Top

Published in:

Open Access 01-12-2022 | Research

RAB4A GTPase regulates epithelial-to-mesenchymal transition by modulating RAC1 activation

Authors: Subbulakshmi Karthikeyan, Patrick J. Casey, Mei Wang

Published in: Breast Cancer Research | Issue 1/2022

Abstract

Epithelial-to-mesenchymal transition (EMT) is a critical underpinning process for cancer progression, recurrence and resistance to drug treatment. Identification of new regulators of EMT could lead to the development of effective therapies to improve the outcome of advanced cancers. In the current study we discovered, using a variety of in vitro and in vivo approaches, that RAB4A function is essential for EMT and related manifestation of stemness and invasive properties. Consistently, RAB4A suppression abolished the cancer cells’ self-renewal and tumor forming ability. In terms of downstream signaling, we found that RAB4A regulation of EMT is achieved through its control of activation of the RAC1 GTPase. Introducing activated RAC1 efficiently rescued EMT gene expression, invasion and tumor formation suppressed by RAB4A knockdown in both the in vitro and in vivo cancer models. In summary, this study identifies a RAB4A-RAC1 signaling axis as a key regulatory mechanism for the process of EMT and cancer progression and suggests a potential therapeutic approach to controlling these processes.

Available only for authorised users

Fares J, Fares MY, Khachfe HH, Salhab HA, Fares Y. Molecular principles of metastasis: a hallmark of cancer revisited. Signal Transduct Target Ther. 2020;5(1):28. https://doi.org/10.1038/s41392-020-0134-x.CrossRefPubMedPubMedCentral

Steeg PS. Tumor metastasis: mechanistic insights and clinical challenges. Nat Med. 2006;12(8):895–904. https://doi.org/10.1038/nm1469.CrossRefPubMed

Mani SA, Guo W, Liao MJ, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells. Cell. 2008;133(4):704–15. https://doi.org/10.1016/j.cell.2008.03.027.CrossRefPubMedPubMedCentral

Hanahan D, Weinberg RA. Hallmarks of cancer: the next generation. Cell. 2011;144(5):646–74. https://doi.org/10.1016/j.cell.2011.02.013.CrossRefPubMed

Fouad YA, Aanei C. Revisiting the hallmarks of cancer. Am J Cancer Res. 2017;7(5):1016–36.PubMedPubMedCentral

Weidenfeld K, Barkan D. EMT and Stemness in Tumor Dormancy and Outgrowth: Are They Intertwined Processes? Front Oncol. 2018;8:381. https://doi.org/10.3389/fonc.2018.00381.CrossRefPubMedPubMedCentral

Wilson MM, Weinberg RA, Lees JA, Guen VJ. Emerging Mechanisms by which EMT Programs Control Stemness. Trends Cancer. 2020;6(9):775–80. https://doi.org/10.1016/j.trecan.2020.03.011.CrossRefPubMed

Kalluri R. EMT: when epithelial cells decide to become mesenchymal-like cells. J Clin Invest. 2009;119(6):1417–9. https://doi.org/10.1172/JCI39675.CrossRefPubMedPubMedCentral

Stenmark H. Rab GTPases as coordinators of vesicle traffic. Nat Rev Mol Cell Biol. 2009;10(8):513–25. https://doi.org/10.1038/nrm2728.CrossRefPubMed

10.

Seabra MC, Mules EH, Hume AN. Rab GTPases, intracellular traffic and disease. Trends Mol Med. 2002;8(1):23–30. https://doi.org/10.1016/s1471-4914(01)02227-4.CrossRefPubMed

11.

van der Sluijs P, Hull M, Webster P, Male P, Goud B, Mellman I. The small GTP-binding protein rab4 controls an early sorting event on the endocytic pathway. Cell. 1992;70(5):729–40.CrossRefPubMed

12.

Goueli BS, Powell MB, Finger EC, Pfeffer SR. TBC1D16 is a Rab4A GTPase activating protein that regulates receptor recycling and EGF receptor signaling. Proc Natl Acad Sci U S A. 2012;109(39):15787–92. https://doi.org/10.1073/pnas.1204540109.CrossRefPubMedPubMedCentral

13.

