Limits...
A high-throughput in vitro drug screen in a genetically engineered mouse model of diffuse intrinsic pontine glioma identifies BMS-754807 as a promising therapeutic agent.

Halvorson KG, Barton KL, Schroeder K, Misuraca KL, Hoeman C, Chung A, Crabtree DM, Cordero FJ, Singh R, Spasojevic I, Berlow N, Pal R, Becher OJ - PLoS ONE (2015)

Bottom Line: In vitro evaluation showed significant cytotoxic effects with an IC50 of 0.13 μM, significant inhibition of proliferation at a concentration of 1.5 μM, as well as inhibition of AKT activation.Interestingly, IGF-1R signaling was absent in serum-free cultures from the PDGF-B; H3.3K27M; p53 deficient model suggesting that the antitumor activity of BMS-754807 in this model is independent of IGF-1R.Pharmacokinetic analysis demonstrated that tumor tissue drug concentrations of BMS-754807 were well below the identified IC50, suggesting that inadequate drug delivery may limit in vivo efficacy.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Duke University Medical Center, Durham, NC, United States of America; Department of Pathology, Duke University Medical Center, Durham, NC, United States of America; Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, NC, United States of America; Department of Surgery, Duke University, Durham, NC, United States of America.

ABSTRACT
Diffuse intrinsic pontine gliomas (DIPGs) represent a particularly lethal type of pediatric brain cancer with no effective therapeutic options. Our laboratory has previously reported the development of genetically engineered DIPG mouse models using the RCAS/tv-a system, including a model driven by PDGF-B, H3.3K27M, and p53 loss. These models can serve as a platform in which to test novel therapeutics prior to the initiation of human clinical trials. In this study, an in vitro high-throughput drug screen as part of the DIPG preclinical consortium using cell-lines derived from our DIPG models identified BMS-754807 as a drug of interest in DIPG. BMS-754807 is a potent and reversible small molecule multi-kinase inhibitor with many targets including IGF-1R, IR, MET, TRKA, TRKB, AURKA, AURKB. In vitro evaluation showed significant cytotoxic effects with an IC50 of 0.13 μM, significant inhibition of proliferation at a concentration of 1.5 μM, as well as inhibition of AKT activation. Interestingly, IGF-1R signaling was absent in serum-free cultures from the PDGF-B; H3.3K27M; p53 deficient model suggesting that the antitumor activity of BMS-754807 in this model is independent of IGF-1R. In vivo, systemic administration of BMS-754807 to DIPG-bearing mice did not prolong survival. Pharmacokinetic analysis demonstrated that tumor tissue drug concentrations of BMS-754807 were well below the identified IC50, suggesting that inadequate drug delivery may limit in vivo efficacy. In summary, an unbiased in vitro drug screen identified BMS-754807 as a potential therapeutic agent in DIPG, but BMS-754807 treatment in vivo by systemic delivery did not significantly prolong survival of DIPG-bearing mice.

No MeSH data available.


Related in: MedlinePlus

BMS-754807 has limited drug delivery to the tumor in the brainstem.Nestin-Tv-a;p53fl/fl mice were injected with DF1 cells expressing RCAS-PDGF-B, RCAS-Cre, and RCAS-H3.3-K27M. Upon appearance of symptoms, mice were treated with either BMS-754807 or vehicle for 3 doses, and sacrificed at 4 hours after the last dose. Tissue concentrations of BMS-754807 were determined by liquid chromatography tandem-mass spectrometry (LC/MS/MS). There was a significant difference between BMS-754807 treated tumor lysates compared to vehicle treated tumor lysates (p = 0.0357 by Mann-Whitney) and between BMS-754807 treated normal brain lysates as compared to vehicle treated normal brain lysates (p = 0.0357 by Mann-Whitney).
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4352073&req=5

pone.0118926.g005: BMS-754807 has limited drug delivery to the tumor in the brainstem.Nestin-Tv-a;p53fl/fl mice were injected with DF1 cells expressing RCAS-PDGF-B, RCAS-Cre, and RCAS-H3.3-K27M. Upon appearance of symptoms, mice were treated with either BMS-754807 or vehicle for 3 doses, and sacrificed at 4 hours after the last dose. Tissue concentrations of BMS-754807 were determined by liquid chromatography tandem-mass spectrometry (LC/MS/MS). There was a significant difference between BMS-754807 treated tumor lysates compared to vehicle treated tumor lysates (p = 0.0357 by Mann-Whitney) and between BMS-754807 treated normal brain lysates as compared to vehicle treated normal brain lysates (p = 0.0357 by Mann-Whitney).

