Sonoporation and Tumor Microenvironment Response in Colorectal Liver Metastases (CEUSON)

Tumor Response to Sonoporation: A Clinical Translational RCT of Contrast-enhanced Ultrasound Induced Changes in Tumor Microenvironment of Colorectal Liver Metastases

This study is an investigator-initiated, randomized controlled trial enrolling patients with colorectal liver metastases. The objectives are to evaluate the safety and efficacy of contrast-enhanced ultrasound (CEUS)-mediated sonoporation as a potential therapeutic intervention and to investigate whether sonoporation can modulate the tumor microenvironment toward a more immune-active state

Study Overview

• Status: Recruiting
• Conditions: Liver Metastases From Colorectal Cancer
• Intervention / Treatment: Other: Low-intensity contrast enhanced ultrasound; Other: High-intensity contrast enhanced ultrasound

Detailed Description

The trial is designed as an investigator-initiated, multicenter, prospective, randomized, controlled, two-arm, intervention trial with a matched historical control group aims to evaluate the therapeutic potential of contrast-enhanced ultrasound (CEUS) in patients with colorectal liver metastases (CLM). The primary objectives are to assess whether CEUS can modulate the tumor microenvironment and to determine the safety, tolerability, and feasibility of CEUS as a therapeutic modality. Eligible patients will be randomized to receive either low-intensity CEUS (LI-CEUS) or high-intensity CEUS (HI-CEUS) focused on a designated study metastasis. Safety, feasibility, and success-rate of the intervention will be registered. Minimum seven days post-intervention a biopsy of the study metastasis will be conducted. Subsequent analyses will compare immune cell infiltration, immune-related gene expression, and stromal and vascular remodeling in LI-CEUS- and HI-CEUS-exposed metastases versus historical controls.

• Study Type: Interventional
• Enrollment (Estimated): 48
• Phase: Not Applicable

Contacts and Locations

• Study Contact: Name: Maja M Jeppesen, M.D; Phone Number: +45 47323011; Email: mjepp@regionsjaelland.dk

Study Locations

• Denmark
  • Copenhagen, Denmark, 2100
    • Recruiting
    • Rigshospitalet
    • Contact: Hans-Christian Pommergaard, MD, DMSc, PhD, FEBS (HPB), Cli; Phone Number: +45 29 93 95 09; Email: hans-christian.pommergaard@regionh.dk
    • Contact: Maja Mønster Jeppesen, MD; Phone Number: +45 29939509; Email: mjepp@regsj.dk
  • Køge, Denmark, 4600
    • Recruiting
    • Surgical Department, Zealand University Hospital
    • Contact: Maja M Jeppesen, MD; Phone Number: +45 47323011; Email: mjepp@regionsjaelland.dk

Participation Criteria

Eligibility Criteria

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

Description

Inclusion Criteria:

• Adult patients (≥18 years)
• Ability to provide written informed consent
• Histopathological confirmed colorectal adenocarcinoma
• Scheduled for liver resection or ablation at the Rigshospital, or admitted to oncologic treatment due to colorectal liver metastases (CLM) at Zealand University Hospital
• Presence of at least one CLM evaluable and accessible by CEUS
• Eastern Cooperative Oncology Group (ECOG) performance status of 0-2.

• Follow the conditions regarding fertility, pregnancy, or lactation:

  • Women of childbearing potential (WOCBP) are defined as women ranging from the period of menarche till the post-menopausal period, unless permanently sterile (e.g. hysterectomy, bilateral salpingectomy and bilateral oophorectomy), Post-menopause is defined as no menses for 12 months without an alternative medical cause.
  • WOCBP should use a secure and highly effective birth control (as stated in the "Recommendations related to contraception and pregnancy testing in clinical trials", version 1.1, section 4.1, from the Clinical Trials Facilitation and Coordination Group) during the entire period of the trial. In cases of uncertainty regarding pregnancy, pregnancy testing either as highly sensitive serum or urine pregnancy test will be used.

Exclusion Criteria:

• Oncologic systemic treatment 2 weeks prior to inclusion
• Prior treatment with an immune checkpoint inhibitor (e.g. anti-PD-L1, anti PD-1, or anti-PD-L2)
• Inability to reliably distinguish the study metastasis from other hepatic metastases using CEUS
• Systemic treatment with either corticosteroids (>10 mg daily prednisolone equivalents) or other immunosuppressive medications within 2 weeks prior inclusion. Inhaled or topical steroids and adrenal replacements doses > 10 mg daily prednisone equivalents are permitted in the absence of active autoimmune disease.
• Known history of human immunodeficiency virus (HIV), active or chronic hepatitis B, or C.
• Pregnancy or lactation.
• Confirmed dMMR positive primary tumors
• Inability to comply with study protocol due to psychological, social, or logistical reasons.

• Contraindications for CEUS:

  • Known hypersensitivity to SonoVue® or any other ultrasound contrast agent
  • Uncontrolled atrial hypertension
  • Known right-to-left intracardiac shunt
  • Severe pulmonary hypertension (SAP >90 mmHg)
  • Adult respiratory distress syndrome.
  • Known hypersensitivity to macrogols
  • Unstable ischemic heart disease or acute coronary syndrome
  • Severe lung disease, e.g. severe chronic obstructive lung disease

Study Plan

How is the study designed?

Design Details

• Primary Purpose: Treatment
• Allocation: Randomized
• Interventional Model: Parallel Assignment
• Masking: Double

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
No Intervention: Historical control group; The historical control group consists of 24 matched patients who underwent liver resection for one or more colorectal liver metastases at Rigshospitalet between 2022 and the end of 2024. These patients serve as a non-intervention comparison cohort.
Experimental: Low-intensity CEUS intervention group; 12 eligible patients will be randomized to receive LI-CEUS intervention	Other: Low-intensity contrast enhanced ultrasound; The LI-CEUS arm will receive a 30-minute contrast-enhanced ultrasound session targeting a designated study metastasis. During the procedure, 1 mL of the ultrasound contrast agent SonoVue will be administered every 3 minutes throughout the entire session. Minimum 7 days after the intervention a biopsy of the study metastasis will be conducted.
Experimental: High-intensity CEUS intervention group; 12 eligible patients will be randomized to receive HI-CEUS intervention	Other: High-intensity contrast enhanced ultrasound; The HI-CEUS intervention group will recieve a 30-minute CEUS procedure targeting a designated study metastasis. During the procedure, 1 mL of the ultrasound contrast agent SonoVue will be administered every 3 minutes throughout the entire session. A high-intensity ultrasound "flash" will be applied every 30 seconds. This "flash" consists of a brief sequence of ultrasound waves with a high mechanical index that causes complete destruction of microbubbles within the imaging field. Minimum 7 days after the intervention a biopsy of the study metastasis will be conducted.

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Time Frame
Immune cell density (CD8+, CD3+) in colorectal liver metastases exposed to LI-CEUS or HI-CEUS, compared to controls.	≥7 days after postinterventional

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Number of participants in the intervention group with treatment-related adverse events assessed by CTCAE v5.0		Daily, until 7 days post-interventional
Procedure success of LI-CEUS and HI-CEUS procedures as assessed by operator-defined completion of the planned intervention		Periinterventional
Differential expression of immune-related genes between CEUS-exposed and control liver metastases	Tissue bulk transcriptional analyses will be evaluated by the Nanostring nCounter platform. Immune-related spatial protein profiling of tumor regions will be evaluated by the GeoMx digital spatial profiler	≥7 days postinterventional
Systemic immune response assessed by changes in the circulating cytokine profiles following LI-CEUS and HI-CEUS		Pre- and ≥7 days postinterventional
Frequency and type of technical issues encountered during LI-CEUS and HI-CEUS procedures assessed by the operator		Periinterventional

Collaborators and Investigators

• Sponsor: Zealand University Hospital
• Collaborators: Rigshospitalet, Denmark
• Investigators: Principal Investigator: Christian P Nolsøe, Professor, Center of Surgical Ultrasound, Surgical department of Zealand University Hospital; Study Director: Ismail Gögenur, Professor, Center of Surgical Science, Surgical department of Zealand University Hospital

Study record dates

Study Major Dates

• Study Start (Actual): February 25, 2026
• Primary Completion (Estimated): January 15, 2027
• Study Completion (Estimated): January 15, 2027

Study Registration Dates

• First Submitted: December 19, 2025
• First Submitted That Met QC Criteria: January 16, 2026
• First Posted (Actual): January 26, 2026

Study Record Updates

• Last Update Posted (Actual): February 27, 2026
• Last Update Submitted That Met QC Criteria: February 24, 2026
• Last Verified: February 1, 2026

More Information

Terms related to this study

• Keywords: colorectal cancer; liver metastasis; liver metastases; tumor microenvironment; CEUS; Contrast enhanced ultrasound; Sonoporation
• Additional Relevant MeSH Terms: Neoplasms by Site; Neoplasms; Intestinal Diseases; Gastrointestinal Neoplasms; Digestive System Neoplasms; Digestive System Diseases; Gastrointestinal Diseases; Intestinal Neoplasms; Rectal Diseases; Colonic Diseases; Colorectal Neoplasms
• Other Study ID Numbers: P-2025-19453

Plan for Individual participant data (IPD)

• Plan to Share Individual Participant Data (IPD)?: NO
• IPD Plan Description: It is not yet decided.

Drug and device information, study documents

• Studies a U.S. FDA-regulated drug product: No
• Studies a U.S. FDA-regulated device product: No