Hellberg C, Schmees C, Karlsson S, Ahgren A, Heldin CH. Activation of protein kinase C alpha is necessary for sorting the PDGF beta-receptor to Rab4a-dependent recycling. Mol Biol Cell. 2009;20(12):2856–63. https://doi.org/10.1091/mbc.E08-12-1228.CrossRefPubMedPubMedCentral

14.

Shin S, Wolgamott L, Yoon SO. Integrin trafficking and tumor progression. Int J Cell Biol. 2012;2012:516789. https://doi.org/10.1155/2012/516789.CrossRefPubMed

15.

Do MT, Chai TF, Casey PJ, Wang M. Isoprenylcysteine carboxylmethyltransferase function is essential for RAB4A-mediated integrin beta3 recycling, cell migration and cancer metastasis. Oncogene. 2017;36(41):5757–67. https://doi.org/10.1038/onc.2017.183.CrossRefPubMedPubMedCentral

16.

Cerami E, Gao J, Dogrusoz U, et al. The cBio cancer genomics portal: an open platform for exploring multidimensional cancer genomics data. Cancer Discov. 2012;2(5):401–4. https://doi.org/10.1158/2159-8290.CD-12-0095.CrossRefPubMed

17.

Gao J, Aksoy BA, Dogrusoz U, et al. Integrative analysis of complex cancer genomics and clinical profiles using the cBioPortal. Sci Signal. 2013;6(269):1. https://doi.org/10.1126/scisignal.2004088.CrossRef

18.

Frittoli E, Palamidessi A, Marighetti P, et al. A RAB5/RAB4 recycling circuitry induces a proteolytic invasive program and promotes tumor dissemination. J Cell Biol. 2014;206(2):307–28. https://doi.org/10.1083/jcb.201403127.CrossRefPubMedPubMedCentral

19.

Shin S, Buel GR, Nagiec MJ, et al. ERK2 regulates epithelial-to-mesenchymal plasticity through DOCK10-dependent Rac1/FoxO1 activation. Proc Natl Acad Sci U S A. 2019;116(8):2967–76. https://doi.org/10.1073/pnas.1811923116.CrossRefPubMedPubMedCentral

20.

Lopez-Haber C, Barrio-Real L, Casado-Medrano V, Kazanietz MG. Heregulin/ErbB3 signaling enhances CXCR4-driven Rac1 activation and breast cancer cell motility via hypoxia-inducible factor 1alpha. Mol Cell Biol. 2016;36(15):2011–26. https://doi.org/10.1128/MCB.00180-16.CrossRefPubMedPubMedCentral

21.

Jamieson C, Lui C, Brocardo MG, Martino-Echarri E, Henderson BR. Rac1 augments Wnt signaling by stimulating beta-catenin-lymphoid enhancer factor-1 complex assembly independent of beta-catenin nuclear import. J Cell Sci. 2015;128(21):3933–46. https://doi.org/10.1242/jcs.167742.CrossRefPubMedPubMedCentral

22.

del Pulgar TG, Benitah SA, Valeron PF, Espina C, Lacal JC. Rho GTPase expression in tumourigenesis: evidence for a significant link. BioEssays. 2005;27(6):602–13. https://doi.org/10.1002/bies.20238.CrossRef

23.

Aznar S, Fernandez-Valeron P, Espina C, Lacal JC. Rho GTPases: potential candidates for anticancer therapy. Cancer Lett. 2004;206(2):181–91. https://doi.org/10.1016/j.canlet.2003.08.035.CrossRefPubMed

24.

Cushman I, Casey PJ. Role of isoprenylcysteine carboxylmethyltransferase-catalyzed methylation in Rho function and migration. J Biol Chem. 2009;284(41):27964–73. https://doi.org/10.1074/jbc.M109.025296.CrossRefPubMedPubMedCentral

25.

