Limits...
DUSP4 deficiency caused by promoter hypermethylation drives JNK signaling and tumor cell survival in diffuse large B cell lymphoma.

Schmid CA, Robinson MD, Scheifinger NA, Müller S, Cogliatti S, Tzankov A, Müller A - J. Exp. Med. (2015)

Bottom Line: The epigenetic dysregulation of tumor suppressor genes is an important driver of human carcinogenesis.We find that a CpG island in the promoter of the dual-specificity phosphatase DUSP4 is aberrantly methylated in nodal and extranodal DLBCL, irrespective of ABC or GCB subtype, resulting in loss of DUSP4 expression in 75% of >200 examined cases.This finding provides a mechanistic basis for the clinical development of JNK inhibitors in DLBCL, ideally in synthetic lethal combinations with inhibitors of chronic active B cell receptor signaling.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Molecular Cancer Research, Institute of Molecular Life Sciences, and Swiss Institute of Bioinformatics (SIB), University of Zürich, 8057 Zürich, Switzerland.

Show MeSH

Related in: MedlinePlus

The JNK inhibitor SP600125 synergizes with ibrutinib in killing ABC-DLBCL cell lines. (A) The viability of DLBCL cell lines was determined by CellTiter-Blue assay after 72 h of treatment with the indicated concentrations of the BTK inhibitor ibrutinib and calculated relative to DMSO-treated cells. Means ± SEM for two to four independent experiments per cell line are shown. (B–G) The viability of the indicated DLBCL cell lines was assessed after 72 h of treatment with increasing concentrations of the JNK inhibitor SP600125 and the BTK inhibitor ibrutinib, either alone or in combination, relative to DMSO-treated cells. In F, Chalice Analyzer inhibition matrices are shown for combination responses to SP600125 and ibrutinib of the indicated cell lines. In G, isobolograms of viability data demonstrating drug synergy are shown for the indicated cell lines. The results presented in G were compiled from B, C, and F. In B, C, F, and G, data from at least three and up to four independent experiments are shown; in D and E, data from two independent experiments (results represent a limited number of repeats) are shown. Results in A–E are presented as means ± SEM.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4419353&req=5

fig8: The JNK inhibitor SP600125 synergizes with ibrutinib in killing ABC-DLBCL cell lines. (A) The viability of DLBCL cell lines was determined by CellTiter-Blue assay after 72 h of treatment with the indicated concentrations of the BTK inhibitor ibrutinib and calculated relative to DMSO-treated cells. Means ± SEM for two to four independent experiments per cell line are shown. (B–G) The viability of the indicated DLBCL cell lines was assessed after 72 h of treatment with increasing concentrations of the JNK inhibitor SP600125 and the BTK inhibitor ibrutinib, either alone or in combination, relative to DMSO-treated cells. In F, Chalice Analyzer inhibition matrices are shown for combination responses to SP600125 and ibrutinib of the indicated cell lines. In G, isobolograms of viability data demonstrating drug synergy are shown for the indicated cell lines. The results presented in G were compiled from B, C, and F. In B, C, F, and G, data from at least three and up to four independent experiments are shown; in D and E, data from two independent experiments (results represent a limited number of repeats) are shown. Results in A–E are presented as means ± SEM.

Mentions: Ibrutinib is a novel inhibitor of BTK that has proven to be effective in ABC-DLBCL with chronic active BCR signaling in various preclinical combination treatments (Mathews Griner et al., 2014). To examine whether simultaneous JNK and BTK inhibition synergizes to kill DLBCL cells, we conducted single and combination treatments with SP600125 and ibrutinib. Ibrutinib treatment alone was insufficient to kill any of our DLBCL cell lines at concentrations up to 1 µM (Fig. 8 A). However, the addition of ibrutinib, even at very low doses of 1–10 nM, strongly boosted the efficacy of SP600125 in two ABC-DLBCL cell lines (Fig. 8, B and C). This effect was not seen in the GCB cell line SU-DHL16 (Fig. 8 D) or in an ABC-DLBCL cell line in which the BCR signaling pathway is constitutively active as the result of a CARD11 mutation (Oci-Ly3; Fig. 8 E). In those cell lines where ibrutinib augmented the effects of JNK inhibition, the two treatments were clearly synergistic, as determined by Chalice matrix analyses and the isobologram method (Fig. 8, F and G). In summary, our data indicate that JNK inhibitors should be considered for assessment as single agents in clinical trials for both subtypes of DLBCL and might be especially promising in combination with ibrutinib in ABC-DLBCL.


