Pleiotropic consequences of Bruton tyrosine kinase deficiency in myeloid lineages lead to poor inflammatory responses

AbstractBruton tyrosine kinase (Btk), a non-receptor-associated tyrosine kinase of the Tec family, appears to participate in many myeloid cell functions. We show that macrophages from X-linked immunodeficient (XID) mice lacking functional Btk cannot generate efficient bursts of reactive oxygen intermediates (ROIs). The induction of apoptotic cell death by inflammatory stimuli is also enhanced in XID macrophages. Phagocytosis of bacterial particles is only marginally affected in them. In vivo, XID mice show reduced severity of inflammatory diseases in models of experimental autoimmune encephalomyelitis (EAE), dextran sulfate sodium (DSS)-induced colitis, and carrageenan-induced acute edema. Also, polymorphonuclear neutrophil granulocytes (PMNs) in XID mice show poor ROI and nitric oxide (NO) induction, along with a reduction in PMN recruitment to peritoneal inflammation. XID mice show reduction in PMN numbers in peripheral blood, and their bone marrow shows a reduction in the numbers of both monocytic and granulocytic lineages, extending to the earliest progenitor populations. Thus, Btk is likely to play a significant role at multiple points during the development and functioning of the myeloid lineages, affecting the outcome of many infectious as well as noninfectious inflammatory events in vivo. (Blood. 2004;104:1191-1197)

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Overexposure to apoptosis via disrupted glial specification perturbs Drosophila macrophage function and reveals roles of the CNS during injury

10.1101/2020.03.04.977546 ◽

2020 ◽

Cited By ~ 1

Author(s):

Emma Louise Armitage ◽

Hannah Grace Roddie ◽

Iwan Robert Evans

Keyword(s):

Inflammatory Responses ◽

Apoptotic Cell ◽

Calcium Waves ◽

Apoptotic Cells ◽

Phagocytic Cells ◽

Macrophage Function ◽

Apoptotic Cell Clearance ◽

Cell Clearance ◽

Wound Responses

AbstractApoptotic cell clearance by phagocytes is a fundamental process during development, homeostasis and the resolution of inflammation. However, the demands placed on phagocytic cells such as macrophages by this process, and the limitations these interactions impose on subsequent cellular behaviours are not yet clear. Here we seek to understand how apoptotic cells affect macrophage function in the context of a genetically-tractable Drosophila model in which macrophages encounter excessive amounts of apoptotic cells. We show that loss of the glial transcription factor repo, and corresponding removal of the contribution these cells make to apoptotic cell clearance, causes macrophages in the developing embryo to be challenged with large numbers of apoptotic cells. As a consequence, macrophages become highly vacuolated with cleared apoptotic cells and their developmental dispersal and migration is perturbed. We also show that the requirement to deal with excess apoptosis caused by a loss of repo function leads to impaired inflammatory responses to injury. However, in contrast to migratory phenotypes, defects in wound responses cannot be rescued by preventing apoptosis from occurring within a repo mutant background. In investigating the underlying cause of these impaired inflammatory responses, we demonstrate that wound-induced calcium waves propagate into surrounding tissues, including neurons and glia of the ventral nerve cord, which exhibit striking calcium waves on wounding, revealing a previously unanticipated contribution of these cells during responses to injury. Taken together these results demonstrate important insights into macrophage biology and how repo mutants can be used to study macrophage-apoptotic cell interactions in the fly embryo.Furthermore, this work shows how these multipurpose cells can be ‘overtasked’ to the detriment of their other functions, alongside providing new insights into which cells govern macrophage responses to injury in vivo.

