) in 10-week samples. Picrosirius red staining (F) and collagen quantification (G) of 10-week LV sections from DSS rats. For (C), (E), and (G), ITF (3) and ITF (30) represent the dose of ITF in mg/kg. For all graphs, mean ± (or +) SEM values are shown and were compared by one-way ANOVA with Newman-Keuls post-test. *P < 0.05 versus NS + vehicle, #P < 0.05 versus HS + vehicle. For (A) and (B), rat numbers for each end point and each time point are provided in table S7; at least eight animals were used per analysis. Scale bars, 20 μm.

  • Fig. 3 Diastolic dysfunction in DSS rats is associated with prolongation of myofibril relaxation that is normalized with HDAC inhibition.

    (A) Schematic representation of the ex vivo myofibril mechanics quantification assay. (B) Representative relaxation traces of myofibrils obtained from LVs of DSS rats fed NS or HS for 10 weeks and gavaged daily with ITF2357 (30 mg/kg) or vehicle control. Duration (tRel, slow) (C) and rate constant (kRel, slow) (D) of the linear relaxation phase of myofibrils from rats at 10 weeks. Exponential relaxation phase of myofibrils (kRel, fast) (E) and myofibril maximal tension generation of myofibrils (F) from rats at 10 weeks. For all graphs, mean + SEM values are shown and were compared by one-way ANOVA with Newman-Keuls post-test. *P < 0.05 versus NS + vehicle. n = 6 rats for NS + vehicle and HS + ITF2357 and n = 5 for HS + vehicle; 7 to 12 myofibrils per animal were analyzed.

  • Fig. 4 Ex vivo deacetylation of cardiac myofibrils prolongs relaxation kinetics.

    (A) Experimental design. Myofibrils from normal Sprague-Dawley rat LVs were incubated with recombinant HDAC2 or p300 before performing mechanics studies. Myofibril relaxation (B) and protein acetylation (C) with and without HDAC2. Myofibril relaxation (D) and protein acetylation (E) with and without p300. (F) Experimental design. Myofibrils isolated from DSS + HS–fed rats were treated with recombinant p300 and acetyl-CoA (coenzyme A). (G) Myofibril relaxation with and without p300. Data are mean + SEM and were compared by Student’s t test. *P < 0.05 versus untreated controls. Data from three separate ARVM preparations were combined; five to nine myofibril bundles were analyzed per ARVM preparation.

  • Fig. 5 HDAC inhibition blocks age-dependent diastolic dysfunction.

    (A) Study overview. Nine-month-old female 129S6/SvEvTac mice were fed normal chow or chow containing ITF2357 (50 mg/kg) for 11 months. Serial echocardiography was performed at 9, 15, 17, and 20 months. (B) Doppler measurement of mitral inflow velocity (E/A) in aged mice treated with vehicle or HDAC inhibitor. Measurements of septal mitral annulus velocities (E′/A′) (C) and IVRT (D). Mouse numbers for each time point are provided in table S7. Mean systemic blood pressure (MSP) (E) and LV-to-tibia length (F) in 20-month-old mice treated with or without ITF2357. (G to J) Ex vivo mechanics analyses of myofibrils from 20-month-old mice fed normal chow or ITF2357-containing chow. Data are mean + SEM and were compared by Student’s t test. *P < 0.05 versus untreated controls. n = 4 mice per group; 9 to 10 myofibril bundles were analyzed per animal.

  • Fig. 6 Human HFpEF is associated with impaired cardiac myofibril relaxation.

    (A) Representative relaxation traces of myofibrils from nonfailing donor heart (n = 1) and hearts from two individuals with HFpEF superimposed on identical time scale. Linear phase duration (tRel, slow) (B) and slope of the linear phase relaxation (kRel, slow) (C) from myofibrils from RCM patients and control nonfailing donor heart. Exponential phase relaxation (kRel, fast) (D) and maximal tension generation (E) from RCM patients and nonfailing donor heart. Data are mean + SEM and were compared by one-way ANOVA with Newman-Keuls post-test. *P < 0.05 versus nonfailing controls. n = 5 nonfailing donors (7 to 16 myofibril bundles per donor). For RCM patient #1, 16 myofibril bundles were analyzed, and for RCM patient #2, 12 myofibril bundles were analyzed.

Supplementary Materials

  • www.sciencetranslationalmedicine.org/cgi/content/full/10/427/eaao0144/DC1

    Materials and Methods

    Fig. S1. DSS rats fed a 4% NaCl–containing diet develop diastolic dysfunction with preserved EF.

