scholarly journals 687 MDSC gene expression analysis in patients with cancer and the response to inhibition of Bruton's tyrosine kinase

2021 ◽  
Vol 9 (Suppl 3) ◽  
pp. A715-A715
Author(s):  
Himanshu Savardekar ◽  
Carter Allen ◽  
Dionisia Quiroga ◽  
Donjun Chung ◽  
Emily Schwarz ◽  
...  
Keyword(s):  
Gene Expression ◽  
Tyrosine Kinase ◽  
Cancer Patients ◽  
Fold Change ◽  
Cell Cluster ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Gene Markers ◽  
Multiple Genes

BackgroundMyeloid-derived suppressor cells (MDSC) are an immunosuppressive immature population of myeloid cells that are elevated in cancer patients. Increased levels of MDSC has been linked to dysregulated anti-tumor responses and reduced efficacy of immune checkpoint therapies thus making them an attractive target. MDSC express Bruton's tyrosine kinase (BTK) and can be depleted using ibrutinib, an FDA-approved irreversible inhibitor of BTK. BTK inhibition leads to reduced MDSC expansion/function in murine models and significantly improved activity of anti-PD-1 antibodies. In this study, single cell RNA-seq (scRNA-seq) was used to characterize the gene expression of MDSC from different cancer types and the effect of ibrutinib on MDSC gene expression.MethodsPeripheral blood mononuclear cells were isolated from patients with melanoma (n=2), head & neck (n=1), and breast cancer (n=1). MDSC were isolated via fluorescence activated cell sorting. MDSC isolated from melanoma patients (n=2) were treated in vitro for 4h with 1 uM ibrutinib or DMSO and scRNA-seq was performed using the Chromium 10x Genomics platform. ScRNA-seq samples were analyzed using the standard integrative workflow of Seurat v3, which addresses the sample heterogeneity. Cell clusters were identified using Seurat and annotated using SingleRversion3.12. Identification of gene markers for each cell cluster and cell-cluster-specific differential expression analyses were conducted using Seurat.ResultsBaseline gene expression of MDSC from patients with breast and head & neck cancer revealed similarities among the top expressed genes (S100A8, VCAN, and LYZ). In vitro ibrutinib treatment of MDSC from patients with melanoma resulted in significant changes in gene expression within the MDSC cluster compared to DMSO treatment. GBP1(-1.72 log fold change), IL 1β(-1.27 log fold change), and CXCL8(-0.63 log fold change) were among the top downregulated genes (p<0.001) and RGS2 (0.68 log fold change) and ABHD5(0.52 log fold change) were among the top upregulated genes (p<0.001). MDSC subset (PMN-MDSC, M-MDSC, early-MDSC, and CD14+/CD15+ double positive) gene expression changes mirrored total MDSC gene changes. Ingenuity pathway analysis revealed significant downregulated pathways including TREM1 (p<0.001), nitric oxide signaling (p<0.003), and IL-6 signaling (p<0.004). Multiple genes associated with cellular movement (CXCL8, CXCL10) and activation of macrophages (CXCL10, CCL3) were downregulated (p<0.001). PCR analysis on isolated melanoma MDSC (n=2) treated in vitro with ibrutinib verified downregulation of CXCL8 (0.42 fold decrease, p<0.05) and CXCL10 (0.40 fold decrease, p<0.001).ConclusionsAnalysis via scRNA-seq revealed similar gene expression patterns for MDSC from different cancer patients. There was downregulation of multiple genes and pathways important to MDSC function and migration after BTK inhibition.Ethics ApprovalThe study obtained ethics approval. IRB# 1999C0348

Abstract 317: Vecabrutinib inhibits C481 mutated Bruton's tyrosine kinase and its downstream signaling in vitro

2019 ◽  
Author(s):  
Burcu Aslan ◽  
Mikhila Mahendra ◽  
Michael D Peoples ◽  
Joe R. Marszalek ◽  
Christopher P Vellano ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Downstream Signaling

scholarly journals Silencing Bruton's tyrosine kinase in alveolar neutrophils protects mice from LPS/immune complex-induced acute lung injury

2014 ◽  
Vol 307 (6) ◽  
pp. L435-L448 ◽  
Author(s):  
Agnieszka Krupa ◽  
Marek Fol ◽  
Moshiur Rahman ◽  
Karen Y. Stokes ◽  
Jon M. Florence ◽  
...  
Keyword(s):  
Acute Lung Injury ◽  
Lung Injury ◽  
Tyrosine Kinase ◽  
Immune Complex ◽  
Distress Syndrome ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Neutrophil Survival

