T cell dysfunction contributes to the susceptibility of patients with the Wiskott-Aldrich syndrome (WAS) to infection, autoimmunity and lymphoid malignancy. This project probes the function of WASP and its partner WASP interacting protein WIP in T cells. Most of WASP is complexed with WIP in T cells. WASP levels of drop to ~10% of normal in T cells from WIP KO mice and from WAS patients with mutations that disrupt WIP binding, suggesting that WIP stabilizes WASP. Our Preliminary data show that the hierarchy of T cell dysfunction is WASP/WIP DKOWIP KOWASP KO, bearing in mind that since WASP levels are low in WIP KO T cells the individual contribution of WIP to the WIP KO phenotype is unknown. Our overall hypothesis is that WIP and WASP function in T cells both as a complex and independently. To test our hypothesis we propose to use genetically engineered mice which selectively lack WIP, WASP or both, or in which the WASP/WIP complex is disrupted. We will: 1. Compare T cells from WASP/WIP DKO mice to T cells from WT mice to define the combined role of WIP and WASP. In addition, comparison with WASP KO T cells would define WIP functions that may be independent of WASP. 2. Reconstitute T cells from WIP KO mice with a peptide that consists of the WASP binding domain (WBD) of WIP in order to restore WASP levels, and thus generate WIP deficient/WASP expressing T cells. We will compare T cells from these mice to WT T cells to define the role of WIP. 3. Disrupt the WASP/WIP complex expressing the WBD in WT T cells to test the hypothesis that there is a subset of signaling functions that is dependent on the WASP/WIP complex. 4. Examine the structure and function of T cells from knockin mice that express a WIP mutant that lacks the actin-binding region to define the role of WIP binding to actin in the subcellular localization and function of the WASP/WIP complex. The results of the proposed experiments in the mouse will aid in our understanding of the pathogenesis of WAS in humans and should lead to novel treatments for this disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL059561-15
Application #
8380169
Study Section
Heart, Lung, and Blood Initial Review Group (HLBP)
Project Start
Project End
2014-08-31
Budget Start
2012-09-01
Budget End
2013-08-31
Support Year
15
Fiscal Year
2012
Total Cost
$410,557
Indirect Cost
$210,557
Name
Children's Hospital Boston
Department
Type
DUNS #
076593722
City
Boston
State
MA
Country
United States
Zip Code
02115
Baptista, Marisa A P; Keszei, Marton; Oliveira, Mariana et al. (2016) Deletion of Wiskott-Aldrich syndrome protein triggers Rac2 activity and increased cross-presentation by dendritic cells. Nat Commun 7:12175
Volpi, Stefano; Santori, Elettra; Abernethy, Katrina et al. (2016) N-WASP is required for B-cell-mediated autoimmunity in Wiskott-Aldrich syndrome. Blood 127:216-20
Shouval, Dror S; Biswas, Amlan; Kang, Yu Hui et al. (2016) Interleukin 1β Mediates Intestinal Inflammation in Mice and Patients With Interleukin 10 Receptor Deficiency. Gastroenterology 151:1100-1104
Lexmond, Willem S; Goettel, Jeremy A; Lyons, Jonathan J et al. (2016) FOXP3+ Tregs require WASP to restrain Th2-mediated food allergy. J Clin Invest 126:4030-4044
Moran, Christopher J; Klein, Christoph; Muise, Aleixo M et al. (2015) Very early-onset inflammatory bowel disease: gaining insight through focused discovery. Inflamm Bowel Dis 21:1166-75
Crestani, Elena; Volpi, Stefano; Candotti, Fabio et al. (2015) Broad spectrum of autoantibodies in patients with Wiskott-Aldrich syndrome and X-linked thrombocytopenia. J Allergy Clin Immunol 136:1401-4.e1-3
Grozovsky, Renata; Begonja, Antonija Jurak; Liu, Kaifeng et al. (2015) The Ashwell-Morell receptor regulates hepatic thrombopoietin production via JAK2-STAT3 signaling. Nat Med 21:47-54
Goettel, Jeremy A; Biswas, Subhabrata; Lexmond, Willem S et al. (2015) Fatal autoimmunity in mice reconstituted with human hematopoietic stem cells encoding defective FOXP3. Blood 125:3886-95
Begonja, Antonija Jurak; Pluthero, Fred G; Suphamungmee, Worawit et al. (2015) FlnA binding to PACSIN2 F-BAR domain regulates membrane tubulation in megakaryocytes and platelets. Blood 126:80-8
Kolhatkar, Nikita S; Brahmandam, Archana; Thouvenel, Christopher D et al. (2015) Altered BCR and TLR signals promote enhanced positive selection of autoreactive transitional B cells in Wiskott-Aldrich syndrome. J Exp Med 212:1663-77

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