Printing: This poster is 48” wide by 36” high. It’s designed to be printed on a large-format printer.

Customizing the Content: The placeholders in this poster are formatted for you. Type in the placeholders to add text, or click an icon to add a table, chart, SmartArt graphic, picture or multimedia file.

To add or remove bullet points from text, click the Bullets button on the Home tab.

If you need more placeholders for titles, content or body text, make a copy of what you need and drag it into place. PowerPoint’s Smart Guides will help you align it with everything else.

Want to use your own pictures instead of ours? No problem! Just click a picture, press the Delete key, then click the icon to add your picture.

Effects of Epigenetic Modifier Inhibitors on AML Cell Sensitivity to Differentiation Therapy

Kalsi K. Heimdal1, Edjay Ralph A. Hernandez1, and Dr. Heidi J. Super1 || 1Department of Biology, Minot State University, Minot, ND, 58707

Background Methods

Proliferation and MTT Assay Results

Cytospin Analysis

Conclusion

• The U937 cells showed that the use of epigenetic inhibitors such as

CI-994 and TCP is effective in sensitizing them to respond to

differentiation therapy.

• Only the combination of CI-994 and ATRA seemed to cause the

MV4;11 cells to respond to differentiation therapy. Using CI-994

alone also showed a slight amount of MV4;11 cell differentiation.

Future Directions and Acknowledgements

• Perform Fluoresce Activated Cell Sorting (FACS) assay to

determine the amount of CD11b expression on the cell surface.

• Research was supported by an Institutional Development Award

(IDeA) from the Nation Institute of General Medical Sciences of the

National Institute of Health under grant number P20GM103442.

• Human acute myeloid leukemia (AML) is associated with specific, recurrent gene alterations, which create leukemogenic oncoproteins that block differentiation and promote proliferation.

• Activation or inhibition of specific genes might be necessary to allow cells to differentiate, which can be done by using epigenetic inhibiting drugs.

• ATRA (all-trans-retinoic acid/Vitamin A) is a differentiation promoting drug, and is also a promising alternative to chemotherapy.

• AML resulting from MLL fusion genes responds poorly to treatment with all-trans-retinoic acid (ATRA), whereas acute myeloid leukemia resulting from a different fusion oncoprotien, fusing the PML gene to the RARA gene, responds extremely well.

• In addition to fusion oncogenes, subtypes of AML exhibit distinct and abnormal epigenetic patterns of histone modification and DNA methylation. Therefore, treatments involving epigenetic modifier inhibitors have become increasingly common.

•

• We are using Tranylcypromine (TCP), and CI-994 on MV4;11 (AML cell line with MLL translocation), and U937 (AML cell line with no MLL translocation) to determine its effects on non-APL AML cells.

• MV4;11 cells were cultured in IMDM with 10% Fetal Bovine Serum,

and 1% penicillin streptomycin.

• U937 cells were cultured in RPMI-1640 with 10% Fetal Bovine

Serum, and 1% penicillin streptomycin.

• Direct cell counts were made using Bio-Rad TC20TM Automated

Cell Counter.

• Viability was determined by trypan blue exclusion.

• MTT assay was conducted following ATCC bioproductsTM MTT Cell

Proliferation Assay Protocol.

• Cytospin analysis was conducted following ThermoShandon

Cytospin Protocol.

Hypothesis

• We hypothesize that changes in epigenetic modifications due to

treatment with TCP or CI-994 could sensitize more types of AML to

differentiation treatments such as ATRA.

• Differentiation will be noted indirectly and directly as reduced cell

proliferation, and metabolic activity, as well as by changes in the

morphology of the cells/nuclei.

Type of Modification Effect on DNA/Gene

Expression

Inhibitor Effect of Inhibitor on

DNA/Gene Expression

Histone deacetylation Transcription repression CI-994 Transcription activation

Histone demethylation Transcription activation TCP Transcription repression

Figure 3.

7-day proliferation

assay of MV4;11 cells

treated with CI-994

and TCP. Cell count is

shown as average of

duplicates of samples

with SEM.

Figure 2.

7-day proliferation

assay of U937 cells

treated with CI-994

and TCP. Cell count is

shown as average of

duplicates of samples

with SEM.

Figure 4.

4-day MTT assay

of MV4;11 cells

treated with CI-994

and TCP. Absorbance

is shown as average of

triplicates of samples

with SEM.

Figure 5. U937 cells treated

with TPA. Figure 6. Untreated U937 cells.

Figure 9. MV4;11 cells

treated with ATRA+CI-994.

Figure 10. MV4;11 cells

treated with ATRA+TCP.

Figure 7. U937 cells treated

with ATRA+CI-994.

Figure 8. U937 cells treated

with ATRA+TCP.

Table 1. Common epigenetic modifications, their affects, and

associated inhibitors.