The Effect of Low-dose Interleukin-2 on the Immune Landscape of Human Atherosclerotic Plaques at Single Cell Resolution.

Study Overview
=================
Brief Summary
-----------------
The goal of this clinical trail is to compare the differences in carotid plaque Treg cells' gene signature for activation, proliferation, and suppressive function using scRNA-seq in patients treated with IL-2 compared to control.

Detailed Description
-----------------
Up till now our Lab has looked at Tregs and immune cells in the blood. The question remains whether low dose IL-2 can have the desired effect on Tregs in atherosclerotic plaques where they could alter the pathophysiology and potentially clinical outcomes for patients. Up until recently, the cellular composition and cell-specific expression patterns of human atherosclerotic plaques remained elusive. However, recent breakthroughs studies using scRNA-seq, CITE-seq, and single-cell ATAC-seq on human carotid plaques have offered important insight into plaque composition, cell heterogeneity, and cell-cell interactions giving new perspectives on mechanisms of disease. The next logical stage is to use this new insight and powerful biological tool to assist in drug development for patients. Therefore, the aims of the study are: To assess if low dose IL-2, given systemically to patients at our proposed dose, can alter Tregs in atherosclerotic plaques (the disease tissue) to exhibit a proliferating, activated, and immunosuppressive phenotype To assess if modulating plaque Tregs can cause a shift in the plaque immune landscape to a less inflammatory phenotype To study the relationship between plaque and circulating immune cells after systemic immune modulation

Official Title
-----------------
The Effect of Low-dose Interleukin-2 on the Immune Landscape of Human Atherosclerotic Plaques at Single Cell Resolution.

Conditions
-----------------
TIA, Carotid Artery Plaque, Carotid Atherosclerosis

Intervention / Treatment
-----------------
* Drug: Interleukin-2 [IL-2]
* Procedure: Standard care - Carotid Endarterectomy


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Presence of carotid stenosis on either ultrasound or CT scan. Planned to undergo carotid endarterectomy. Exclusion Criteria: Autoimmune disease Any regular immunosuppressive treatment [Inhaled or topical steroids are permissible] Modified Rankin Scale score of ≥4 at screening Known active hepatic disease or alanine aminotransferase (ALT) > 2xULN Severe chronic kidney disease (defined as eGFR < 30 ml/min/1.73m2) Allergy or intolerance to aldesleukin Signs or symptoms of active infection History of human immunodeficiency virus (HIV), hepatitis B or C Current malignancy requiring active treatment Vaccine within 4 weeks prior to screening or plans for vaccination during study period Women of child-bearing potential and pregnancy Women who are breast-feeding Clinically relevant medical or surgical conditions that, in the opinion of the

Ages Eligible for Study
-----------------
Minimum Age: 18 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
No

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Low dose interleukin-2<br>Commercially available aldesleukin with a UK marketing authorisation will be used and will be initially prepared as per SmPC. | Drug: Interleukin-2 [IL-2]<br>* 5 sequential days of treatment (1.5MIU/day subcutaneously)<br>Procedure: Standard care - Carotid Endarterectomy<br>* Standard care for patients with carotid stenosis undergoing carotid endarterectomy<br>|
| Active Comparator: Control<br>Standard of care treatment | Procedure: Standard care - Carotid Endarterectomy<br>* Standard care for patients with carotid stenosis undergoing carotid endarterectomy<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Difference in gene expression in Tregs | Comparing differential gene expression using scRNA-seq technologies, from isolated Tregs from carotid plaques from the two patient groups (IL-2 treatment and control). Differential gene expression will be assessed using standard techniques (Z-Score) | Time of surgery | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Difference in gene expression in Teff cells. | Comparing differential gene expression using scRNA-seq technologies, from isolated Teffs from carotid plaques from the two patient groups (IL-2 treatment and control). Differential gene expression will be assessed using standard techniques (Z-Score) | Time of surgery | 
| Immune cell gene signature, in plaque and blood. | Difference in immune cell gene signature will be compared using scRNA-seq in both plaque and blood samples, and further compared between IL-2 and control groups. Comparative analysis will be completed using gene set enrichment analysis (Enrichment Score). | Time of surgery | 
| Difference in gene expression patterns in Treg & Teff cells. | Comparison between Treg and Teff cells collected at baseline and day of surgery will be compared between IL-2 and control groups. Differential gene expression will be assessed using standard techniques (Z-Score) | Baseline and at time of surgery | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
Interleukin-2, IL-2, TIA, Carotid Stenosis