Previous observations made by our laboratory indicate that Bruton's tyrosine kinase (Btk) may play an important role in the pathophysiology of local inflammation in acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). We have shown that there is cross talk between FcγRIIa and TLR4 in alveolar neutrophils from patients with ALI/ARDS and that Btk mediates the molecular cooperation between these two receptors. To study the function of Btk in vivo we have developed a unique two-hit model of ALI: LPS/immune complex (IC)-induced ALI. Furthermore, we conjugated F(ab)2 fragments of anti-neutrophil antibodies (Ly6G1A8) with specific siRNA for Btk to silence Btk specifically in alveolar neutrophils. It should be stressed that we are the first group to perform noninvasive transfections of neutrophils, both in vitro and in vivo. Importantly, our present findings indicate that silencing Btk in alveolar neutrophils has a dramatic protective effect in mice with LPS/IC-induced ALI, and that Btk regulates neutrophil survival and clearance of apoptotic neutrophils in this model. In conclusion, we put forward a hypothesis that Btk-targeted neutrophil specific therapy is a valid goal of research geared toward restoring homeostasis in lungs of patients with ALI/ARDS.

Abstract 317: Vecabrutinib inhibits C481 mutated Bruton's tyrosine kinase and its downstream signaling in vitro

2019 ◽  
Author(s):  
Burcu Aslan ◽  
Mikhila Mahendra ◽  
Michael D Peoples ◽  
Joe R. Marszalek ◽  
Christopher P Vellano ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Downstream Signaling

scholarly journals Direct and two-step bioorthogonal probes for Bruton's tyrosine kinase based on ibrutinib: a comparative study

2015 ◽  
Vol 13 (18) ◽  
pp. 5147-5157 ◽  
Author(s):  
Nora Liu ◽  
Sascha Hoogendoorn ◽  
Bas van de Kar ◽  
Allard Kaptein ◽  
Tjeerd Barf ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Comparative Study ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase

Direct and two-step activity-based probes allow for profiling of Bruton's tyrosine kinase in vitro and in situ.

scholarly journals Bruton’s Tyrosine Kinase Inhibitors Ibrutinib and Acalabrutinib Counteract Anthracycline Resistance in Cancer Cells Expressing AKR1C3

Cancers ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 3731
Author(s):  
Anselm Morell ◽  
Lucie Čermáková ◽  
Eva Novotná ◽  
Lenka Laštovičková ◽  
Melodie Haddad ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Cancer Cells ◽  
Kinase Inhibitors ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Anthracycline Resistance ◽  
Target Action ◽  
Combination Regimens ◽  
Btk Inhibitors

Over the last few years, aldo-keto reductase family 1 member C3 (AKR1C3) has been associated with the emergence of multidrug resistance (MDR), thereby hindering chemotherapy against cancer. In particular, impaired efficacy of the gold standards of induction therapy in acute myeloid leukaemia (AML) has been correlated with AKR1C3 expression, as this enzyme metabolises several drugs including anthracyclines. Therefore, the development of selective AKR1C3 inhibitors may help to overcome chemoresistance in clinical practice. In this regard, we demonstrated that Bruton’s tyrosine kinase (BTK) inhibitors ibrutinib and acalabrutinib efficiently prevented daunorubicin (Dau) inactivation mediated by AKR1C3 in both its recombinant form as well as during its overexpression in cancer cells. This revealed a synergistic effect of BTK inhibitors on Dau cytotoxicity in cancer cells expressing AKR1C3 both exogenously and endogenously, thus reverting anthracycline resistance in vitro. These findings suggest that BTK inhibitors have a novel off-target action, which can be exploited against leukaemia through combination regimens with standard chemotherapeutics like anthracyclines.

scholarly journals Identification of Bruton’s tyrosine kinase as a therapeutic target in acute myeloid leukemia

Blood ◽  
2014 ◽  
Vol 123 (8) ◽  
pp. 1229-1238 ◽  
Author(s):  
Stuart A. Rushworth ◽  
Megan Y. Murray ◽  
Lyubov Zaitseva ◽  
Kristian M. Bowles ◽  
David J. MacEwan
Keyword(s):  
Acute Myeloid Leukemia ◽  
Chronic Lymphocytic Leukemia ◽  
Tyrosine Kinase ◽  
Myeloid Leukemia ◽  
Lymphocytic Leukemia ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Key Points ◽  
Acute Myeloid

