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Functionally defined therapeutic targets in diffuse intrinsic pontine glioma.

Grasso CS, Tang Y, Truffaux N, Berlow NE, Liu L, Debily MA, Quist MJ, Davis LE, Huang EC, Woo PJ, Ponnuswami A, Chen S, Johung TB, Sun W, Kogiso M, Du Y, Qi L, Huang Y, Hütt-Cabezas M, Warren KE, Le Dret L, Meltzer PS, Mao H, Quezado M, van Vuurden DG, Abraham J, Fouladi M, Svalina MN, Wang N, Hawkins C, Nazarian J, Alonso MM, Raabe EH, Hulleman E, Spellman PT, Li XN, Keller C, Pal R, Grill J, Monje M - Nat. Med. (2015)

Bottom Line: Diffuse intrinsic pontine glioma (DIPG) is a fatal childhood cancer.The multi-histone deacetylase inhibitor panobinostat demonstrated therapeutic efficacy both in vitro and in DIPG orthotopic xenograft models.Combination testing of panobinostat and the histone demethylase inhibitor GSK-J4 revealed that the two had synergistic effects.

View Article: PubMed Central - PubMed

Affiliation: Center for Spatial Systems Biomedicine, Department of Molecular and Medical Genetics, Oregon Health &Science University (OHSU), Portland, Oregon, USA.

ABSTRACT
Diffuse intrinsic pontine glioma (DIPG) is a fatal childhood cancer. We performed a chemical screen in patient-derived DIPG cultures along with RNA-seq analyses and integrated computational modeling to identify potentially effective therapeutic strategies. The multi-histone deacetylase inhibitor panobinostat demonstrated therapeutic efficacy both in vitro and in DIPG orthotopic xenograft models. Combination testing of panobinostat and the histone demethylase inhibitor GSK-J4 revealed that the two had synergistic effects. Together, these data suggest a promising therapeutic strategy for DIPG.

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Related in: MedlinePlus

Functionally-defined targets in DIPG therapy(a) Chemical screen of 14 patient-derived DIPG cell cultures. Heatmap demonstrating DIPG cell line sensitivity to each of the 83 agents tested. The values shown are the absolute IC50 divided by the maximum dose. All drugs had a maximum dose of 10 µM except for vismodegib, sodium butyrate, pazopanib, alisertib, and vemurafenib, which had a maximum dose of 100 µM. Values are shown as gradations of red to white, with red representing submicromolar IC50 values, white indicating IC50 greater than the maximum dose for that drug (i.e. 10 µM or 100 µM), and pink showing the range in between. Grey boxes indicate those drugs not included in the screen for that cell line. Numbers corresponding to the drug names in the key below the heatmap are listed on the horizontal axis, culture IDs listed on the vertical axis. The histone status of each culture used in the screen is indicated by green (wild type, WT), yellow (H3.3K27M, H3F3A-K27M) or blue (H3.1K27M, HIST1H3B-K27M); see also Supplementary Table 1. Recurrent “hits” are visualized as a column of red or pink. (b) Dose-response curves: Patient-derived DIPG lines (SU-DIPG IV, SU-DIPG-VI, SU-DIPG-XIII, JHH-DIPG1, SF77613) were treated with the indicated drugs at 0.001/0.01/0.1/1/10 µM or 0.1% DMSO control in at least triplicate (n = 3 wells) and cell viabilities were assessed at 72 hr. Data are expressed as relative to the 0.1% DMSO control values. A pediatric cortical GBM line (SU-pcGBM2; histone WT; orange curves) was treated in parallel for comparison in a subset. Data are shown as mean ± SD. (c) Panobinostat time course: DIPG cells were treated with panobinostat in quadruplicate (n = 4 wells) at indicated concentrations (25 nM – 500 nM) or 0.1% DMSO vehicle control. Cell viabilities were assessed at 0, 24, 48 and 72 hrs of treatment. Data are shown as mean ± SD. **P < 0.01, ***P < 0.001 (two-tailed t test results shown for the lowest concentration to reveal a significant difference at 48 or 72 hours).
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Figure 1: Functionally-defined targets in DIPG therapy(a) Chemical screen of 14 patient-derived DIPG cell cultures. Heatmap demonstrating DIPG cell line sensitivity to each of the 83 agents tested. The values shown are the absolute IC50 divided by the maximum dose. All drugs had a maximum dose of 10 µM except for vismodegib, sodium butyrate, pazopanib, alisertib, and vemurafenib, which had a maximum dose of 100 µM. Values are shown as gradations of red to white, with red representing submicromolar IC50 values, white indicating IC50 greater than the maximum dose for that drug (i.e. 10 µM or 100 µM), and pink showing the range in between. Grey boxes indicate those drugs not included in the screen for that cell line. Numbers corresponding to the drug names in the key below the heatmap are listed on the horizontal axis, culture IDs listed on the vertical axis. The histone status of each culture used in the screen is indicated by green (wild type, WT), yellow (H3.3K27M, H3F3A-K27M) or blue (H3.1K27M, HIST1H3B-K27M); see also Supplementary Table 1. Recurrent “hits” are visualized as a column of red or pink. (b) Dose-response curves: Patient-derived DIPG lines (SU-DIPG IV, SU-DIPG-VI, SU-DIPG-XIII, JHH-DIPG1, SF77613) were treated with the indicated drugs at 0.001/0.01/0.1/1/10 µM or 0.1% DMSO control in at least triplicate (n = 3 wells) and cell viabilities were assessed at 72 hr. Data are expressed as relative to the 0.1% DMSO control values. A pediatric cortical GBM line (SU-pcGBM2; histone WT; orange curves) was treated in parallel for comparison in a subset. Data are shown as mean ± SD. (c) Panobinostat time course: DIPG cells were treated with panobinostat in quadruplicate (n = 4 wells) at indicated concentrations (25 nM – 500 nM) or 0.1% DMSO vehicle control. Cell viabilities were assessed at 0, 24, 48 and 72 hrs of treatment. Data are shown as mean ± SD. **P < 0.01, ***P < 0.001 (two-tailed t test results shown for the lowest concentration to reveal a significant difference at 48 or 72 hours).

