2017 — 2018 |
Berry, James Dale Butovsky, Oleg |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Identification of Microrna and Proteomics in Immune Cells and Plasma in Als @ Brigham and Women's Hospital
Project Summary The exact pathogenetic process that triggers neurodegeneration in ALS is currently unknown, but it is likely to be multifactorial, as for other neurodegenerative diseases. Recently, growing attention has been focused on events related to innate and adaptive immune responses, and there is increasing evidence of a significant involvement of inflammatory and immune pathways in the disease. However, their exact cellular identity, molecular and functional phenotypes, and their protective or detrimental roles in ALS progression are poorly understood. We previously found that both peripheral Ly6CHi monocytes in SOD1 mice and their analogous CD14+/CD16? monocytes from ALS patients express proinflammatory profiles. These studies lead us to begin a series of investigations to define markers of disease onset and progression in ALS. We started a prospective, longitudinal study collecting detailed clinical information, peripheral blood mononuclear cells (PBMCs) and plasma from people with ALS. Two cohorts of ALS subjects, sporadic (sALS) and familial (fALS), are followed up in time, from early until the late stages of the disease. The proposed project will identify the micro-RNA and proteomic signatures in blood-derived inflammatory monocytes and plasma in ALS. In Aim 1, we will investigate miRNA and proteomic changes in blood CD14+/CD16? monocytes and plasma from the onset and during disease progression of sALS and fALS (with SOD1 or C9orf72 gene mutations). The Aim design allows a comparative analysis of miRNA and protein expression in multiple dimensions utilizing 1) small RNA-seq to compare temporal changes of miRNAs expression within the ALS and healthy controls; 2) correlated clinical information in the variability over the time course; and 3) Integrated network data analysis between miRNA and proteomics. We leverage our unique longitudinal collection of sALS and fALS for which we have multipoint blood draws during their disease progression In Aim 2, we will characterize and validate the main hits in an independent validation cohort of ALS subjects. Acknowledging that the biomarker discovery in any field suffers from lack of replication, we believe that replication of our findings in an independent cohort is of highest importance. The independent replication, along with the validation of the main hits using different technologies will identify only robust potential biomarkers to further test for clinical utility.
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0.906 |
2018 — 2019 |
Berry, James Dale Butovsky, Oleg Ikezu, Tsuneya |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Identification of Exosome Signature in Serum From Als Patients @ Brigham and Women's Hospital
PROJECT SUMMARY The main objective of this application is to explore the biological potential of serum exosomal miRNA and protein as biomarkers of amyotrophic lateral sclerosis (ALS). It is critical to develop a reliable biomarker for disease progression that would enable us to monitor the effect of treatment and improve the success rate of drug development.The incomplete knowledge of ALS pathophysiology and the absence of reliable biomarkers are major gaps in our understanding of the disease. We previously identified a unique microRNA and immune gene expression signature in blood monocytes from ALS mice and ALS patients. microRNAs are uniquely enriched in exosomes, which are extracellular vesicles and have been isolated from biological fluids such as plasma and serum. Exosomal microRNA can provide insight into the state of the cell in which the exosomes are released. Exosomes not only carry miRNA but also pathogenic molecules of ALS, such as TDP-43 and SOD1. Thus, altered miRNA and protein in exosomes may result in the dysregulation of biological pathways that contribute to the development and/or progression of ALS. There is so far no systematic investigation of blood exosome miRNA and protein profiles before and after the onset of ALS. We hypothesize that abnormalities in miRNA and proteomic signatures in serum exosomes in ALS may serve as novel biomarkers for the prognosis and diagnosis of ALS. This study is supported by our highly unique longitudinal studies of subjects with sporadic (sALS) and familial ALS (fALS) and controls. In these studies, we collected serum samples over a four-year period at regular time points, while tracking disease progression clinically. Here, we propose a comprehensive profiling of miRNA and the proteins in serum exosomes. In Aim 1, we will identify serum exosome-based RNA and proteomic signature in sALS and fALS progression. In Aim 2, we will replicate and validate the identified biomarkers in another cohort of sALS and fALS. We believe that our findings, in combination with the clinical information collected at each time-point, will allow us to define an ALS miRNA and proteomic signature in serum exosomes and serve as a potential biomarker of ALS progression. !
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0.906 |