Mentions: Due to its efficacy in vitro, we were interested to determine if BMS-754807 can significantly prolong survival of DIPG-bearing mice as a single agent in vivo. Nestin expressing brainstem progenitors of neonatal Ntv-a; p53fl/fl mice were infected with PDGFB, H3.3 K27M, and Cre at postnatal day 3–5. At 3 weeks post injection, mice were randomized to treatment with either BMS-754807 (50 mg/kg oral gavage) or vehicle once a day for three weeks (or until euthanasia criteria were met) and monitored for symptoms of tumor formation. No significant survival benefit was observed with BMS-754807 treatment, as the median survival for the vehicle group was 34 days vs. 31 days for the BMS-754807 group (n = 7 for BMS-754807 and n = 8 for vehicle; p = 0.26 by Log-Rank) (Fig. 4). To investigate achievable intra-tumor drug concentrations, tumor-bearing mice were treated with BMS-754807 for 3 days at the same dose used in the survival study and sacrificed 4 hours after the last dose. Based on liquid chromatography tandem-mass spectrometry (LC/MS/MS), we observed significant levels of drug present within the brain tissue of these animals compared with vehicle-treated mice, however the levels were well below the previously determined IC50 values observed in vitro (Fig. 5). This suggests that limited drug delivery of BMS-754807 to the tumor in the brainstem may explain why daily BMS-754807 treatment to DIPG-bearing mice did not significantly prolong survival.


A high-throughput in vitro drug screen in a genetically engineered mouse model of diffuse intrinsic pontine glioma identifies BMS-754807 as a promising therapeutic agent.

Halvorson KG, Barton KL, Schroeder K, Misuraca KL, Hoeman C, Chung A, Crabtree DM, Cordero FJ, Singh R, Spasojevic I, Berlow N, Pal R, Becher OJ - PLoS ONE (2015)

BMS-754807 has limited drug delivery to the tumor in the brainstem.Nestin-Tv-a;p53fl/fl mice were injected with DF1 cells expressing RCAS-PDGF-B, RCAS-Cre, and RCAS-H3.3-K27M. Upon appearance of symptoms, mice were treated with either BMS-754807 or vehicle for 3 doses, and sacrificed at 4 hours after the last dose. Tissue concentrations of BMS-754807 were determined by liquid chromatography tandem-mass spectrometry (LC/MS/MS). There was a significant difference between BMS-754807 treated tumor lysates compared to vehicle treated tumor lysates (p = 0.0357 by Mann-Whitney) and between BMS-754807 treated normal brain lysates as compared to vehicle treated normal brain lysates (p = 0.0357 by Mann-Whitney).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4352073&req=5

