Developing an engineered bacterial probiotic via targeted conjugative killing of fungal pathogen nakaseomyces glabratus - 1 project summary/abstract 2 despite killing millions of people every year,…

The National Institute of Allergy and Infectious Diseases awarded the University of Wisconsin System $393,413 on September 1, 2026, under the Allergy and Infectious Diseases Research program (CFDA 93.855) to develop an engineered bacterial probiotic strategy for targeting the fungal pathogen Nakaseomyces glabratus.

The research aims to demonstrate species-specific killing of N. glabratus through interdomain conjugation—the delivery of engineered DNA from bacteria to fungal cells—using a conjugatively delivered CRISPR-Cas9 system that targets only N. glabratus cells while sparing benign fungal strains. The project will characterize control mechanisms for interdomain conjugation via mannoprotein adhesion and develop tools to control and tailor the approach to infectious N. glabratus cells. The long-term objective is to establish a novel antifungal treatment platform readily adaptable to other infectious species, requiring substantially less time and resources than conventional drug development. N. glabratus causes candidiasis and candidemia with mortality rates as high as 74 percent, and drug-resistant strains are rising faster than related fungal infections, with few new antifungals in development.

Work is performed in Stevens Point, Wisconsin, with a period of performance from September 1, 2026, through August 31, 2029. The award is classified as a Project Grant, a standard assistance type under the program supporting biomedical research and research training in infectious diseases and related areas at public and private nonprofit institutions.

Developing an engineered bacterial probiotic via targeted conjugative killing of fungal pathogen nakaseomyces glabratus - 1 project summary/abstract 2 despite killing millions of people every year, fungal infections receive markedly less attention than 3 bacterial infections. These fungal infections are also predicted to rise, due chiefly to changes in 4 climate and overuse of antifungals, and drug-resistance fungi are also increasing. Nakaseomyces 5 glabratus is a leading cause of candidiasis-infection of organs such as the mouth, vagina, or 6 skin-often leading to candidemia of the blood, which carries with IT mortality rates as high as 7 74% as cells attack the heart, central nervous system, and other internal organs. Worse, cases of 8 N. Glabratus infections are rising faster than other related fungal infections, as are drug-resistant 9 strains, and new antifungals to combat this growing threat are few and slow coming. Thus, we 10 propose an innovative strategy that uses engineered DNA, delivered by bacterial conjugation to 11 infectious fungal cells (interdomain conjugation, or IDC), as way to reliably and specifically treat 12 against N. Glabratus. The objective of this research is to cause depression and/or collapse of N. 13 glabratus populations via IDC, by developing tools to control IDC and tailoring these tools to 14 infectious N. Glabratus cells. The long-term goal of this work is to demonstrate a novel platform 15 for antifungal treatment that could be readily adapted to other infectious species, with much less 16 time and fewer resources than required for drug development. Thus, we propose three specific 17 aims to develop and demonstrate this strategy: 1) demonstrate species-specific killing of N. 18 glabratus by IDC, using a conjugatively delivered crispr-CAS9 system that targets only N. 19 glabratus cells (VS. A benign saccharomyces cerevisiae strain). 2) characterize control 20 mechanisms for IDC via mannoprotein adhesion. Because bacterial escherichia coli cells used to 21 deliver DNA through IDC are naturally able to adhere to mannoproteins in fungal cell walls, and 22 because previous research has demonstrated the importance of this adhesion for IDC efficiency, 23 we will put this adhesion under engineered control, to test the limits of its effect on IDC-mediated 24 killing. 3) determine infection-specific binding modalities for IDC, by testing different adhesins 25 against both benign yeast cells and fungal biofilm cells, to determine which conditions favor 26 bacterial binding to biofilm cells specifically, and thus greater IDC to those cells. This proposal, 27 in addition to providing rural and low-income students experience with innovative 28 bioengineering, will expose them to techniques used in cutting edge biotech and medical research.