LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancer
Abstract Background Lemur Tail Kinase 3 (LMTK3) promotes cell proliferation, invasiveness and therapy resistance, and its expression correlates with poor survival in several different malignancies, including breast cancer. Crosstalk through extracellular vesicles (EVs) is an increasingly appreciated...
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2025-05-01
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| Online Access: | https://doi.org/10.1186/s12943-025-02346-2 |
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| author | Mark Samuels Christos Karakostas Simoni Besta Andrea Lauer Betrán Katerina Tsilingiri Charlotte Turner Reza Shirazi Nia Niloufar Poudine Richard Goodyear William Jones Apostolos Klinakis Georgios Giamas |
| author_facet | Mark Samuels Christos Karakostas Simoni Besta Andrea Lauer Betrán Katerina Tsilingiri Charlotte Turner Reza Shirazi Nia Niloufar Poudine Richard Goodyear William Jones Apostolos Klinakis Georgios Giamas |
| author_sort | Mark Samuels |
| collection | DOAJ |
| description | Abstract Background Lemur Tail Kinase 3 (LMTK3) promotes cell proliferation, invasiveness and therapy resistance, and its expression correlates with poor survival in several different malignancies, including breast cancer. Crosstalk through extracellular vesicles (EVs) is an increasingly appreciated mechanism of cell communication within the tumour immune microenvironment, which contributes to different aspects of cancer progression and plays a pivotal role in shaping tumour fate. Methods Nanoparticle tracking analysis and transmission electron microscopy were used to study the effects of LMTK3 on EV size, while single particle interferometry allowed us to examine LMTK3-dependent effects on the subpopulation distribution of EVs. Quantitative mass spectrometry was used to profile LMTK3-dependent proteomics changes in breast cancer-derived EVs. Bioinformatics analysis of clinical data along with in vitro and cell-based assays were implemented to explore the effects of LMTK3-dependent EV protein cargo on the tumour immune microenvironment. To elucidate the mechanism through which LMTK3 impacts endosomal trafficking and regulates EV biogenesis, we used a variety of approaches, including in vitro kinase assays, confocal and electron microscopy, as well as in vivo subcutaneous and orthotopic breast cancer mouse models. Results Here, we report that LMTK3 increases the average size of EVs, modulates immunoregulatory EV proteomic cargo and alters the subpopulation distribution of EVs released by breast cancer cells. Mechanistically, we provide evidence that LMTK3 phosphorylates Rab7, a key regulator of multivesicular body (MVB) trafficking, thereby reducing the fusion of MVBs with lysosomes and subsequent degradation of intralumenal vesicles, resulting in altered EV release. Moreover, LMTK3 causes increased packaging of phosphoserine aminotransferase 1 (PSAT1) in EVs, leading to a paracrine upregulation of phosphoglycerate dehydrogenase (PHGDH) in monocytes when these EVs are taken up. PSAT1 and PHGDH play key roles in the serine biosynthesis pathway, which is closely linked to cancer progression and regulation of monocyte behaviour. LMTK3 EV-induced elevated PHGDH expression in monocytes reduces their infiltration into breast cancer 3D spheroids and in vivo breast cancer mouse models. Furthermore, these infiltrating monocytes preferentially differentiate into pro-tumourigenic M2-like macrophages. Additional breast cancer mouse studies highlight the contribution of LMTK3-dependent EVs in the observed immunosuppressive macrophage phenotype. Finally, in vitro experiments show that pharmacological inhibition of LMTK3 reverses the pro-tumourigenic and immunomodulatory effects mediated by EVs derived from LMTK3 overexpressing cells. Conclusion Overall, this study advances our knowledge on the mechanisms of EV biogenesis and highlights a novel oncogenic role of LMTK3 in the breast TME, further supporting it as a target for cancer therapy. |
| format | Article |
| id | doaj-art-6385a63d06ab46feb104afa55a80327d |
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| publishDate | 2025-05-01 |
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| series | Molecular Cancer |
| spelling | doaj-art-6385a63d06ab46feb104afa55a80327d2025-08-20T03:08:40ZengBMCMolecular Cancer1476-45982025-05-0124112310.1186/s12943-025-02346-2LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancerMark Samuels0Christos Karakostas1Simoni Besta2Andrea Lauer Betrán3Katerina Tsilingiri4Charlotte Turner5Reza Shirazi Nia6Niloufar Poudine7Richard Goodyear8William Jones9Apostolos Klinakis10Georgios Giamas11International Oncology Institute, The First Affiliated Hospital of Zhejiang Chinese Medical University. Oncology department of the first affiliated Hospital of Zhejiang Chinese Medical UniversityCenter of Basic Research Biomedical Research Foundation of the Academy of AthensInternational Oncology Institute, The First Affiliated Hospital of Zhejiang Chinese Medical University. Oncology department of the first affiliated Hospital of Zhejiang Chinese Medical UniversityDepartment of Biochemistry and Biomedicine, School of Life Sciences, University of Sussex, JMS BuildingCenter of