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Combination treatment for myeloproliferative neoplasms using JAK and pan-class I PI3K inhibitors.

Choong ML, Pecquet C, Pendharkar V, Diaconu CC, Yong JW, Tai SJ, Wang SF, Defour JP, Sangthongpitag K, Villeval JL, Vainchenker W, Constantinescu SN, Lee MA - J. Cell. Mol. Med. (2013)

Bottom Line: Synergy was not observed in Bcr-Abl transformed cells.The best JAK2/JAK1 and PI3K inhibitor combination pair (ruxolitinib and GDC0941) reduces spleen weight in nude mice inoculated with Ba/F3 cells expressing TpoR and JAK2 V617F.Our data support the use of a combination of JAK2 and pan-class I PI3K inhibitors in the treatment of MPNs.

View Article: PubMed Central - PubMed

Affiliation: Experimental Therapeutics Centre, Agency for Science Technology and Research, Singapore.

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Related in: MedlinePlus

Cell lines and small molecules used in combination for detection of synergy with JAK2 inhibitors in inhibiting proliferation of model myeloproliferative neoplasm cells. (A) Ba/F3 cell lines used for inhibitor screens. Ba/F3 parental and Ba/F3 TpoR JAK2 wild-type (WT) cells were maintained in medium supplemented with IL3, while Ba/F3 JAK2 V617F, TpoR V617F, TpoR W515L and Bcr-Abl were maintained in medium without cytokines. (B) A schematic representation of an 8 × 8 constant ratio design for combination treatment. The two combination drugs were used at their equipotent concentration ratio (IC50 of drug A to IC50 of drug B is 1:1) in the centre column. The concentration ratio of drug A to drug B is progressively increased towards the left, while the concentration ratio of drug B to drug A is progressively increased towards the right. Each column in the design is a dose–response curve with constant concentration ratio between drug A and drug B. Although usually C.I. <0.8 is considered significant, we selected for combinations showing C.I. <0.5. (C) Combination study using JAK2/JAK1 inhibitor ruxolitinib with various kinase inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells. (D) Combination study using JAK2/JAK1 inhibitor ruxolitinib with several other PI3K inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells.
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fig01: Cell lines and small molecules used in combination for detection of synergy with JAK2 inhibitors in inhibiting proliferation of model myeloproliferative neoplasm cells. (A) Ba/F3 cell lines used for inhibitor screens. Ba/F3 parental and Ba/F3 TpoR JAK2 wild-type (WT) cells were maintained in medium supplemented with IL3, while Ba/F3 JAK2 V617F, TpoR V617F, TpoR W515L and Bcr-Abl were maintained in medium without cytokines. (B) A schematic representation of an 8 × 8 constant ratio design for combination treatment. The two combination drugs were used at their equipotent concentration ratio (IC50 of drug A to IC50 of drug B is 1:1) in the centre column. The concentration ratio of drug A to drug B is progressively increased towards the left, while the concentration ratio of drug B to drug A is progressively increased towards the right. Each column in the design is a dose–response curve with constant concentration ratio between drug A and drug B. Although usually C.I. <0.8 is considered significant, we selected for combinations showing C.I. <0.5. (C) Combination study using JAK2/JAK1 inhibitor ruxolitinib with various kinase inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells. (D) Combination study using JAK2/JAK1 inhibitor ruxolitinib with several other PI3K inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells.

Mentions: The JAK2/JAK1 inhibitor ruxolitinib (also known as INC424 or INCB018424) (Albany Molecular Research Inc., Albany, NY, USA) and the JAK2 inhibitor TG101348 (SYNthesis Med Chem, San Diego, CA, USA) were used. All compounds were dissolved in 100% dimethyl sulfoxide (Sigma-Aldrich, St. Louis, MO, USA) to prepare 20 mM stocks except for NVP-BEZ235, which was dissolved to prepare 10 mM stock. The identity of compounds used in this study is shown in Figure 1. All compounds were synthesized by SynMedChem except AZD6244 and XL147 (Selleck Chemicals, Houstan, TX, USA), Rapamycin and Temsirolimus (Tocris Bioscience, Bristol, UK), LY294002 from Sigma-Aldrich and SB1518 and CC401 from AMRI (Albany Molecular Research Inc.).


