Evaluation of Low-Dose Peginterferon Alfa-2a on Perioperative Immune Modulation in Patients with Colon Cancer

The primary objective of this study is to evaluate the **effect** of low-dose **Interferon-alfa2a** on perioperative immune suppression in patients with colon cancer. Specifically, the study aims to assess differences in measures of lymphocytic subpopulations between the intervention and placebo groups, with a focus on a higher number of CD3, CD4, CD8, and HLA-positive cells in the intervention group on the day of and the day after surgery. Additionally, the study seeks to determine differences in tumor-infiltrating lymphocytes in the resected specimen at the tumor center and invasive margin between the two groups, analyzed via immunohistochemistry for CD3+, CD4+, and CD8+ T-cells. These objectives are clinically relevant as they may provide insights into the modulation of immune responses during the perioperative period, potentially impacting patient outcomes.

The secondary objectives include:

• Assessing differences in Quality of Recovery (QoR-15) scores between the intervention and placebo groups, with a higher mean QoR expected in the intervention group on the third and 12-16 post-operative day.
• Evaluating differences in key systemic immune responses in blood, analyzed via a multiplex gene assay, focusing on pathways related to antigen presentation and innate cytotoxic and inflammatory activity.
• Investigating differences in the tumor microenvironment in the resected specimen, analyzed via a multiplex gene assay.
• Measuring differences in circulating cell-free DNA (cfDNA) at various time points before and after surgery.
• Examining intragroup differences in the tumor microenvironment between biopsy before surgery and the resected specimen.
• Comparing standard blood sample measures, with a focus on lower inflammation-grade in the intervention group prior to surgery.
• Analyzing differences in cytokines and interleukins related to post-operative inflammation, with lower inflammatory cytokines expected in the intervention group.
• Exploring differences in phenotype and clonality of tumor-infiltrating and circulating T cells, analyzed via combined single-cell transcriptome and TCR sequencing.

The clinical trial involves participants diagnosed with **colon cancer**, specifically those with pMMR colonic adenocarcinoma who are scheduled for laparoscopic hemicolectomy. The study population includes both male and female subjects, aged 18 years and older, classified under ASA class I-III according to the American Society of Anesthesiology. The trial does not include a vulnerable population. Participants were selected based on their diagnosis and surgical schedule, with no specific lifestyle considerations such as diet or physical activity mentioned. The sponsor has not provided information regarding the total number of participants in the trial.

The clinical trial is designed to evaluate the effect of low-dose **peginterferon alfa-2a** on perioperative immune suppression in patients with **colon cancer**. This is a Phase 4, randomized, double-blind, controlled trial. The trial will involve two groups: an intervention group receiving peginterferon alfa-2a and a placebo group receiving **sodium chloride**. The trial is expected to commence on August 1, 2023, and conclude by December 31, 2026. The maximum treatment period for each participant is 14 days.

Participants will undergo a series of study visits, beginning with an inclusion (screening) visit to confirm eligibility based on criteria such as age (above 18 years), diagnosis of pMMR colonic adenocarcinoma, and scheduled laparoscopic hemicolectomy. Follow-up visits will be conducted to assess primary endpoints, including differences in lymphocytic subpopulations and tumor-infiltrating lymphocytes between the intervention and placebo groups. Secondary endpoints will evaluate quality of recovery, T-cell phenotype and clonality, and tumor microenvironment differences. The end-of-study visit will mark the completion of the trial for each participant.

Participant involvement is expected to last for the duration of the treatment period, with additional time allocated for follow-up assessments. Conditions that may lead to early termination from the study include adverse reactions to the investigational product or withdrawal of consent. The trial aims to provide valuable insights into the immunomodulatory effects of peginterferon alfa-2a in the perioperative setting for colon cancer patients.

The clinical trial involves the administration of two treatments. The first treatment is **Sodium Chloride Fresenius Kabi Italia 0.9% Solution for Infusion**. This is a **solution for infusion** containing the active substance **sodium chloride**, which is of chemical origin. The pharmaceutical form is a solution intended for subcutaneous injection. The maximum daily dose is 1 ml, with a total maximum dose of 2 ml over a treatment period of up to 14 days. This product is manufactured by Fresenius Kabi Italia S.R.L. and is used as a placebo in the trial.

The second treatment is **Pegasys 135 micrograms Solution for Injection in Pre-filled Syringe**. This is a **solution for injection** containing the active substance **peginterferon alfa-2a**, which is a protein of other origin. The pharmaceutical form is a solution intended for subcutaneous injection. The maximum daily dose is 45 micrograms, with a total maximum dose of 90 micrograms over a treatment period of up to 14 days. This product is manufactured by Pharmaand GmbH and serves as the test treatment in the trial.

Efficacy in this clinical trial will be assessed through both primary and secondary endpoints. The primary endpoints focus on the differences in measures of lymphocytic subpopulations between the intervention and placebo groups. This will be evaluated by observing a higher number of CD3, CD4, CD8, and HLA-positive cells in the intervention group on the day of and the day after surgery. Additionally, differences in **tumor-infiltrating lymphocytes** in the resected specimen at the tumor center and invasive margin will be analyzed using immunohistochemistry with staining for CD3+, CD4+, and CD8+ T-cells.

Secondary endpoints include differences in measures of Quality of Recovery (QoR-15) between the intervention and placebo groups, with a higher mean QoR expected in the intervention group on the third and 12-16 post-operative days. Furthermore, differences in the phenotype and clonality of tumor-infiltrating and circulating T cells will be analyzed via combined single-cell transcriptome and TCR sequencing. The trial will also assess differences in the tumor microenvironment in the resected specimen. These assessments will provide comprehensive insights into the efficacy of the intervention in modulating immune responses in patients undergoing surgery for colon cancer.