Journal: PLoS ONE

Article Title: The JAK2/STAT3 inhibitor pacritinib effectively inhibits patient-derived GBM brain tumor initiating cells in vitro and when used in combination with temozolomide increases survival in an orthotopic xenograft model

doi: 10.1371/journal.pone.0189670

Figure Lengend Snippet: (A) Pacritinib dramatically decreased cell viability in a dose dependent manner in eleven molecularly diverse patient-derived BTIC cultures with IC 50 values from 0.62 μM to 1.66 μM. Pacritinib decreased sphere formation in a dose dependent manner. Representative images (B) and quantifications (C) are shown for two representative BTIC cultures (BT69 and BT147). Pacritinib completely abolished sphere formation at 3 μM in BT69 and BT147 (**** denotes p < 0.0001 vs untreated; ANOVA). Error bars represent SEM. (D) Pacritinib had on-target activity as seen by a decrease in the phosphorylation of tyrosine 705 at 3 hours. It also resulted in increased cell death as seen by an increase in the cleaved PARP at 24 hours (representative line BT69 shown).

Article Snippet: BTICs were treated with suboptimal doses of 1 μM pacritinib (CTI Biopharma), 10 μg/mL TMZ (Sigma), or a combination of pacritinib and TMZ.

Techniques: Derivative Assay, Activity Assay

Journal: PLoS ONE

Article Title: The JAK2/STAT3 inhibitor pacritinib effectively inhibits patient-derived GBM brain tumor initiating cells in vitro and when used in combination with temozolomide increases survival in an orthotopic xenograft model

doi: 10.1371/journal.pone.0189670

Figure Lengend Snippet: (A) The MGMT methylated BTIC cultures were highly sensitive to TMZ. Pacritinib did not reverse the effectiveness of TMZ in these methylated BTICs (representative BTIC cultures, BT67 and BT69, shown; **** denotes p < 0.0001; ANOVA). Error bars represent SEM. (B) MGMT unmethylated BTIC cultures were largely resistant to TMZ, but responded to the combination of pacritinib and TMZ (representative BTIC culture BT12 shown; ** denotes p < 0.003; ANOVA). Error bars represent SEM. (C) The combination of pacritinib and TMZ had no effect on normal human astrocytes. (D) Bliss independence analysis shows that suboptimal doses of pacritinib and TMZ are synergistic in BTICs (representative BTIC cultures, BT12 and BT53, shown). (E) 1 μM pacritinib dramatically reduced phosphorylation of tyrosine 705 of STAT3 (p-STAT3 Y705) in BT53 at 48 hours. Treatment with 10 μg/mL TMZ resulted in a dramatic increase in p-STAT3 Y705. Combinatorial treatment with pacritinib and TMZ resulted in the abrogation of activated STAT3.

Article Snippet: BTICs were treated with suboptimal doses of 1 μM pacritinib (CTI Biopharma), 10 μg/mL TMZ (Sigma), or a combination of pacritinib and TMZ.

Techniques: Methylation

Journal: PLoS ONE

Article Title: The JAK2/STAT3 inhibitor pacritinib effectively inhibits patient-derived GBM brain tumor initiating cells in vitro and when used in combination with temozolomide increases survival in an orthotopic xenograft model

doi: 10.1371/journal.pone.0189670

Figure Lengend Snippet: Low micromolar concentrations of pacritinib reduced phosphorylation of tyrosine 705 of STAT3 in BT147 and BT53 at 3 hours, but did not affect phosphorylation of p44/42 MAPK or AKT S473. Pacritinib reduced phosphorylation of AKT S473 and p44/42 MAPK at higher concentrations (5 μM and 10 μM).

Article Snippet: BTICs were treated with suboptimal doses of 1 μM pacritinib (CTI Biopharma), 10 μg/mL TMZ (Sigma), or a combination of pacritinib and TMZ.

Techniques:

Journal: PLoS ONE

Article Title: The JAK2/STAT3 inhibitor pacritinib effectively inhibits patient-derived GBM brain tumor initiating cells in vitro and when used in combination with temozolomide increases survival in an orthotopic xenograft model

doi: 10.1371/journal.pone.0189670

Figure Lengend Snippet: Five-day pharmacokinetic properties of pacritinib.

Article Snippet: BTICs were treated with suboptimal doses of 1 μM pacritinib (CTI Biopharma), 10 μg/mL TMZ (Sigma), or a combination of pacritinib and TMZ.

Techniques:

Journal: PLoS ONE

Article Title: The JAK2/STAT3 inhibitor pacritinib effectively inhibits patient-derived GBM brain tumor initiating cells in vitro and when used in combination with temozolomide increases survival in an orthotopic xenograft model

doi: 10.1371/journal.pone.0189670

Figure Lengend Snippet: Kaplan-Meier survival curves showing no survival benefit from treatment with 100 mg/kg pacritinib in mouse orthotopic xenograft models of (A) BT53 (p < 0.7237; log-rank test) and (B) BT147 (p < 0.4649; log-rank test). (C) In mouse liver microsomes (MLM), pacritinib is rapidly metabolized, while in human liver microsomes (HLM), 76% of pacritinib remains after the 30-minute incubation period. Error bars represent SD.

Article Snippet: BTICs were treated with suboptimal doses of 1 μM pacritinib (CTI Biopharma), 10 μg/mL TMZ (Sigma), or a combination of pacritinib and TMZ.

Techniques: Incubation

Journal: PLoS ONE

Article Title: The JAK2/STAT3 inhibitor pacritinib effectively inhibits patient-derived GBM brain tumor initiating cells in vitro and when used in combination with temozolomide increases survival in an orthotopic xenograft model

doi: 10.1371/journal.pone.0189670

Figure Lengend Snippet: (A) Systemic administration of 100 mg/kg pacritinib in combination with 30 mg/kg TMZ thrice weekly for five consecutive weeks significantly increased median overall survival of BT147 xenografted animals compared to either agent alone. The combination of pacritinib and TMZ resulted in a median survival of 62.5 days compared to a median survival of 52 days in the control cohort, 48 days in the pacritinib cohort, and 58 days in the TMZ cohort. (B) For BT53, 100 mg/kg of pacritinib was administered twice per day alone or in combination with 50 mg/kg TMZ for one week. The following two weeks, mice received 100 mg/kg pacritinib twice per day in combination with 10 mg/kg TMZ. This three-week treatment regimen significantly increased median overall survival of BT53 xenografted animals. The combination of pacritinib and TMZ resulted in a median survival of 60 days compared to a median survival of 52 days in the control cohort.

Article Snippet: BTICs were treated with suboptimal doses of 1 μM pacritinib (CTI Biopharma), 10 μg/mL TMZ (Sigma), or a combination of pacritinib and TMZ.

Techniques: