Abstract 2224: Impact of NT/NTR1 signaling on metabolic pathways and tumor-associated microbiomes in colorectal cancer

Background: Colorectal cancer (CRC) is a leading cause of cancer-related deaths worldwide with substantial heterogeneity in patient outcomes. Neurotensin (NT) and its high-affinity receptor (NTR1) have been implicated in CRC progression, particularly in modulating tumor metabolism and immune responses. NT/NTR1 expression has been linked to CRC progression and outcomes, while gut microbiota is increasingly recognized for its role in cancer development and therapy. The purpose of current study was to explore the relationship between NT/NTR1 expression, microbial communities, and transcriptomic changes in tumor versus adjacent normal tissues, focusing on their role in shaping tumor-associated microbiomes and influencing cancer progression. Methods: We performed: (i) RNA sequencing of tumor samples from 96 Appalachian Kentucky CRC patients to assess NT/NTR1 expression and its association with transcriptomic pathways; (ii) paired RNA sequencing and 16S rRNA-based microbiome profiling from tumor and adjacent normal tissues of 15 patients to evaluate transcriptomic and microbial differences. Functional gene prediction was performed using Picrust2 based on 16S rRNA profiles, and differential expression analysis was conducted using DESeq2. NT/NTR1 expression and bacterial abundance were validated by qPCR. Results: (i) RNA sequencing of the 96-patient cohort showed that high NT/NTR1 expression was associated with dysregulation of multiple metabolic pathways (e.g., lipid, carbohydrate, and amino acid metabolism) and immune-related pathways (e.g., "immune disease" and "infection disease: bacteria"). (ii) In paired tumor and adjacent normal tissues, NT/NTR1 expression was significantly elevated in tumor tissues. Microbial profiling revealed higher microbial abundance and diversity in tumor tissues, with notable enrichment of Campylobacter and Chlamydia species. Functional gene prediction using Picrust2 identified significant upregulation of flagellar assembly-related genes, such as K13626, in tumor microbiomes compared to normal tissues (P = 3.66 × 10-3). These findings suggest enhanced microbial motility and interaction in the tumor microenvironment facilitated by NT/NTR1-driven metabolic changes. Conclusions: NT/NTR1 signaling remodels metabolic pathways in CRCs, enhances nutrient absorption and creates a more favorable environment for microbial colonization. The observed enrichment of motility-related bacterial species and functional genes highlights the role of the NT/NTR1 signaling pathway in promoting tumor-microbe interactions. Importantly, these findings provide potential avenues for CRC diagnosis and therapeutic strategies targeting the NT/NTR1 axis. Citation Format: Haoming Wu, Yang Wang, Moumita Banerjee, Jinpeng Liu, Chi Wang, Dahe He, Thèrése Bocklage, Jing Li, B.Mark Evers. Impact of NT/NTR1 signaling on metabolic pathways and tumor-associated microbiomes in colorectal cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 2224.