Serum exosomal protein profiling for the non-invasive detection of cardiac allograft rejection

doi:10.1016/j.healun.2017.07.012

The Journal of Heart and Lung Transplantation

Volume 37, Issue 3, March 2018, Pages 409-417

https://doi.org/10.1016/j.healun.2017.07.012 Get rights and content

Background

Exosomes are cell-derived circulating vesicles that play an important role in cell–cell communication. Exosomes are actively assembled and carry messenger RNAs, microRNAs and proteins. The “gold standard” for cardiac allograft surveillance is endomyocardial biopsy (EMB), an invasive technique with a distinct complication profile. The development of novel, non-invasive methods for the early diagnosis of allograft rejection is warranted. We hypothesized that the exosomal proteome is altered in acute rejection, allowing for a distinction between non-rejection and rejection episodes.

Methods

Serum samples were collected from heart transplant (HTx) recipients with no rejection, acute cellular rejection (ACR) and antibody-mediated rejection (AMR). Liquid chromatography–tandem mass spectrometry (LC-MS/MS) analysis of serum exosome was performed using a mass spectrometer (Orbitrap Fusion Tribrid).

Results

Principal component analysis (PCA) revealed a clustering of 3 groups: (1) control and heart failure (HF); (2) HTx without rejection; and (3) ACR and AMR. A total of 45 proteins were identified that could distinguish between groups (q < 0.05). Comparison of serum exosomal proteins from control, HF and non-rejection HTx revealed 17 differentially expressed proteins in at least 1 group (q < 0.05). Finally, comparisons of non-rejection HTx, ACR and AMR serum exosomes revealed 15 differentially expressed proteins in at least 1 group (q < 0.05). Of these 15 proteins, 8 proteins are known to play a role in the immune response. Of note, the majority of proteins identified were associated with complement activation, adaptive immunity such as immunoglobulin components and coagulation.

Conclusions

Characterizing of circulating exosomal proteome in different cardiac disease states reveals unique protein expression patterns indicative of the respective pathologies. Our data suggest that HTx and allograft rejection alter the circulating exosomal protein content. Exosomal protein analysis could be a novel approach to detect and monitor acute transplant rejection and lead to the development of predictive and prognostic biomarkers.

Section snippets

Patient enrollment and baseline demographics

Study participants were divided into 5 groups: healthy controls (n = 10); HF patients without allograft (n = 10); HTx patients without rejection (n = 10); and HTx patients undergoing ACR (n = 10) or AMR (n = 8). Control serum samples were collected from healthy volunteers (see Table S1 in the Supplementary Material available online at www.jhltonline.org). Non-allograft HF patients were recruited during visits to the outpatient HF Clinic at New York–Presbyterian Hospital/Columbia University

Exosomal protein profiling distinguishes between various cardiac pathologies

A total of 3,537 proteins were identified based on a 1% FDR at the peptide level. LIMMA was applied to the semi-quantitative values (spectral counts) of the entire data set and differentially expressed proteins were identified with the FDR threshold at 5% (q < 0.05). PCA applied to the data set identified an exosomal protein signature, which distinguished the following 3 patient groups: (1) control and HF; (2) HTx, no rejection; and (3) ACR and AMR. A total of 45 proteins were identified that

Discussion

Exosomes are secretory vesicles that are now known to play an increasingly important role in intercellular signaling.17, 19 Exosomes have been shown to modulate antigen presentation, cytokine production and cell proliferation both in vitro and in vivo.29, 30, 31, 32 Our study has shown that cardiac allograft rejection is linked to significant changes in the serum exosomal proteome, especially in proteins controlling immunity and hemostasis, in a comparison with HTx patients not experiencing

Disclosure statement

The authors have no conflicts of interest to disclose. This work was supported by the National Heart Lung and Blood Institute (R01HL114813 to P.C.S. and 5T35HL007616-35 to A.H.).

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Original Pre-Clinical ScienceSerum exosomal protein profiling for the non-invasive detection of cardiac allograft rejection

Background

Methods

Results

Conclusions

Section snippets

Patient enrollment and baseline demographics

Exosomal protein profiling distinguishes between various cardiac pathologies

Discussion

Disclosure statement

Lancet

Transplant Proc

J Heart Lung Transplant

Lung Cancer

Mol Cell Proteomics

J Heart Lung Transplant

Biochem Pharmacol

Curr Opin Cell Biol

Blood

Immunity

Blood

Genomics

J Biol Chem

Adv Immunol

Pathol Biol (Paris)

Int J Cardiol

Hum Immunol

Mayo Clin Proc

J Heart Lung Transplant

Cardiovasc Pathol

Am J Transplant

J Am Coll Cardiol

Biochim Biophys Acta

Forecasting the impact of heart failure in the United States: a policy statement from the American Heart Association

Circ Heart Fail

The failing heart—an engine out of fuel

N Engl J Med

Heart disease and stroke statistics—2008 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee

Circulation

Percutaneous transvenous endomyocardial biopsy

JAMA

Long-term care of the heart transplant recipient

Curr Opin Organ Transplant

Increased RIPK4 expression is associated with progression and poor prognosis in cervical squamous cell carcinoma patients

Sci Rep

Applying precision medicine to the active surveillance of prostate cancer

Cancer

Negative expression of CPSF2 predicts a poorer clinical outcome in patients with papillary thyroid carcinoma

Thyroid

Original Pre-Clinical Science
Serum exosomal protein profiling for the non-invasive detection of cardiac allograft rejection