Limits...
Marine compounds inhibit growth of multiple myeloma in vitro and in vivo.

Steiner N, Ribatti D, Willenbacher W, Jöhrer K, Kern J, Marinaccio C, Aracil M, García-Fernández LF, Gastl G, Untergasser G, Gunsilius E - Oncotarget (2015)

Bottom Line: We identified a subset of marine compounds with strong anti-myeloma activity in vitro and in vivo.Moreover, some of the compounds inhibited myeloma-related angiogenesis in the in vivo gelatin sponge assay.They merit further drug development to improve treatment options for MM.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Tumor Biology & Angiogenesis, Innsbruck Medical University, Innsbruck, Austria.

ABSTRACT

Purpose: The prognosis of patients with multiple myeloma (MM) is still dismal despite recent improvements achieved by introducing new therapeutic agents. However, there remains an urgent need for progress in myeloma drug development. We here show that novel marine-derived compounds can exert potent anti-myeloma activity.

Experimental design: Nine marine-derived compounds were applied at low nM concentrations (0.1-100 nM) to MM cell lines (OPM-2, NCI-H929, U266, RPMI-8226), to primary human myeloma cells and to peripheral blood mononuclear cells. Apoptosis was determined by flow cytometry. In addition, eGFP-transgenic MM cell lines growing with mesenchymal cells from bone marrow were used to visualize tumors by fluorescence stereomicroscopy. Anti-myelomaactivities were studied in vitro in 3D spheroids and in vivo in myeloma xenografts on chicken embryos. Tumor size was analyzed by measuring GFP content with a GFP ELISA. Anti-angiogenic activities of compounds were tested in an in vivo gelatin sponge assay with conditioned media from primary bone marrow-derived endothelial cells.

Results: We identified a subset of marine compounds with strong anti-myeloma activity in vitro and in vivo. Moreover, some of the compounds inhibited myeloma-related angiogenesis in the in vivo gelatin sponge assay. They merit further drug development to improve treatment options for MM.

No MeSH data available.


Related in: MedlinePlus

3D Multiple Myeloma Model(A) Human multiple myeloma cell lines were lentivirally transfected and selected to stably express eGFP. OPM-2eGFP and RPMI-8226eGFP cells were cultivated in hanging drops with primary human bone marrow mesenchymal cells and collagen type I as extracellular matrix component. Spheroids were incubated with culture medium (co.), and respective concentrations (1-100 nM) of all target compounds effective on primary MM cells. MM cell lines were grown for 72h in spheroids and photographed daily using a stereo-fluorescence microscope. Bars indicate 500 μm. (B) Single spheroids were homogenized in lysis buffer and subsequently measured in a GFP ELISA. GFP concentrations of single spheroids were calculated (n=6, Mean ± SEM). Stars indicate p values <0.05. Con.: control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4480745&req=5

Figure 1: 3D Multiple Myeloma Model(A) Human multiple myeloma cell lines were lentivirally transfected and selected to stably express eGFP. OPM-2eGFP and RPMI-8226eGFP cells were cultivated in hanging drops with primary human bone marrow mesenchymal cells and collagen type I as extracellular matrix component. Spheroids were incubated with culture medium (co.), and respective concentrations (1-100 nM) of all target compounds effective on primary MM cells. MM cell lines were grown for 72h in spheroids and photographed daily using a stereo-fluorescence microscope. Bars indicate 500 μm. (B) Single spheroids were homogenized in lysis buffer and subsequently measured in a GFP ELISA. GFP concentrations of single spheroids were calculated (n=6, Mean ± SEM). Stars indicate p values <0.05. Con.: control.

Mentions: Due to the limitations of culturing primary human MM cells in vitro we established a 3D in vitro culture model of human MM cell lines using a collagen extracellular growth matrix and supportive primary human mesenchymal cells from bone marrow. MM cell lines transgenic for eGFP allow visualization and quantification of MM tumor mass after 3D growth in spheroids (Figure 1A). Both MM cell lines OPM-2eGFP and RPMI-8226eGFP were cultivated for three days in the presence of increasing concentrations (1-100 nM) of the respective target compounds, and tumors were visualized by GFP expression (Figure 1A). All target compounds significantly inhibited MM growth at concentrations of 10 and 100 nM in OPM-2eGFP and RPMI-8226eGFP spheroids.


