T cell receptor (TCR) ligation induces intracellular signaling cascades including the Ca2+-NFAT (nuclear factor of activated T cells) and MAPK (mitogen-activated protein kinase) pathways that play crucial roles in activation and differentiation of T cells. Defects in these pathways lead to immune deficiency or hypersensitive immune responses in humans and mice. The small GTPases, Ras, Rac and Rho are known to play a central role in activation of TCR signaling pathways and are therapeutic targets in autoimmune diseases. These small GTPases are recruited into the immunological synapse (IS), the interface between an antigen-presenting cell and a T cell via interactions with signaling adaptors and activate the downstream MAPK pathway. However, in contrast to Ras, Rac and Rho GTPases, little is known about the role of more than 60 members of the Rab GTPase family in T cell activation. Our long-term goal is to uncover the role of Rab GTPases in TCR signaling to identify novel therapeutic targets to alleviate autoimmunity. We have recently identified a novel Rab GTPase, CRACR2A GTPase that plays a key role in TCR signaling. Our preliminary data show that CRACR2A GTPase is a lymphocyte-specific large Rab GTPase with many functional domains in contrast to small GTPases. We found that CRACR2A GTPase translocates into the IS to activate the downstream JNK (c- Jun N-terminal kinase) MAPK pathway. Furthermore, it is highly expressed in TH1 cells and plays an important role in expression of IFN-? and T-bet, and thus in autoimmunity. Our central hypothesis is that CRACR2A GTPase is recruited into the IS via protein interactions and regulates the JNK pathway to drive TH1 differentiation. The objective of this proposal is to identify 1) interacting partners, 2) GTP binding and prenylation-mediated regulation, and 3) the physiological role of CRACR2A GTPase in T cells using high resolution imaging techniques, genomics and conditionally targeted knockout mice. CRACR2A GTPase is the first example of a large molecule containing the small Rab GTPase domain. We expect to reveal a hitherto unknown mechanism of regulation of the JNK pathway in T cell differentiation. We are confident that we will also identify a unique signaling function of Rab GTPases, which are otherwise known to mediate only intracellular protein trafficking. These mechanistic studies will have significant impact on therapeutic exploitation of this novel GTPase to treat autoimmune diseases.

Public Health Relevance

The goal of this proposal is to understand the fundamental questions on immunology, the mechanism of how T cells become activated and once activated, how this stimulation is terminated on time to avoid deleterious consequences. We will address the urgent need for inflammatory disease patients by taking advantage of the discovery of a novel signaling mechanism that plays a crucial for activation of type-1 helper T cells. The proposed studies are novel and innovative, and they will have a broad impact on basic understanding of the onset mechanism of T cell-mediated inflammation, and on therapeutic exploitation of this novel signaling molecule to treat autoimmune diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
2R01AI083432-06A1
Application #
9036856
Study Section
Cellular and Molecular Immunology - A Study Section (CMIA)
Program Officer
Mallia, Conrad M
Project Start
2009-07-01
Project End
2020-10-31
Budget Start
2015-11-10
Budget End
2016-10-31
Support Year
6
Fiscal Year
2016
Total Cost
$385,000
Indirect Cost
$135,000
Name
University of California Los Angeles
Department
Physiology
Type
Schools of Medicine
DUNS #
092530369
City
Los Angeles
State
CA
Country
United States
Zip Code
90095
Srikanth, Sonal; Kim, Kyun-Do; Gao, Yuanyuan et al. (2016) A large Rab GTPase encoded by CRACR2A is a component of subsynaptic vesicles that transmit T cell activation signals. Sci Signal 9:ra31
Srikanth, Sonal; Woo, Jin Seok; Gwack, Yousang (2016) A large Rab GTPase family in a small GTPase world. Small GTPases :1-6
Woo, Jin Seok; Srikanth, Sonal; Nishi, Miyuki et al. (2016) Junctophilin-4, a component of the endoplasmic reticulum-plasma membrane junctions, regulates Ca2+ dynamics in T cells. Proc Natl Acad Sci U S A 113:2762-7
Nurbaeva, Meerim K; Eckstein, Miriam; Concepcion, Axel R et al. (2015) Dental enamel cells express functional SOCE channels. Sci Rep 5:15803
Sohn, S; Park, Y; Srikanth, S et al. (2015) The Role of ORAI1 in the Odontogenic Differentiation of Human Dental Pulp Stem Cells. J Dent Res 94:1560-7
Bertin, Samuel; Aoki-Nonaka, Yukari; de Jong, Petrus Rudolf et al. (2014) The ion channel TRPV1 regulates the activation and proinflammatory properties of CD4⁺ T cells. Nat Immunol 15:1055-63
Kim, Kyun-Do; Srikanth, Sonal; Tan, Yossan-Var et al. (2014) Calcium signaling via Orai1 is essential for induction of the nuclear orphan receptor pathway to drive Th17 differentiation. J Immunol 192:110-22
Srikanth, Sonal; Kim, Kyun-Do; Gwack, Yousang (2014) Methods to measure cytoplasmic and mitochondrial Ca(2+) concentration using Ca(2+)-sensitive dyes. Methods Enzymol 543:1-20
Groenendyk, Jody; Peng, Zhenling; Dudek, Elzbieta et al. (2014) Interplay between the oxidoreductase PDIA6 and microRNA-322 controls the response to disrupted endoplasmic reticulum calcium homeostasis. Sci Signal 7:ra54
Srikanth, Sonal; Gwack, Yousang (2013) Orai1-NFAT signalling pathway triggered by T cell receptor stimulation. Mol Cells 35:182-94

Showing the most recent 10 out of 20 publications