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The Role of the Adrenergic System in Hypoglycaemia Induced Inflammatory Response in People With Type 1 Diabetes and People Without Type 1 Diabetes-RAID-II

Not Applicable
Recruiting
Conditions
Diabetes Mellitus, Type 1
Inflammation
Interventions
Drug: hyperinsulinaemic hypoglycaemic clamp
Registration Number
NCT06422494
Lead Sponsor
Radboud University Medical Center
Brief Summary

The goal of this trial is to study the effect that adrenaline has on the immune reaction seen during a low blood sugar. People with type 1 diabetes do not produce their own insulin. The cells in the pancreas that produce insulin are destroyed. People with type 1 diabetes require daily insulin administration. As a consequence of this insulin therapy the blood sugar can dip too low, causing symptoms such as confusion, irritation and tiredness. This is called hypoglycaemia. Hypoglycaemia has been associated with an increased risk for cardiovascular disease such as heart attacks. During hypoglycaemia the immune system is activated. The immune system consists of white blood cells which produce cytokines, these are proteins used to kill pathogens such as bacteria. During hypoglycaemia there are no pathogens but the cytokines are still produced, leading to unwanted damage. A previous study performed by our research group showed that the immune system activation caused by hypoglycaemia is associated with the stress hormone adrenaline. Adrenaline is released by the body in moments of stress such as during running or bungee jumping. Adrenaline is also released by the body during hypoglycaemia to increase the sugar level. Our hypothesis is that adrenaline activates the immune system during hypoglycaemia. Adrenaline acts in the body through two receivers, these are called alpha and beta receptors. These are present on almost all cells in the body especially on the immune cells. With the study we want to study the situation where there is a hypoglycaemia without the adrenaline. We will achieve this by lowering the blood sugar in participants. During the low blood sugar we will administer two drugs, which will attach themselves to the adrenaline receivers, the alpha and beta receptor. With this method we hope to block the adrenaline effects and with that block the immune response caused by adrenaline.

Detailed Description

Rationale: Hypoglycaemia has shown to cause a sustained pro-inflammatory response which could promote a pro-atherogenic state and explain the association between hypoglycaemia and cardiovascular events. This pro-inflammatory response has been linked to the adrenaline response to hypoglycaemia. Adrenergic blockade with α and β adrenergic receptor antagonists (ARA) has shown to blunt the leukocyte response after hypoglycaemia induction and adrenaline administration. Whether and to what degree a combined blockade blunts the hypoglycaemia induced pro-inflammatory response is unknown.

Objective: to examine the effect of adrenergic inhibition on the hypoglycaemia induced inflammatory response (e.g. leukocyte phenotype, cytokines, inflammatory proteins) by performing a hyperinsulinaemic hypoglycaemic glucose clamp alongside infusion of α-ARA and β-ARA. Secondary objectives consist of the effect of adrenergic blockade during hypoglycaemia on atherogenic parameters and glucose metrics ( e.g. time in range).

Study design: Intervention study with a cross-over design

Study population: Potentially eligible adult ( 16 - 75 years) participants will be recruited through social media, the Radboudumc outpatient clinic and other advertisements. We will recruit a total of 24 individuals, i.e. 12 healthy participants and 12 participants with type 1 diabetes. Participants with type 1 diabetes will be twice ( as there are two investigational days) equipped with a blinded continuous glucose monitoring device (CGM) during the test, which will measure interstitial glucose levels for a total of 10 days.

Intervention: All participants will undergo a hyperinsulinaemic hypoglycaemic glucose clamp ( nadir 2.8 mmol/L). During the clamp the participants will be randomized to receive an infusion of saline or an infusion of phentolamine and propranolol. This will be done using a cross-over design. The participants will undergo both the saline and adrenergic blockade.

Main study parameters/endpoints: The main study parameter will be the monocyte count after 60 minutes hyperinsulinaemic hypoglycaemic clamp and adrenergic blockade during the clamp.

Recruitment & Eligibility

Status
RECRUITING
Sex
All
Target Recruitment
24
Inclusion Criteria
  • Overall inclusion criteria:

    • Ability to provide written informed consent
    • Body-Mass Index: 18,5-35 kg/m2
    • Age ≥16 years, ≤ 75 years
    • Blood pressure: <140/90 mmHg
    • Non-smoking
    • Electrocardiogram not showing any serious arrythmias (premature ventricular complexes and premature atrial complexes accepted)

Diabetes group specific criteria:

  • Insulin treatment according to basal-bolus insulin regimen (injections or insulin pump)
  • Duration of diabetes > 1 year
  • HbA1c < 100 mmol/mol,
Exclusion Criteria
  • Any event of cardiovascular disease in the past 5 years (e.g. myocardial infarction, stroke, symptomatic peripheral arterial disease)
  • Pregnancy or breastfeeding or unwillingness to undertake measures for birth control
  • Active epilepsy ( with the need for treatment)
  • Allergy for sulphite
  • Active asthma with use of β2-bronchodilators or obstructive lung disease
  • Current treatment with Alpha- or beta-blockers (e.g. doxazosin, propranolol)
  • History of clinical significant Arrhythmias
  • Use of immune-modifying drugs or antibiotics
  • Use of antidepressants ( Including monoamine oxidase inhibitors, tricyclic antidepressants and serotonin-reuptake inhibitors)
  • Use of antipsychotics
  • Use of statins with the inability to stop statins >2 weeks before the investigational day.
  • Proliferative retinopathy
  • Nephropathy with an estimated glomerular filtration rate (by Chronic Kidney Disease Epidemiology Collaboration equation, CKD-EPI) ˂60ml/min/1.73m2