Combination treatment for myeloproliferative neoplasms using JAK and pan-class I PI3K inhibitors.

Choong ML, Pecquet C, Pendharkar V, Diaconu CC, Yong JW, Tai SJ, Wang SF, Defour JP, Sangthongpitag K, Villeval JL, Vainchenker W, Constantinescu SN, Lee MA - J. Cell. Mol. Med. (2013)

Cell lines and small molecules used in combination for detection of synergy with JAK2 inhibitors in inhibiting proliferation of model myeloproliferative neoplasm cells. (A) Ba/F3 cell lines used for inhibitor screens. Ba/F3 parental and Ba/F3 TpoR JAK2 wild-type (WT) cells were maintained in medium supplemented with IL3, while Ba/F3 JAK2 V617F, TpoR V617F, TpoR W515L and Bcr-Abl were maintained in medium without cytokines. (B) A schematic representation of an 8 × 8 constant ratio design for combination treatment. The two combination drugs were used at their equipotent concentration ratio (IC50 of drug A to IC50 of drug B is 1:1) in the centre column. The concentration ratio of drug A to drug B is progressively increased towards the left, while the concentration ratio of drug B to drug A is progressively increased towards the right. Each column in the design is a dose–response curve with constant concentration ratio between drug A and drug B. Although usually C.I. <0.8 is considered significant, we selected for combinations showing C.I. <0.5. (C) Combination study using JAK2/JAK1 inhibitor ruxolitinib with various kinase inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells. (D) Combination study using JAK2/JAK1 inhibitor ruxolitinib with several other PI3K inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4117552&req=5

fig01: Cell lines and small molecules used in combination for detection of synergy with JAK2 inhibitors in inhibiting proliferation of model myeloproliferative neoplasm cells. (A) Ba/F3 cell lines used for inhibitor screens. Ba/F3 parental and Ba/F3 TpoR JAK2 wild-type (WT) cells were maintained in medium supplemented with IL3, while Ba/F3 JAK2 V617F, TpoR V617F, TpoR W515L and Bcr-Abl were maintained in medium without cytokines. (B) A schematic representation of an 8 × 8 constant ratio design for combination treatment. The two combination drugs were used at their equipotent concentration ratio (IC50 of drug A to IC50 of drug B is 1:1) in the centre column. The concentration ratio of drug A to drug B is progressively increased towards the left, while the concentration ratio of drug B to drug A is progressively increased towards the right. Each column in the design is a dose–response curve with constant concentration ratio between drug A and drug B. Although usually C.I. <0.8 is considered significant, we selected for combinations showing C.I. <0.5. (C) Combination study using JAK2/JAK1 inhibitor ruxolitinib with various kinase inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells. (D) Combination study using JAK2/JAK1 inhibitor ruxolitinib with several other PI3K inhibitors at equipotent concentration ratio on TpoR JAK2 V617F cells.
Mentions: The JAK2/JAK1 inhibitor ruxolitinib (also known as INC424 or INCB018424) (Albany Molecular Research Inc., Albany, NY, USA) and the JAK2 inhibitor TG101348 (SYNthesis Med Chem, San Diego, CA, USA) were used. All compounds were dissolved in 100% dimethyl sulfoxide (Sigma-Aldrich, St. Louis, MO, USA) to prepare 20 mM stocks except for NVP-BEZ235, which was dissolved to prepare 10 mM stock. The identity of compounds used in this study is shown in Figure 1. All compounds were synthesized by SynMedChem except AZD6244 and XL147 (Selleck Chemicals, Houstan, TX, USA), Rapamycin and Temsirolimus (Tocris Bioscience, Bristol, UK), LY294002 from Sigma-Aldrich and SB1518 and CC401 from AMRI (Albany Molecular Research Inc.).

Bottom Line: Synergy was not observed in Bcr-Abl transformed cells.The best JAK2/JAK1 and PI3K inhibitor combination pair (ruxolitinib and GDC0941) reduces spleen weight in nude mice inoculated with Ba/F3 cells expressing TpoR and JAK2 V617F.Our data support the use of a combination of JAK2 and pan-class I PI3K inhibitors in the treatment of MPNs.

View Article: PubMed Central - PubMed

Affiliation: Experimental Therapeutics Centre, Agency for Science Technology and Research, Singapore.

Show MeSH
Related in: MedlinePlus