scholarly journals Gene expression signatures but not cell cycle checkpoint functions distinguish AT carriers from normal individuals

2013 ◽  
Vol 45 (19) ◽  
pp. 907-916
Author(s):  
Liwen Zhang ◽  
Dennis A. Simpson ◽  
Cynthia L. Innes ◽  
Jeff Chou ◽  
Pierre R. Bushel ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Lines ◽  
Cell Cycle Checkpoint ◽  
G2 Checkpoint ◽  
P53 Signaling ◽  
Normal Individuals ◽  
Gene Expression Signatures ◽  
Cycle Checkpoint ◽  
Atm Cell

Ataxia telangiectasia (AT) is a rare autosomal recessive disease caused by mutations in the ataxia telangiectasia-mutated gene ( ATM). AT carriers with one mutant ATM allele are usually not severely affected although they carry an increased risk of developing cancer. There has not been an easy and reliable diagnostic method to identify AT carriers. Cell cycle checkpoint functions upon ionizing radiation (IR)-induced DNA damage and gene expression signatures were analyzed in the current study to test for differential responses in human lymphoblastoid cell lines with different ATM genotypes. While both dose- and time-dependent G1 and G2 checkpoint functions were highly attenuated in ATM−/− cell lines, these functions were preserved in ATM+/− cell lines equivalent to ATM+/+ cell lines. However, gene expression signatures at both baseline (consisting of 203 probes) and post-IR treatment (consisting of 126 probes) were able to distinguish ATM+/− cell lines from ATM+/+ and ATM−/− cell lines. Gene ontology (GO) and pathway analysis of the genes in the baseline signature indicate that ATM function-related categories, DNA metabolism, cell cycle, cell death control, and the p53 signaling pathway, were overrepresented. The same analyses of the genes in the IR-responsive signature revealed that biological categories including response to DNA damage stimulus, p53 signaling, and cell cycle pathways were overrepresented, which again confirmed involvement of ATM functions. The results indicate that AT carriers who have unaffected G1 and G2 checkpoint functions can be distinguished from normal individuals and AT patients by expression signatures of genes related to ATM functions.

scholarly journals Defective Cell Cycle Checkpoint Functions in Melanoma Are Associated with Altered Patterns of Gene Expression

2008 ◽  
Vol 128 (1) ◽  
pp. 175-187 ◽  
Author(s):  
William K. Kaufmann ◽  
Kathleen R. Nevis ◽  
Pingping Qu ◽  
Joseph G. Ibrahim ◽  
Tong Zhou ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Cycle Checkpoint ◽  
Cycle Checkpoint

scholarly journals Exploring the ATR-CHK1 pathway in the response of doxorubicin-induced DNA damages in acute lymphoblastic leukemia cells

2021 ◽  
Author(s):  
Andrea Ghelli Luserna Di Rorà ◽  
Martina Ghetti ◽  
Lorenzo Ledda ◽  
Anna Ferrari ◽  
Matteo Bocconcelli ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Acute Lymphoblastic Leukemia ◽  
T Cell ◽  
Cell Lines ◽  
Lymphoblastic Leukemia ◽  
Cell Cycle Checkpoint ◽  
M Cell ◽  
Dna Damages ◽  
Cycle Checkpoint ◽  
Dna Double Helix

AbstractDoxorubicin (Dox) is one of the most commonly used anthracyclines for the treatment of solid and hematological tumors such as B−/T cell acute lymphoblastic leukemia (ALL). Dox compromises topoisomerase II enzyme functionality, thus inducing structural damages during DNA replication and causes direct damages intercalating into DNA double helix. Eukaryotic cells respond to DNA damages by activating the ATM-CHK2 and/or ATR-CHK1 pathway, whose function is to regulate cell cycle progression, to promote damage repair, and to control apoptosis. We evaluated the efficacy of a new drug schedule combining Dox and specific ATR (VE-821) or CHK1 (prexasertib, PX) inhibitors in the treatment of human B−/T cell precursor ALL cell lines and primary ALL leukemic cells. We found that ALL cell lines respond to Dox activating the G2/M cell cycle checkpoint. Exposure of Dox-pretreated ALL cell lines to VE-821 or PX enhanced Dox cytotoxic effect. This phenomenon was associated with the abrogation of the G2/M cell cycle checkpoint with changes in the expression pCDK1 and cyclin B1, and cell entry in mitosis, followed by the induction of apoptosis. Indeed, the inhibition of the G2/M checkpoint led to a significant increment of normal and aberrant mitotic cells, including those showing tripolar spindles, metaphases with lagging chromosomes, and massive chromosomes fragmentation. In conclusion, we found that the ATR-CHK1 pathway is involved in the response to Dox-induced DNA damages and we demonstrated that our new in vitro drug schedule that combines Dox followed by ATR/CHK1 inhibitors can increase Dox cytotoxicity against ALL cells, while using lower drug doses. Graphical abstract • Doxorubicin activates the G2/M cell cycle checkpoint in acute lymphoblastic leukemia (ALL) cells. • ALL cells respond to doxorubicin-induced DNA damages by activating the ATR-CHK1 pathway. • The inhibition of the ATR-CHK1 pathway synergizes with doxorubicin in the induction of cytotoxicity in ALL cells. • The inhibition of ATR-CHK1 pathway induces aberrant chromosome segregation and mitotic spindle defects in doxorubicin-pretreated ALL cells.

