Shear stress-exposed pulmonary artery endothelial cells fail to upregulate HSP70 in chronic thromboembolic pulmonary hypertension

PLoS One. 2020 Dec 3;15(12):e0242960. doi: 10.1371/journal.pone.0242960. eCollection 2020.

Abstract

The pathophysiological mechanisms underlying chronic thromboembolic pulmonary hypertension (CTEPH) are still unclear. Endothelial cell (EC) remodeling is believed to contribute to this pulmonary disease triggered by thrombus and hemodynamic forces disbalance. Recently, we showed that HSP70 levels decrease by proatherogenic shear stress. Molecular chaperones play a major role in proteostasis in neurological, cancer and inflammatory/ infectious diseases. To shed light on microvascular responses in CTEPH, we characterized the expression of molecular chaperones and annexin A2, a component of the fibrinolytic system. There is no animal model that reproduces microvascular changes in CTEPH, and this fact led us to isolated endothelial cells from patients with CTEPH undergoing pulmonary endarterectomy (PEA). We exposed CTEPH-EC and control human pulmonary endothelial cells (HPAEC) to high- (15 dynes/cm2) or low- (5 dynes/cm2) shear stress. After high-magnitude shear stress HPAEC upregulated heat shock protein 70kDa (HSP70) and the HSP ER paralogs 78 and 94kDa glucose-regulated protein (GRP78 and 94), whereas CTEPH-ECs failed to exhibit this response. At static conditions, both HSP70 and HSP90 families in CTEPH-EC are decreased. Importantly, immunohistochemistry analysis showed that HSP70 expression was downregulated in vivo, and annexin A2 was upregulated. Interestingly, wound healing and angiogenesis assays revealed that HSP70 inhibition with VER-155008 further impaired CTEPH-EC migratory responses. These results implicate HSP70 as a novel master regulator of endothelial dysfunction in type 4 PH. Overall, we first show that global failure of HSP upregulation is a hallmark of CTEPH pathogenesis and propose HSP70 as a potential biomarker of this condition.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomechanical Phenomena
  • Chronic Disease
  • Endoplasmic Reticulum Chaperone BiP
  • Endothelial Cells / pathology*
  • HSP70 Heat-Shock Proteins / metabolism*
  • Humans
  • Hypertension, Pulmonary / complications
  • Hypertension, Pulmonary / metabolism
  • Hypertension, Pulmonary / pathology*
  • Pulmonary Artery / pathology*
  • Shear Strength
  • Stress, Mechanical*
  • Thromboembolism / complications*
  • Up-Regulation*

Substances

  • Endoplasmic Reticulum Chaperone BiP
  • HSP70 Heat-Shock Proteins
  • HSPA5 protein, human

Grants and funding

This work was supported by Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP) in the form of a research grant awarded to TLSA (18/13739-8) and fellowships awarded to TLSA (19/20435-8, 15/06210-2), LBCTC (20/11249-3) and EGM (19/25503-9), Centro de Pesquisa, Inovação e Difusão FAPESP (CEPID “Processos Redox em Biomedicina”) in the form of a research grant awarded to FRML (13/07937-8), and Fundação Zerbini and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - Brasil (CAPES) (Finance Code 001) in the form of funds used for the purchase of materials and reagents. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.