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Single-Cell Analysis Reveals Distinct Immune and Smooth Muscle Cell Populations that Contribute to Chronic Thromboembolic Pulmonary Hypertension.

Publication ,  Journal Article
Viswanathan, G; Kirshner, HF; Nazo, N; Ali, S; Ganapathi, A; Cumming, I; Zhuang, Y; Choi, I; Warman, A; Jassal, C; Almeida-Peters, S; Haney, J ...
Published in: Am J Respir Crit Care Med
May 15, 2023

Rationale: Chronic thromboembolic pulmonary hypertension (CTEPH) is a sequela of acute pulmonary embolism (PE) in which the PE remodels into a chronic scar in the pulmonary arteries. This results in vascular obstruction, pulmonary microvasculopathy, and pulmonary hypertension. Objectives: Our current understanding of CTEPH pathobiology is primarily derived from cell-based studies limited by the use of specific cell markers or phenotypic modulation in cell culture. Therefore, our main objective was to identify the multiple cell types that constitute CTEPH thrombusy and to study their dysfunction. Methods: Here we used single-cell RNA sequencing of tissue removed at the time of pulmonary endarterectomy surgery from five patients to identify the multiple cell types. Using in vitro assays, we analyzed differences in phenotype between CTEPH thrombus and healthy pulmonary vascular cells. We studied potential therapeutic targets in cells isolated from CTEPH thrombus. Measurements and Main Results: Single-cell RNA sequencing identified multiple cell types, including macrophages, T cells, and smooth muscle cells (SMCs), that constitute CTEPH thrombus. Notably, multiple macrophage subclusters were identified but broadly split into two categories, with the larger group characterized by an upregulation of inflammatory signaling predicted to promote pulmonary vascular remodeling. CD4+ and CD8+ T cells were identified and likely contribute to chronic inflammation in CTEPH. SMCs were a heterogeneous population, with a cluster of myofibroblasts that express markers of fibrosis and are predicted to arise from other SMC clusters based on pseudotime analysis. Additionally, cultured endothelial, smooth muscle, and myofibroblast cells isolated from CTEPH fibrothrombotic material have distinct phenotypes from control cells with regard to angiogenic potential and rates of proliferation and apoptosis. Last, our analysis identified PAR1 (protease-activated receptor 1) as a potential therapeutic target that links thrombosis to chronic PE in CTEPH, with PAR1 inhibition decreasing SMC and myofibroblast proliferation and migration. Conclusions: These findings suggest a model for CTEPH similar to atherosclerosis, with chronic inflammation promoted by macrophages and T cells driving vascular remodeling through SMC modulation, and suggest new approaches for pharmacologically targeting this disease.

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Published In

Am J Respir Crit Care Med

DOI

EISSN

1535-4970

Publication Date

May 15, 2023

Volume

207

Issue

10

Start / End Page

1358 / 1375

Location

United States

Related Subject Headings

  • Vascular Remodeling
  • Thrombosis
  • Single-Cell Analysis
  • Respiratory System
  • Receptor, PAR-1
  • Pulmonary Embolism
  • Pulmonary Artery
  • Myocytes, Smooth Muscle
  • Inflammation
  • Hypertension, Pulmonary
 

Citation

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MLA
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Viswanathan, G., Kirshner, H. F., Nazo, N., Ali, S., Ganapathi, A., Cumming, I., … Rajagopal, S. (2023). Single-Cell Analysis Reveals Distinct Immune and Smooth Muscle Cell Populations that Contribute to Chronic Thromboembolic Pulmonary Hypertension. Am J Respir Crit Care Med, 207(10), 1358–1375. https://doi.org/10.1164/rccm.202203-0441OC
Viswanathan, Gayathri, Hélène Fradin Kirshner, Nour Nazo, Saba Ali, Asvin Ganapathi, Ian Cumming, Yonghua Zhuang, et al. “Single-Cell Analysis Reveals Distinct Immune and Smooth Muscle Cell Populations that Contribute to Chronic Thromboembolic Pulmonary Hypertension.Am J Respir Crit Care Med 207, no. 10 (May 15, 2023): 1358–75. https://doi.org/10.1164/rccm.202203-0441OC.
Viswanathan G, Kirshner HF, Nazo N, Ali S, Ganapathi A, Cumming I, et al. Single-Cell Analysis Reveals Distinct Immune and Smooth Muscle Cell Populations that Contribute to Chronic Thromboembolic Pulmonary Hypertension. Am J Respir Crit Care Med. 2023 May 15;207(10):1358–75.
Viswanathan, Gayathri, et al. “Single-Cell Analysis Reveals Distinct Immune and Smooth Muscle Cell Populations that Contribute to Chronic Thromboembolic Pulmonary Hypertension.Am J Respir Crit Care Med, vol. 207, no. 10, May 2023, pp. 1358–75. Pubmed, doi:10.1164/rccm.202203-0441OC.
Viswanathan G, Kirshner HF, Nazo N, Ali S, Ganapathi A, Cumming I, Zhuang Y, Choi I, Warman A, Jassal C, Almeida-Peters S, Haney J, Corcoran D, Yu Y-R, Rajagopal S. Single-Cell Analysis Reveals Distinct Immune and Smooth Muscle Cell Populations that Contribute to Chronic Thromboembolic Pulmonary Hypertension. Am J Respir Crit Care Med. 2023 May 15;207(10):1358–1375.

Published In

Am J Respir Crit Care Med

DOI

EISSN

1535-4970

Publication Date

May 15, 2023

Volume

207

Issue

10

Start / End Page

1358 / 1375

Location

United States

Related Subject Headings

  • Vascular Remodeling
  • Thrombosis
  • Single-Cell Analysis
  • Respiratory System
  • Receptor, PAR-1
  • Pulmonary Embolism
  • Pulmonary Artery
  • Myocytes, Smooth Muscle
  • Inflammation
  • Hypertension, Pulmonary