# Multiplex Epitope Editing to Enable Novel Immunotherapies for Acute Myeloid Leukemia

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 2024 · $780,389

## Abstract

PROJECT SUMMARY
While immunotherapies have proved compelling efficacy against other leukemias, their application for acute
myeloid leukemia (AML) is still hampered by the absence of tumor-restricted targets. The most suitable AML
targets are shared with healthy hematopoietic stem/progenitor cells (HSPCs) or mature myeloid cells, leading to
on-target/off-tumor toxicity and impairment of hematopoietic reconstitution. To address this issue, we
hypothesized that epitope-engineering of donor HSPCs used for conventional bone marrow transplantation can
endow hematopoietic lineages with selective resistance to CAR-T or monoclonal antibodies (mAb), without
affecting protein function or regulation. This strategy allows targeting genes essential for leukemia survival
regardless of shared expression on HSPCs, thus reducing the risk of tumor immune escape by antigen
downregulation/loss. We have already identified single amino-acid (aa) changes that abrogate the binding of
therapeutic mAb targeting FLT3, CD123, and KIT and optimized a base-editing approach to introduce them into
CD34+ HSPCs, which retain long-term engraftment and multilineage differentiation capacity. We confirmed the
in vivo resistance of epitope-edited hematopoiesis to CAR-T treatment and the concomitant eradication of
patient-derived AML xenografts (Casirati et al., Nature 2023). Here, we will capitalize on these achievements
and exploit state-of-the-art genetic engineering tools and in vivo modeling with the objectives to i) generate and
functionally validate “stealth” FLT3, KIT, and IL3RA genes by multiplex base-editing; ii) identify the best-
performing CAR configuration for multi-Ag targeting on AML samples and iii) validate resistance of edited HSPC
to new immunotherapies directed against these targets. This project will provide fundamental advancement of
new and more effective immunotherapy approaches for AML that should additionally have broad applicability to
several other hematopoietic malignancies.

## Key facts

- **NIH application ID:** 10980798
- **Project number:** 1R01CA286036-01A1
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **Principal Investigator:** Pietro Genovese
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $780,389
- **Award type:** 1
- **Project period:** 2024-07-03 → 2029-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10980798

## Citation

> US National Institutes of Health, RePORTER application 10980798, Multiplex Epitope Editing to Enable Novel Immunotherapies for Acute Myeloid Leukemia (1R01CA286036-01A1). Retrieved via AI Analytics 2026-06-02 from https://api.ai-analytics.org/grant/nih/10980798. Licensed CC0.

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