Heterogeneity at the invasion front of triple negative breast cancer cells

Yong, Koh Meng Aw; Ulintz, Peter; Cáceres, Sara; Cheng, Xu; Bao, Liwei; Wu, Zhifen; Jiagge, Evelyn; Merajver, Sofía D.

doi:10.1038/s41598-020-62516-8

Cited by 17 publications

(12 citation statements)

References 27 publications

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“…This model was further tested in the patient-derived early passage triple negative breast cancer cell line VARI068 with robust EGFR expression. 40,41 VARI068 cells exhibit approximately 2-fold upregulation of Med25 relative to normal-like non-tumorigenic MCF10A cells. Treatment of VARI068 cells with NA blocks the Med25•ETV5 complex (Figure 4D).…”

Section: Resultsmentioning

confidence: 96%

Norstictic acid is a selective allosteric transcriptional regulator

Garlick

Sturlis

Bruno

et al. 2021

Preprint

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Inhibitors of transcriptional protein-protein interactions (PPIs) have high value both as tools and for therapeutic applications. The PPI network mediated by the transcriptional coactivator Med25, for example, regulates stress-response and motility pathways and dysregulation of the PPI networks contributes to oncogenesis and metastasis. The canonical transcription factor binding sites within Med25 are large (~900 square angstroms) and have little topology, and thus do not present an array of attractive small-molecule binding sites for inhibitor discovery. Here we demonstrate that the depsidone natural product norstictic acid functions through an alternative binding site to block Med25-transcriptional activator PPIs in vitro and in cell culture. Norstictic acid targets a binding site comprised of a highly dynamic loop flanking one canonical binding surface and in doing so, it both orthosterically and allosterically alters Med25-driven transcription in a patient-derived model of triple negative breast cancer. These results highlight the potential of Med25 as a therapeutic target as well as the inhibitor discovery opportunities presented by structurally dynamic loops within otherwise challenging proteins.

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Section: Resultsmentioning

confidence: 96%

Norstictic acid is a selective allosteric transcriptional regulator

Garlick

Sturlis

Bruno

et al. 2021

Preprint

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“…In support of our observation, several reports have suggested that the KRT14 + cells have metastatic advantage in compared to KRT14 - cells in TNBC heterogeneous population and regulate the function of several genes involved in different stages of metastatic cascade. (Cheung et al, 2016; Granit et al, 2018; Yong et al, 2020). Further, KRT14 acts as a stem cell for the natural or injury induced bladder regeneration and typically lead the origin of bladder cancer (Papafotiou et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

H3K27me3 mediated KRT14 upregulation promotes TNBC peritoneal metastasis

Verma

Singh

Nengroo

et al. 2021

Preprint

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Triple Negative Breast Cancer (TNBC) is known to have poor prognosis and adverse clinical outcome among all breast cancer subtypes due to the absence of available targeted therapy for it. Emerging literature indicates that epigenetic reprogramming is now appreciated as a driving force for TNBC pathophysiology. High expression of epigenetic modulator EZH2 (Enhancer of zeste homolog 2) has been shown to correlate with TNBC poor prognosis but the contribution of EZH2 catalytic (H3K27me3) versus non-catalytic EZH2 (NC-EZH2) function in TNBC growth and progression remains elusive. In the process of dissecting the impact of H3K27me3 versus NC-EZH2 function in TNBC pathogenesis, we reveal that selective hyper-activation of H3K27me3 over NC-EZH2 not only promotes TNBC metastasis but also alters the metastatic landscape of TNBC. Using extensive in- vivo live animal imaging, we present conclusive evidence that peritoneal metastasis, particularly splenic metastasis of TNBC is governed by H3K27me3. Transcriptome analyses of hyperactive H3K27me3 cells lead us to discover Cytokeratin-14 (KRT14) as a new target of H3K27me3. Unlike classical H3K27me3 mediated suppression of gene expression, here; we observe that H3K27me3 enhances KRT14 transcription by attenuating the binding of transcriptional repressor Sp1 to its promoter. Further, loss of KRT14 significantly reduces TNBC migration, invasion and splenic metastasis. Finally, genetic ablation of EZH2 or pharmacological inhibition of EZH2 catalytic function by FDA approved drug tazemetostat (EPZ6438) robustly inhibits TNBC peritoneal metastasis. Altogether, our preclinical findings posit a rational insight for the clinical development of H3K27me3 inhibitor like tazemetostat as a targeted therapy against TNBC.

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“…130-104-694) was used to enrich the digest for PCa human cells by depleting murine cell populations including fibroblasts, immune cells, and endothelial cells from the PDX digests. Previous studies have used this kit for similar applications 31 , 32 . Approximately 5 × 10 7 dissociated PDX cells were resuspended with 80 µL of cold separation buffer (0.5% BSA, 100 U/mL PenStrep in PBS) and 20 µL of Mouse Cell Depletion cocktail, and then incubated for 15 min at 4 °C, with occasional agitation.…”

Section: Methodsmentioning

confidence: 99%

Using the Microwell-mesh to culture microtissues in vitro and as a carrier to implant microtissues in vivo into mice

Monterosso

Futrega

Lott

et al. 2021

Sci Rep

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Prostate cancer (PCa) patient-derived xenografts (PDXs) are commonly propagated by serial transplantation of “pieces” of tumour in mice, but the cellular composition of pieces is not standardised. Herein, we optimised a microwell platform, the Microwell-mesh, to aggregate precise numbers of cells into arrays of microtissues, and then implanted the Microwell-mesh into NOD-scid IL2γ−/− (NSG) mice to study microtissue growth. First, mesh pore size was optimised using microtissues assembled from bone marrow-derived stromal cells, with mesh opening dimensions of 100×100 μm achieving superior microtissue vascularisation relative to mesh with 36×36 μm mesh openings. The optimised Microwell-mesh was used to assemble and implant PCa cell microtissue arrays (hereafter microtissues formed from cancer cells are referred to as microtumours) into mice. PCa cells were enriched from three different PDX lines, LuCaP35, LuCaP141, and BM18. 3D microtumours showed greater in vitro viability than 2D cultures, but neither proliferated. Microtumours were successfully established in mice 81% (57 of 70), 67% (4 of 6), 76% (19 of 25) for LuCaP35, LuCaP141, and BM18 PCa cells, respectively. Microtumour growth was tracked using live animal imaging for size or bioluminescence signal. If augmented with further imaging advances and cell bar coding, this microtumour model could enable greater resolution of PCa PDX drug response, and lead to the more efficient use of animals. The concept of microtissue assembly in the Microwell-mesh, and implantation in vivo may also have utility in implantation of islets, hair follicles or other organ-specific cells that self-assemble into 3D structures, providing an important bridge between in vitro assembly of mini-organs and in vivo implantation.

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Heterogeneity at the invasion front of triple negative breast cancer cells

Cited by 17 publications

References 27 publications

Norstictic acid is a selective allosteric transcriptional regulator

Norstictic acid is a selective allosteric transcriptional regulator

H3K27me3 mediated KRT14 upregulation promotes TNBC peritoneal metastasis

Using the Microwell-mesh to culture microtissues in vitro and as a carrier to implant microtissues in vivo into mice

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