The Potential Role of Lysosomal Sequestration in Sunitinib Resistance of Renal Cell Cancer

Azijli, Kaamar; Gotink, Kristy J.; Verheul, Henk M.W.

doi:10.15586/jkcvhl.2015.44

Cited by 22 publications

(19 citation statements)

References 26 publications

(39 reference statements)

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“…Additionally, nuclear translocation of TFEB was observed after MEK inhibition and a corresponding upregulation of its target genes. This may point to the fact that MEK inhibition in PDAC induce lysosomal biogenesis and foster lysosomal drug accumulation and subsequent exocytosis as previously hypothesized 32 . To verify this hypothesis, we isolated lysosomes from trametinib-treated cells and using mass spectrometry,we were able to detect the presence of trametinib in these organelle.…”

Section: Discussionmentioning

confidence: 55%

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TFEB-mediated lysosomal biogenesis and lysosomal drug sequestration confer resistance to MEK inhibition in pancreatic cancer

Zhao

Dierichs

et al. 2020

Cell Death Discov.

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Oncogenic KRAS mutations are encountered in more than 90% of pancreatic ductal adenocarcinomas. MEK inhibition has failed to procure any clinical benefits in mutant RAS-driven cancers including pancreatic ductal adenocarcinoma (PDAC). To identify potential resistance mechanisms underlying MEK inhibitor (MEKi) resistance in PDAC, we investigated lysosomal drug accumulation in PDAC models both in vitro and in vivo. Mouse PDAC models and human PDAC cell lines as well as human PDAC xenografts treated with the MEK inhibitor trametinib or refametinib led to an enhanced expression of lysosomal markers and enrichment of lysosomal gene sets. A time-dependent, increase in lysosomal content was observed upon MEK inhibition. Strikingly, there was a strong activation of lysosomal biogenesis in cell lines of the classical compared to the basal-like molecular subtype. Increase in lysosomal content was associated with nuclear translocation of the Transcription Factor EB (TFEB) and upregulation of TFEB target genes. siRNA-mediated depletion of TFEB led to a decreased lysosomal biogenesis upon MEK inhibition and potentiated sensitivity. Using LC-MS, we show accumulation of MEKi in the lysosomes of treated cells. Therefore, MEK inhibition triggers lysosomal biogenesis and subsequent drug sequestration. Combined targeting of MEK and lysosomal function may improve sensitivity to MEK inhibition in PDAC.

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Section: Discussionmentioning

confidence: 55%

“…Malignant cancer cells may adopt such mechanisms to guard against therapeutic assaults. Indeed, lysosomal drug sequestration has been reported for other anticancer agents 36 and is responsible, at least in part for drug resistance 32,37,38 .…”

Section: Discussionmentioning

confidence: 99%

TFEB-mediated lysosomal biogenesis and lysosomal drug sequestration confer resistance to MEK inhibition in pancreatic cancer

Zhao

Dierichs

et al. 2020

Cell Death Discov.

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“…This effect was however found to be additive at the highest concentration tested, indicating that the mechanism of sunitinib resistance is not related to lysosomal sequestration. Lysosomal sequestering of sunitinib has been associated with resistance by others [7,29,31]. To our knowledge, direct experiments addressing the cytotoxic impact of lysosomal accumulation of sunitinib in resistant cells has only been included in two reports where only modest increase in sunitinib toxicity was reported by combining sunitinib with Leu-Leu-O-Methyl in vitro [29] or chloroquine in vivo in sunitinib-resistant HT-29 xenografts [30].…”

Section: Discussionmentioning

confidence: 99%

“…Lysosomal accumulation of sunitinib has been reported as a mechanism of resistance [7,[28][29][30][31][32] and we tested if PCI could counteract this mechanism by releasing endosomal sunitinib sequestered during long-term exposure. Sunitinib resistant HT-29 cells, HT-29/SR, was generated by continuous exposure to 2 µM.…”

Section: Acquired Resistance and Endo/lysosomal Accumulation Followinmentioning

confidence: 99%

Photochemically-Induced Release of Lysosomal Sequestered Sunitinib: Obstacles for Therapeutic Efficacy

