Selectively targeting an inactive conformation of interleukin-2-inducible T-cell kinase by allosteric inhibitors

2014 ◽  
Vol 460 (2) ◽  
pp. 211-222 ◽  
Author(s):  
Seungil Han ◽  
Robert M. Czerwinski ◽  
Nicole L. Caspers ◽  
David C. Limburg ◽  
WeiDong Ding ◽  
...  
Keyword(s):  
T Cell ◽  
Drug Design ◽  
Kinase Inhibitor ◽  
Interleukin 2 ◽  
Allosteric Inhibitors ◽  
Structure Based Drug Design ◽  
Inactive Conformation ◽  
Allosteric Inhibitor ◽  
Inactive Form ◽  
Inducible T Cell Kinase

Using structure-based drug design, a promiscuous ATP site kinase inhibitor scaffold was transformed into a highly-selective allosteric inhibitor that preferentially binds to an inactive form of interleukin-2-inducible T-cell kinase (ITK).

Discovery of potent inhibitors of interleukin-2 inducible T-cell kinase (ITK) through structure-based drug design

2009 ◽  
Vol 19 (3) ◽  
pp. 773-777 ◽  
Author(s):  
Brian N. Cook ◽  
Jörg Bentzien ◽  
Andre White ◽  
Peter A. Nemoto ◽  
Ji Wang ◽  
...  
Keyword(s):  
T Cell ◽  
Drug Design ◽  
Interleukin 2 ◽  
Structure Based Drug Design ◽  
Inducible T Cell Kinase

scholarly journals Identification of a New Inhibitor That Stabilizes Interleukin-2-Inducible T-Cell Kinase in Its Inactive Conformation

2019 ◽  
Vol 24 (8) ◽  
pp. 854-862
Author(s):  
Rie Hantani ◽  
Saya Hanawa ◽  
Shohei Oie ◽  
Kayo Umetani ◽  
Toshihiro Sato ◽  
...  
Keyword(s):  
T Cell ◽  
Conformational Changes ◽  
Kinase Inhibitors ◽  
Kinase Inhibitor ◽  
Interleukin 2 ◽  
Fluorescence Emission ◽  
T Cell Signaling ◽  
Inactive Conformation ◽  
Fluorescent Biosensor ◽  
Inducible T Cell Kinase

Interleukin-2-inducible T-cell kinase (ITK) plays an important role in T-cell signaling and is considered a promising drug target. As the ATP binding sites of protein kinases are highly conserved, the design of selective kinase inhibitors remains a challenge. Targeting inactive kinase conformations can address the issue of kinase inhibitor selectivity. It is important for selectivity considerations to identify compounds that stabilize inactive conformations from the primary screen hits. Here we screened a library of 390,000 compounds with an ADP-Glo assay using dephosphorylated ITK. After a surface plasmon resonance (SPR) assay was used to filter out promiscuous inhibitors, 105 hits were confirmed. Next, we used a fluorescent biosensor to enable the detection of conformational changes to identify inactive conformation inhibitors. A single-cysteine-substituted ITK mutant was labeled with acrylodan, and fluorescence emission was monitored. Using a fluorescent biosensor assay, we identified 34 inactive conformation inhibitors from SPR hits. Among them, one compound was bound to a site other than the ATP pocket and exhibited excellent selectivity against a kinase panel. Overall, (1) biochemical screening using dephosphorylated kinase, (2) hit confirmation by SPR assay, and (3) fluorescent biosensor assay that can distinguish inactive compounds provide a useful platform and offer opportunities to identify selective kinase inhibitors.

scholarly journals A negative role for the interleukin-2-inducible T-cell kinase (ITK) in human Foxp3+ TREG differentiation

2018 ◽  
Author(s):  
Polina Mamontov ◽  
Ryan A. Eberwine ◽  
Jackie Perrigoue ◽  
Anuk Das ◽  
Joshua R. Friedman ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Cell Activation ◽  
Kinase Inhibitor ◽  
Interleukin 2 ◽  
Tec Kinases ◽  
Human T Cells ◽  
Inducible T Cell Kinase

ABSTRACTThe Tec kinases ITK (interleukin-2-inducible T-cell kinase) and RLK (resting lymphocyte kinase) are critical components of the proximal TCR/CD3 signal transduction machinery, and data in mice suggest that ITK negatively regulates TREG differentiation. However, whether Tec kinases modulate TREG development and/or function in human T cells remains unknown. Using a novel self-delivery siRNA platform (sdRNA), we found that ITK knockdown in primary human naïve peripheral blood CD4 T cells increased Foxp3+ TREG differentiation under both TREG and T effector (Teff) cell priming conditions. ITK knockdown also enhanced the expression of the co-inhibitory receptor PD-1 on FoxP3+ T cells. TREGS differentiated in vitro (iTREG) after ITK knockdown displayed suppressive capacity against effector CD4+ T cell proliferation. ITK knockdown decreased IL-17A production in T cells primed under Th17 conditions and increased Th1 differentiation. Finally, a dual ITK/RLK Tec kinase inhibitor blocked TREG differentiation and T cell activation in general. Our data suggest that targeting ITK in human T cells may be an effective approach to boost TREG in the context of autoimmune diseases, but non-specific inhibition of other Tec family kinases may broadly inhibit T cell activation.

scholarly journals Targeting Interleukin-2-Inducible T-Cell Kinase (ITK) Differentiates GVL and GVHD in Allo-HSCT

2020 ◽  
Vol 11 ◽  
Author(s):  
Mahinbanu Mammadli ◽  
Weishan Huang ◽  
Rebecca Harris ◽  
Aisha Sultana ◽  
Ying Cheng ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Lymphoblastic Leukemia ◽  
Interleukin 2 ◽  
Treatment Strategies ◽  
Receptor Expression ◽  
Hematopoietic Stem ◽  
Target Organs ◽  
Donor T Cells ◽  
Inducible T Cell Kinase

Allogeneic hematopoietic stem cell transplantation is a potentially curative procedure for many malignant diseases. Donor T cells prevent disease recurrence via graft-versus-leukemia (GVL) effect. Donor T cells also contribute to graft-versus-host disease (GVHD), a debilitating and potentially fatal complication. Novel treatment strategies are needed which allow preservation of GVL effects without causing GVHD. Using murine models, we show that targeting IL-2-inducible T cell kinase (ITK) in donor T cells reduces GVHD while preserving GVL effects. Both CD8+ and CD4+ donor T cells from Itk-/- mice produce less inflammatory cytokines and show decrease migration to GVHD target organs such as the liver and small intestine, while maintaining GVL efficacy against primary B-cell acute lymphoblastic leukemia (B-ALL). Itk-/- T cells exhibit reduced expression of IRF4 and decreased JAK/STAT signaling activity but upregulating expression of Eomesodermin (Eomes) and preserve cytotoxicity, necessary for GVL effect. Transcriptome analysis indicates that ITK signaling controls chemokine receptor expression during alloactivation, which in turn affects the ability of donor T cells to migrate to GVHD target organs. Our data suggest that inhibiting ITK could be a therapeutic strategy to reduce GVHD while preserving the beneficial GVL effects following allo-HSCT treatment.

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