pone.0118926.g005: BMS-754807 has limited drug delivery to the tumor in the brainstem.Nestin-Tv-a;p53fl/fl mice were injected with DF1 cells expressing RCAS-PDGF-B, RCAS-Cre, and RCAS-H3.3-K27M. Upon appearance of symptoms, mice were treated with either BMS-754807 or vehicle for 3 doses, and sacrificed at 4 hours after the last dose. Tissue concentrations of BMS-754807 were determined by liquid chromatography tandem-mass spectrometry (LC/MS/MS). There was a significant difference between BMS-754807 treated tumor lysates compared to vehicle treated tumor lysates (p = 0.0357 by Mann-Whitney) and between BMS-754807 treated normal brain lysates as compared to vehicle treated normal brain lysates (p = 0.0357 by Mann-Whitney).
Mentions: Due to its efficacy in vitro, we were interested to determine if BMS-754807 can significantly prolong survival of DIPG-bearing mice as a single agent in vivo. Nestin expressing brainstem progenitors of neonatal Ntv-a; p53fl/fl mice were infected with PDGFB, H3.3 K27M, and Cre at postnatal day 3–5. At 3 weeks post injection, mice were randomized to treatment with either BMS-754807 (50 mg/kg oral gavage) or vehicle once a day for three weeks (or until euthanasia criteria were met) and monitored for symptoms of tumor formation. No significant survival benefit was observed with BMS-754807 treatment, as the median survival for the vehicle group was 34 days vs. 31 days for the BMS-754807 group (n = 7 for BMS-754807 and n = 8 for vehicle; p = 0.26 by Log-Rank) (Fig. 4). To investigate achievable intra-tumor drug concentrations, tumor-bearing mice were treated with BMS-754807 for 3 days at the same dose used in the survival study and sacrificed 4 hours after the last dose. Based on liquid chromatography tandem-mass spectrometry (LC/MS/MS), we observed significant levels of drug present within the brain tissue of these animals compared with vehicle-treated mice, however the levels were well below the previously determined IC50 values observed in vitro (Fig. 5). This suggests that limited drug delivery of BMS-754807 to the tumor in the brainstem may explain why daily BMS-754807 treatment to DIPG-bearing mice did not significantly prolong survival.

Bottom Line: In vitro evaluation showed significant cytotoxic effects with an IC50 of 0.13 μM, significant inhibition of proliferation at a concentration of 1.5 μM, as well as inhibition of AKT activation.Interestingly, IGF-1R signaling was absent in serum-free cultures from the PDGF-B; H3.3K27M; p53 deficient model suggesting that the antitumor activity of BMS-754807 in this model is independent of IGF-1R.Pharmacokinetic analysis demonstrated that tumor tissue drug concentrations of BMS-754807 were well below the identified IC50, suggesting that inadequate drug delivery may limit in vivo efficacy.

View Article: PubMed Central - PubMed

Affiliation: Department of Pediatrics, Duke University Medical Center, Durham, NC, United States of America; Department of Pathology, Duke University Medical Center, Durham, NC, United States of America; Preston Robert Tisch Brain Tumor Center, Duke University Medical Center, Durham, NC, United States of America; Department of Surgery, Duke University, Durham, NC, United States of America.

ABSTRACT
Diffuse intrinsic pontine gliomas (DIPGs) represent a particularly lethal type of pediatric brain cancer with no effective therapeutic options. Our laboratory has previously reported the development of genetically engineered DIPG mouse models using the RCAS/tv-a system, including a model driven by PDGF-B, H3.3K27M, and p53 loss. These models can serve as a platform in which to test novel therapeutics prior to the initiation of human clinical trials. In this study, an in vitro high-throughput drug screen as part of the DIPG preclinical consortium using cell-lines derived from our DIPG models identified BMS-754807 as a drug of interest in DIPG. BMS-754807 is a potent and reversible small molecule multi-kinase inhibitor with many targets including IGF-1R, IR, MET, TRKA, TRKB, AURKA, AURKB. In vitro evaluation showed significant cytotoxic effects with an IC50 of 0.13 μM, significant inhibition of proliferation at a concentration of 1.5 μM, as well as inhibition of AKT activation. Interestingly, IGF-1R signaling was absent in serum-free cultures from the PDGF-B; H3.3K27M; p53 deficient model suggesting that the antitumor activity of BMS-754807 in this model is independent of IGF-1R. In vivo, systemic administration of BMS-754807 to DIPG-bearing mice did not prolong survival. Pharmacokinetic analysis demonstrated that tumor tissue drug concentrations of BMS-754807 were well below the identified IC50, suggesting that inadequate drug delivery may limit in vivo efficacy. In summary, an unbiased in vitro drug screen identified BMS-754807 as a potential therapeutic agent in DIPG, but BMS-754807 treatment in vivo by systemic delivery did not significantly prolong survival of DIPG-bearing mice.

No MeSH data available.


Related in: MedlinePlus