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Resolvins as Regulators of the Immune System

The Scientific World JOURNAL ◽

10.1100/tsw.2010.72 ◽

2010 ◽

Vol 10 ◽

pp. 818-831 ◽

Cited By ~ 20

Author(s):

Hiroyuki Seki ◽

Takaharu Sasaki ◽

Tomomi Ueda ◽

Makoto Arita

Keyword(s):

Immune System ◽

Acute Inflammation ◽

Immune Cell ◽

Inflammatory Responses ◽

Bioactive Metabolites ◽

Cell Functions ◽

First Response ◽

Beneficial Impact

Inflammation is the first response of the immune system to infection or injury, but excessive or inappropriate inflammatory responses contribute to a range of acute and chronic human diseases. Clinical assessment of dietary supplementation of ω-3 polyunsaturated fatty acids (i.e., eicosapentaenoic acid [EPA] and docosahexaenoic acid [DHA]) indicate that they have beneficial impact on these diseases, although the mechanisms are poorly understood at the molecular level. In this decade, it has been revealed that EPA and DHA are enzymatically converted to bioactive metabolites in the course of acute inflammation and resolution. These metabolites were shown to regulate immune cell functions and to display potent anti-inflammatory actions bothin vitroandin vivo. Because of their ability to resolve an acute inflammatory response, they are referred to as proresolving mediators, or resolvins. In this review, we provide an overview of the formation and actions of these lipid mediators.

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A Novel Mutation In Bruton Tyrosine Kinase Confers Acquired Resistance To Ibrutinib (PCI-32765) In CLL

Blood ◽

10.1182/blood.v122.21.4914.4914 ◽

2013 ◽

Vol 122 (21) ◽

pp. 4914-4914 ◽

Cited By ~ 2

Author(s):

Richard R. Furman ◽

Shuhua Cheng ◽

Pin Lu ◽

Menu Setty ◽

Alexandar Perez ◽

...

Keyword(s):

Tyrosine Kinase ◽

Ex Vivo ◽

Novel Mutation ◽

Expression Profiles ◽

Covalent Binding ◽

Lymphocytic Leukemia ◽

Brdu Incorporation ◽

Bruton Tyrosine Kinase ◽

Pre Treatment

Abstract The Bruton tyrosine kinase (BTK) inhibitor, ibrutinib has produced durable remissions in chronic lymphocytic leukemia (CLL). We describe a CLL patient who progressed while receiving ibrutinib following 20 months of once daily dosing. A cysteine-to-serine amino acid replacement was identified in BTK at position 481 that disrupts the covalent, but not non-covalent, binding of ibrutinib to BTK in silico structural modeling1. The mutation was present in relapsed samples while absent in the pre-treatment and responding samples. Following the mutation, the B cell receptor (BCR) pathway was reactivated as evidenced by increased cell signaling activities and gene expression profiles. Comparing the relapsed samples with the pre-treatment and responding samples, at the cellular level, mutated CLL cells displayed higher levels of the cell proliferation marker Ki67 in vivo and higher levels of ex-vivo BrdU incorporation. Transfection of the C481S mutant construct into a sensitive lymphoma cell line rendered it much more resistant to ibrutinib treatment demonstrating the cellular impact of the mutation (see attached graph). Interestingly, the ibrutinib-resistant CLL cells remained sensitive to other BCR inhibitors such as dasatinib and SYK inhibitors. These results confirm BTK as an important pharmacologic target of ibrutinib. Further, a mechanism of resistance was revealed, and alternative therapeutic options for ibrutinib resistance were explored. (First three authors with equal contribution) Disclosures: Furman: Genentech: Consultancy, Speakers Bureau; GlaxoSmithKline: Consultancy, Speakers Bureau; Pharmacyclics: Consultancy; Gilead: Consultancy. Chang:Pharmacyclics: Employment, Equity Ownership.

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Bruton tyrosine kinase is essential for botrocetin/VWF-induced signaling and GPIb-dependent thrombus formation in vivo

Blood ◽

10.1182/blood-2006-01-011817 ◽

2006 ◽

Vol 108 (8) ◽

pp. 2596-2603 ◽

Cited By ~ 88

Author(s):

Junling Liu ◽

Malinda E. Fitzgerald ◽

Michael C. Berndt ◽

Carl W. Jackson ◽

T. Kent Gartner

Keyword(s):

