ML098 enhances intracellular bacterial clearance in bladder epithelial cells by activating Rab7 and promoting lysosomal biogenesis
DOI:
https://doi.org/10.3329/bjp.v19i4.77475Keywords:
Intracellular bacteria; ML098; Active Rab7; LysosomeAbstract
The aim of this study was to investigate the effect of ML098 on the intra-cellular bacterial count of bladder epithelial cells infected with uropathogenic Escherichia coli. The number of intracellular bacteria were counted after cell lysis present in coated Luria-Bertani agar plates. Lysotracker red was used to detect the numbers of lysosomes in cells. The expression of lysosome associated membrane protein 1 within cells was detected by western blot and q-PCR methods. Rab7 pulldown activation assay was used to detect the expression of intracellular active Rab7. The results showed that ML098 (1 µM) could reduce the number of intracellular bacteria in bladder epithelial cells 6 hours after E. coli infection (p<0.05), increased the number of lysosomes in cells (p<0.05), and increased the expression of intracellular active Rab7 (p<0.05). Therefore, ML098 can promote intracellular bacterial clearance, possibly by increasing the expression of active Rab7 in cells, thereby increasing the number of lysosomes in bladder epithelial cells.
Downloads
72
61
References
Beebout CJ, Robertson GL, Reinfeld BI, Blee AM, Morales GH, Brannon JR, Chazin WJ, Rathmell WK, Rathmell JC, Gama V, Hadjifrangiskou M. Uropathogenic Escherichia coli subverts mitochondrial metabolism to enable intracellular bacterial pathogenesis in urinary tract infection. Nat Microbiol. 2022; 7: 1348-60.
Bhattacharya A, Mukherjee R, Kuncha SK, Brunstein ME, Rathore R, Junek S, Münch C, Dikic I. A lysosome mem-brane regeneration pathway depends on TBC1D15 and autophagic lysosomal reformation proteins. Nat Cell Biol. 2023; 25: 685-98.
Cai CZ, Yang C, Zhuang XX, Yuan NN, Wu MY, Tan JQ, Song JX, Cheung KH, Su H, Wang YT, Tang BS, Behrends C, Durairajan SSK, Yue Z, Li M, Lu JH. NRBF2 is a RAB7 effector required for autophagosome maturation and mediates the association of APP-CTFs with active form of RAB7 for degradation. Autophagy 2021; 17: 1112-30.
Chan H, Li Q, Wang X, Liu WY, Hu W, Zeng J, Xie C, Kwong TNY, Ho IHT, Liu X, Chen H, Yu J, Ko H, Chan RCY, Ip M, Gin T, Cheng ASL, Zhang L, Chan MTV, Wong SH, Wu WKK. Vitamin D3 and carbamazepine protect against Clostridioides difficile infection in mice by restoring macrophage lysosome acidification. Autophagy 2022; 18: 2050-67.
Dong X, He Y, An J, He L, Zheng Y, Wang X, Wang J, Chen S, Zhang Y. Increased apoptosis of gingival epithelium is associated with impaired autophagic flux in medication-related osteonecrosis of the jaw. Autophagy 2023; 19: 2899-911.
Ellis MJ, Lekka C, Holden KL, Tulmin H, Seedat F, O'Brien DP, Dhayal S, Zeissler ML, Knudsen JG, Kessler BM, Morgan NG, Todd JA, Richardson SJ, Stefana MI. Identification of high-performing antibodies for the reliable detection of Tau proteoforms by Western blotting and immunohistochemistry. Acta Neuropathol. 2024; 147: 87.
Engel DR, Wagenlehner FME, Shevchuk O. Scientific advances in understanding the pathogenesis, diagnosis, and prevention of urinary tract infection in the past 10 years. Infect Dis Clin North Am. 2024; 38: 229-40.
Fisher H, Oluboyede Y, Chadwick T, Abdel-Fattah M, Brennand C, Fader M, Harrison S, Hilton P, Larcombe J, Little P, McClurg D, McColl E, N'Dow J, Ternent L, Thiruchelvam N, Timoney A, Vale L, Walton K, von Wilamowitz-Moellendorff A, Wilkinson J, Wood R, Pickard R. Continuous low-dose antibiotic prophylaxis for adults with repeated urinary tract infections (AnTIC): A randomised, open-label trial. Lancet Infect Dis. 2018; 18: 957-68.
Ghosh S, Dellibovi-Ragheb TA, Kerviel A, Pak E, Qiu Q, Fisher M, Takvorian PM, Bleck C, Hsu VW, Fehr AR, Perlman S, Achar SR, Straus MR, Whittaker GR, de Haan CAM, Kehrl J, Altan-Bonnet G, Altan-Bonnet N. β-Coronaviruses use lysosomes for egress instead of the biosynthetic secretory pathway. Cell 2020; 183: 1520-35.