Basic Research Biomedical Research Foundation of the Academy of AthensDepartment of Biochemistry and Biomedicine, School of Life Sciences, University of Sussex, JMS BuildingInternational Oncology Institute, The First Affiliated Hospital of Zhejiang Chinese Medical University. Oncology department of the first affiliated Hospital of Zhejiang Chinese Medical UniversityInternational Oncology Institute, The First Affiliated Hospital of Zhejiang Chinese Medical University. Oncology department of the first affiliated Hospital of Zhejiang Chinese Medical UniversitySussex Neuroscience, School of Life Sciences, University of SussexDepartment of Biochemistry and Biomedicine, School of Life Sciences, University of Sussex, JMS BuildingCenter of Basic Research Biomedical Research Foundation of the Academy of AthensInternational Oncology Institute, The First Affiliated Hospital of Zhejiang Chinese Medical University. Oncology department of the first affiliated Hospital of Zhejiang Chinese Medical UniversityAbstract Background Lemur Tail Kinase 3 (LMTK3) promotes cell proliferation, invasiveness and therapy resistance, and its expression correlates with poor survival in several different malignancies, including breast cancer. Crosstalk through extracellular vesicles (EVs) is an increasingly appreciated mechanism of cell communication within the tumour immune microenvironment, which contributes to different aspects of cancer progression and plays a pivotal role in shaping tumour fate. Methods Nanoparticle tracking analysis and transmission electron microscopy were used to study the effects of LMTK3 on EV size, while single particle interferometry allowed us to examine LMTK3-dependent effects on the subpopulation distribution of EVs. Quantitative mass spectrometry was used to profile LMTK3-dependent proteomics changes in breast cancer-derived EVs. Bioinformatics analysis of clinical data along with in vitro and cell-based assays were implemented to explore the effects of LMTK3-dependent EV protein cargo on the tumour immune microenvironment. To elucidate the mechanism through which LMTK3 impacts endosomal trafficking and regulates EV biogenesis, we used a variety of approaches, including in vitro kinase assays, confocal and electron microscopy, as well as in vivo subcutaneous and orthotopic breast cancer mouse models. Results Here, we report that LMTK3 increases the average size of EVs, modulates immunoregulatory EV proteomic cargo and alters the subpopulation distribution of EVs released by breast cancer cells. Mechanistically, we provide evidence that LMTK3 phosphorylates Rab7, a key regulator of multivesicular body (MVB) trafficking, thereby reducing the fusion of MVBs with lysosomes and subsequent degradation of intralumenal vesicles, resulting in altered EV release. Moreover, LMTK3 causes increased packaging of phosphoserine aminotransferase 1 (PSAT1) in EVs, leading to a paracrine upregulation of phosphoglycerate dehydrogenase (PHGDH) in monocytes when these EVs are taken up. PSAT1 and PHGDH play key roles in the serine biosynthesis pathway, which is closely linked to cancer progression and regulation of monocyte behaviour. LMTK3 EV-induced elevated PHGDH expression in monocytes reduces their infiltration into breast cancer 3D spheroids and in vivo breast cancer mouse models. Furthermore, these infiltrating monocytes preferentially differentiate into pro-tumourigenic M2-like macrophages. Additional breast cancer mouse studies highlight the contribution of LMTK3-dependent EVs in the observed immunosuppressive macrophage phenotype. Finally, in vitro experiments show that pharmacological inhibition of LMTK3 reverses the pro-tumourigenic and immunomodulatory effects mediated by EVs derived from LMTK3 overexpressing cells. Conclusion Overall, this study advances our knowledge on the mechanisms of EV biogenesis and highlights a novel oncogenic role of LMTK3 in the breast TME, further supporting it as a target for cancer therapy.https://doi.org/10.1186/s12943-025-02346-2LMTK3Rab7Extracellular VesiclesBreast CancerMonocytesMacrophages |
| spellingShingle | Mark Samuels Christos Karakostas Simoni Besta Andrea Lauer Betrán Katerina Tsilingiri Charlotte Turner Reza Shirazi Nia Niloufar Poudine Richard Goodyear William Jones Apostolos Klinakis Georgios Giamas LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancer Molecular Cancer LMTK3 Rab7 Extracellular Vesicles Breast Cancer Monocytes Macrophages |
| title | LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancer |
| title_full | LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancer |
| title_fullStr | LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancer |
| title_full_unstemmed | LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancer |
| title_short | LMTK3 regulation of EV biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro-tumourigenic macrophage polarisation in breast cancer |
| title_sort | lmtk3 regulation of ev biogenesis and cargo sorting promotes tumour growth by reducing monocyte infiltration and driving pro tumourigenic macrophage polarisation in breast cancer |
| topic | LMTK3 Rab7 Extracellular Vesicles Breast Cancer Monocytes Macrophages |
| url | https://doi.org/10.1186/s12943-025-02346-2 |
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