当前位置: X-MOL 学术Autophagy › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
LRRK2 is required for CD38-mediated NAADP-Ca2+ signaling and the downstream activation of TFEB (transcription factor EB) in immune cells
Autophagy ( IF 13.3 ) Pub Date : 2021-07-27 , DOI: 10.1080/15548627.2021.1954779
Neel R Nabar 1 , Christopher N Heijjer 1 , Chong-Shan Shi 1 , Il-Young Hwang 1 , Sundar Ganesan 2 , Mikael C I Karlsson 3 , John H Kehrl 1
Affiliation  

ABSTRACT

CD38 is a cell surface receptor capable of generating calcium-mobilizing second messengers. It has been implicated in host defense and cancer biology, but signaling mechanisms downstream of CD38 remain unclear. Mutations in LRRK2 (leucine-rich repeat kinase 2) are the most common genetic cause of Parkinson disease; it is also a risk factor for Crohn disease, leprosy, and certain types of cancers. The pathogenesis of these diseases involves inflammation and macroautophagy/autophagy, processes both CD38 and LRRK2 are implicated in. Here, we mechanistically and functionally link CD38 and LRRK2 as upstream activators of TFEB (transcription factor EB), a host defense transcription factor and the master transcriptional regulator of the autophagy/lysosome machinery. In B-lymphocytes and macrophages, we show that CD38 and LRRK2 exist in a complex on the plasma membrane. Ligation of CD38 with the monoclonal antibody clone 90 results in internalization of the CD38-LRRK2 complex and its targeting to the endolysosomal system. This generates an NAADP-dependent calcium signal, which requires LRRK2 kinase activity, and results in the downstream activation of TFEB. lrrk2 KO macrophages accordingly have TFEB activation defects following CD38 or LPS stimulation and fail to switch to glycolytic metabolism after LPS treatment. In overexpression models, the pathogenic LRRK2G2019S mutant promotes hyperactivation of TFEB even in the absence of CD38, both by stabilizing TFEB and promoting its nuclear translocation via aberrant calcium signaling. In sum, we have identified a physiological CD38-LRRK2-TFEB signaling axis in immune cells. The common pathogenic mutant, LRRK2G2019S, appears to hijack this pathway.

Abbreviations:ADPR: ADP-ribose; AMPK: AMP-activated protein kinase; BMDM: bone marrow-derived macrophage; cADPR: cyclic-ADP-ribose; COR: C-terminal of ROC; CTSD: cathepsin D; ECAR: extracellular acidification rate; EDTA: ethylenediaminetetraacetic acid; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; GFP: green fluorescent protein; GPN: Gly-Phe β-naphthylamide; GSK3B/GSK3β: glycogen synthase kinase 3 beta; GTP: guanosine triphosphate; KD: knockdown; LAMP1: lysosomal-associated membrane protein 1; LRR: leucine rich repeat; LRRK2: leucine rich repeat kinase 2; mAb: monoclonal antibody; MAP1LC3B/LC3B: microtubule-associated protein 1 light chain 3 beta; MAPK/ERK: mitogen-activated protein kinase; MCOLN1: mucolipin 1; MFI: mean fluorescence intensity; mRNA: messenger RNA; MTOR: mechanistic target of rapamycin kinase; NAADP: nicotinic acid adenine dinucleotide phosphate; NAD: nicotinamide adenine dinucleotide; NADP: nicotinamide adenine dinucleotide phosphate; PD: Parkinson disease; PPP3CB: protein phosphatase 3, catalytic subunit, beta isoform; q-RT-PCR: quantitative reverse transcription polymerase chain reaction; ROC: Ras of complex; siRNA: small interfering RNA; SQSTM1/p62: sequestome 1; TFEB: transcription factor EB; TPCN: two pore channel; TRPM2: transient receptor potential cation channel, subfamily M, member 2; ZKSCAN3: zinc finger with KRAB and SCAN domains 3



中文翻译:

