Investigation of coagulation and proteomics profiles in symptomatic feline hypertrophic cardiomyopathy and healthy control cats

Abstract

Background: Hypertrophic cardiomyopathy (HCM) is a crucial heart disease in cats. The clinical manifestations of HCM comprise pulmonary edema, dyspnea, syncope, arterial thromboembolism (ATE), and sudden cardiac death. D-dimer and prothrombin time (PT) are powerful biomarkers used to assess coagulation function. Dysregulation in these two biomarkers may be associated with HCM in cats. This study aims to assess D-dimer levels, PT, and proteomic profiling in healthy cats in comparison to cats with symptomatic HCM.

Results: Twenty-nine client-owned cats with HCM were enrolled, including 15 healthy control and 14 symptomatic HCM cats. The D-dimer concentration and PT were examined. Proteomic analysis was conducted by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry and liquid chromatography-tandem mass spectrometry (LC-MS/MS). In symptomatic cats, D-dimer levels were statistically significantly higher (mean ± SEM: 372.19 ng/ml ± 58.28) than in healthy cats (mean ± SEM: 208.54 ng/ml ± 10.92) with P-value of less than 0.01, while PT was statistically significantly lower in symptomatic cats (mean ± SEM: 9.8 s ± 0.15) compared to healthy cats (mean ± SEM: 11.08 s ± 0.23) with P-value of less than 0.0001. The proteomics analysis revealed upregulation of integrin subunit alpha M (ITGAM), elongin B (ELOB), and fibrillin 2 (FBN2) and downregulation of zinc finger protein 316 (ZNF316) and ectonucleoside triphosphate diphosphohydrolase 8 (ENTPD8) in symptomatic HCM cats. In addition, protein-drug interaction analysis identified the Ras signaling pathway and PI3K-Akt signaling pathway.

Conclusions: Cats with symptomatic HCM have higher D-dimer and lower PT than healthy cats. Proteomic profiles may be used as potential biomarkers for the detection and management of HCM in cats. The use of D-dimer as a biomarker for HCM detection and the use of proteomic profiling for a better understanding of disease mechanisms remain to be further studied in cats.

Keywords: Cats; D-dimer; Hypertrophic cardiomyopathy; Proteomics; Prothrombin time.

Fig. 1

The results of coagulation markers, including (A) D-dimer level and (B) Prothrombin time (PT). All data was analyzed by t-test. The mean ± standard error of the mean (SEM) of coagulation markers levels of healthy and symptomatic HCM is illustrated. The area behind both bar graphs showed the standard ranges of each coagulation marker. ** p < 0. 01, ****p < 0.0001 were considered statistically significant

Fig. 2

Peptide mass fingerprints (PMFs) from MALDI-TOF mass spectrometry with (A) healthy control cats and (B) Symptomatic HCM cats

Fig. 3

Partial least squares discriminant analysis (PLS-DA) score plot of the proteins in healthy cats (red) and symptomatic HCM (green) demonstrated the protein expression in both groups that completely separated

Fig. 4

Volcano plots representing the results of comparisons between healthy control cats and symptomatic HCM cats, including downregulation (blue), non-significant (gray), and upregulation (red)

Fig. 5

The protein and drug interaction networks form LC-MS/MS. The circle indicates the protein associated with pathways for cardiac hypertrophy