Tyrosine Kinase ◽

Thrombus Formation ◽

Dependent Manner ◽

Akt Phosphorylation ◽

Atp Secretion ◽

Arterial Thrombus ◽

Bruton Tyrosine Kinase ◽

Von Willebrand ◽

Willebrand Factor

AbstractBotrocetin (bt)-facilitated binding of von Willebrand factor (VWF) to the platelet membrane glycoprotein (GP) Ib-IX-V complex on platelets in suspension initiates a signaling cascade that causes αIIbβ3 activation and platelet aggregation. Previous work has demonstrated that bt/VWF-mediated agglutination activates αIIbβ3 and elicits ATP secretion in a thromboxane A2 (TxA2)-dependent manner. The signaling that results in TxA2 production was shown to be initiated by Lyn, enhanced by Src, and propagated through Syk, SLP-76, PI3K, PLCγ2, and PKC. Here, we demonstrate that the signaling elicited by GPIb-mediated agglutination that results in TxA2 production is dependent on Bruton tyrosine kinase (Btk). The results demonstrate that Btk is downstream of Lyn, Syk, SLP-76, and PI3K; upstream of ERK1/2, PLCγ2, and PKC; and greatly enhances Akt phosphorylation. The relationship(s), if any, between ERK1/2, PLCγ2, and PKC were not elucidated. The requirement for Btk and TxA2 receptor function in GPIb-dependent arterial thrombosis was confirmed in vivo by characterizing blood flow in ferric chloride-treated mouse carotid arteries. These results demonstrate that the Btk family kinase, Tec, cannot provide the function(s) missing because of the absence of Btk and that Btk is essential for both bt/VWF-mediated agglutination-induced TxA2 production and GPIb-dependent stable arterial thrombus formation in vivo.

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Analysis of combinatorial chemokine receptor expression dynamics using multi-receptor reporter mice

10.1101/2021.08.11.455927 ◽

2021 ◽

Author(s):

Laura Medina-Ruiz ◽

Robin Bartolini ◽

Douglas P Dyer ◽

Francesca Vidler ◽

Catherine Hughes ◽

...

Keyword(s):

Chemokine Receptor ◽

Inflammatory Responses ◽

Expression Patterns ◽

Myeloid Cell ◽

Receptor Expression ◽

In Vivo Models ◽

Chemokine Receptor Expression ◽

Reporter Mice ◽

Inflammatory Chemokines

Inflammatory chemokines and their receptors are central to the development of inflammatory/immune pathologies. The apparent complexity of this system, coupled with lack of appropriate in vivo models, has limited our understanding of how chemokines orchestrate inflammatory responses and has hampered attempts at targeting this system in inflammatory disease. Novel approaches are therefore needed to provide crucial biological, and therapeutic, insights into the chemokine-chemokine receptor family. Here, we report the generation of transgenic multi-chemokine receptor reporter mice in which spectrally-distinct fluorescent reporters mark expression of CCRs 1, 2, 3 and 5, key receptors for myeloid cell recruitment in inflammation. Analysis of these animals has allowed us to define, for the first time, individual and combinatorial receptor expression patterns on myeloid cells in resting and inflamed conditions. Our results demonstrate that chemokine receptor expression is highly specific, and more selective than previously anticipated.

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In vivo inhibition of Bruton Tyrosine Kinase (BTK) protects against alcoholic liver disease

Alcohol ◽

10.1016/j.alcohol.2021.09.023 ◽

2021 ◽

Vol 96 ◽

pp. 104

Author(s):

P.T. Nagesh ◽

R. Joshi ◽

A.R. Amigo ◽

Y. Zhuang ◽

Y. Cho ◽

...

Keyword(s):

Liver Disease ◽

Tyrosine Kinase ◽

Alcoholic Liver Disease ◽

Bruton Tyrosine Kinase

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Genetic analysis of Parkinson's disease-linked leucine-rich repeat kinase 2

Biochemical Society Transactions ◽

10.1042/bst20120112 ◽

2012 ◽

Vol 40 (5) ◽

pp. 1042-1046 ◽

Cited By ~ 11

Author(s):