Jin X, Wang K, Wang L, Liu W, Zhang C, Qiu Y, Liu W, Zhang H, Zhang D, Yang Z, Wu T, Li J. RAB7 activity is required for the regulation of mitophagy in oocyte meiosis and oocyte quality control during ovarian aging. Autophagy 2022; 18: 643-60.
Kitagawa K, Shigemura K, Yamamichi F, Alimsardjono L, Rahardjo D, Kuntaman K, Shirakawa T, Fujisawa M. International comparison of causative bacteria and antimicrobial susceptibilities of urinary tract infections between Kobe, Japan, and Surabaya, Indonesia. Jpn J Infect Dis. 2018; 71: 8-13.
Kretschmer-Kazemi Far R, Frank K, Sczakiel G. Sampling, logistics, and analytics of urine for RT-qPCR-based diagnostics. Cancers (Basel). 2021; 13: 4381.
Langemeyer L, Frohlich F, Ungermann C. Rab GTPase function in endosome and lysosome biogenesis. Trends Cell Biol. 2018; 28: 957-70.
Li X, Jiang L, Zhang S, Zhou J, Liu L, Jin C, Sun H, Wang Q, Liu Y, Pang Y. Uropathogenic Escherichia coli subverts host autophagic defenses by stalling preautophagosomal structures to escape lysosome exocytosis. J Infect Dis. 2024; 230: e548-58.
Liu M, Shao J, Zhao Y, Ma B, Ge S. Porphyromonas gingivalis evades immune clearance by regulating lysosome efflux. J Dent Res. 2023; 102: 555-64.
Miao Y, Li G, Zhang X, Xu H, Abraham SN. A TRP channel senses lysosome neutralization by pathogens to trigger their expulsion. Cell 2015; 161: 1306-19.
Pang Y, Cheng Z, Zhang S, Li S, Li X, Li X, Zhang X, Li X, Feng Y, Cui H, Chen Z, Liu L, Li Q, Huang J, Zhang M, Zhu S, Wang L, Feng L. Bladder epithelial cell phosphate trans-porter inhibition protects mice against uropathogenic Escherichia coli infection. Cell Rep. 2022; 39: 110698.
Pillai-Kastoori L, Schutz-Geschwender AR, Harford JA. A systematic approach to quantitative western blot analysis. Anal Biochem. 2020; 593: 113608.
Seaman MNJ, Mukadam AS, Breusegem SY. Inhibition of TBC1D5 activates Rab7a and can enhance the function of the retromer cargo-selective complex. J Cell Sci. 2018; 131: jcs217398.
Vanlandingham PA, Ceresa BP. Rab7 regulates late endocytic trafficking downstream of multivesicular body biogenesis and cargo sequestration. J Biol Chem. 2009; 284: 12110-24.
Wang C, Bauckman KA, Ross ASB, Symington JW, Ligon MM, Scholtes G, Kumar A, Chang HW, Twentyman J, Fashemi BE, Xavier RJ, Mysorekar IU. A non-canonical autophagy-dependent role of the ATG16L1T300A variant in urothelial vesicular trafficking and uropathogenic Escherichia coli persistence. Autophagy 2019; 15: 527-42.
Wong YC, Ysselstein D, Krainc D. Mitochondria-lysosome contacts regulate mitochondrial fission via RAB7 GTP hydrolysis. Nature 2018; 554: 382-86.
Xu Y, Zhou P, Cheng S, Lu Q, Nowak K, Hopp AK, Li L, Shi X, Zhou Z, Gao W, Li D, He H, Liu X, Ding J, Hottiger MO, Shao F. A bacterial effector reveals the V-ATPase-ATG16L1 axis that initiates xenophagy. Cell 2019; 178:552-66.
Yim WWY, Mizushima N. Lysosome biology in autophagy. Cell Discov. 2020; 6: 6.
Yin H, Zhu J, Jiang Y, Mao Y, Tang C, Cao H, Huang Y, Zhu H, Luo J, Jin Q, Jin Q. Shionone relieves urinary tract infections by removing bacteria from bladder epithelial cells. Cell Microbiol. 2023; 2023.
Zhao YG, Codogno P, Zhang H. Machinery, regulation and pathophysiological implications of autophagosome maturation. Nat Rev Mol Cell Biol. 2021; 22: 733-50.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Hao Yin, Jiaoli Zhu, Jiamin Yin, Yi Xue, Xin Wang
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
