Supplementary Materials

Supplementary Material for:

PPARγ agonist pioglitazone reverses pulmonary hypertension and prevents right heart failure via fatty acid oxidation

Ekaterina Legchenko, Philippe Chouvarine, Paul Borchert, Angeles Fernandez-Gonzalez, Erin Snay, Martin Meier, Lavinia Maegel, S. Alex Mitsialis, Eva A. Rog-Zielinska, Stella Kourembanas, Danny Jonigk, Georg Hansmann*

*Corresponding author. Email: georg.hansmann{at}gmail.com

Published 25 April 2018, Sci. Transl. Med. 10, eaao0303 (2018)
DOI: 10.1126/scitranslmed.aao0303

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Mice with targeted deletion of PPARγ in cardiomyocytes (cmPPARγ−/−), in the absence of PAH, do not develop cardiac hypertrophy or fibrosis at the age of 12 to 16 weeks.
  • Fig. S2. Schematic depicting the vicious cycle of RV failure in PAH.
  • Fig. S3. PAH but no RV failure is evident 1 week after the end of hypoxia (3 + 1 weeks), and RV failure develops by week 6 after the end of hypoxia in SuHx-exposed rats.
  • Fig. S4. RV glucose uptake increases with chronic RV pressure afterload and correlates with RV systolic function 6 weeks after the end of hypoxia in SuHx-exposed rats.
  • Fig. S5. Decrease in minimal mitochondrial diameter, presence of autophagosomes and cytoplasmic vacuoles, and collagen deposits indicating RV failure and fibrosis are present in SuHx RVs but not SuHx + Pio RVs.
  • Fig. S6. In silico predicted miRNA/mRNA pairing of miR-491 with the mRNA of monoacylglycerol lipase (MGLL).
  • Fig. S7. Expression of miRNAs that are altered in rat and human RV failure is not changed by hypoxia in the RV of FVB mice.
  • Fig. S8. mRNA/miRNA expression signatures and networks in the failing RV (SuHx, on the left) and the PPARγ-mediated effects in the RV of the SuHx rat PAH model (SuHx + Pio, on the right).
  • Fig. S9. Important molecular interactions based on differential gene expression (mRNA) analysis in the RV of SuHx + Pio versus SuHx rats.
  • Fig. S10. Pioglitazone has no negative effects on survival, GO, or FAO in human PAECs and no impact on cardiomyocyte survival.
  • Fig. S11. Neither VEGFR2 blockade nor oral pioglitazone treatment cause any significant changes in blood glucose in rats.
  • Table S1. Cardiac MRI, ECHO, and cardiac catheterization hemodynamic and morphological data obtained in cmPPARγ−/− mice and littermate controls at 12 to 16 weeks of age.
  • Table S2. Echocardiographic measurements in rats 1 week after the end of hypoxia (=SuHx rat with PAH but no RV failure yet, 3 + 1 weeks).
  • Table S3A. Invasive hemodynamic and echocardiographic measurements in rats at 3 + 6 weeks (SuHx rat PAH/RV failure model).
  • Table S3B. Cardiac MRI, 18FDG-PET/CT, and heart weight measurements in rats of the SuHx study at 3 + 6 weeks.
  • Table S3C. Complete blood count, plasma NT-proBNP, and plasma APN in rats of the SuHx study at 3 + 6 weeks.
  • Table S4. Human heart and lung tissue specimens used for laser capture microdissection or whole-tissue gene expression assays.
  • Table S5. List of significantly differentially expressed genes (FDR < 5%) based on inversion of RV mRNA expression patterns in the SuHx PAH/RV failure model (RNA-seq) with PPARγ agonist pioglitazone, first up- or down-regulated in PAH/RV failure (SuHx) and inversely regulated with pioglitazone treatment (SuHx + Pio).
  • Table S6. List of mRNA transcripts related to angiogenesis identified by RNA-seq in the RV of control rats (ConHx), rats with PAH and RV failure (SuHx), and Pio-treated rats (SuHx + Pio).
  • Table S7. Fresh human lung tissue specimens obtained during lung transplantation.
  • Legends for movies S1 to S4
  • References (6681)

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Other Supplementary Material for this manuscript includes the following:

  • Movie S1 (.mov format). Cardiac MRI ConNx.
  • Movie S2 (.mov format). Cardiac MRI ConHx.
  • Movie S3 (.mov format). Cardiac MRI SuHx.
  • Movie S4 (.mov format). Cardiac MRI SuHx + Pio.

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