Youren Tong ◽

Jie Shen

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Oxidative Damage ◽

Inflammatory Responses ◽

Dopamine D2 Receptor ◽

Apoptotic Cell ◽

Protein Carbonyls ◽

Mutant Form ◽

Leucine Rich Repeat

Mutations in LRRK2 (leucine-rich repeat kinase 2) are the most common genetic cause of PD (Parkinson's disease). To investigate how mutations in LRRK2 cause PD, we generated LRRK2 mutant mice either lacking its expression or expressing the R1441C mutant form. Homozygous R1441C knockin mice exhibit no dopaminergic neurodegeneration or alterations in steady-state levels of striatal dopamine, but they show impaired dopamine neurotransmission, as was evident from reductions in amphetamine-induced locomotor activity and stimulated catecholamine release in cultured chromaffin cells as well as impaired dopamine D2 receptor-mediated functions. Whereas LRRK2−/− brains are normal, LRRK2−/− kidneys at 20 months of age develop striking accumulation and aggregation of α-synuclein and ubiquitinated proteins, impairment of the autophagy–lysosomal pathway, and increases in apoptotic cell death, inflammatory responses and oxidative damage. Our further analysis of LRRK2−/− kidneys at multiple ages revealed unique age-dependent biphasic alterations of the autophagic activity, which is unchanged at 1 month of age, enhanced at 7 months, but reduced at 20 months. Levels of α-synuclein and protein carbonyls, a general oxidative damage marker, are also decreased in LRRK2−/− kidneys at 7 months of age. Interestingly, this biphasic alteration is associated with increased levels of lysosomal proteins and proteases as well as progressive accumulation of autolysosomes and lipofuscin granules. We conclude that pathogenic mutations in LRRK2 impair the nigrostriatal dopaminergic pathway, and LRRK2 plays an essential role in the dynamic regulation of autophagy function in vivo.

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FLT3-ITD up-regulates MCL-1 to promote survival of stem cells in acute myeloid leukemia via FLT3-ITD–specific STAT5 activation

Blood ◽

10.1182/blood-2008-12-196055 ◽

2009 ◽

Vol 114 (24) ◽

pp. 5034-5043 ◽

Cited By ~ 127

Author(s):

Goichi Yoshimoto ◽

Toshihiro Miyamoto ◽

Siamak Jabbarzadeh-Tabrizi ◽

Tadafumi Iino ◽

Jennifer L. Rocnik ◽

...

Keyword(s):

Stem Cells ◽

Acute Myeloid Leukemia ◽

Tyrosine Kinase ◽

Myeloid Leukemia ◽

Kinase Inhibitor ◽

Apoptotic Cell ◽

Critical Role ◽

Myeloid Cell ◽

Hematopoietic Stem ◽

Acute Myeloid

Abstract Myeloid cell leukemia-1 (MCL-1) is an essential survival factor for hematopoiesis. In humans, hematopoietic stem cells (HSCs) express MCL-1 at the highest level in response to FMS-like tyrosine kinase-3 (FLT3) signaling. We here show that this FLT3-dependent stem cell maintenance system also plays a critical role in survival of leukemic stem cells (LSCs) in acute myeloid leukemia (AML). The CD34+CD38− LSC fraction expresses high levels of FLT3 as well as MCL-1, even compared with normal HSCs. Treatment with FLT3 ligand induced further MCL-1 up-regulation in LSCs in all AML cases tested. Interestingly, the group of samples expressing the highest levels of MCL-1 constituted AML with FLT3–internal tandem duplications (ITD). In FLT3-ITD AML cell lines, cells expressed a high level of MCL-1, and an inhibition of MCL-1 induced their apoptotic cell death. A tyrosine kinase inhibitor suppressed MCL-1 expression, and induced apoptosis that was reversed by the enforced MCL-1 expression. Finally, transduction of FLT3-ITD into HSCs strongly activated MCL-1 expression through its signal transducer and activator of transcription 5 (STAT5)–docking domains. This effect was completely abrogated when STAT5 activation was blocked. Thus, the acquisition of FLT3-ITD ensures LSC survival by up-regulating MCL-1 via constitutive STAT5 activation that is independent of wild-type FLT3 signaling.

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Protein synthesis rates of human PBMC and PMN can be determined simultaneously in vivo by using small blood samples

AJP Cell Physiology ◽

10.1152/ajpcell.00563.2002 ◽

2004 ◽

Vol 286 (6) ◽

pp. C1474-C1478 ◽

Cited By ~ 12

Author(s):

Stéphane Walrand ◽

Christelle Guillet ◽

Pierre Gachon ◽

Paulette Rousset ◽

Christophe Giraudet ◽

...

