Pandor, S. | bioRxiv Science

bioRxiv2025

An automated metabolite extraction workflow for global metabolomics analysis using the Agilent Bravo liquid handling platform

Metabolomics is the comprehensive study of small molecules that provides a snapshot of an organisms physiological state. Reflecting phenotype more closely than genes or proteins, metabolites reveal changes linked to diseases, mutations, genetic interventions, and environmental stimuli. Recent technological advancements in metabolomics analysis through the application of ion mobility mass spectrometry have enhanced the comprehensive analysis of complex metabolic mixtures. However, pre-analytical bottlenecks in throughput and consistent extraction persist. We developed an automated, sample-agnostic metabolite extraction workflow for diverse liquid samples using an Agilent Bravo liquid handling platform. Here, we provide device, protocol, and form files for efficient sample processing to extract metabolites for global metabolomics analysis using liquid chromatography - mass spectrometry techniques.

biochemistry↗

bioRxiv2024

Mitochondrial metabolism is a key determinant of chemotherapy sensitivity in Colorectal Cancer

Therapy resistance is attributed to over 80% of cancer deaths per year emphasizing the urgent need to overcome this challenge for improved patient outcomes. Despite its widespread use in colorectal cancer (CRC) treatment, resistance to 5-fluorouracil (5FU) remains poorly understood. Here, we investigate the transcriptional responses of CRC cells to 5FU treatment, revealing significant metabolic reprogramming towards heightened mitochondrial activity. Utilizing CRC models, we demonstrate sustained enhancement of mitochondrial biogenesis and function following 5FU treatment, leading to resistance in both in vitro and in vivo settings. Furthermore, we show that targeting mitochondrial metabolism, specifically by inhibiting Complex I (CI), sensitizes CRC cells to 5FU, resulting in delayed tumour growth and prolonged survival in preclinical models. Additionally, our analysis of patient data suggests that oxidative metabolism signatures may predict responses to 5FU-based chemotherapy. These findings shed light on mechanisms underlying 5FU resistance and propose a rational strategy for combination therapy in CRC, emphasizing the potential clinical benefit of targeting mitochondrial metabolism to overcome resistance and enhance patient outcomes.

cancer biology↗