DUSP4 deficiency caused by promoter hypermethylation drives JNK signaling and tumor cell survival in diffuse large B cell lymphoma.

Schmid CA, Robinson MD, Scheifinger NA, Müller S, Cogliatti S, Tzankov A, Müller A - J. Exp. Med. (2015)

The JNK inhibitor SP600125 synergizes with ibrutinib in killing ABC-DLBCL cell lines. (A) The viability of DLBCL cell lines was determined by CellTiter-Blue assay after 72 h of treatment with the indicated concentrations of the BTK inhibitor ibrutinib and calculated relative to DMSO-treated cells. Means ± SEM for two to four independent experiments per cell line are shown. (B–G) The viability of the indicated DLBCL cell lines was assessed after 72 h of treatment with increasing concentrations of the JNK inhibitor SP600125 and the BTK inhibitor ibrutinib, either alone or in combination, relative to DMSO-treated cells. In F, Chalice Analyzer inhibition matrices are shown for combination responses to SP600125 and ibrutinib of the indicated cell lines. In G, isobolograms of viability data demonstrating drug synergy are shown for the indicated cell lines. The results presented in G were compiled from B, C, and F. In B, C, F, and G, data from at least three and up to four independent experiments are shown; in D and E, data from two independent experiments (results represent a limited number of repeats) are shown. Results in A–E are presented as means ± SEM.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4419353&req=5

fig8: The JNK inhibitor SP600125 synergizes with ibrutinib in killing ABC-DLBCL cell lines. (A) The viability of DLBCL cell lines was determined by CellTiter-Blue assay after 72 h of treatment with the indicated concentrations of the BTK inhibitor ibrutinib and calculated relative to DMSO-treated cells. Means ± SEM for two to four independent experiments per cell line are shown. (B–G) The viability of the indicated DLBCL cell lines was assessed after 72 h of treatment with increasing concentrations of the JNK inhibitor SP600125 and the BTK inhibitor ibrutinib, either alone or in combination, relative to DMSO-treated cells. In F, Chalice Analyzer inhibition matrices are shown for combination responses to SP600125 and ibrutinib of the indicated cell lines. In G, isobolograms of viability data demonstrating drug synergy are shown for the indicated cell lines. The results presented in G were compiled from B, C, and F. In B, C, F, and G, data from at least three and up to four independent experiments are shown; in D and E, data from two independent experiments (results represent a limited number of repeats) are shown. Results in A–E are presented as means ± SEM.
Mentions: Ibrutinib is a novel inhibitor of BTK that has proven to be effective in ABC-DLBCL with chronic active BCR signaling in various preclinical combination treatments (Mathews Griner et al., 2014). To examine whether simultaneous JNK and BTK inhibition synergizes to kill DLBCL cells, we conducted single and combination treatments with SP600125 and ibrutinib. Ibrutinib treatment alone was insufficient to kill any of our DLBCL cell lines at concentrations up to 1 µM (Fig. 8 A). However, the addition of ibrutinib, even at very low doses of 1–10 nM, strongly boosted the efficacy of SP600125 in two ABC-DLBCL cell lines (Fig. 8, B and C). This effect was not seen in the GCB cell line SU-DHL16 (Fig. 8 D) or in an ABC-DLBCL cell line in which the BCR signaling pathway is constitutively active as the result of a CARD11 mutation (Oci-Ly3; Fig. 8 E). In those cell lines where ibrutinib augmented the effects of JNK inhibition, the two treatments were clearly synergistic, as determined by Chalice matrix analyses and the isobologram method (Fig. 8, F and G). In summary, our data indicate that JNK inhibitors should be considered for assessment as single agents in clinical trials for both subtypes of DLBCL and might be especially promising in combination with ibrutinib in ABC-DLBCL.

Bottom Line: The epigenetic dysregulation of tumor suppressor genes is an important driver of human carcinogenesis.We find that a CpG island in the promoter of the dual-specificity phosphatase DUSP4 is aberrantly methylated in nodal and extranodal DLBCL, irrespective of ABC or GCB subtype, resulting in loss of DUSP4 expression in 75% of >200 examined cases.This finding provides a mechanistic basis for the clinical development of JNK inhibitors in DLBCL, ideally in synthetic lethal combinations with inhibitors of chronic active B cell receptor signaling.

View Article: PubMed Central - HTML - PubMed

Affiliation: Institute of Molecular Cancer Research, Institute of Molecular Life Sciences, and Swiss Institute of Bioinformatics (SIB), University of Zürich, 8057 Zürich, Switzerland.

Show MeSH
Related in: MedlinePlus