Keyword(s):

Amino Acid ◽

Immune Cell ◽

Gradient Centrifugation ◽

Polymorphonuclear Neutrophil ◽

Healthy Human ◽

Peripheral Blood Mononuclear ◽

Blood Samples ◽

Precursor Pool ◽

Cell Functions

Immune cell functions can be evaluated in vivo by measuring their specific protein fractional synthesis rates (FSR). Using stable isotope dilution techniques, we describe a new method allowing simultaneous in vivo assessment of FSR in two leukocyte populations in healthy human subjects, using small blood samples. Peripheral blood mononuclear cell (PBMC) and polymorphonuclear neutrophil (PMN) FSR were measured during primed continuous intravenous infusion of l-[1-13C]leucine. Immune cells from 6 ml of whole blood were isolated by density gradient centrifugation. In a first study, we calculated the FSR using plasma [13C]leucine or α-[13C]ketoisocaproate (KIC) enrichments as precursor pools. In a second study, we compared protein FSR in leukocytes, using enrichments of either intracellular or plasma free [13C]leucine as immediate precursor pools. The present approach showed a steady-state enrichment of plasma and circulating immune cell free [13C]leucine precursor pools. The linearity of labeled amino acid incorporation rate within mixed PBMC and PMN proteins also was verified. Postabsorptive protein FSR was 4.09 ± 0.39%/day in PBMC and 1.44 ± 0.08%/day in PMN when plasma [13C]KIC was the precursor pool. The difference between PBMC and PMN FSR was statistically significant, whatever the precursor pool used, suggesting large differences in their synthetic activities and functions. Use of the plasma [13C]KIC pool led to an underestimation of leukocyte FSR compared with the intracellular pool (PBMC: 6.04 ± 0.94%/day; PMN: 2.98 ± 0.30%/day). Hence, the intracellular free amino acid pool must be used as precursor to obtain reliable results. In conclusion, it is possible to assess immune cell metabolism in vivo in humans by using small blood samples to directly indicate their metabolic activity in various clinical situations and in response to regulating factors.

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Neutrophil development and function critically depend on Bruton tyrosine kinase in a mouse model of X-linked agammaglobulinemia

Blood ◽

10.1182/blood-2010-04-281170 ◽

2011 ◽

Vol 117 (4) ◽

pp. 1329-1339 ◽

Cited By ~ 54

Author(s):

Katja Fiedler ◽

Anca Sindrilaru ◽

Grzegorz Terszowski ◽

Enikö Kokai ◽

Thorsten B. Feyerabend ◽

...

Keyword(s):

Bone Marrow ◽

Tyrosine Kinase ◽

Neutrophil Migration ◽

Edema Formation ◽

Gm Csf ◽

Bruton Tyrosine Kinase ◽

And Function ◽

Bone Marrow Chimeras ◽

Deficient Mice

Abstract Bruton tyrosine kinase (Btk) is essential for B cell development and function and also appears to be important for myeloid cells. The bone marrow of Btk-deficient mice shows enhanced granulopoiesis compared with that of wild-type mice. In purified granulocyte-monocyte-progenitors (GMP) from Btk-deficient mice, the development of granulocytes is favored at the expense of monocytes. However, Btk-deficient neutrophils are impaired in maturation and function. Using bone marrow chimeras, we show that this defect is cell-intrinsic to neutrophils. In GMP and neutrophils, Btk plays a role in GM-CSF– and Toll-like receptor–induced differentiation. Molecular analyses revealed that expression of the lineage-determining transcription factors C/EBPα, C/EBPβ, and PU.1, depends on Btk. In addition, expression of several granule proteins, including myeloperoxidase, neutrophilic granule protein, gelatinase and neutrophil elastase, is Btk-dependent. In the Arthus reaction, an acute inflammatory response, neutrophil migration into tissues, edema formation, and hemorrhage are significantly reduced in Btk-deficient animals. Together, our findings implicate Btk as an important regulator of neutrophilic granulocyte maturation and function in vivo.

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