Teh JT, Zhu WL, Ilkayeva OR, et al. Isoprenylcysteine carboxylmethyltransferase regulates mitochondrial respiration and cancer cell metabolism. Oncogene. 2015;34(25):3296–304. https://doi.org/10.1038/onc.2014.260.CrossRefPubMed

26.

Manu KA, Chai TF, Teh JT, Zhu WL, Casey PJ, Wang M. Inhibition of isoprenylcysteine carboxylmethyltransferase induces cell-cycle arrest and apoptosis through p21 and p21-regulated BNIP3 induction in pancreatic cancer. Mol Cancer Ther. 2017;16(5):914–23. https://doi.org/10.1158/1535-7163.MCT-16-0703.CrossRefPubMed

27.

Wang M, Tan W, Zhou J, et al. A small molecule inhibitor of isoprenylcysteine carboxymethyltransferase induces autophagic cell death in PC3 prostate cancer cells. J Biol Chem. 2008;283(27):18678–84. https://doi.org/10.1074/jbc.M801855200.CrossRefPubMed

28.

Wang M, Hossain MS, Tan W, et al. Inhibition of isoprenylcysteine carboxylmethyltransferase induces autophagic-dependent apoptosis and impairs tumor growth. Oncogene. 2010;29(35):4959–70. https://doi.org/10.1038/onc.2010.247.CrossRefPubMed

29.

Karthikeyan S, Lantvit DD, Chae DH, Burdette JE. Cadherin-6 type 2, K-cadherin (CDH6) is regulated by mutant p53 in the fallopian tube but is not expressed in the ovarian surface. Oncotarget. 2016;7(43):69871–82. https://doi.org/10.18632/oncotarget.11499.CrossRefPubMedPubMedCentral

30.

Zhu WL, Hossain MS, Guo DY, et al. A role for Rac3 GTPase in the regulation of autophagy. J Biol Chem. 2011;286(40):35291–8. https://doi.org/10.1074/jbc.M111.280990.CrossRefPubMedPubMedCentral

31.

Burleigh A, McKinney S, Brimhall J, et al. A co-culture genome-wide RNAi screen with mammary epithelial cells reveals transmembrane signals required for growth and differentiation. Breast Cancer Res. 2015;17:4. https://doi.org/10.1186/s13058-014-0510-y.CrossRefPubMedPubMedCentral

32.

Chai TF, Manu KA, Casey PJ, Wang M. Isoprenylcysteine carboxylmethyltransferase is required for the impact of mutant KRAS on TAZ protein level and cancer cell self-renewal. Oncogene. 2020;39(31):5373–89. https://doi.org/10.1038/s41388-020-1364-7.CrossRefPubMedPubMedCentral

33.

Chia CY, Kumari U, Casey PJ. Breast cancer cell invasion mediated by Galpha12 signaling involves expression of interleukins-6 and -8, and matrix metalloproteinase-2. J Mol Signal. 2014;9:6. https://doi.org/10.1186/1750-2187-9-6.CrossRefPubMedPubMedCentral

34.

Ellerbroek SM, Wu YI, Overall CM, Stack MS. Functional interplay between type I collagen and cell surface matrix metalloproteinase activity. J Biol Chem. 2001;276(27):24833–42. https://doi.org/10.1074/jbc.M005631200.CrossRefPubMed

35.

Kelly P, Moeller BJ, Juneja J, et al. The G12 family of heterotrimeric G proteins promotes breast cancer invasion and metastasis. Proc Natl Acad Sci U S A. 2006;103(21):8173–8. https://doi.org/10.1073/pnas.0510254103.CrossRefPubMedPubMedCentral

36.

Lau HY, Tang J, Casey PJ, Wang M. Evaluating the epithelial-mesenchymal program in human breast epithelial cells cultured in soft agar using a novel macromolecule extraction protocol. Cancers (Basel). 2021. https://doi.org/10.3390/cancers13040807.CrossRefPubMedCentral

37.

