Global proteomics and pathway analysis of pressure-overload-induced heart failure and its attenuation by mitochondrial-targeted peptides - PubMed (original) (raw)

. 2013 Sep 1;6(5):1067-76.

doi: 10.1161/CIRCHEARTFAILURE.113.000406. Epub 2013 Aug 9.

Edward J Hsieh, Tony Chen, Lorena G Menendez, Nathan B Basisty, Lauren Tsai, Richard P Beyer, David A Crispin, Nicholas J Shulman, Hazel H Szeto, Rong Tian, Michael J MacCoss, Peter S Rabinovitch

Affiliations

Global proteomics and pathway analysis of pressure-overload-induced heart failure and its attenuation by mitochondrial-targeted peptides

Dao-Fu Dai et al. Circ Heart Fail. 2013.

Abstract

Background: We investigated the protective effects of mitochondrial-targeted antioxidant and protective peptides, Szeto-Schiller (SS) 31 and SS20, on cardiac function, proteomic remodeling, and signaling pathways.

Methods and results: We applied an improved label-free shotgun proteomics approach to evaluate the global proteomics changes in transverse aortic constriction (TAC)-induced heart failure and the associated signaling pathway changes using ingenuity pathway analysis. We found that 538 proteins significantly changed after TAC, which mapped to 53 pathways. The top pathways were in the categories of actin cytoskeleton, mitochondrial function, intermediate metabolism, glycolysis/gluconeogenesis, and citrate cycle. Concomitant treatment with SS31 ameliorated the congestive heart failure phenotypes and mitochondrial damage induced by TAC, in parallel with global attenuation of mitochondrial proteome changes, with an average of 84% protection of mitochondrial and 69% of nonmitochondrial protein changes. This included significant amelioration of all the ingenuity pathway analysis noted above. SS20 had only modest effects on heart failure and this tracked with only partial attenuation of global proteomics changes; furthermore, actin cytoskeleton pathways were significantly protected in SS20, whereas mitochondrial and metabolic pathways essentially were not.

Conclusions: This study elucidates the signaling pathways significantly changed in pressure-overload-induced heart failure. The global attenuation of TAC-induced proteomic alterations by the mitochondrial-targeted peptide SS31 suggests that perturbed mitochondrial function may be an upstream signal to many of the pathway alterations in TAC and supports the potential clinical application of mitochondrial-targeted peptide drugs for the treatment heart failure.

Keywords: heart failure; mitochondria; proteomics; signal transduction.

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Figures

Figure 1

Figure 1. Echocardiography measurement after 4 weeks of TAC

(A) Left ventricular mass index (LVMI). (B) Fractional shortening significantly declined in saline-TAC; this was significantly ameliorated by SS20 and more so by SS31, but not by SS48. (C) Interventricular septum (IVS), left ventricular posterior wall (LVPW) thickness and left ventricular end-diastolic dimension (LVEDD). *p<0.05 compared with sham.

Figure 2

Figure 2. Pathology of TAC-induced heart failure

(A) Representative images of sham, saline-TAC and SS31-TAC treated hearts. (B) Heart weight and lung weight normalized to tibia length (mg/mm). (C) Representative Masson trichrome stain of the heart displayed marked interstitial fibrosis (blue) in saline-TAC hearts. (D) Quantitative analysis of fibrosis (% myocardial area) revealed that fibrosis in saline-TAC was significantly attenuated by SS31, but not by SS20 or SS48. (E) Quantitative PCR of collagen1a2 normalized to 18s. *p<0.05 for sham vs. saline-TAC, #p<0.05 TAC-SS vs. saline-TAC.

Figure 3

Figure 3. Mitochondrial damage in TAC-induced heart failure

(A) Mitochondrial protein carbonyl content (nmol/mg) measured by enzyme immunoassay. (B) Western blots of microtubule associated protein light chain 3 (LC3) demonstrated significant increase in LC3-II, a marker of activated autophagosomes, in saline-TAC hearts, which was significantly attenuated in SS31-TAC and partially attenuated in SS20-TAC hearts. Representative electron micrograph of cardiac apices in sham (C), saline-TAC (D) and SS31-TAC (E) demonstrated increased numbers of damaged mitochondria and autophagosomes (arrows) in saline-TAC hearts. (F) Higher magnification of mitochondria showed disrupted cristae in saline-TAC hearts (left) compared to normal cristae in SS31-TAC hearts (right panel). (G) Quantitative analysis of the number of autophagosomes (per 100μm2) and mitochondrial cristae density (cristae/μm). *p<0.05 for sham vs. saline-TAC, #p<0.05 SS31-TAC vs. saline-TAC.

Figure 4

Figure 4. Scatter plots of protein changes by proteomics analysis

The fold change (log2) in 538 significantly altered proteins in sham vs. saline-TAC hearts is shown on the x-axis (A–C). The y axis shows the corresponding change of each protein in each TAC-SS-peptide vs. saline-TAC comparison: SS31 (A) or SS20 (B). Proteins are shown as mitochondrial (red) or non-mitochondrial (blue) in panels A–D. (C) shows the fold changes in 277 proteins significantly altered only in SS31-TAC vs. saline-TAC that are not significant in the sham vs. saline-TAC comparison.

Figure 5

Figure 5. Heat map of abundance ratio of the top canonical pathways from Ingenuity Pathway Analysis

The abundance ratio of each component proteins of a pathway is colored red when higher in the SS-20, sham or SS-31treatment (numerator) than in the saline-TAC (denominator) in the ratio comparison. The color is blue for the converse. The top 15 canonical pathways are ordered from most significant (top of left set) to least (bottom of right set) Any protein that was significant in any one or more of the three ratio comparisons is included in the heat map. The intensity of color indicates the magnitude of change, with the scale shown at the bottom.

Figure 6

Figure 6. Network of mitochondrial oxidative phosphorylation (A, B) and citrate cycle (C, D), derived from Ingenuity Pathway Analysis

Symbols are colored red when significantly down regulated and green when significantly up-regulated; the intensity of color indicates the magnitude of change, with the scale shown in Figure 4A. (A) and (C) Sham vs. saline-TAC, (B) and (D) SS31-TAC vs. saline-TAC. Red in A and C indicates downregulation in TAC; red in B and D indicates SS31 attenuation of downregulation in TAC.

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