Mentions: We assembled a panel of 14 patient-derived DIPG cell cultures, representing neurosphere and adherent models, obtained from both biopsy or autopsy samples (methods and 9–11; Fig. 1a), representing the breadth of DIPG cell cultures available worldwide at the initiation of the study. Two additional cell cultures (methods and 3) were subsequently added (Fig. 1). Clinical, histopathological and molecular characteristics of the cultures are summarized in Supplementary Tables 1–2. We performed whole exome sequencing (WES) of culture models used for drug screening when primary tumor DNA was available (WES for 7/9 of tumors not previously reported10 are described here); additional DIPG tumors not associated with cultures (n = 15) were also sequenced to add to publically available DIPG genomic data (n = 22 DIPG tumors with WES data; Supplementary Fig. 1 and Supplementary Table 3).


Functionally defined therapeutic targets in diffuse intrinsic pontine glioma.

Grasso CS, Tang Y, Truffaux N, Berlow NE, Liu L, Debily MA, Quist MJ, Davis LE, Huang EC, Woo PJ, Ponnuswami A, Chen S, Johung TB, Sun W, Kogiso M, Du Y, Qi L, Huang Y, Hütt-Cabezas M, Warren KE, Le Dret L, Meltzer PS, Mao H, Quezado M, van Vuurden DG, Abraham J, Fouladi M, Svalina MN, Wang N, Hawkins C, Nazarian J, Alonso MM, Raabe EH, Hulleman E, Spellman PT, Li XN, Keller C, Pal R, Grill J, Monje M - Nat. Med. (2015)

Functionally-defined targets in DIPG therapy(a) Chemical screen of 14 patient-derived DIPG cell cultures. Heatmap demonstrating DIPG cell line sensitivity to each of the 83 agents tested. The values shown are the absolute IC50 divided by the maximum dose. All drugs had a maximum dose of 10 µM except for vismodegib, sodium butyrate, pazopanib, alisertib, and vemurafenib, which had a maximum dose of 100 µM. Values are shown as gradations of red to white, with red representing submicromolar IC50 values, white indicating IC50 greater than the maximum dose for that drug (i.e. 10 µM or 100 µM), and pink showing the range in between. Grey boxes indicate those drugs not included in the screen for that cell line. Numbers corresponding to the drug names in the key below the heatmap are listed on the horizontal axis, culture IDs listed on the vertical axis. The histone status of each culture used in the screen is indicated by green (wild type, WT), yellow (H3.3K27M, H3F3A-K27M) or blue (H3.1K27M, HIST1H3B-K27M); see also Supplementary Table 1. Recurrent “hits” are visualized as a column of red or pink. (b) Dose-response curves: Patient-derived DIPG lines (SU-DIPG IV, SU-DIPG-VI, SU-DIPG-XIII, JHH-DIPG1, SF77613) were treated with the indicated drugs at 0.001/0.01/0.1/1/10 µM or 0.1% DMSO control in at least triplicate (n = 3 wells) and cell viabilities were assessed at 72 hr. Data are expressed as relative to the 0.1% DMSO control values. A pediatric cortical GBM line (SU-pcGBM2; histone WT; orange curves) was treated in parallel for comparison in a subset. Data are shown as mean ± SD. (c) Panobinostat time course: DIPG cells were treated with panobinostat in quadruplicate (n = 4 wells) at indicated concentrations (25 nM – 500 nM) or 0.1% DMSO vehicle control. Cell viabilities were assessed at 0, 24, 48 and 72 hrs of treatment. Data are shown as mean ± SD. **P < 0.01, ***P < 0.001 (two-tailed t test results shown for the lowest concentration to reveal a significant difference at 48 or 72 hours).
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Related In: Results  -  Collection

