Natthakan Thongon Department of Leukemia Colla’s Lab
Targeting DNA2 Overcomes Metabolic Reprogramming in 1q21 Multiple Myeloma
CRISPR/Cas9-based screening to identify DNA repair effectors whose loss of function suppresses MM cells’ resistance to ILF2 ASO-induced DNA damage
Targeting DNA2 enhances ILF2 ASO-induced apoptosis in JJN3 cells.
NT ASOs
ILF2 ASOs Veh NSC
90
Veh NSC
2
✱✱✱
✱✱✱✱
Vinculin
60
✱✱✱
0
ILF2 Cleaved caspase 3
30
DNA2 DNA2
-2
MMS19 DDB1 PRPF19 DNA2
g -H2AX
0
0
50
100
150
200
Rank
Western blot (left) and apoptosis (right) analyses were performed in JJN3 cells treated with vehicle (Veh) or the DNA2 inhibitor NSC105808 (NSC; 1µM) for 48 hours after long-term exposure to NT or ILF2 ASOs.
sgRNAs targeting MMS19 , DNA2 , and DDB1 genes were significantly depleted in ILF2 ASO-treated JJN3 cells but not KMS11 cells after 3 weeks of ASO-treatment. DNA2 is the only druggable target.
Aim #2: To dissect the mechanisms of DNA2 inhibition-induced synthetic lethality in MM cells undergoing metabolic reprogramming in the context of ILF2 depletion DNA2 inhibition decreases the oxygen consumption rate and increases ROS production in ILF2-depleted cells.
75
NT ASOs NT ASOs + NSC ILF2 ASOs ILF2 ASOs + NSC
500
✱✱✱
Oligomycin FCCP R/A
✱✱✱
60
400
45
300
✱✱✱
30
✱✱✱
200
Seahorse experiments (left) were performed in JJN3 cells treated with ASOs for 7 days prior to receiving NSC for 72 hours. Quantification of ROS production (middle) and transmission electron microscopy (right) were performed in JJN3 cells treated with ASOs for 7 days prior to receiving NSC for 48 hours.
15
100
0
0
0
20
40
60
80
Time (minutes)
Made with FlippingBook Online newsletter maker