Optimisation of the Chicken Chorioallantoic Membrane Assay in Uveal Melanoma Research

Sokolenko, Ekaterina; Berchner‐Pfannschmidt, Utta; Ting, Saskia; Schmid, Kurt Werner; Bechrakis, Nikolaos E.; Seitz, Berthold; Tsimpaki, Theodora; Kraemer, Miriam M.; Fiorentzis, Miltiadis

doi:10.3390/pharmaceutics14010013

Cited by 27 publications

(7 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The chicken CAM has been previously used as a successful in vivo system to model UM xenograft growth [ 18 , 31 ]. To test the effect of IRS-1 inhibition on tumor progression in vivo, we engrafted luciferase-tagged 92.1 cells (a UM cell line) on the chicken CAM to generate UM xenografts.…”

Section: Resultsmentioning

confidence: 99%

Targeting IRS-1/2 in Uveal Melanoma Inhibits In Vitro Cell Growth, Survival and Migration, and In Vivo Tumor Growth

Chattopadhyay

Bhattacharya

Roszik

et al. 2022

Cancers

View full text Add to dashboard Cite

Uveal melanoma originating in the eye and metastasizing to the liver is associated with poor prognosis and has only one approved therapeutic option. We hypothesized that liver-borne growth factors may contribute to UM growth. Therefore, we investigated the role of IGF-1/IGF-1R signaling in UM. Here, we found that IRS-1, the insulin receptor substrate, is overexpressed in both UM cells and tumors. Since we previously observed that IGF-1R antibody therapy was not clinically effective in UM, we investigated the potential of NT157, a small molecule inhibitor of IRS-1/2, in blocking this pathway in UM. NT157 treatment of multiple UM cell lines resulted in reduced cell growth and migration and increased apoptosis. This treatment also significantly inhibited UM tumor growth in vivo, in the chicken egg chorioallantoic membrane (CAM) and subcutaneous mouse models, validating the in vitro effect. Mechanistically, through reverse phase protein array (RPPA), we identified significant proteomic changes in the PI3K/AKT pathway, a downstream mediator of IGF-1 signaling, with NT157 treatment. Together, these results suggest that NT157 inhibits cell growth, survival, and migration in vitro, and tumor growth in vivo via inhibiting IGF-1 signaling in UM.

show abstract

Section: Resultsmentioning

confidence: 99%

Targeting IRS-1/2 in Uveal Melanoma Inhibits In Vitro Cell Growth, Survival and Migration, and In Vivo Tumor Growth

Chattopadhyay

Bhattacharya

Roszik

et al. 2022

Cancers

View full text Add to dashboard Cite

show abstract

“…The chicken CAM has been previously used as a successful in vivo system to model UM xenograft growth [15, 29]. To test the effect of IRS-1 inhibition on tumor progression in vivo , we engrafted luciferase-tagged 92.1 cells (a UM cell line) on the chicken CAM to generate UM xenografts.…”

Section: Resultsmentioning

confidence: 99%

Targeting IRS-1/2 in uveal melanoma inhibitsin vitrocell growth, survival and migration, andin vivotumor growth

Chattopadhyay¹,

Bhattacharya²,

Roszik³

et al. 2022

Preprint

View full text Add to dashboard Cite

Uveal melanoma (UM) originating in the eye and metastasizing to the liver is associated with poor prognosis. Here, we investigated whether the IGF-1/IGF-1R signaling axis is involved in UM growth and metastasis. TCGA dataset analysis reveals that UM has high IRS-1 expression, which is the first substrate of IGF-1R. Furthermore, IRS-1 is over-expressed in all UM cell lines tested (relative to non -cancer/normal cells) and in matched eye and liver UM tumors. Therefore, we targeted IRS-1/2 in UM cells as well as UM tumors developed on a chicken egg chorioallantoic membrane (CAM) model, and subcutaneous (subQ) UM tumors grown in mice using NT157, a small molecule inhibitor of IRS-1/2. NT157 treatment in UM cells resulted in reduced cell survival and cell migration, and increased apoptosis. NT157 treatment also significantly inhibited UM tumor growth in the in vivo chicken egg CAM and subQ mouse models, validating the in vitro effect. Moreover, NT157 appears more effective than a monoclonal antibody-based approach to block IGF-1R signaling. Mechanistically, through reverse phase protein array (RPPA) analysis, we identified significant proteomic changes in the PI3K/AKT pathway with NT157 treatment. Together, these results suggest that NT157 inhibits cell survival, migration in vitro and tumor growth in vivo via inhibiting IGF-1 signaling in UM cells.

show abstract

“…Our preclinical data indicate that certain β-blockers such as nebivolol may be suitable for co-adjuvant treatment of UM to support local tumor control and to prevent recurrence or metastasis. Although nebivolol is an already FDA-approved drug, the tumor control potential of nebivolol for UM needs to be further assessed by subsequent in vivo studies using orthotopic mouse models or the chicken chorioallantoic membrane assay, which we have recently optimized for UM 3D tumor spheroids [ 54 ].…”

Section: Discussionmentioning

confidence: 99%

Blockade of ß-Adrenergic Receptors by Nebivolol Enables Tumor Control Potential for Uveal Melanoma in 3D Tumor Spheroids and 2D Cultures

Farhoumand

Liu

Tsimpaki

et al. 2023

IJMS

Self Cite

View full text Add to dashboard Cite

Uveal melanoma (UM) is the most common primary cancer of the eye in adults. A new systemic therapy is needed to reduce the high metastasis and mortality rate. As β-blockers are known to have anti-tumor effects on various cancer entities, this study focuses on investigating the effect of β1-selective blockers atenolol, celiprolol, bisoprolol, metoprolol, esmolol, betaxolol, and in particular, nebivolol on UM. The study was performed on 3D tumor spheroids as well as 2D cell cultures, testing tumor viability, morphological changes, long-term survival, and apoptosis. Flow cytometry revealed the presence of all three β-adrenoceptors with a dominance of β2-receptors on cell surfaces. Among the blockers tested, solely nebivolol concentration-dependently decreased viability and altered 3D tumor spheroid structure. Nebivolol blocked the repopulation of cells spreading from 3D tumor spheroids, indicating a tumor control potential at a concentration of ≥20 µM. Mechanistically, nebivolol induced ATP depletion and caspase-3/7 activity, indicating that mitochondria-dependent signaling is involved. D-nebivolol or nebivolol combined with the β2-antagonist ICI 118.551 displayed the highest anti-tumor effects, suggesting a contribution of both β1- and β2-receptors. Thus, the present study reveals the tumor control potential of nebivolol in UM, which may offer a perspective for co-adjuvant therapy to reduce recurrence or metastasis.

show abstract

Optimisation of the Chicken Chorioallantoic Membrane Assay in Uveal Melanoma Research

Cited by 27 publications

References 23 publications

Targeting IRS-1/2 in Uveal Melanoma Inhibits In Vitro Cell Growth, Survival and Migration, and In Vivo Tumor Growth

Targeting IRS-1/2 in Uveal Melanoma Inhibits In Vitro Cell Growth, Survival and Migration, and In Vivo Tumor Growth

Targeting IRS-1/2 in uveal melanoma inhibitsin vitrocell growth, survival and migration, andin vivotumor growth

Blockade of ß-Adrenergic Receptors by Nebivolol Enables Tumor Control Potential for Uveal Melanoma in 3D Tumor Spheroids and 2D Cultures

Contact Info

Product

Resources

About