Engineering car-neutrophils as a novel therapeutic modality for aspergillus fumigatus infection

The National Institute of Allergy and Infectious Diseases awarded the University of Wisconsin–Madison $213,811 on May 1, 2026, under the Allergy and Infectious Diseases Research program (CFDA 93.855) to develop chimeric antigen receptor-engineered neutrophils as a therapeutic for Aspergillus fumigatus infection.

The award funds research to engineer induced pluripotent stem cell-derived neutrophils armed with anti-Aspergillus CARs to overcome the limited efficacy of conventional antifungal agents and neutrophil transfusions in immunocompromised patients. The R21 phase aims to establish proof of principle that CAR-engineered neutrophils possess superior fungicidal properties. Work includes identifying single-chain variable fragments most effective for targeting A. fumigatus in induced pluripotent stem cell-derived neutrophils and characterizing their antifungal potential in vitro. If R21 milestones are achieved, the research will advance into an R33 phase to enhance antifungal potential and demonstrate efficacy and safety in vivo.

The period of performance runs from May 1, 2026, through April 30, 2028. Work is performed in Madison, Wisconsin.

Abstract

Invasive fungal infection by aspergillus species, most commonly by aspergillus fumigatus, remains one of the leading causes of mortality in immunocompromised patients undergoing solid organ or HSC transplantation because antifungal agents currently used in clinics are poorly effective in treating aspergillus infection in patients with impaired immunity and neutropenia due to drug toxicities, drug-drug interactions, and the emergence of drug-resistant strains. Neutrophils provide the first line of defense against fungal infection. They effectively kill fungi by fungicidal oxidative bursts, presentation of fungal antigens to t cells, and by increasing fungal clearance by folding their hyphae and decreasing the spreading of infection with extracellular traps (nets). However, the efficacy of neutrophil transfusions for a. Fumigatus has been limited. Therefore, enhancing the antifungal properties of neutrophils is essential for advancing adoptive neutrophil transfer for treating aspergillus infections. In this application, we aim to develop a novel therapeutic approach to effectively target a. Fumigatus infections using induced pluripotent stem cell (ipsc)-derived neutrophils (ineutrophils) armed with anti-aspergillus cars. The major goal of the R21 phase is to provide proof of principle that ineutrophils equipped with anti-aspergillus cars possess superior fungicidal properties. In aim 1, we will identify the single-chain variable fragment (scfv) that is most effective in ipsc-derived neutrophils for targeting a. Fumigatus. In aim 2, we will characterize the anti-fungal potential of anti-a. Fumigatus car-ineutrophils in vitro. If the R21 milestones are achieved, we will advance the development of car-ineutrophil therapies into the R33 phase by enhancing their antifungal potential and demonstrating their efficacy and safety in vivo. In aim 3, we will identify the most effective car configuration and genetic modifications that enhance the fungicidal properties of ineutrophils. In aim 4, we will assess the efficacy and toxicity of car-ineutrophils in vivo using zebrafish larvae and invasive pulmonary aspergillosis mouse models. Overall, generating car-ineutrophils that directly target fungal species will enable the development of a new class of antifungal therapies. These therapies will employ the adoptive transfer of readily available neutrophils with enhanced antifungal functions to treat life-threatening drug-resistant a. Fumigatus infections in patients with neutropenia or dysfunctional neutrophils.