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Figure 1: Functionally-defined targets in DIPG therapy(a) Chemical screen of 14 patient-derived DIPG cell cultures. Heatmap demonstrating DIPG cell line sensitivity to each of the 83 agents tested. The values shown are the absolute IC50 divided by the maximum dose. All drugs had a maximum dose of 10 µM except for vismodegib, sodium butyrate, pazopanib, alisertib, and vemurafenib, which had a maximum dose of 100 µM. Values are shown as gradations of red to white, with red representing submicromolar IC50 values, white indicating IC50 greater than the maximum dose for that drug (i.e. 10 µM or 100 µM), and pink showing the range in between. Grey boxes indicate those drugs not included in the screen for that cell line. Numbers corresponding to the drug names in the key below the heatmap are listed on the horizontal axis, culture IDs listed on the vertical axis. The histone status of each culture used in the screen is indicated by green (wild type, WT), yellow (H3.3K27M, H3F3A-K27M) or blue (H3.1K27M, HIST1H3B-K27M); see also Supplementary Table 1. Recurrent “hits” are visualized as a column of red or pink. (b) Dose-response curves: Patient-derived DIPG lines (SU-DIPG IV, SU-DIPG-VI, SU-DIPG-XIII, JHH-DIPG1, SF77613) were treated with the indicated drugs at 0.001/0.01/0.1/1/10 µM or 0.1% DMSO control in at least triplicate (n = 3 wells) and cell viabilities were assessed at 72 hr. Data are expressed as relative to the 0.1% DMSO control values. A pediatric cortical GBM line (SU-pcGBM2; histone WT; orange curves) was treated in parallel for comparison in a subset. Data are shown as mean ± SD. (c) Panobinostat time course: DIPG cells were treated with panobinostat in quadruplicate (n = 4 wells) at indicated concentrations (25 nM – 500 nM) or 0.1% DMSO vehicle control. Cell viabilities were assessed at 0, 24, 48 and 72 hrs of treatment. Data are shown as mean ± SD. **P < 0.01, ***P < 0.001 (two-tailed t test results shown for the lowest concentration to reveal a significant difference at 48 or 72 hours).
Mentions: We assembled a panel of 14 patient-derived DIPG cell cultures, representing neurosphere and adherent models, obtained from both biopsy or autopsy samples (methods and 9–11; Fig. 1a), representing the breadth of DIPG cell cultures available worldwide at the initiation of the study. Two additional cell cultures (methods and 3) were subsequently added (Fig. 1). Clinical, histopathological and molecular characteristics of the cultures are summarized in Supplementary Tables 1–2. We performed whole exome sequencing (WES) of culture models used for drug screening when primary tumor DNA was available (WES for 7/9 of tumors not previously reported10 are described here); additional DIPG tumors not associated with cultures (n = 15) were also sequenced to add to publically available DIPG genomic data (n = 22 DIPG tumors with WES data; Supplementary Fig. 1 and Supplementary Table 3).

Bottom Line: Diffuse intrinsic pontine glioma (DIPG) is a fatal childhood cancer.The multi-histone deacetylase inhibitor panobinostat demonstrated therapeutic efficacy both in vitro and in DIPG orthotopic xenograft models.Combination testing of panobinostat and the histone demethylase inhibitor GSK-J4 revealed that the two had synergistic effects.

View Article: PubMed Central - PubMed

Affiliation: Center for Spatial Systems Biomedicine, Department of Molecular and Medical Genetics, Oregon Health &Science University (OHSU), Portland, Oregon, USA.

ABSTRACT
Diffuse intrinsic pontine glioma (DIPG) is a fatal childhood cancer. We performed a chemical screen in patient-derived DIPG cultures along with RNA-seq analyses and integrated computational modeling to identify potentially effective therapeutic strategies. The multi-histone deacetylase inhibitor panobinostat demonstrated therapeutic efficacy both in vitro and in DIPG orthotopic xenograft models. Combination testing of panobinostat and the histone demethylase inhibitor GSK-J4 revealed that the two had synergistic effects. Together, these data suggest a promising therapeutic strategy for DIPG.

Show MeSH
Related in: MedlinePlus