scholarly journals Array CGH and gene-expression profiling reveals distinct genomic instability patterns associated with DNA repair and cell-cycle checkpoint pathways in Ewing's sarcoma

Oncogene ◽  
2007 ◽  
Vol 27 (14) ◽  
pp. 2084-2090 ◽  
Author(s):  
B I Ferreira ◽  
J Alonso ◽  
J Carrillo ◽  
F Acquadro ◽  
C Largo ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Dna Repair ◽  
Gene Expression Profiling ◽  
Genomic Instability ◽  
Expression Profiling ◽  
Array Cgh ◽  
Ewing's Sarcoma ◽  
Cell Cycle Checkpoint ◽  
Cycle Checkpoint

Rapid construction of transgene-amplified CHO cell lines by cell cycle checkpoint engineering

2013 ◽  
Vol 97 (13) ◽  
pp. 5731-5741 ◽  
Author(s):  
Kyoung Ho Lee ◽  
Masayoshi Onitsuka ◽  
Kohsuke Honda ◽  
Hisao Ohtake ◽  
Takeshi Omasa
Keyword(s):  
Cell Cycle ◽  
Cell Lines ◽  
Cell Cycle Checkpoint ◽  
Cho Cell ◽  
Rapid Construction ◽  
Cycle Checkpoint

scholarly journals Cell cycle checkpoint status in human malignant mesothelioma cell lines: response to gamma radiation

2003 ◽  
Vol 88 (3) ◽  
pp. 388-395 ◽  
Author(s):  
C Vivo ◽  
C Lecomte ◽  
F Levy ◽  
K Leroy ◽  
Y Kirova ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Gamma Radiation ◽  
Cell Lines ◽  
Malignant Mesothelioma ◽  
Cell Cycle Checkpoint ◽  
Cycle Checkpoint ◽  
Mesothelioma Cell

scholarly journals Heterogeneities in Cell Cycle Checkpoint Activation Following Doxorubicin Treatment Reveal Targetable Vulnerabilities in TP53 Mutated Ultra High-Risk Neuroblastoma Cell Lines

2021 ◽  
Vol 22 (7) ◽  
pp. 3664
Author(s):  
Linnéa Ödborn Jönsson ◽  
Maryam Sahi ◽  
Ximena Lopez-Lorenzo ◽  
Faye Leilah Keller ◽  
Ourania N. Kostopoulou ◽  
...  
Keyword(s):  
Cell Cycle ◽  
High Risk ◽  
Cell Lines ◽  
Neuroblastoma Cell ◽  
Cell Cycle Checkpoint ◽  
Cell Cycle Checkpoints ◽  
Dna Integrity ◽  
Checkpoint Activation ◽  
Ultra High Risk ◽  
Cycle Checkpoint

Most chemotherapeutics target DNA integrity and thereby trigger tumour cell death through activation of DNA damage responses that are tightly coupled to the cell cycle. Disturbances in cell cycle regulation can therefore lead to treatment resistance. Here, a comprehensive analysis of cell cycle checkpoint activation following doxorubicin (doxo) treatment was performed using flow cytometry, immunofluorescence and live-cell imaging in a panel of TP53 mutated ultra high-risk neuroblastoma (NB) cell lines, SK-N-DZ, Kelly, SK-N-AS, SK-N-FI, and BE(2)-C. Following treatment, a dose-dependent accumulation in either S- and/or G2/M-phase was observed. This coincided with a heterogeneous increase of cell cycle checkpoint proteins, i.e., phos-ATM, phos-CHK1, phos-CHK2, Wee1, p21Cip1/Waf1, and p27Kip among the cell lines. Combination treatment with doxo and a small-molecule inhibitor of ATM showed a delay in regrowth in SK-N-DZ, of CHK1 in BE(2)-C, of Wee1 in SK-N-FI and BE(2)-C, and of p21 in Kelly and BE(2)-C. Further investigation revealed, in all tested cell lines, a subset of cells arrested in mitosis, indicating independence on the intra-S- and/or G2/M-checkpoints. Taken together, we mapped distinct cell cycle checkpoints in ultra high-risk NB cell lines and identified checkpoint dependent and independent druggable targets.

