The surprising discovery that blood and other body fluids harbor abundant and stable extracellular microRNAs (ex-miRNAs) set off a boom of studies to exploit them for diagnostic purposes and to understand their function in health and disease. To realize the potential of this research opportunity, studies are urgently needed to fill gaps in our knowledge regarding the source cells that secrete ex-miRNAs, the forms in which they are stabilized in body fluids, the regulation and specificity of their secretion, and the functional effects of ex-miRNAs in both source and target cells. In this project, we will test the central hypothesis that immune cells release ex-mi RNAs in response to inflammatory stimuli, and that this process is critical for their immune function. Published findings suggest that circulating immune cells may be the biggest source of ex-miRNAs in blood.
In Specific Aim 1, we will use mouse genetic models of immunodeficiency and protocols for immune cell depletion together with a high throughput qRT-PCR platform for ex-mlRNA profiling to define the immune cell contribution and extracellular form of blood and airway lining fluid ex-miRNAs. These studies will be extended to mouse models of asthma in Aim 2 to determine how inflammation regulates ex-mlRNA release from immune cells.
The second aim will also encompass in vitro experiments that will define the form and molecular pathway of T cell ex-miRNA and Ago2 secretion in response to antigen-driven activation. And in Aim 3 we will examine the functional consequences of this induced ex-miRNA release for the source T cells. Specifically, we will determine whether ex-miRNA secretion is involved in the global down regulation of cellular miRNAs and Ago proteins and rapid resetting ofthe miRNA repertoire that supports activated T cell proliferation and differentiation into immune effector cells.
The Study of extracellular miRNAs (ex-miRNAs) presents tremendous opportunities for the development of noninvasive diagnostics for the detection of cancer, inflammation, infection, and more. In this proposal, we focus our efforts on uncovering the cellular sources and biochemical form of ex-miRNAs, and how inflammation and ex-miRNA secretion regulate one another.
|Li, Kang; Wong, David K; Hong, King Yeung et al. (2018) Cushioned-Density Gradient Ultracentrifugation (C-DGUC): A Refined and High Performance Method for the Isolation, Characterization, and Use of Exosomes. Methods Mol Biol 1740:69-83|
|Raffai, Robert L (2018) MicroRNA-146a & hematopoiesis: friend or foe in atherosclerosis. Noncoding RNA Investig 2:|
|Roy, Sanchari; Hooiveld, Guido J; Seehawer, Marco et al. (2018) microRNA 193a-5p Regulates Levels of Nucleolar- and Spindle-Associated Protein 1 to Suppress Hepatocarcinogenesis. Gastroenterology 155:1951-1966.e26|
|Chen, Baohui; Zou, Wei; Xu, Haiyue et al. (2018) Efficient labeling and imaging of protein-coding genes in living cells using CRISPR-Tag. Nat Commun 9:5065|
|Li, Kang; Wong, David K; Luk, Fu Sang et al. (2018) Isolation of Plasma Lipoproteins as a Source of Extracellular RNA. Methods Mol Biol 1740:139-153|
|Ong, Wei Qiang; Citron, Y Rose; Sekine, Sayaka et al. (2017) Live Cell Imaging of Endogenous mRNA Using RNA-Based Fluorescence ""Turn-On"" Probe. ACS Chem Biol 12:200-205|
|Anderton, Brittany; Camarda, Roman; Balakrishnan, Sanjeev et al. (2017) MYC-driven inhibition of the glutamate-cysteine ligase promotes glutathione depletion in liver cancer. EMBO Rep 18:569-585|
|Singh, Priti B; Pua, Heather H; Happ, Hannah C et al. (2017) MicroRNA regulation of type 2 innate lymphoid cell homeostasis and function in allergic inflammation. J Exp Med 214:3627-3643|
|Montoya, Misty M; Ansel, K Mark (2017) Small RNA Transfection in Primary Human Th17 Cells by Next Generation Electroporation. J Vis Exp :|
|Montoya, Misty M; Maul, Julia; Singh, Priti B et al. (2017) A Distinct Inhibitory Function for miR-18a in Th17 Cell Differentiation. J Immunol 199:559-569|
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