Pyroptosis leads to loss of centrosomal integrity in macrophages
Pyroptosis leads to loss of centrosomal integrity in macrophages
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DOI：
10.1101/2023.11.22.568260
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发表时间：
2023-11
期刊：
影响因子：
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通讯作者：
Siyi Bai;Fátima Martín-Sánchez;D. Brough;G. López-Castejón
中科院分区：
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作者：
Siyi Bai;Fátima Martín-Sánchez;D. Brough;G. López-Castejón
当巨噬细胞感知感染或组织损伤时，NLRP 3形成多蛋白炎性体复合物以引发炎症反应，这导致半胱天冬酶-1活化和成熟以及炎性细胞因子白细胞介素-1 β（IL-1β）和IL-18以及Gasdermin-D（GSDMD）介导的焦亡的释放。NLRP 3炎性体活性必须得到控制，因为不受调节的慢性炎症是炎性和自身免疫性疾病的基础。一些研究发现，NLRP 3炎性小体活性受中心体定位蛋白如NEK 7和HDAC 6的调节，然而，在炎性小体激活期间中心体组成或结构是否改变尚不清楚。我们的数据显示，在人类和小鼠巨噬细胞中，中心体支架蛋白pericentrin（PCNT）的水平在NLRP 3炎性小体活化后通过不同的活化剂降低。PCNT损失发生在膜稳定剂安石榴苷的存在下，表明这不是膜破裂的结果。我们发现PCNT的丢失依赖于NLRP 3和活性半胱天冬酶，因为MCC 950和泛半胱天冬酶抑制剂ZVAD阻止其降解。此外，胱天蛋白酶-1和GSDMD都是这种NLRP 3介导的PCNT损失所需的，因为胱天蛋白酶-1或GSDMD的缺乏通过响应尼日利亚菌素刺激的胱天蛋白酶-3的裂解触发PCNT的替代调节。PCNT降解响应于尼日利亚菌素而发生，但也响应于其他NLRP 3激活剂，包括促溶素剂L-亮氨酰-L-亮氨酸甲酯（LLOMe）和低渗性而发生。我们的工作揭示了NLRP 3炎性小体激活影响中心体的组成和结构，这可能加深我们对激活的NLRP 3炎性小体如何参与炎症性疾病发病机制的理解。
NLRP3 forms a multiprotein inflammasome complex to initiate the inflammatory response when macrophages sense infection or tissue damage, which leads to caspase-1 activation and maturation and release of the inflammatory cytokines interleukin-1β (IL-1β) and IL-18, and Gasdermin-D (GSDMD) mediated pyroptosis. NLRP3 inflammasome activity must be controlled as unregulated and chronic inflammation underlies inflammatory and autoimmune diseases. Several findings uncovered that NLRP3 inflammasome activity is under the regulation of centrosome localized proteins such as NEK7 and HDAC6, however, whether the centrosome composition or structure is altered during the inflammasome activation is not known. Our data show that levels of the centrosomal scaffold protein pericentrin (PCNT) are reduced upon NLRP3 inflammasome activation via different activators in human and murine macrophages. PCNT loss occurs in the presence of membrane stabilizer punicalagin, suggesting this is not a consequence of membrane rupture. We found that PCNT loss is dependent on NLRP3 and active caspases as MCC950 and pan caspase inhibitor ZVAD prevent its degradation. Moreover, caspase-1 and GSDMD are both required for this NLRP3-mediated PCNT loss because absence of caspase-1 or GSDMD triggers an alternative regulation of PCNT via its cleavage by caspase-3 in response to nigericin stimulation. PCNT degradation occurs in response to nigericin, but also other NLRP3 activators including lysomotropic agent L-Leucyl-L-Leucine methyl ester (LLOMe) and hypotonicity. Our work reveals that the NLRP3 inflammasome activation affects centrosome composition and structure which may deepen our understandings of how activated NLRP3 inflammasomes are involved in the pathogenesis of inflammatory diseases.