    Fig. S2. Treatment of ARVMs in culture with ITF2357 led to acceleration of myocyte relaxation kinetics without affecting contractility.

    Fig. S3. HDAC inhibition does not affect HS diet–induced increases in β-MyHC expression or function.

    Fig. S4. Myofibril calcium sensitivity is unaffected by HS feeding or ITF2357 treatment of DSS rats.

    Fig. S5. Titin expression and function are unaffected by HS feeding or ITF2357 treatment of DSS rats.

    Fig. S6. Myofibrillar protein phosphorylation and expression are unaffected by HS feeding or ITF2357 treatment of DSS rats.

    Fig. S7. Site-specific phosphorylation of myofibrillar protein and myofibrillar protein expression are unaffected by HS feeding or ITF2357 treatment of DSS rats.

    Fig. S8. HDAC2 co-purifies with cardiac myofibrils.

    Fig. S9. Ex vivo acetylation/deacetylation does not alter myofibril contraction.

    Fig. S10. Anti–acetyl-lysine immunoblotting of cardiac myofibrils from DSS rats.

    Fig. S11. A model for HDAC inhibitor–mediated improvement in diastolic function and treatment of HFpEF.

    Table S1. Hematological profiles of DSS rats.

    Table S2. LV hemodynamic and echocardiographic parameters of DSS rats.

    Table S3. Quantification of myocyte cross-sectional area and interstitial fibrosis in DSS rats.

    Table S4. Echocardiographic parameters of aging mice.

    Table S5. Echocardiographic, hemodynamic, and hypertrophy parameters of aging mice.

    Table S6. Characteristics of the RCM patients and nonfailing donor controls.

    Table S7. Group sizes of animal models.

  • Supplementary Material for:

    Histone deacetylase activity governs diastolic dysfunction through a nongenomic mechanism

    Mark Y. Jeong, Ying H. Lin, Sara A. Wennersten, Kimberly M. Demos-Davies, Maria A. Cavasin, Jennifer H. Mahaffey, Valmen Monzani, Chandrasekhar Saripalli, Paolo Mascagni, T. Brett Reece, Amrut V. Ambardekar, Henk L. Granzier, Charles A. Dinarello, Timothy A. McKinsey*

    *Corresponding author. Email: timothy.mckinsey{at}ucdenver.edu

    Published 7 February 2018, Sci. Transl. Med. 10, eaao0144 (2018)
    DOI: 10.1126/scitranslmed.aao0144

    This PDF file includes:

    • Materials and Methods
    • Fig. S1. DSS rats fed a 4% NaCl–containing diet develop diastolic dysfunction with preserved EF.
    • Fig. S2. Treatment of ARVMs in culture with ITF2357 led to acceleration of myocyte relaxation kinetics without affecting contractility.
    • Fig. S3. HDAC inhibition does not affect HS diet–induced increases in β-MyHC expression or function.
    • Fig. S4. Myofibril calcium sensitivity is unaffected by HS feeding or ITF2357 treatment of DSS rats.
    • Fig. S5. Titin expression and function are unaffected by HS feeding or ITF2357 treatment of DSS rats.
    • Fig. S6. Myofibrillar protein phosphorylation and expression are unaffected by HS feeding or ITF2357 treatment of DSS rats.
    • Fig. S7. Site-specific phosphorylation of myofibrillar protein and myofibrillar protein expression are unaffected by HS feeding or ITF2357 treatment of DSS rats.
    • Fig. S8. HDAC2 co-purifies with cardiac myofibrils.
    • Fig. S9. Ex vivo acetylation/deacetylation does not alter myofibril contraction.
    • Fig. S10. Anti–acetyl-lysine immunoblotting of cardiac myofibrils from DSS rats.
    • Fig. S11. A model for HDAC inhibitor–mediated improvement in diastolic function and treatment of HFpEF.
    • Table S1. Hematological profiles of DSS rats.
    • Table S2. LV hemodynamic and echocardiographic parameters of DSS rats.
    • Table S3. Quantification of myocyte cross-sectional area and interstitial fibrosis in DSS rats.
    • Table S4. Echocardiographic parameters of aging mice.
    • Legend for table S5
    • Table S6. Characteristics of the RCM patients and nonfailing donor controls.
    • Table S7. Group sizes of animal models.

    [Download PDF]

    Other Supplementary Material for this manuscript includes the following:

    • Table S5. Echocardiographic, hemodynamic, and hypertrophy parameters of aging mice. (Excel file format)

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