Key Points Inhibition of Bruton’s tyrosine kinase is as effective in vitro against AML as chronic lymphocytic leukemia. Ibrutinib shows activity in AML because Bruton’s tyrosine kinase is constitutively active.

scholarly journals Oral administration of Bruton's tyrosine kinase inhibitors impairs GPVI-mediated platelet function

2016 ◽  
Vol 310 (5) ◽  
pp. C373-C380 ◽  
Author(s):  
Rachel A. Rigg ◽  
Joseph E. Aslan ◽  
Laura D. Healy ◽  
Michael Wallisch ◽  
Marisa L. D. Thierheimer ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Platelet Activation ◽  
Platelet Function ◽  
Lymphocytic Leukemia ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Glycoprotein Vi ◽  
Activation Spreading

The Tec family kinase Bruton's tyrosine kinase (Btk) plays an important signaling role downstream of immunoreceptor tyrosine-based activation motifs in hematopoietic cells. Mutations in Btk are involved in impaired B-cell maturation in X-linked agammaglobulinemia, and Btk has been investigated for its role in platelet activation via activation of the effector protein phospholipase Cγ2 downstream of the platelet membrane glycoprotein VI (GPVI). Because of its role in hematopoietic cell signaling, Btk has become a target in the treatment of chronic lymphocytic leukemia and mantle cell lymphoma; the covalent Btk inhibitor ibrutinib was recently approved by the US Food and Drug Administration for treatment of these conditions. Antihemostatic events have been reported in some patients taking ibrutinib, although the mechanism of these events remains unknown. We sought to determine the effects of Btk inhibition on platelet function in a series of in vitro studies of platelet activation, spreading, and aggregation. Our results show that irreversible inhibition of Btk with two ibrutinib analogs in vitro decreased human platelet activation, phosphorylation of Btk, P-selectin exposure, spreading on fibrinogen, and aggregation under shear flow conditions. Short-term studies of ibrutinib analogs administered in vivo also showed abrogation of platelet aggregation in vitro, but without measurable effects on plasma clotting times or on bleeding in vivo. Taken together, our results suggest that inhibition of Btk significantly decreased GPVI-mediated platelet activation, spreading, and aggregation in vitro; however, prolonged bleeding was not observed in a model of bleeding.

scholarly journals Bruton’s Tyrosine Kinase Inhibition Promotes Myelin Repair

2020 ◽  
Vol 5 (2) ◽  
pp. 123-133
Author(s):  
Elodie Martin ◽  
Marie-Stéphane Aigrot ◽  
Roland Grenningloh ◽  
Bruno Stankoff ◽  
Catherine Lubetzki ◽  
...  
Keyword(s):  
Tyrosine Kinase ◽  
Ex Vivo ◽  
Fold Increase ◽  
Experimental Models ◽  
Bruton’S Tyrosine Kinase ◽  
Bruton's Tyrosine Kinase ◽  
Myelin Repair ◽  
Before And After

Background: Microglia are the resident macrophages of the central nervous system (CNS). In multiple sclerosis (MS) and related experimental models, microglia have either a pro-inflammatory or a pro-regenerative/pro-remyelinating function. Inhibition of Bruton’s tyrosine kinase (BTK), a member of the Tec family of kinases, has been shown to block differentiation of pro-inflammatory macrophages in response to granulocyte–macrophage colony-stimulating factor in vitro. However, the role of BTK in the CNS is unknown. Methods: Our aim was to investigate the effect of BTK inhibition on myelin repair in ex vivo and in vivo experimental models of demyelination and remyelination. The remyelination effect of a BTK inhibitor (BTKi; BTKi-1) was then investigated in LPC-induced demyelinated cerebellar organotypic slice cultures and metronidazole-induced demyelinated Xenopus MBP-GFP-NTR transgenic tadpoles. Results: Cellular detection of BTK and its activated form BTK-phospho-Y223 (p-BTK) was determined by immunohistochemistry in organotypic cerebellar slice cultures, before and after lysophosphatidylcholine (LPC)-induced demyelination. A low BTK signal detected by immunolabeling under normal conditions in cerebellar slices was in sharp contrast to an 8.5-fold increase in the number of BTK-positive cells observed in LPC-demyelinated slice cultures. Under both conditions, approximately 75% of cells expressing BTK and p-BTK were microglia and 25% were astrocytes. Compared with spontaneous recovery, treatment of demyelinated slice cultures and MTZ-demyelinated transgenic tadpoles with BTKi resulted in at least a 1.7-fold improvement of remyelination. Conclusion: Our data demonstrate that BTK inhibition is a promising therapeutic strategy for myelin repair.

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