Karthikeyan S, Russo A, Dean M, Lantvit DD, Endsley M, Burdette JE. Prolactin signaling drives tumorigenesis in human high grade serous ovarian cancer cells and in a spontaneous fallopian tube derived model. Cancer Lett. 2018;433:221–31. https://doi.org/10.1016/j.canlet.2018.07.003.CrossRefPubMedPubMedCentral

38.

Russo A, Czarnecki AA, Dean M, et al. PTEN loss in the fallopian tube induces hyperplasia and ovarian tumor formation. Oncogene. 2018;37(15):1976–90. https://doi.org/10.1038/s41388-017-0097-8.CrossRefPubMedPubMedCentral

39.

Jahn SC, Law ME, Corsino PE, et al. An in vivo model of epithelial to mesenchymal transition reveals a mitogenic switch. Cancer Lett. 2012;326(2):183–90. https://doi.org/10.1016/j.canlet.2012.08.013.CrossRefPubMedPubMedCentral

40.

Jung AR, Jung CH, Noh JK, Lee YC, Eun YG. Epithelial-mesenchymal transition gene signature is associated with prognosis and tumor microenvironment in head and neck squamous cell carcinoma. Sci Rep. 2020;10(1):3652. https://doi.org/10.1038/s41598-020-60707-x.CrossRefPubMedPubMedCentral

41.

Stemmler MP, Eccles RL, Brabletz S, Brabletz T. Non-redundant functions of EMT transcription factors. Nat Cell Biol. 2019;21(1):102–12. https://doi.org/10.1038/s41556-018-0196-y.CrossRefPubMed

42.

Thompson JC, Hwang WT, Davis C, et al. Gene signatures of tumor inflammation and epithelial-to-mesenchymal transition (EMT) predict responses to immune checkpoint blockade in lung cancer with high accuracy. Lung Cancer. 2020;139:1–8. https://doi.org/10.1016/j.lungcan.2019.10.012.CrossRefPubMed

43.

Marston DJ, Anderson KL, Swift MF, et al. High Rac1 activity is functionally translated into cytosolic structures with unique nanoscale cytoskeletal architecture. Proc Natl Acad Sci U S A. 2019;116(4):1267–72. https://doi.org/10.1073/pnas.1808830116.CrossRefPubMedPubMedCentral

44.

Papaharalambus C, Sajjad W, Syed A, et al. Tumor necrosis factor alpha stimulation of Rac1 activity. Role of isoprenylcysteine carboxylmethyltransferase. J Biol Chem. 2005;280(19):18790–6. https://doi.org/10.1074/jbc.M410081200.CrossRefPubMed

45.

Mehlen P, Puisieux A. Metastasis: a question of life or death. Nat Rev Cancer. 2006;6(6):449–58. https://doi.org/10.1038/nrc1886.CrossRefPubMed

46.

Mittal V. Epithelial Mesenchymal Transition in Tumor Metastasis. Annu Rev Pathol. 2018;13:395–412. https://doi.org/10.1146/annurev-pathol-020117-043854.CrossRefPubMed

Title: RAB4A GTPase regulates epithelial-to-mesenchymal transition by modulating RAC1 activation
Authors: Subbulakshmi Karthikeyan
Patrick J. Casey
Mei Wang
Publication date: 01-12-2022
Publisher: BioMed Central
Published in: Breast Cancer Research / Issue 1/2022
Electronic ISSN: 1465-542X
DOI: https://doi.org/10.1186/s13058-022-01564-6

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Please log in to get access to this content

Other articles of this Issue 1/2022

SETDB1 interactions with PELP1 contributes to breast cancer endocrine therapy resistance

LncRNA IPW inhibits growth of ductal carcinoma in situ by downregulating ID2 through miR-29c

Quantitative glycoproteomics analysis identifies novel FUT8 targets and signaling networks critical for breast cancer cell invasiveness

Elevated NRAS expression during DCIS is a potential driver for progression to basal-like properties and local invasiveness

Artificial intelligence in mammographic phenotyping of breast cancer risk: a narrative review

The impact of breast density notification on rescreening rates within a population-based mammographic screening program

Keynote webinar | Spotlight on antibody–drug conjugates in cancer