Genome-Wide shRNA Screen Identifies WEE1 As a Critical Mediator of Cell Fate and Novel Therapeutic Target in AML,

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 3503-3503
Author(s):  
Christopher C. Porter ◽  
Jihye Kim ◽  
Susan Fosmire ◽  
Christy M. Gearheart ◽  
Annemie van Linden ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Fate ◽  
Cell Lines ◽  
Therapeutic Target ◽  
Cellular Proliferation ◽  
Conflicts Of Interest ◽  
Cell Cycle Checkpoint ◽  
Phase Ii Trials ◽  
Genome Wide ◽  
Cycle Checkpoint

Abstract Abstract 3503 Acute myeloid leukemia (AML) remains a therapeutic challenge despite increasing knowledge of the molecular origins of the disease, as the mechanisms of AML cell escape from chemotherapy remain poorly defined. We hypothesized that AML cells are addicted to specific molecular pathways in the context of chemotherapy and used complementary approaches to identify these addictions. Using novel molecular and computational strategies, we performed genome-wide shRNA screens to identify proteins which mediate AML cell fate in cytarabine in 2 AML cell lines (Molm13 and MV4-11). Over 55,000 shRNAs targeting over 11,000 genes were quantified by deep sequencing to identify shRNAs under-represented in the context of cytarabine as compared to no treatment. Complementary analyses identified 125 genes as mediators of AML cell fate in cytarabine. In addition we performed gene expression profiling of AML cells exposed to cytarabine to identify genes with induced expression in this context and examined existing gene expression data from primary patient samples. The integration of these independent analyses strongly implicates cell cycle checkpoint proteins, particularly WEE1, as critical mediators of AML cell fate in cytarabine. Knockdown of WEE1 in a secondary screen confirmed its role in AML cell survival in cytarabine. Pharmacologic inhibition of WEE1 in several, but not all, AML cell lines is synergistic with cytarabine, suggesting underlying molecular susceptibility to this combination of drugs. A WEE1 inhibitor is in Phase II trials in solid tumors, primarily as a means to abrogate the G2/M checkpoint in tumors with TP53 dysfunction. Further experiments demonstrate that inhibition of WEE1 prevents slowed S-phase progression induced by cytarabine in AML cells, broadening the functions of WEE1 that may be exploited therapeutically. Preliminary experiments indicate synergistic inhibition of AML cellular proliferation with daunorubicin in some AML cell lines. Experiments to determine whether WEE1 inhibition in combination with chemotherapy prolongs survival of mice with leukemia are underway. These data highlight the power of integrating functional and descriptive genomics, and identify WEE1 as potential therapeutic target in AML. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Sensitivity of cells to ATR and CHK1 inhibitors requires hyperactivation of CDK2 rather than endogenous replication stress or ATM dysfunction

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Jennifer P. Ditano ◽  
Katelyn L. Donahue ◽  
Laura J. Tafe ◽  
Charlotte F. McCleery ◽  
Alan Eastman
Keyword(s):  
Cell Cycle ◽  
Dna Damage ◽  
Cell Lines ◽  
Cell Cycle Progression ◽  
Single Agent ◽  
Replication Stress ◽  
Cell Cycle Checkpoint ◽  
High Threshold ◽  
Cycle Checkpoint ◽  
Atm Signaling

AbstractDNA damage activates cell cycle checkpoint proteins ATR and CHK1 to arrest cell cycle progression, providing time for repair and recovery. Consequently, inhibitors of ATR (ATRi) and CHK1 (CHK1i) enhance damage-induced cell death. Intriguingly, both CHK1i and ATRi alone elicit cytotoxicity in some cell lines. Sensitivity has been attributed to endogenous replications stress, but many more cell lines are sensitive to ATRi than CHK1i. Endogenous activation of the DNA damage response also did not correlate with drug sensitivity. Sensitivity correlated with the appearance of γH2AX, a marker of DNA damage, but without phosphorylation of mitotic markers, contradicting suggestions that the damage is due to premature mitosis. Sensitivity to ATRi has been associated with ATM mutations, but dysfunction in ATM signaling did not correlate with sensitivity. CHK1i and ATRi circumvent replication stress by reactivating stalled replicons, a process requiring a low threshold activity of CDK2. In contrast, γH2AX induced by single agent ATRi and CHK1i requires a high threshold activity CDK2. Hence, phosphorylation of different CDK2 substrates is required for cytotoxicity induced by replication stress plus ATRi/CHK1i as compared to their single agent activity. In summary, sensitivity to ATRi and CHK1i as single agents is elicited by premature hyper-activation of CDK2.

Defective cell cycle checkpoint response to DNA double strand breaks is associated with altered patterns of gene expression

2006 ◽  
Vol 16 (Supplement 1) ◽  
pp. S6
Author(s):  
W. Kaufmann ◽  
K. Nevis ◽  
P. Qu ◽  
J. Ibrahim ◽  
T. Zhou ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Cell Cycle Checkpoint ◽  
Double Strand Breaks ◽  
Dna Double Strand Breaks ◽  
Checkpoint Response ◽  
Strand Breaks ◽  
Cycle Checkpoint
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