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

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.

Study Overview

• Status: Not yet recruiting
• Conditions: Carotid Artery Plaque; Carotid Atherosclerosis; TIA
• Intervention / Treatment: Drug: Interleukin-2 [IL-2]; Procedure: Standard care - Carotid Endarterectomy

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:

1. 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
2. To assess if modulating plaque Tregs can cause a shift in the plaque immune landscape to a less inflammatory phenotype
3. To study the relationship between plaque and circulating immune cells after systemic immune modulation

• Study Type: Interventional
• Enrollment (Estimated): 30
• Phase: Not Applicable

Contacts and Locations

Study Locations

• United Kingdom
  • Cambridgeshire
    • Cambridge, Cambridgeshire, United Kingdom, CB2 0QQ
      • Addenbrookes Hospital
      • Contact: Tian Zhao, MBBS, PhD; Phone Number: 01223768678; Email: Txz20@cam.ac.uk

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: Adult; Older Adult
• Accepts Healthy Volunteers: No

Description

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

Study Plan

How is the study designed?

Design Details

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

Number of Arms

2

Arms and Interventions

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

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

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Other immune cell and vascular smooth muscle cell gene expression	As per primary outcome measure but in immune cells and vascular smooth muscle cells (excluding Treg & Teff cells). Differential gene expression will be assessed using standard techniques (Z-Score)	Time of surgery
Ligand-receptor interactions.	Differential regulation of ligand-receptor interactions between control and IL-2 treated cells will be identified by comparing single-cell/single-cell ligand-receptor interaction scores for the two groups. This measure is descriptive in nature.	Time of surgery
T-Cell receptor profile	Difference in TCR clonality in plaques between the two groups will be assessed from clonotypes based on identical V-J gene usage and identical CDR3 junctions.	Time of surgery
Inflammatory pathway activation.	Activation of inflammatory pathways both at plaque and systemic level. Assess for enrichment of genes involved in inflammatory pathways - enrichment score.	Time of surgery

Collaborators and Investigators

• Sponsor: Cambridge University Hospitals NHS Foundation Trust

Study record dates

Study Major Dates

• Study Start (Estimated): August 26, 2023
• Primary Completion (Estimated): August 1, 2025
• Study Completion (Estimated): September 1, 2025

Study Registration Dates

• First Submitted: June 14, 2023
• First Submitted That Met QC Criteria: August 2, 2023
• First Posted (Actual): August 4, 2023

Study Record Updates

• Last Update Posted (Actual): August 4, 2023
• Last Update Submitted That Met QC Criteria: August 2, 2023
• Last Verified: August 1, 2023

More Information

Terms related to this study

• Keywords: Carotid Stenosis; IL-2; TIA; Interleukin-2
• Additional Relevant MeSH Terms: Cardiovascular Diseases; Vascular Diseases; Cerebrovascular Disorders; Brain Diseases; Central Nervous System Diseases; Nervous System Diseases; Arteriosclerosis; Arterial Occlusive Diseases; Pathological Conditions, Anatomical; Carotid Stenosis; Carotid Artery Diseases; Atherosclerosis; Plaque, Atherosclerotic; Physiological Effects of Drugs; Peripheral Nervous System Agents; Analgesics; Sensory System Agents; Analgesics, Non-Narcotic; Antineoplastic Agents; Interleukin-2
• Other Study ID Numbers: A096090

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No
• product manufactured in and exported from the U.S.: No