Marine compounds inhibit growth of multiple myeloma in vitro and in vivo.

Steiner N, Ribatti D, Willenbacher W, Jöhrer K, Kern J, Marinaccio C, Aracil M, García-Fernández LF, Gastl G, Untergasser G, Gunsilius E - Oncotarget (2015)

3D Multiple Myeloma Model(A) Human multiple myeloma cell lines were lentivirally transfected and selected to stably express eGFP. OPM-2eGFP and RPMI-8226eGFP cells were cultivated in hanging drops with primary human bone marrow mesenchymal cells and collagen type I as extracellular matrix component. Spheroids were incubated with culture medium (co.), and respective concentrations (1-100 nM) of all target compounds effective on primary MM cells. MM cell lines were grown for 72h in spheroids and photographed daily using a stereo-fluorescence microscope. Bars indicate 500 μm. (B) Single spheroids were homogenized in lysis buffer and subsequently measured in a GFP ELISA. GFP concentrations of single spheroids were calculated (n=6, Mean ± SEM). Stars indicate p values <0.05. Con.: control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4480745&req=5

Figure 1: 3D Multiple Myeloma Model(A) Human multiple myeloma cell lines were lentivirally transfected and selected to stably express eGFP. OPM-2eGFP and RPMI-8226eGFP cells were cultivated in hanging drops with primary human bone marrow mesenchymal cells and collagen type I as extracellular matrix component. Spheroids were incubated with culture medium (co.), and respective concentrations (1-100 nM) of all target compounds effective on primary MM cells. MM cell lines were grown for 72h in spheroids and photographed daily using a stereo-fluorescence microscope. Bars indicate 500 μm. (B) Single spheroids were homogenized in lysis buffer and subsequently measured in a GFP ELISA. GFP concentrations of single spheroids were calculated (n=6, Mean ± SEM). Stars indicate p values <0.05. Con.: control.
Mentions: Due to the limitations of culturing primary human MM cells in vitro we established a 3D in vitro culture model of human MM cell lines using a collagen extracellular growth matrix and supportive primary human mesenchymal cells from bone marrow. MM cell lines transgenic for eGFP allow visualization and quantification of MM tumor mass after 3D growth in spheroids (Figure 1A). Both MM cell lines OPM-2eGFP and RPMI-8226eGFP were cultivated for three days in the presence of increasing concentrations (1-100 nM) of the respective target compounds, and tumors were visualized by GFP expression (Figure 1A). All target compounds significantly inhibited MM growth at concentrations of 10 and 100 nM in OPM-2eGFP and RPMI-8226eGFP spheroids.

Bottom Line: We identified a subset of marine compounds with strong anti-myeloma activity in vitro and in vivo.Moreover, some of the compounds inhibited myeloma-related angiogenesis in the in vivo gelatin sponge assay.They merit further drug development to improve treatment options for MM.

View Article: PubMed Central - PubMed

Affiliation: Laboratory for Tumor Biology & Angiogenesis, Innsbruck Medical University, Innsbruck, Austria.

ABSTRACT

Purpose: The prognosis of patients with multiple myeloma (MM) is still dismal despite recent improvements achieved by introducing new therapeutic agents. However, there remains an urgent need for progress in myeloma drug development. We here show that novel marine-derived compounds can exert potent anti-myeloma activity.

Experimental design: Nine marine-derived compounds were applied at low nM concentrations (0.1-100 nM) to MM cell lines (OPM-2, NCI-H929, U266, RPMI-8226), to primary human myeloma cells and to peripheral blood mononuclear cells. Apoptosis was determined by flow cytometry. In addition, eGFP-transgenic MM cell lines growing with mesenchymal cells from bone marrow were used to visualize tumors by fluorescence stereomicroscopy. Anti-myelomaactivities were studied in vitro in 3D spheroids and in vivo in myeloma xenografts on chicken embryos. Tumor size was analyzed by measuring GFP content with a GFP ELISA. Anti-angiogenic activities of compounds were tested in an in vivo gelatin sponge assay with conditioned media from primary bone marrow-derived endothelial cells.

Results: We identified a subset of marine compounds with strong anti-myeloma activity in vitro and in vivo. Moreover, some of the compounds inhibited myeloma-related angiogenesis in the in vivo gelatin sponge assay. They merit further drug development to improve treatment options for MM.

No MeSH data available.


Related in: MedlinePlus