ACKR3 agonism induces heterodimerization with chemokine receptor CXCR4 and attenuates platelet function

IF 4.4 3区医学 Q1 MEDICINE, GENERAL & INTERNAL

European Journal of Clinical Investigation Pub Date : 2024-10-07 DOI:10.1111/eci.14327

Valerie Dicenta-Baunach, Zoi Laspa, David Schaale, Manuel Sigle, Alp Bayrak, Tatsiana Castor, Thanigaimalai Pillaiyar, Stefan Laufer, Meinrad Paul Gawaz, Anne-Katrin Rohlfing

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引用次数: 0

Abstract

Background

Platelet receptors ACKR3 and CXCR4 play a crucial role in a variety of cardiovascular diseases. Like most chemokine receptors, CXCR4 is a G protein coupled receptor that induces platelet activation. In contrast, the atypical chemokine receptor 3 (ACKR3) lacks the ability to activate heterotrimeric G proteins and its activation leads to platelet inhibition and attenuates thrombus formation. In nucleated cells, heterodimerization of ACKR3 with CXCR4 regulates CXCL12-dependent signalling. The aim of our study was to investigate the formation of ACKR3/CXCR4 heterodimers in platelets and the subsequent consequences for platelet function.

Methods and Results

Using a proximity ligation assay (PLA, Duolink®) to screen for CXCR4/ACKR3 heterodimerization inducing compounds, we found that ACKR3 agonism but not conventional platelet agonists or endogen ligands lead to heterodimer formation. To further characterize the formation of ACKR3/CXCR4 heterodimers, we studied the CXCL12-dependent platelet activation via CXCR4. Both, CXCL12-dependent platelet aggregation and collagen-dependent ex vivo thrombus formation were significantly downregulated by ACKR3 agonism. Moreover, platelet intracellular calcium and Akt signalling were increased by CXCL12 and again suppressed by ACKR3-specific agonists. Previously, CXCL12 was shown to decrease platelet cAMP levels via CXCR4. Treatment with a specific ACKR3 agonist counteracted this CXCL12/CXCR4-dependent cAMP decrease.

Conclusion

Our results reveal that the formation of platelet ACKR3/CXCR4 heterodimers is dependent on ACKR3 rather than CXCR4. Furthermore, ACKR3 agonism induced heterodimerization is associated with mitigating CXCL12/CXCR4-dependent platelet activation possibly by modulating CXCR4-dependent G protein signalling. Our results indicate possible ACKR3 agonist functions and reinforce the potential therapeutic applications of ACKR3 agonists.

Abstract Image

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ACKR3 激动可诱导与趋化因子受体 CXCR4 异源二聚化，并削弱血小板功能。

背景：血小板受体 ACKR3 和 CXCR4 在多种心血管疾病中起着至关重要的作用。与大多数趋化因子受体一样，CXCR4 是一种诱导血小板活化的 G 蛋白偶联受体。与此相反，非典型趋化因子受体 3（ACKR3）缺乏激活异源三聚 G 蛋白的能力，它的激活会导致血小板抑制并减少血栓形成。在有核细胞中，ACKR3 与 CXCR4 的异源二聚化调节 CXCL12 依赖性信号。我们的研究旨在调查血小板中 ACKR3/CXCR4 异源二聚体的形成及其对血小板功能的影响：我们发现 ACKR3 激动剂而非传统血小板激动剂或内源性配体会导致异二聚体的形成。为了进一步描述 ACKR3/CXCR4 异源二聚体形成的特点，我们研究了通过 CXCR4 激活 CXCL12 依赖性血小板的情况。在 ACKR3 的激动下，CXCL12 依赖性血小板聚集和胶原依赖性体外血栓形成均显著下调。此外，CXCL12 会增加血小板细胞内钙离子和 Akt 信号，ACKR3 特异性激动剂会再次抑制这些信号。以前的研究表明，CXCL12 可通过 CXCR4 降低血小板的 cAMP 水平。使用特异性 ACKR3 激动剂可抵消 CXCL12/CXCR4 依赖性 cAMP 的降低：我们的研究结果表明，血小板 ACKR3/CXCR4 异源二聚体的形成依赖于 ACKR3 而非 CXCR4。此外，ACKR3 激动诱导的异二聚体与减轻 CXCL12/CXCR4 依赖性血小板活化有关，可能是通过调节 CXCR4 依赖性 G 蛋白信号。我们的研究结果表明了 ACKR3 激动剂可能具有的功能，并加强了 ACKR3 激动剂的潜在治疗应用。

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来源期刊

European Journal of Clinical Investigation 医学-医学：内科

CiteScore

9.50

自引率

3.60%

发文量

192

审稿时长

1 months

期刊介绍： EJCI considers any original contribution from the most sophisticated basic molecular sciences to applied clinical and translational research and evidence-based medicine across a broad range of subspecialties. The EJCI publishes reports of high-quality research that pertain to the genetic, molecular, cellular, or physiological basis of human biology and disease, as well as research that addresses prevalence, diagnosis, course, treatment, and prevention of disease. We are primarily interested in studies directly pertinent to humans, but submission of robust in vitro and animal work is also encouraged. Interdisciplinary work and research using innovative methods and combinations of laboratory, clinical, and epidemiological methodologies and techniques is of great interest to the journal. Several categories of manuscripts (for detailed description see below) are considered: editorials, original articles (also including randomized clinical trials, systematic reviews and meta-analyses), reviews (narrative reviews), opinion articles (including debates, perspectives and commentaries); and letters to the Editor.