Role of immune-checkpoint LAG3 as a biomarker finding tool in patient-derived organoid cultures of breast cancer

Sep 21, 2026

Uploaded by Theodora

Role of immune-checkpoint LAG3 as a biomarker finding tool in patient-derived organoid cultures of breast cancer explores LAG3’s immunoregulatory function and evaluates its potential as a biomarker for selecting patients for α-LAG3 therapy. 3D breast cancer organoid models are used to assess LAG3 dysregulation, demonstrating markedly higher expression in tumor tissue versus non-cancer controls. LAG3 blockade suppresses organoid proliferation and remodels the immune micro-environment by reducing PD-L1, TIM-3, CTLA-4 and increasing pro-inflammatory cytokines and EMT markers, promoting a more permissive anti-tumor immune response.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages10

Reading Time25 min

AccessFree to download

Introduction

Immune checkpoints and LAG3 background

Methods

Patients’ selection

Results

LAG3 expression and therapeutic impact

[www. nature.com/scientificreports](www. nature.com/scientificreports) OPEN Role of immune-checkpoint LAG3 as a biomarker finding tool inpatient-derived organoid cultures of breast cancer Barbara Carrese1, Luigi Coppola1, Giovanni Smaldone1, Massimiliano D’Aiuto2, Gennaro Mossetti3, Antonio Febbraro4, Andrea Soricelli1, Marco Salvatore1 & Vincenza Ciaramella1􀀍 LAG3 plays a regulatory role in immunity and emerged as an inhibitory immune checkpoint molecule comparable to PD-L1 and CTLA-4 and a potential target for enhancing anti-cancer immune responses. We generated 3D cancer cultures as a model to identify novel molecular biomarkers for the selection of patients suitable for α-LAG3 treatment and simultaneously the possibility to perform an early diagnosis due to its higher presence in breast cancer, also to achieve a theragnostic approach. Our data confirm the extreme dysregulation of LAG3 in breast cancer with significantly higher expression in tumor tissue specimens, compared to non-cancerous tissue controls. LAG3 blockade inhibited proliferation of in vitro and ex vivo 3D human organoids and immune micro-environment through both a decrease of PD-L1, TIM-3 and CTLA4 expression and an increased production of several proinflammatory cytokines (IFNγ, IL-12, IL-6, IL-1β, TNFα) and EMT markers. These effects trigger a more permissive anti-tumor immune reaction, recruiting immune cells to the tumor sites, boosting the antitumor response. LAG3 acts as an immunosuppressive molecule in breast cancer, inhibiting T CD8 +cell proliferation and cytokine production by T cells. We proposed the modulation of a novel checkpoint molecule, such as LAG3, as potential biomarkers associated to a rapid diagnosis. Keywords Breast cancer, Organoids, LAG3, Biomarkers, Immune micro-environment Abbreviations CTL Cytotoxic CD8 T cells EMT Epithelial to mesenchymal transition LAG3 Lymphocyte activation gene-3 CLL Lymphocytic leukemia NK Natural killer PDOs Patient’s derived organoids Treg Regulatory T Th T helper 3D Three-dimensional TME Tumor microenvironment CTRL Untreated control Tumors are a major public health concern and, currently, immunotherapy is the most promising tumor treatment. Indeed, the immune system has an essential role in the surveillance against tumors, but cancer cells develop several strategies to escape immune surveillance1. Escaping the surveillance of the immune system, through which proliferation and growth of cancer cells can be successfully achieved, is one of the main mechanisms of development and progression of a tumor2. In this context, the immune checkpoints are crucial molecules expressed on the surface of cancer cells or immune cells, particularly on T cells. These molecules can negatively or positively regulate immune responses, as well as mitigate immune injury and prevent the development of 1IRCCS SYNLAB SDN, Naples 80143, Italy. 2Clinica Villa Fiorita, Aversa 81031, Italy. 3Pathological Anatomy Service, Maria Rosaria Clinic, Pompei, Naples 80045, Italy. 4Oncology Unit, Casa di Cura Cobellis, Vallo della Lucania, Italy. 􀀍 email: [vincenza.ciaramella@synlab.it](vincenza.ciaramella@synlab.it) [www. nature.com/scientificreports/](www. nature.com/scientificreports/) autoimmunity to minimize the tissue damage. Several evidence showed that inhibitory or stimulatory immune checkpoint molecules are expressed on a sizeable fraction of tumor types. Furthermore, immune checkpoint molecules regulate not only the epithelial-mesenchymal transition (EMT) and consequently the formation of metastases, but also other processes such as drug resistance, anti-apoptosis, angiogenesis, or increased energy metabolism3. Back in 1990, Dr. Frédéric Triebel and colleagues identified Lymphocyte activation gene-3 (LAG3) as a novel lymphocyte activation gene closely related to CD4, resulting in approximately 26% amino acid homology. The authors found a 2-kb mRNA to be selectively transcribed in activated human T

FAQs

Why is LAG3 considered relevant in breast cancer immunotherapy?

LAG3 functions as an inhibitory immune checkpoint molecule and is proposed as a target to enhance anti-cancer immune responses. Prior work also links dysregulated LAG3 to clinical outcomes across multiple cancer types.

How do the study’s 3D organoid models support biomarker discovery?

The work generates patient-derived 3D cancer cultures to identify molecular biomarkers for patient selection for α-LAG3 treatment. It also evaluates whether LAG3 shows higher presence in breast cancer for possible early diagnosis.

What changes occur after LAG3 blockade in organoid experiments?

LAG3 blockade inhibits proliferation in vitro and ex vivo and affects the immune micro-environment. It decreases PD-L1, TIM-3, and CTLA4 expression while increasing pro-inflammatory cytokines and EMT markers, which supports a stronger anti-tumor immune reaction.

Sep 21, 2026

Uploaded by Theodora