THE LAMOND LAB
Proteomics, Splicing, Insight
News
. Screening for variable drug responses using human iPSC cohorts has just been published in PLOS one demonstrating that a cohort of hiPSC lines from different donors can be screened efficiently in their pluripotent state, using high-throughput Cell Painting assays. Variable phenotypic responses between hiPSC lines were detected with a wide range of clinically approved drugs, in use across multiple disease areas
Systematic characterization of site-specific proline hydroxylation using hydrophilic interaction chromatography and mass spectrometry is in Reviewed Preprint on eLife describing a robust and novel method of identifying Proline Hydroxylation, an important post translational modification crucial in hypoxia research. It involves a robust workflow for the identification of proline hydroxylation sites in proteins, using a combination of hydrophilic interaction chromatography (HILIC) enrichment and high-resolution nano-Liquid Chromatography-Mass Spectrometry (LC-MS) together with refining and filtering parameters during data analysis
RESEARCH
Our Current Focus
Proteomics
We use mass spectrometry to perform quantitative proteomics analyses in our studies on cell biology, RNA processing and on mechanisms of disease and gene regulation. At the University of Dundee, School of Life Sciences, we have access to one of the largest MS facilities in Europe. We use MS methods to study cell phenotypes, including identifying protein expression levels and post-translational modifications (PTMs), identifying protein targets of small molecule drugs using Thermal Proteome Profiling, mapping protein and peptide QTLs and measuring protein turnover in cells and tissues.
RNA Processing & Alternative Splicing
We are studying mechanisms involved in the control of RNA processing in the nuclei of mammalian cells, including the dynamic organisation of nuclear bodies containing proteins and RNPs involved in the processing of pre-mRNA and pre-rRNA. A major aim of our work is characterising the mechanisms of action and protein targets of novel small molecule modulators we have identified that specifically alter either splice site selection (i.e. causing changes in alternative pre-mRNA splicing patterns), and/or change the morphology and composition of specific nuclear bodies. We are also studying how modulation of alternative splicing can contribute to mechanisms of human disease, with a focus on studying Huntington’s Disease and other inherited neurodegenerative disorders.
Human Genetics & iPS Cells
We are studying human phenotypic diversity with a major focus on the impact of how genetics affects protein expression in human populations and how this in turn affects cellular phenotypes and biological responses. These studies include mapping quantitative trait loci by comparative measurements of levels of protein expression across different donors, both using libraries of iPS cells and in human tissues. Our studies include detection of alternatively spliced protein isoforms and post-translational modifications. We aim to detect how genetic effects on protein expression can influence variable sensitivity to disease and variable responses by patients to different drugs and therapeutic treatments.
