Dual intron-targeted CRISPR-Cas9-mediated disruption of the AML RUNX1-RUNX1T1 fusion gene effectively inhibits proliferation and decreases tumor volume in vitro and in vivo

TY - JOUR

T1 - Dual intron-targeted CRISPR-Cas9-mediated disruption of the AML RUNX1-RUNX1T1 fusion gene effectively inhibits proliferation and decreases tumor volume in vitro and in vivo

AU - Neldeborg, Signe

AU - Soerensen, Johannes Frasez

AU - Møller, Charlotte Thornild

AU - Bill, Marie

AU - Gao, Zongliang

AU - Bak, Rasmus O

AU - Holm, Kasper

AU - Sorensen, Boe

AU - Nyegaard, Mette

AU - Luo, Yonglun

AU - Hokland, Peter

AU - Stougaard, Magnus

AU - Ludvigsen, Maja

AU - Holm, Christian Kanstrup

PY - 2023/9

Y1 - 2023/9

N2 - Oncogenic fusion drivers are common in hematological cancers and are thus relevant targets of future CRISPR-Cas9-based treatment strategies. However, breakpoint-location variation in patients pose a challenge to traditional breakpoint-targeting CRISPR-Cas9-mediated disruption strategies. Here we present a new dual intron-targeting CRISPR-Cas9 treatment strategy, for targeting t(8;21) found in 5-10% of de novo acute myeloid leukemia (AML), which efficiently disrupts fusion genes without prior identification of breakpoint location. We show in vitro growth rate and proliferation reduction by 69 and 94% in AML t(8;21) Kasumi-1 cells, following dual intron-targeted disruption of RUNX1-RUNX1T1 compared to a non t(8;21) AML control. Furthermore, mice injected with RUNX1-RUNX1T1-disrupted Kasumi-1 cells had in vivo tumor growth reduction by 69 and 91% compared to controls. Demonstrating the feasibility of RUNX1-RUNX1T1 disruption, these findings were substantiated in isolated primary cells from a patient diagnosed with AML t(8;21). In conclusion, we demonstrate proof-of-principle of a dual intron-targeting CRISPR-Cas9 treatment strategy in AML t(8;21) without need for precise knowledge of the breakpoint location.

AB - Oncogenic fusion drivers are common in hematological cancers and are thus relevant targets of future CRISPR-Cas9-based treatment strategies. However, breakpoint-location variation in patients pose a challenge to traditional breakpoint-targeting CRISPR-Cas9-mediated disruption strategies. Here we present a new dual intron-targeting CRISPR-Cas9 treatment strategy, for targeting t(8;21) found in 5-10% of de novo acute myeloid leukemia (AML), which efficiently disrupts fusion genes without prior identification of breakpoint location. We show in vitro growth rate and proliferation reduction by 69 and 94% in AML t(8;21) Kasumi-1 cells, following dual intron-targeted disruption of RUNX1-RUNX1T1 compared to a non t(8;21) AML control. Furthermore, mice injected with RUNX1-RUNX1T1-disrupted Kasumi-1 cells had in vivo tumor growth reduction by 69 and 91% compared to controls. Demonstrating the feasibility of RUNX1-RUNX1T1 disruption, these findings were substantiated in isolated primary cells from a patient diagnosed with AML t(8;21). In conclusion, we demonstrate proof-of-principle of a dual intron-targeting CRISPR-Cas9 treatment strategy in AML t(8;21) without need for precise knowledge of the breakpoint location.

KW - Animals

KW - Mice

KW - RUNX1 Translocation Partner 1 Protein/genetics

KW - Translocation, Genetic

KW - Introns/genetics

KW - Core Binding Factor Alpha 2 Subunit/genetics

KW - Tumor Burden

KW - CRISPR-Cas Systems

KW - Leukemia, Myeloid, Acute/genetics

KW - Cell Proliferation

KW - Oncogene Proteins, Fusion/genetics

UR - http://www.scopus.com/inward/record.url?scp=85164995645&partnerID=8YFLogxK

U2 - 10.1038/s41375-023-01950-9

DO - 10.1038/s41375-023-01950-9

M3 - Journal article

C2 - 37464068

SN - 0887-6924

VL - 37

SP - 1792

EP - 1801

JO - Leukemia

JF - Leukemia

IS - 9

ER -