PMID- 35990647
OWN - NLM
STAT- MEDLINE
DCOM- 20220823
LR  - 20220825
IS  - 1664-3224 (Electronic)
IS  - 1664-3224 (Linking)
VI  - 13
DP  - 2022
TI  - Multifunctional regulation of VAMP3 in exocytic and endocytic pathways of RBL-2H3 
      cells.
PG  - 885868
LID - 10.3389/fimmu.2022.885868 [doi]
LID - 885868
AB  - Mast cells (MCs) are inflammatory cells involved in allergic reactions. 
      Crosslinking of the high-affinity receptor for IgE (FcϵRI) with multivalent 
      antigens (Ags) induces secretory responses to release various inflammatory 
      mediators. These responses are largely mediated by soluble 
      N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs). 
      Vesicle-associated membrane protein 3 (VAMP3) is a vesicular-SNARE that interacts 
      with targeted SNARE counterparts, driving the fusion of MC secretory granules 
      with the membrane and affecting subsequent assembly of the plasma membrane. 
      However, the role of VAMP3 in FcϵRI-mediated MC function remains unclear. In this 
      study, we comprehensively examined the role of VAMP3 and the molecular mechanisms 
      underlying VAMP3-mediated MC function upon FcϵRI activation. VAMP3 shRNA 
      transduction considerably decreased VAMP3 expression compared with non-target 
      shRNA-transduced (NT) cells. VAMP3 knockdown (KD) cells were sensitized with an 
      anti-DNP IgE antibody and subsequently stimulated with Ag. The VAMP3 KD cells 
      showed decreased degranulation response upon Ag stimulation. Next, we observed 
      intracellular granule formation using CD63-GFP fluorescence. The VAMP3 KD cells 
      were considerably impaired in their capacity to increase the size of granules 
      when compared to NT cells, suggesting that VAMP3 mediates granule fusion and 
      therefore promotes granule exocytosis in MCs. Analysis of FcϵRI-mediated 
      activation of signaling events (FcϵRI, Lyn, Syk, and intracellular Ca(2+) 
      response) revealed that signaling molecule activation was enhanced in VAMP3 KD 
      cells. We also found that FcϵRI expression on the cell surface decreased 
      considerably in VAMP3 KD cells, although the amount of total protein did not 
      vary. VAMP3 KD cells also showed dysregulation of plasma membrane homeostasis, 
      such as endocytosis and lipid raft formation. The difference in the plasma 
      membrane environment in VAMP3 KD cells might affect FcϵRI membrane dynamics and 
      the subsequent signalosome formation. Furthermore, IgE/Ag-mediated secretion of 
      TNF-alpha and IL-6 is oppositely regulated in the absence of VAMP3, which appears to 
      be attributed to both the activation of FcϵRI and defects in VAMP3-mediated 
      membrane fusion. Taken together, these results suggest that enhanced 
      FcϵRI-mediated signal transduction in VAMP3 KD cells occurs due to the disruption 
      of plasma membrane homeostasis. Hence, a multifunctional regulation of VAMP3 is 
      involved in complex secretory responses in MCs.
CI  - Copyright (c) 2022 Mishima, Sakamoto, Kioka, Nagata and Suzuki.
FAU - Mishima, Satomi
AU  - Mishima S
AD  - Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical and 
      Health Sciences, Kanazawa University, Kanazawa, Japan.
FAU - Sakamoto, Marin
AU  - Sakamoto M
AD  - Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical and 
      Health Sciences, Kanazawa University, Kanazawa, Japan.
FAU - Kioka, Hikaru
AU  - Kioka H
AD  - Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical and 
      Health Sciences, Kanazawa University, Kanazawa, Japan.
FAU - Nagata, Yuka
AU  - Nagata Y
AD  - Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical and 
      Health Sciences, Kanazawa University, Kanazawa, Japan.
FAU - Suzuki, Ryo
AU  - Suzuki R
AD  - Faculty of Pharmaceutical Sciences, Institute of Medical, Pharmaceutical and 
      Health Sciences, Kanazawa University, Kanazawa, Japan.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20220805
PL  - Switzerland
TA  - Front Immunol
JT  - Frontiers in immunology
JID - 101560960
RN  - 0 (RNA, Small Interfering)
RN  - 0 (Receptors, IgE)
RN  - 0 (SNARE Proteins)
RN  - 0 (Vesicle-Associated Membrane Protein 3)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - *Exocytosis
MH  - Immunoglobulin E
MH  - RNA, Small Interfering
MH  - *Receptors, IgE/metabolism
MH  - SNARE Proteins
MH  - Vesicle-Associated Membrane Protein 3/genetics/metabolism
PMC - PMC9388853
OTO - NOTNLM
OT  - FcϵRI
OT  - SNAREs
OT  - VAMP3
OT  - allergy
OT  - exocytosis
OT  - mast cell (MC)
EDAT- 2022/08/23 06:00
MHDA- 2022/08/24 06:00
PMCR- 2022/01/01
CRDT- 2022/08/22 03:58
PHST- 2022/02/28 00:00 [received]
PHST- 2022/07/18 00:00 [accepted]
PHST- 2022/08/22 03:58 [entrez]
PHST- 2022/08/23 06:00 [pubmed]
PHST- 2022/08/24 06:00 [medline]
PHST- 2022/01/01 00:00 [pmc-release]
AID - 10.3389/fimmu.2022.885868 [doi]
PST - epublish
SO  - Front Immunol. 2022 Aug 5;13:885868. doi: 10.3389/fimmu.2022.885868. eCollection 
      2022.