Study & Design

Study Type
INTERVENTIONAL
Study Design
CROSSOVER
Arm && Interventions
GroupInterventionDescription
Participants without type 1 diabeteshyperinsulinaemic hypoglycaemic clampThe participants without type 1 diabetes
Participants without type 1 diabetesPhentolamineThe participants without type 1 diabetes
Participants with type 1 diabeteshyperinsulinaemic hypoglycaemic clampParticipants with type 1 diabetes
Participants with type 1 diabetesPhentolamineParticipants with type 1 diabetes
Participants without type 1 diabetesPropranolol Hydrochloride 1 MG/MLThe participants without type 1 diabetes
Participants with type 1 diabetesPropranolol Hydrochloride 1 MG/MLParticipants with type 1 diabetes
Primary Outcome Measures
NameTimeMethod
Monocyte count after 60 minutes of hypoglycaemia and adrenergic blockadeAfter 60 minutes of hypoglycaemia and adrenergic blockade

The number of monocytes following 60 minutes hypoglycaemia and adrenergic blockade compared to baseline. Adrenergic blockade using Phentolamine and Propranolol intravenously. Expressed in 10\^3/µl measured using a sysmex machine.

Secondary Outcome Measures
NameTimeMethod
Variability measured by the blinded continuous glucose monitorDuring the full study, 3 days before and 7 days after each investigational day

Variability of glucose expressed as a standard deviation of the mean glucose

Average glucose measured by the blinded continuous glucose monitorDuring the full study, 3 days before and 7 days after each investigational day

Average glucose during the 10 days of measuring expressed as mmol/L

Amount of plasma glycerol0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Amount of plasma glycerol during and after hypoglycaemia

Amount of Non-esterified fatty acids0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Amount of Non-esterified fatty acids (NEFAs) during and after hypoglycaemia

Untargeted metabolomics profiling0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Measuring a panel of amino acids

Functional changes in monocytes0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Functional changes in monocytes (e.g. using adhesion assays, differentiation experiments)

Adrenergic symptoms assessed using the validated Edinburgh Hypoglycaemia Score0, 30 minutes after euglycaemia, 30 minutes and 60 minutes during hypoglycaemia
Hypoglycaemia awareness using the modified Clarke scoreAt screening
Plasma levels of hormones0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Plasma levels of hormones ( Cortisol, insulin, glucagon, growth-hormone, adrenaline, noradrenaline)

Amount of hypoglycaemic events measured by the blinded continuous glucose monitorDuring the full study, 3 days before and 7 days after each investigational day

Amount of events

Time in range measured by the blinded continuous glucose monitorDuring the full study, 3 days before and 7 days after each investigational day

Amount of time that glucose is between 3.8 and 10 mmol/L expressed as a percentage

Ex vivo production of pro- and anti-inflammatory cytokines and chemokines0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia, +1 day, +3 days and 1 week after of hypoglycaemia

Ex vivo production of pro- and anti-inflammatory cytokines and chemokines after ex vivo stimulation of isolated leukocytes, including Tumor necrosis factor-α, Interleukin-6, Interleukin-10 and Interleukin-1β, 1β

Inflammatory plasma protein ( e.g. high-sensitive crp)0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Inflammatory plasma protein using ELISA,(e.g high sensitive-crp)

Leukocyte count at the time points0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia, +1 day, +3 days and 1 week after of hypoglycaemia

Leukocyte count at the time points 0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia, +1 day, +3 days and 1 week after of hypoglycaemia (e.g. Monocytes, granulocytes, lymphocytes).

92 circulating inflammatory proteins0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

92 circulating inflammatory proteins using Olink Proteomics inflammation panel

Atherogenic parameters0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Atherogenic parameters using ELISA including but not limited to, vascular endothelial cell adhesion molecule-1, vascular endothelial cell adhesion molecule-1, E-Selectin, P-selectin, Plasminogen activator inhibitor-1, Plasma Endothelin

Gene expression changes in leukocytes0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Gene expression changes in leukocytes (e.g. using RNA sequencing, quantitative PCR)

Epigenetic changes in leukocytes0, 30 minutes after euglycaemia, 60 minutes during hypoglycaemia

Epigenetic changes in leukocytes (e.g. using Assay for Transposase- Accessible Chromatin using sequencing (ATACseq), DNA methylation analysis)

Trial Locations

Locations (1)

Radboud University Medical Center, Nijmegen, Netherlands

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Nijmegen, Gelderland, Netherlands

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