ABSTRACT: To investigate differential protein and phospho-protein changes in the signaling cascades related to mutant G2019S LRRK2 using peripheral blood mononuclear cells (PBMCs)
Project description:Supported by the Michael J Fox Foundation we established a biorepository of blood cells from G2019S LRRK2-PD patients recruited at the Hospital Clínic de Barcelona (Barcelona). Using this cohort, we performed a phospho-proteomic pilot study by mass spectrometry and identified a differential combination of phosphorylated proteins associated with the G2019S mutation. Here, we aim to validate and expand these findings using additional G2019S and R1441G cohorts (PMID: 35049090 By state-of-the-art DIA phospho-proteomics we aim to validate and expand our preliminary findings in additional G2019S and R1441G LRRK2 cohorts of similar design, size, and blood cell collection methods collected at additional centers in Spain (Hospital de Valdecilla in Santander, Hospital de Donostia in San Sebastian). We expect to identify differential protein phosphorylation changes in LRRK2-PD patients affected by the G2019S and R1441G mutations that could be useful as LRRK2 pharmacodynamic biomarkers. In asymptomatic LRRK2 mutation carriers, we will explore the presence of these phosphorylation changes and evaluate their applicability as early disease biomarkers.
Project description:Supported by the Michael J Fox Foundation we established a biorepository of blood cells from G2019S LRRK2-PD patients recruited at the Hospital Clínic de Barcelona (Barcelona). Using this cohort, we performed a phospho-proteomic pilot study by mass spectrometry and identified a differential combination of phosphorylated proteins associated with the G2019S mutation. Here, we aim to validate and expand these findings using additional G2019S and R1441G cohorts (PMID: 35049090 By state-of-the-art DIA phospho-proteomics we aim to validate and expand our preliminary findings in additional G2019S and R1441G LRRK2 cohorts of similar design, size, and blood cell collection methods collected at additional centers in Spain (Hospital de Valdecilla in Santander, Hospital de Donostia in San Sebastian). We expect to identify differential protein phosphorylation changes in LRRK2-PD patients affected by the G2019S and R1441G mutations that could be useful as LRRK2 pharmacodynamic biomarkers. In asymptomatic LRRK2 mutation carriers, we will explore the presence of these phosphorylation changes and evaluate their applicability as early disease biomarkers.
Project description:Genome-Scale draft model for Human Peripheral Blood Mononuclear Cells (PBMCs). A GEM for PBMCs was developed by applying the INIT
algorithm on Human Metabolic Reconstruction (HMR 2.0) as a template model. GEMs were contextualised/ constrained for different conditions using expression datasets. The gene/transcript expression data obtained from PBMCs of Type 1 Diabetes progressors, non-progressors, and healthy controls were employed to score each reaction of HMR 2.0. For further detail please refer to Electronic Supplementary Information of Sen et.al, Metabolic alterations in immune cells associate with progression to type 1 diabetes, Diabetologia, 15/01/2020, (https://doi.org/10.1007/s00125-020-05107-6).
Project description:Differential gene expression profiling in peripheral blood mononuclear cells (PBMCs) was performed using Human Transcriptome Array 2 (HTA2)
Project description:We performed a systematic analysis of differential gene expression in a wide set of peripheral human immune cells of MPO-deficient patients using single cell RNA-sequencing (scRNAseq) of peripheral blood mononuclear cells (PBMCs) in stable disease state.
Project description:Gain-of kinase function variants in LRRK2 (leucine-rich repeat kinase 2) cause Parkinson’s disease (PD), albeit with incomplete and age-dependent penetrance, offering the prospect of disease-modifying treatment strategies via LRRK2 kinase inhibition. LRRK2 phosphorylates a subgroup of RabGTPases including Rab10 and pathogenic mutations enhance LRRK2-mediated phosphorylation of Rab10 at Thr73. In this study we analyse LRRK2 dependent Rab10Thr73 phosphorylation in human peripheral blood neutrophils isolated from 101 individuals using quantitative immunoblotting and mass spectrometry. Our cohort includes 42 LRRK2 mutation carriers (21 with the G2019S mutation that resides in the kinase domain and 21 with the R1441G mutation that lies within the ROC-COR domain), 27 patients with idiopathic PD, and 32 controls. We show that LRRK2 dependent Rab10 Thr73 phosphorylation is significantly elevated in all R1441G LRRKR2 mutation carriers irrespective of disease status. PD manifesting and non-manifesting G2019S mutation carriers as well as idiopathic PD samples did not display elevated Rab10 Thr73 phosphorylation. Furthermore, we analysed brain samples of 10 G2019S and 1 R1441H mutation carriers as well as 10 individuals with idiopathic PD and 10 controls. We find high variability for pRab10Thr73 phosphorylation amongst donors irrespective of genetic and disease state. We conclude that in vivo LRRK2 dependent pRab10Thr73 analysis in human peripheral blood neutrophils is a specific and robust biomarker for LRRK2 kinase activation for individuals with mutations such as R1441G that enhance pRab10Thr73 phosphorylation over 2-fold. We provide the first evidence that the LRRK2 R1441G mutation enhances LRRK2 kinase activity in a primary human cell.
Project description:The SCOT (Scleroderma: Cyclophosphamide or Transplantation) trial demonstrated clinical benefit of autologous hematopoietic stem cell transplant (HSCT) compared to cyclophosphamide (CYC) in scleroderma. We analyzed gene expression from peripheral blood mononuclear cells (PBMCs) of SCOT participants to identify those with differential treatment response.
Project description:Omics technologies and biomolecules included genomics (DNA), whole transcriptomics (miRNAs, lncRNAs, circRNAs and mRNAs), proteomics (proteins) were used for collecting high throughput data from peripheral blood mononuclear cells (PBMCs) of subjects in our study. The joint analysis method of multi-omics was taken advantage of in our studies for uncovering some SLE disease-related molecular regulatory networks. For WGBS study, PBMCs of blood samples from the people recruited by our study were collected and used for whole-genome bisulphite sequencing (WGBS). There were 10322 genes presenting differential methylation levels in PBMCs of SLE patients (6305 hypermethylation genes and 4017 hypomethylation genes).
Project description:To investigate the differential expression profile of genes and microRNA (miRNA) in peripheral blood mononuclear cells (PBMCs) from patients infected with Human T-lymphotropic Virus 1 (HTLV-1), as well as their impact on progression to Tropical Spastic Paraparesis /HTLV Associated Myelopathy (PET/HAM)