Integrating Human Genetics and Protective Genome Editing to Enable ADGRE2-Directed AML Therapy

Keschner, Y.; Etchin, J.; Silva, M.; Mager, H.; Hoyt, H.; Xavier-Ferrucio, J.; Manalo, N.; Halfond, A.; Qiu, H.; Wang, R.; Lin, M. I.; Ge, H. G.; Scherer, J.; Chakraborty, T.; Lydeard, J. R.

doi:10.1101/2025.08.21.671614

bioRxiv · 10.1101/2025.08.21.671614

Integrating Human Genetics and Protective Genome Editing to Enable ADGRE2-Directed AML Therapy

Keschner, Y.·Etchin, J.·Silva, M.·Mager, H.·Hoyt, H.·Xavier-Ferrucio, J.·Manalo, N.·Halfond, A.·Qiu, H.·Wang, R.·Lin, M. I.·Ge, H. G.·Scherer, J.·Chakraborty, T.·Lydeard, J. R.

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Abstract

Acute myeloid leukemia (AML) remains a major therapeutic challenge due to extensive disease heterogeneity and lack of cancer-specific antigens. ADGRE2 has emerged as a promising AML target with broad expression in AML patient blast and leukemic stem cell-enriched populations. However, comparable expression in healthy hematopoietic stem and progenitor cells (HSPCs) and myeloid lineages suggests a high susceptibility to on-target, off-tumor myelotoxicity with ADGRE2-targeted therapies. Guided by human genetics data identifying loss-of-function variants, we evaluated whether ADGRE2 is dispensable in hematopoietic stem cells as a protective approach for transplant-based shielding from ADGRE2-directed therapies. Using CRISPR-Cas9 and adenine base editors, we achieved high-efficiency ADGRE2 knockout (>94%) in HSPCs with corresponding protein loss without impairing cell viability, differentiation, and cytokine release in vitro, or long-term engraftment, multilineage differentiation, and persistence of gene editing in mouse xenografts. We also developed novel ADGRE2-specific chimeric antigen receptor (CAR) T cells that demonstrated potent cytotoxicity against AML cells, even at low antigen levels. Together, these findings establish ADGRE2 as a compelling AML target and provide a framework for hematopoietic stem cell transplant with protective gene editing to enable ADGRE2-directed immunotherapies while minimizing myelotoxicity.

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Keschner, Y., Etchin, J., Silva, M., Mager, H., Hoyt, H., Xavier-Ferrucio, J., Manalo, N., Halfond, A., Qiu, H., Wang, R., Lin, M. I., Ge, H. G., Scherer, J., Chakraborty, T., Lydeard, J. R.. 2025-08-22. Integrating Human Genetics and Protective Genome Editing to Enable ADGRE2-Directed AML Therapy. https://doi.org/10.1101/2025.08.21.671614

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