Cheuk Hong Leung

The Supplementary File contains figures S1 to S15 and Table S1, which contain supporting data for the experiments reported in the main manuscript.

Purpose:The anti–cytotoxic T-lymphocyte antigen 4 (CTLA-4) monoclonal antibody, ipilimumab (IPI), has shown clinical benefit across multiple tumor types, both as monotherapy and in combination with nivolumab or chemotherapy. However, not all tumors respond, and peripheral effects can lead to immune-related adverse events. We characterized three novel anti–CTLA-4 antibodies: peptide-masked [PROBODY conditionally activatable therapeutic (PB)], non-fucosylated (NF), and combined NF-PB (BMS-986288). Experimental Design:We evaluated the preclinical characteristics, including pharmacodynamics, tolerability, antitumor activity and efficacy, and peripheral immune responses, of these novel anti–CTLA-4 antibodies using in vitro systems, animal models, and human data. This includes data from preclinical mouse models of colorectal cancer as well as non–small cell lung cancer. Results:NF demonstrated greater T-cell priming and antitumor activity than both IPI and the unmasked PB antibody in cell-based assays and mouse models. Whereas the intact PB antibody showed minimal CTLA-4 binding and peripheral immune activation, unmasking restored its functional activity to levels comparable with those of IPI. Unmasked anti–CTLA-4 NF-PB retained the effectiveness of anti–CTLA-4 NF, and both molecules demonstrated more profound antitumor activity, increased effector memory T-cell response, and prolonged survival in mouse models compared with IPI. Anti–CTLA-4 NF-PB demonstrated reduced peripheral immune responses compared with anti–CTLA-4 NF or IPI in non-human primates and patients with solid tumors. Conclusions:Anti–CTLA-4 NF-PB has enhanced antitumor activity, efficacy, and reduced peripheral activity in preclinical models, and has the potential to provide therapeutic benefit in solid tumors.See related commentary by Galvez-Cancino et al., p. 3112

Supplementary figure S1. Effects of fecal microbiota transfer from Lcn2-deficient mice in various mouse models.

Differentially expressed genes in the myeloid cluster of the colonic lamina propria.

Differentially expressed genes in the tumor microenvironment.

Supplementary figure S2. scRNA-seq of the colonic lamina propria in the syngeneic mice following FMT.

Table S1 from Inflammation Mediated by Gut Microbiome Alterations Promotes Lung Cancer Development and an Immunosuppressed Tumor Microenvironment

Comparative analysis of microbial abundance in murine fecal samples across FMT groups.

Supplementary Figure S4. Immune changes within the TME induced by inflammation related to gut microbiome alterations.

Supplementary figure S5. Different species belonging to the genus Alistipes promote the development of LUAD.