Aberrant endothelial and smooth muscle progenitor cell frequency and function in Type 2 diabetes: a predisposing factor for increased cardiovascular risk?

Aberrant endothelial and smooth muscle progenitor cell frequency and function in Type 2 diabetes: a predisposing factor for increased cardiovascular risk? - Vascular progenitor cells in Type 2 diabetes

Status

Active, not recruiting

Phases

Unknown

Study type

Observational

Source

NL-OMON

Registry ID

NL-OMON35549

Enrollment

180

Registered

2009-02-13

Start date

2010-01-16

Completion date

Unknown

Last updated

2024-05-06

For informational purposes only — not medical advice. Sourced from public registries and may not reflect the latest updates. Terms

Conditions

atherosclerosis cardiovascular disease

Interventions

None listed

Eligibility

Age

18 Years to 99 Years

Inclusion criteria

Inclusion criteria: Patients eligible for participation are diagnosed with T2DM (duration 10-15 yrs) and divided into diabetic patients with PAD, with CAD or without cardiovascular disease. Patients eligible for participation in the various control groups are patients without a history of T2DM but with PAD or CAD, respectively. In addition, healthy individuals who have neither a history of diabetes, cardiovascular disease, inflammatory or autoimmune disease, nor obesity nor any other chronic disease are will be included.;Definitions PAD and CAD:;Peripheral artery disease (PAD) PAD is defined by the presence of carotid stenosis or lower extremity atherosclerosis obliterans. Severity of atherosclerosis of carotid vessels will be assessed by ultrasonography as a continuous variable. Diagnosis of atherosclerotic involvement of the lower extremities will be assessed by history of claudication or rest pain, bilateral pulses examination (dorsal pedal, posterior tibial, popliteal, and femoral arteries), ultrasonography performed bilaterally at levels of popliteal and femoral arteries, and, eventually, angiography. Patients are then classified according to the Leriche/Fontaine clinical classification of lower limb atherosclerosis obliterans.;Coronary artery disease (CAD) CAD is defined as previous myocardial infarction, resting ECG indicative of past MI, positive ECG at exercise stress test, echocardiography stress-test positive for inducible ischemia, evidence of significant coronary artery stenose on angiography with or without typical chest pain. ;Participants allocated to the different groups will be matched for: - gender - age - smoking - BMI - arterial hypertension - HbA1c (diabetics) - lipid profile

Exclusion criteria

Exclusion criteria: Predefined exclusion criteria for T2DM include diagnosis of T1DM and presence of any of the following (self-reported) medical conditions: - microvascular disease (diabetic retinopathy and nephropathy) - auto-immune diseases - current or prior cancers - acute or chonic infection - recent surgery or vascular intervention - age >80yrs - recent myocardial infarction - hemodialysis - immunosuppression - any other unrelated disease;Exclusion criteria for non-diabetics include: - diagnosis of T1DM or T2DM - auto-immune diseases - current or prior cancers - acute or chonic infection - recent surgery or vascular intervention - age >80yrs - recent myocardial infarction - hemodialysis - immunosuppression - any other unrelated disease

Design outcomes

Primary

Measure	Time frame
From the peripheral blood obtained PBMCs will be isolated using Ficoll density gradient centrifugation. Isolated cells will then be subjected to: 1) flowcytometric analyses: These phenotypic analyses allows quantification circulating endothelial and smooth muscle progenitor cells (EPCs and SMPCs, respectively) in the peripheral blood of diabetic and non-diabetic subjects and correlate frequencies of these specific cell subsets to the presence of cardiovascular disease. 2) flowcytometric cell sorting: These analyses allow purification of specific EPC and SMPC subsets which will be used for more in-depth functional and molecular analyses as described below. 3) in vitro EPC and SMPC culture: These analyses allow functional studies on EPCs and SMPCs like tube formation, anti-thrombotic activity, differentiation into mature ECs and SMCs, SMC contractility, and matrix production. 4) molecular analyses: In order to gain insight into the molecular pathways involved in aberrant EPC and SMPC function as determined under point 3, mRNA and proteins will be isolated from purified cells susbsets and/or after expansion/differentiation in vitro. Isolated mRNA and proteins will then be subjected to gene expression analysis (qRT-PCR and microarray analysis [Illumina platform] and protein expression analysis (Western-blotting and kinome profiling [PepChip platform]), respectively. 5) histological analyses From a subpopulation of patients with moderate to severe peripheral artery disease the endarterectomy specimen will be collected and processed for histological and moelcular biological analyses. Results of these analyses will be correlated with the in vitro generated data on cultured cells using the methods described above (1 to 4). Collectively, the results of these analyses will contribute to the determination of the level of vascular progenitor cell dysfunction in T2D and to the identification of novel molecular targets for therapeutic interventio	—

Measure

Time frame

From the peripheral blood obtained PBMCs will be isolated using Ficoll density gradient centrifugation. Isolated cells will then be subjected to: 1) flowcytometric analyses: These phenotypic analyses allows quantification circulating endothelial and smooth muscle progenitor cells (EPCs and SMPCs, respectively) in the peripheral blood of diabetic and non-diabetic subjects and correlate frequencies of these specific cell subsets to the presence of cardiovascular disease. 2) flowcytometric cell sorting: These analyses allow purification of specific EPC and SMPC subsets which will be used for more in-depth functional and molecular analyses as described below. 3) in vitro EPC and SMPC culture: These analyses allow functional studies on EPCs and SMPCs like tube formation, anti-thrombotic activity, differentiation into mature ECs and SMCs, SMC contractility, and matrix production. 4) molecular analyses: In order to gain insight into the molecular pathways involved in aberrant EPC and SMPC function as determined under point 3, mRNA and proteins will be isolated from purified cells susbsets and/or after expansion/differentiation in vitro. Isolated mRNA and proteins will then be subjected to gene expression analysis (qRT-PCR and microarray analysis [Illumina platform] and protein expression analysis (Western-blotting and kinome profiling [PepChip platform]), respectively. 5) histological analyses From a subpopulation of patients with moderate to severe peripheral artery disease the endarterectomy specimen will be collected and processed for histological and moelcular biological analyses. Results of these analyses will be correlated with the in vitro generated data on cultured cells using the methods described above (1 to 4). Collectively, the results of these analyses will contribute to the determination of the level of vascular progenitor cell dysfunction in T2D and to the identification of novel molecular targets for therapeutic interventio

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Secondary

Measure	Time frame
n.a.	—

Countries

Netherlands

Outcome results