Arcos Mariella, Goodla Lavanya, Kim Hyeoncheol, Desai Sharina P, Liu Rui, Yin Kunlun, Liu Zhaoli, Martin David R, Xue Xiang
Department of Biochemistry and Molecular Biology, University of New Mexico, Albuquerque, NM, USA.
Department of Molecular Genetics Microbiology, University of New Mexico, Albuquerque, NM, USA.
Autophagy. 2025 Apr;21(4):737-753. doi: 10.1080/15548627.2024.2425594. Epub 2024 Nov 16.
Mitophagy, the process by which cells eliminate damaged mitochondria, is mediated by PINK1 (PTEN induced kinase 1). Our recent research indicates that PINK1 functions as a tumor suppressor in colorectal cancer by regulating cellular metabolism. Interestingly, PINK1 ablation activated the NLRP3 (NLR family pyrin domain containing 3) inflammasome, releasing IL1B (interleukin 1 beta). However, inhibiting the NLRP3-IL1B signaling pathway with an IL1R (interleukin 1 receptor) antagonist or NLRP3 inhibitor did not hinder colon tumor growth after PINK1 loss. To identify druggable targets in PINK1-deficient tumors, ribonucleic acid sequencing analysis was performed on colon tumors from knockout and wild-type mice. Gene Set Enrichment Analysis highlighted the enrichment of iron ion transmembrane transporter activity. Subsequent qualitative polymerase chain reaction and western blot analysis revealed an increase in mitochondrial iron transporters, including mitochondrial calcium uniporter, in PINK1-deficient colon tumor cells and tissues. Live-cell iron staining demonstrated elevated cellular and mitochondrial iron levels in PINK1-deficient cells. Clinically used drugs deferiprone and minocycline reduced mitochondrial iron and superoxide levels, resulting in decreased colon tumor cell growth and . Manipulating the mitochondrial iron uptake protein MCU (mitochondrial calcium uniporter) also affected cell and xenograft tumor growth. This study suggests that therapies aimed at reducing mitochondrial iron levels may effectively inhibit colon tumor growth, particularly in patients with low PINK1 expression.: ANOVA: analysis of variance; APC: adenomatous polyposis coli; cAMP: cyclic adenosine monophosphate; CDX2: caudal type homeobox 2; CGAS: cyclic GMP-AMP synthase; CRC: colorectal cancer; DNA: deoxyribonucleic acid; DFP: deferiprone; DMEM: Dulbecco's modified Eagle medium; DSS: dextran sodium sulfate; ERT2-Cre: Cre recombinase fused to a triple mutant form of the human estrogen receptor; EV: empty vector; GLB: glybenclamide/glyburide; H&E: hematoxylin and eosin; ICP-MS: inductively coupled plasma mass spectrometer; IL1B: interleukin 1 beta; kDa: kilodalton; MCU: mitochondrial calcium uniporter; MKI67: marker of proliferation Ki-67; mRNA: messenger ribonucleic acid; MTT: 3-(4,5-dimethylthiazol-2-Yl)-2,5-diphenyltetrazolium bromide; NLRP3: NLR family pyrin domain containing 3; OE: overexpression; PBS: phosphate-buffered saline; p-CREB: phosphorylated cAMP responsive element binding protein; PINK1: PTEN induced kinase 1; p-PRKAA/AMPK: phosphorylated protein kinase AMP-activated catalytic subunit alpha; qPCR: qualitative polymerase chain reaction; RNA-seq: ribonucleic acid sequencing; ROS: reactive oxygen species; sg: single guide; sh: short hairpin; SLC25A28: solute carrier family 25 member 28; SLC25A37/MFRN: solute carrier family 25 member 37; STING1: stimulator of interferon response cGAMP interactor 1; TP53/p53: tumor protein p53; TUBA: tubulin alpha; µL: microliter; µm: micrometer; µM: micromolar; mm: millimeter.
线粒体自噬是细胞清除受损线粒体的过程，由PINK1（PTEN诱导激酶1）介导。我们最近的研究表明，PINK1在结直肠癌中通过调节细胞代谢发挥肿瘤抑制作用。有趣的是，PINK1缺失激活了NLRP3（含NLR家族pyrin结构域3）炎性小体，释放IL1B（白细胞介素1β）。然而，用IL1R（白细胞介素1受体）拮抗剂或NLRP3抑制剂抑制NLRP3 - IL1B信号通路并不能阻碍PINK1缺失后的结肠肿瘤生长。为了确定PINK1缺陷肿瘤中可成药的靶点，对敲除小鼠和野生型小鼠的结肠肿瘤进行了核糖核酸测序分析。基因集富集分析突出了铁离子跨膜转运蛋白活性的富集。随后的定性聚合酶链反应和蛋白质印迹分析显示，PINK1缺陷的结肠肿瘤细胞和组织中线粒体铁转运蛋白增加，包括线粒体钙单向转运体。活细胞铁染色显示PINK1缺陷细胞中细胞和线粒体铁水平升高。临床使用的药物去铁酮和米诺环素降低了线粒体铁和超氧化物水平，导致结肠肿瘤细胞生长减少。操纵线粒体铁摄取蛋白MCU（线粒体钙单向转运体）也影响细胞和异种移植肿瘤的生长。这项研究表明，旨在降低线粒体铁水平的疗法可能有效抑制结肠肿瘤生长，特别是在PINK1表达低的患者中。：ANOVA：方差分析；APC：腺瘤性息肉病 coli；cAMP：环磷酸腺苷；CDX2：尾型同源盒2；CGAS：环鸟苷酸 - 腺苷酸合酶；CRC：结直肠癌；DNA：脱氧核糖核酸；DFP：去铁酮；DMEM：杜氏改良 Eagle培养基；DSS：葡聚糖硫酸钠；ERT2 - Cre：与人雌激素受体三重突变形式融合的Cre重组酶；EV：空载体；GLB：格列本脲/优降糖；H&E：苏木精和伊红；ICP - MS：电感耦合等离子体质谱仪；IL1B：白细胞介素1β；kDa：千道尔顿；MCU：线粒体钙单向转运体；MKI67：增殖标志物Ki - 67；mRNA：信使核糖核酸；MTT：3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐；NLRP3：含NLR家族pyrin结构域3；OE：过表达；PBS：磷酸盐缓冲盐水；p - CREB：磷酸化的环磷酸腺苷反应元件结合蛋白；PINK1：PTEN诱导激酶1；p - PRKAA/AMPK：磷酸化的蛋白激酶AMP激活催化亚基α；qPCR：定性聚合酶链反应；RNA - seq：核糖核酸测序；ROS：活性氧；sg：单向导；sh：短发夹；SLC25A28：溶质载体家族25成员28；SLC25A37/MFRN：溶质载体家族25成员37；STING1：干扰素反应cGAMP相互作用蛋白1；TP53/p53：肿瘤蛋白p53；TUBA：微管蛋白α；µL：微升；µm：微米；µM：微摩尔；mm：毫米。
