PB0008 - Increased levels of plasma cell-free DNA are associated with symptomatology in patients with carotid stenosis and are independent of plasma DNaseI activity

Sunday, June 25, 2023

18:30 – 19:30 ET

Room: Exhibition

Co-Author(s)

EP

Emma Plana

Postdoctoral Researcher
Haemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain; Angiology and Vascular Surgery Service, La Fe University and Polytechnic Hospital, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
Julia Oto

Postdoctoral Researcher
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain

Presenting Author(s)

Raquel Herranz, MSc (she/her/hers)

PhD Candidate
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain

Co-Author(s)

FM

Francesca Maria Mursia

Medical doctor
Angiology and Vascular Surgery Service. University and Polytechnic Hospital La Fe. Valencia. Spain
Valencia, Comunidad Valenciana, Spain
NG

Noelia Garzon

Medical doctor
Angiology and Vascular Surgery Service. University and Polytechnic Hospital La Fe. Valencia. Spain
Valencia, Comunidad Valenciana, Spain
FC

Fernando Cana

Laboratory Technician
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
PM

Pilar Medina

PI
Hemostasis, Thrombosis, Arteriosclerosis and Vascular Biology Research Group, Medical Research Institute Hospital La Fe, Valencia, Spain
Valencia, Comunidad Valenciana, Spain
MM

Manuel Miralles

Head of Angiology and Vascular Surgery Service
Angiology and Vascular Surgery Service. University and Polytechnic Hospital La Fe. Valencia. Spain
Valencia, Comunidad Valenciana, Spain

Background: Plasma cell-free DNA (cfDNA) is increased in several disorders and may represent a biomarker for stroke, but it is presently unknown whether could be a marker of plaque instability. Plasma DNaseI activity modulates cfDNA concentration, but in carotid stenosis has not been assessed.

Aims: To evaluate the performance of plasma cfDNA as biomarker of plaque vulnerability in patients with carotid stenosis, and to ascertain its regulation by plasma DNaseI activity.

Methods: Twenty-six patients with symptomatic carotid stenosis and 40 asymptomatic sex and age matched were recruited. Symptomatology was considered when stroke, transient ischaemic attack or amaurosis fugax occurred within < 21 days before surgery. cfDNA was quantified using the Quant-iT™ PicoGreen™ dsDNA Assay kit (Invitrogen). Fluorescence was interpolated in a standard curve prepared with known concentrations of λ DNA.
DNaseI activity was measured with the single radial enzyme diffusion (SRED) assay in 1% agarose gels prepared in 10mM Tris-HCl, 5mM MgCl, 1mM CaCl2 containing 22. µg/ml salmon testis DNA (Sigma) and Safeview (NBSbiological). DNaseI activity of 2µl plasma was estimated as the halo area formed by DNA degradation during 17h at 37ºC in a humid chamber, measured with the ImageJ software.

Results: cfDNA concentration was higher in patients with symptomatic than asymptomatic carotid stenosis (1289 vs. 1190 ng/ml; p=0.047) (Figure 1A). No differences in DNaseI activity were observed between symptomatic and asymptomatic patients (Figure 1B). Moreover, plasma cfDNA concentration was not correlated with DNaseI activity in these patients.

Conclusion(s): Patients with symptomatic carotid stenosis have an increased plasma cfDNA concentration, which may be caused by tissue damage and inflammation. This increase appears to be, at least in part, independent from plasma DNaseI activity. Additional studies would clarify the utility of cfDNA as a marker of plaque vulnerability and scoring of patient´s prognosis and diagnosis. ISCIII-FEDER (PI20/01171, PI20/00075, FI21/00171).