LRRK2 是 CD38 介导的 NAADP-Ca2+ 信号传导和免疫细胞中 TFEB(转录因子 EB)下游激活所必需的

摘要

CD38 是一种细胞表面受体,能够产生调动钙的第二信使。它与宿主防御和癌症生物学有关,但 CD38 下游的信号传导机制仍不清楚。LRRK2(富含亮氨酸的重复激酶 2)突变是帕金森病最常见的遗传原因;它也是克罗恩病、麻风病和某些类型癌症的危险因素。这些疾病的发病机制涉及炎症和巨自噬/自噬,涉及 CD38 和 LRRK2 的过程。在这里,我们在机制和功能上将 CD38 和 LRRK2 连接为 TFEB(转录因子 EB)的上游激活因子,宿主防御转录因子和主自噬/溶酶体机制的转录调节因子。在 B 淋巴细胞和巨噬细胞中,我们表明 CD38 和 LRRK2 存在于质膜上的复合物中。CD38 与单克隆抗体克隆 90 的连接导致 CD38-LRRK2 复合物内化并靶向内溶酶体系统。这会产生一个依赖于 NAADP 的钙信号,这需要 LRRK2 激酶活性,并导致 TFEB 的下游激活。因此, lrrk2 KO 巨噬细胞在 CD38 或 LPS 刺激后具有 TFEB 活化缺陷,并且在 LPS 处理后无法转换为糖酵解代谢。在过表达模型中,即使在没有 CD38 的情况下,致病性 LRRK2 G2019S突变体也会通过稳定 TFEB 和通过异常钙信号传导促进其核转位来促进 TFEB 的过度活化。总之,我们已经确定了免疫细胞中的生理 CD38-LRRK2-TFEB 信号轴。常见的致病突变体 LRRK2 G2019S似乎劫持了这一途径。

缩写:ADPR:ADP-核糖;AMPK:AMP激活的蛋白激酶;BMDM:骨髓来源的巨噬细胞;cADPR:环状-ADP-核糖;COR:ROC的C端;CTSD:组织蛋白酶 D;ECAR:细胞外酸化率;EDTA:乙二胺四乙酸;GAPDH:3-磷酸甘油醛脱氢酶;GFP:绿色荧光蛋白;GPN:Gly-Phe β-萘酰胺;GSK3B/GSK3β:糖原合酶激酶 3 β;GTP:鸟苷三磷酸;KD:击倒;LAMP1:溶酶体相关膜蛋白 1;LRR:富含亮氨酸的重复;LRRK2:富含亮氨酸的重复激酶 2;mAb:单克隆抗体;MAP1LC3B/LC3B:微管相关蛋白1轻链3β;MAPK/ERK:丝裂原活化蛋白激酶;MCOLN1:粘磷脂1;MFI:平均荧光强度;mRNA:信使RNA;MTOR:雷帕霉素激酶的机制靶点;全国有色人种计划:烟酸腺嘌呤二核苷酸磷酸盐;NAD:烟酰胺腺嘌呤二核苷酸;NADP:烟酰胺腺嘌呤二核苷酸磷酸盐;PD:帕金森病;PPP3CB:蛋白磷酸酶 3,催化亚基,β 异构体;q-RT-PCR:定量逆转录聚合酶链反应;ROC:复数Ras;siRNA:小干扰RNA;SQSTM1/p62:sequestome 1;TFEB:转录因子EB;TPCN:二孔通道;TRPM2:瞬时受体电位阳离子通道,亚家族 M,成员 2;ZKSCAN3:具有 KRAB 和 SCAN 结构域 3 的锌指 SQSTM1/p62:sequestome 1;TFEB:转录因子EB;TPCN:二孔通道;TRPM2:瞬时受体电位阳离子通道,亚家族 M,成员 2;ZKSCAN3:具有 KRAB 和 SCAN 结构域 3 的锌指 SQSTM1/p62:sequestome 1;TFEB:转录因子EB;TPCN:二孔通道;TRPM2:瞬时受体电位阳离子通道,亚家族 M,成员 2;ZKSCAN3:具有 KRAB 和 SCAN 结构域 3 的锌指

更新日期:2021-07-27
down
wechat
bug