Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
[Preprint]. 2025 May 13:rs.3.rs-6528748.
doi: 10.21203/rs.3.rs-6528748/v1.

Identifying Targeted Therapies for CBFA2T3::GLIS2 Acute Myeloid Leukemia

Fanny Gonzales  1 , Constanze Schneider  2 , Gabriela Alexe  3 , Shan Lin  4 , Delan Khalid  2 , Montserrat Alvarez  1 , Allen Basanthakumar  2 , Jana Ellegast  5 , Lucy Merickel  2 , Silvi Salhotra  2 , Audrey Taillon  2 , Mark Wunderlich  6 , Marc Ansari  7 , Jennifer Perry  2 , Barbara Degar  2 , Yana Pikman  2 , Kimberly Stegmaier  2
Affiliations

Identifying Targeted Therapies for CBFA2T3::GLIS2 Acute Myeloid Leukemia

Fanny Gonzales et al. Res Sq. .

Abstract

CBFA2T3::GLIS2 fusion positive pediatric acute myeloid leukemia (AML) remains one of the worst prognostic AML subgroups. To uncover innovative targeted therapeutic approaches in this disease subtype we performed genome-scale CRISPR-Cas9 screening that highlighted a strong, selective dependency on JAK2 compared to other types of cancer. Using a doxycycline-inducible JAK2 knockout (KO) system, we validated JAK2 dependency in CBFA2T3::GLIS2 cell lines, observing impaired proliferation in vitro and in vivo and induced apoptosis with JAK2 KO. Both type I (ruxolitinib) and type II (CHZ868) JAK2 inhibitors showed selective in vitro activity in CBFA2T3::GLIS2 positive AML models. To identify resistance and sensitizer mechanisms to JAK2 inhibitors, we used CRISPR-Cas9 ruxolitinib anchor screening in CBFA2T3::GLIS2 AML. sgRNAs targeting negative regulators of the MAPK pathway were enriched in the ruxolitinib-treated cells. Similarly, CBFA2T3::GLIS2 AML sublines grown to resistance under chronic ruxolitinib treatment expressed pathogenic NRAS mutations. Both approaches converged on MAPK pathway activation as a resistance mechanism to ruxolitinib treatment. Combining ruxolitinib with MEK inhibitors showed a synergistic effect in cell lines and patient-derived xenograft (PDX) cells expressing the fusion and in vivo activity in a CBFA2T3::GLIS2 AML PDX, suggesting a potential approach to target this signaling circuitry in this poor outcome AML subtype.

PubMed Disclaimer

Conflict of interest statement

Additional Declarations: Yes there is potential conflict of interest.

Figures

Figure 1
Figure 1
Legend not included with this version
Figure 2
Figure 2
Legend not included with this version
Figure 3
Figure 3
Legend not included with this version
Figure 4
Figure 4
Legend not included with this version
Figure 5
Figure 5
Legend not included with this version
Figure 6
Figure 6
Legend not included with this version
Figure 7
Figure 7
Legend not included with this version
See this image and copyright information in PMC

References

    1. Lonetti A, Pession A, Masetti R. Targeted Therapies for Pediatric AML: Gaps and Perspective. Front Pediatr. 2019;7:463. - PMC - PubMed
    1. Athale UH, Razzouk BI, Raimondi SC, Tong X, Behm FG, Head DR, et al. Biology and outcome of childhood acute megakaryoblastic leukemia: a single institution’s experience. Blood. 2001. Jun 15;97(12):3727–32. - PubMed
    1. Barnard DR, Alonzo TA, Gerbing RB, Lange B, Woods WG, Children’s Oncology Group. Comparison of childhood myelodysplastic syndrome, AML FAB M6 or M7, CCG 2891: report from the Children’s Oncology Group. Pediatr Blood Cancer. 2007. Jul;49(1):17–22. - PubMed
    1. Gruber TA, Larson Gedman A, Zhang J, Koss CS, Marada S, Ta HQ, et al. An Inv(16)(p13.3q24.3)-encoded CBFA2T3-GLIS2 fusion protein defines an aggressive subtype of pediatric acute megakaryoblastic leukemia. Cancer Cell. 2012. Nov 13;22(5):683–97. - PMC - PubMed
    1. Thirant C, Ignacimouttou C, Lopez CK, Diop M, Le Mouël L, Thiollier C, et al. ETO2-GLIS2 Hijacks Transcriptional Complexes to Drive Cellular Identity and Self-Renewal in Pediatric Acute Megakaryoblastic Leukemia. Cancer Cell. 2017. Mar 13;31(3):452–65. - PubMed

Publication types

LinkOut - more resources