Wong

Berstad

Fremstedal

et al. 2020

Cancers

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Lysosomal accumulation of sunitinib has been suggested as an underlying mechanism of resistance. Here, we investigated if photochemical internalization (PCI), a technology for cytosolic release of drugs entrapped in endosomes and lysosomes, would activate lysosomal sequestered sunitinib. By super-resolution fluorescence microscopy, sunitinib was found to accumulate in the membrane of endo/lysosomal compartments together with the photosensitizer disulfonated tetraphenylchlorin (TPCS2a). Furthermore, the treatment effect was potentiated by PCI in the human HT-29 and the mouse CT26.WT colon cancer cell lines. The cytotoxic outcome of sunitinib-PCI was, however, highly dependent on the treatment protocol. Thus, neoadjuvant PCI inhibited lysosomal accumulation of sunitinib. PCI also inhibited lysosomal sequestering of sunitinib in HT29/SR cells with acquired sunitinib resistance, but did not reverse the resistance. The mechanism of acquired sunitinib resistance in HT29/SR cells was therefore not related to lysosomal sequestering. Sunitinib-PCI was further evaluated on HT-29 xenografts in athymic mice, but was found to induce only a minor effect on tumor growth delay. In immunocompetent mice sunitinib-PCI enhanced areas of treatment-induced necrosis compared to the monotherapy groups. However, the tumor growth was not delayed, and decreased infiltration of CD3-positive T cells was indicated as a possible mechanism behind the failed overall response.

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“…For sunitinib, the lysosomal accumulation has also been demonstrated by using its fluorescent properties which co-localized with the Lysotracker, a marker for lysosomes [7]. Moreover, pre-incubation with Bafilomycin-A prevented trapping of sunitinib in the lysosomes [7,16]. Sunitinib achieves very high cellular concentrations since it has been shown that 90% of accumulated drug is held within lysosomes within the cell structure.…”

Section: Discussionmentioning

confidence: 99%

Subcellular localization of several structurally different tyrosine kinase inhibitors

Honeywell¹,

Hitzerd²,

Kathmann³

et al. 2018

ADMET DMPK

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<p>Protein tyrosine kinases form an important target for a new class of anticancer drugs, the tyrosine kinase inhibitors (TKIs). Recently we demonstrated that sunitinib, an inhibitor of the membrane-associated vascular endothelial growth factor receptor (VEGFR), is trapped in lysosomes which isolates the drug from its intended target. Therefore we investigated whether this also holds for other TKIs, targeted against different protein kinases. For this purpose we used the ProteoExtractR kit, which enables a subcellular extraction separating cellular proteins into four distinct fractions covering the cytosol, membranes and membrane organelles (including lysosomes), nuclear proteins and the cytoskeleton. Since TKIs are 98-100 % protein bound we used this property to study their subcellular distribution and used Caco-2 cells as a model. As expected after 2 hours exposure sunitinib was trapped in cytosol (58 %) and organelles (42 % including lysosomes). Crizotinib, an inhibitor of ALK-EML4, showed a similar distribution. However, erlotinib, an inhibitor of the epidermal growth factor receptor (EGFR) showed a very low cellular accumulation and was limited to the organelle fraction. In contrast, the other EGFR inhibitor, gefitinib was predominantly located in the cytosolic (39 %) and membrane fraction (44 %). Sorafenib, another VEGFR inhibitor was predominantly located in the organelle fraction (85 %) and cytosol (15 %) after 2 hours, while after 24 hours distribution decreased (9.9 fold) with a slight shift. Dasatinib, an inhibitor of BCR-Abl was located only in the cytosol (100 %). In general localization after 24 hours was comparable, albeit several small changes were seen. In conclusion protein fractionation with the ProteoExtractR Subcellular Proteome Extraction kit demonstrated large differences in TKI levels in various cellular organelles, with a pattern in agreement with lysosomal accumulation of sunitinib.</p>

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The Potential Role of Lysosomal Sequestration in Sunitinib Resistance of Renal Cell Cancer

Cited by 22 publications

References 26 publications

TFEB-mediated lysosomal biogenesis and lysosomal drug sequestration confer resistance to MEK inhibition in pancreatic cancer

TFEB-mediated lysosomal biogenesis and lysosomal drug sequestration confer resistance to MEK inhibition in pancreatic cancer

Photochemically-Induced Release of Lysosomal Sequestered Sunitinib: Obstacles for Therapeutic Efficacy

Subcellular localization of several structurally different tyrosine kinase inhibitors

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