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Single-agent CHK1 inhibition in CLL
nM, respectively) (Figure 3B). Four cell lines were tested within the project of DepMap and lethal phenotype with the CHEK1 elimination using CRISPR was noted (Online Supplementary Figure S1). Although some cell lines mani- fested a high baseline RS level (see phosphorylated CHK1 isoforms and/or gH2AX in Online Supplementary Figure S3A), we did not observe an apparent correlation to the viability decrease (Online Supplementary Figure S3B).
Essentially, non-cancerous cells were much less sensitive to MU380, specifically: immortalized epithelial cells (RPE- 1 cell line) (IC50 > 10 μM), immortalized bone
Table 1. Effects of MU380 in cancer cell lines and non-cancerous cells.
marrow/stromal fibroblasts (HS-5 cell line) (IC50 = 3.7 μM), primary skin fibroblasts (IC50 = 3.7 μM), and primary skin fibroblasts from a patient with ataxia telangiectasia har- boring complete ATM gene inactivation (IC50 > 10 μM) (Online Supplementary Figure S3).
To elucidate MU380 mechanistic effects, we analyzed cell cycle profile and apoptosis in six selected cell lines. The inhibitor (400 nM, 24 h treatment) significantly affect- ed the cell cycle profile in CLL-derived cell lines: TP53- mutated MEC-1 and MEC-2 exhibited profound S-phase accumulation together with G2/M phase decrease, while
Cell lines / Cells
MEC-1
MEC-2
BL-41
SU-DHL-4
Cancer
CLL/PL
CLL/PL
BL
DLBCL
TP53 status c.949dupC
p.Q317fs
c.949dupC
Other genetics
NA
NA
t(8;14) (MYC/IGH)
t(14;18)
(IGH/BCL2) t(11;14)
(CCND1/IGH)
NA
t(8;14) (MYC/IGH)
t(11;14)
(CCND1/IGH) t(12;21)
(TEL/AML1) t(11;14)
(CCND1/IGH) t(11;14)
(CCND1/IGH)
NA
t(11;14) (CCND1/IGH)
NA t(14;18)
(IGH/BCL2) t(8;14;18)
(MYC/IGH/BCL2) t(5;12)
(ETV6/PDGFRB) t(11;14)
(CCND1/IGH)
NA
(MLL/AF9)
NA
NA
NA
NA
Synergy with Gem
+++
+++
+++
+++++
++++
++++ +++
+++
ND
++++
+++
+++ +++
+++ ++++
+++
+++
+++
ND
ND
ND
ND
ND
ND
Single-agent MU380 (IC50; μM)
0.20
0.41
0.38
0.40
0.39
0.20 0.50
0.48
0.25
0.25
0.46
0.22 0.22
0.34 0.32
0.43
0.38
ND
0.34 0.14 >10 3.7 3.7 >10
c.743G˃A
p.Q317fs
c.817C˃T p.R273C
c.173delC p.P58X
p.R248Q
JEKO-1 MCL
NALM-16 ALL RAJI BL
REC-1 MCL
REH ALL
MAVER-1 MCL
MINO MCL
OSU-CLL CLL
JVM-2 B-PLL
JVM-3 B-PLL
WSU-NHL DLBCL
c.868_869insTC p.R290fs c.638G˃A; p.R213Q
c.700T˃C; p.Y234H c.734G˃A; p.G245D
c.949C˃T; p.Q317X c.541C˃T
c.843C˃G p.D281E
p.R181C
c.440T˃G p.V147G
DOHH-2
NALM-6
GRANTA-519
OCI-AML3
MOLM-13
RPE-1 Non-cancerous HS-5 Non-cancerous Fibroblasts normal Non-cancerous
Fibroblasts AT Non-cancerous
WT WT
WT WT
WT
WT
WT
(del 17p) WT WT NA NA NA
NA
FL
ALL
MCL
AML AML
Genetic characteristics of the cell lines were adopted from the German Collection of Microorganisms and Cell Cultures (DSMZ). Statistical evaluation of the synergy between MU380 and gemcitabine (Gem) was done by Chou-Talalay test; +++++ very strong synergism; ++++ strong synergism; +++ synergism. NA: not applicable; ND: not determined. CLL/PL: chronic lymphocytic leukemia in prolymphocytoid transformation; BL: Burkitt lymphoma; DLBCL: diffuse large B-cell lymphoma; MCL: mantle cell lymphoma; ALL: acute lymphoblastic leukemia; B-PLL: B-cell prolymphocytic leukemia; FL: follicular lymphoma; AML: acute myeloid leukemia. Characterization of the non-cancerous cells and cell lines is provided in Online Supplementary Appendix.
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