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Tool development for Anaplasma phagocytophilum to understand determinants of infection

Award Number: R61AI179933
ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 01/01/2024
PERIOD OF PERFORMANCE END DATE: 12/31/2026

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Tool development for Anaplasma phagocytophilum to understand determinants of infection - PROJECT SUMMARY Rickettsiales are a diverse order of obligate, intracellular bacteria often transmitted by arthropods and pose a substantial threat to public health. Rickettsial organisms manipulate a wide array of host cell processes to gain entry and establish a replicative niche, while simultaneously avoiding host cell defenses. How rickettsial pathogens mediate this complex network of interactions is not well understood. A central feature of all rickettsial pathogens is the Type 4 Secretion System (T4SS), which secretes effector proteins into the cytosol and manipulates biological processes in the host cell. However, few T4SS translocated effectors have been identified or characterized from rickettsial organisms owing, in part, to the lack of genetic tools available among these bacteria. Their obligate intracellular nature has historically hindered genetic manipulation of Rickettsiales. Recently, we achieved targeted gene deletion in the rickettsial pathogen A. phagocytophilum using allelic exchange by homologous recombination. In the R61 portion of this proposal we will leverage this technique to add to the A. phagocytophilum genetic toolbox by developing 1) a T4SS effector translocation assay for use in the organism of study, 2) conditional expression systems, and 3) luminescent reporters. Using these tools in the R33 phase we will: 4) characterize the A. phagocytophilum T4SS effector repertoire, 5) evaluate the contributions of essential A. phagocytophilum effectors to infection, and 6) monitor the expression kinetics of effectors in vivo. The objectives of this proposal will expand the tractability of A. phagocytophilum and our understanding of the host-pathogen interactions it navigates during infection through elucidation to the T4 effector repertoire, and an understanding of how, when and where these genes are regulated. The impact of this proposal will extend to other rickettsial organisms, as these approaches can be adapted for use in related intracellular pathogens.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2026 ( Subtotal = $251,560 )
2026	2026	WASHINGTON STATE UNIVERSITY	240 FRENCH ADMINISTRATION BLDG	PULLMAN	WA	99164	WHITMAN	USA	Allergy and Infectious Diseases Research	000	3	1/9/2026	NON-COMPETING CONTINUATION	$251,560
														Subtotal = $251,560

Issue Date FY: 2025 ( Subtotal = $259,814 )
2025	2025	WASHINGTON STATE UNIVERSITY	240 FRENCH ADMINISTRATION BLDG	PULLMAN	WA	99164	WHITMAN	USA	Allergy and Infectious Diseases Research	000	2	12/11/2024	NON-COMPETING CONTINUATION	$259,814
														Subtotal = $259,814

Issue Date FY: 2024 ( Subtotal = $267,750 )
2024	2024	WASHINGTON STATE UNIVERSITY	240 FRENCH ADMINISTRATION BLDG	PULLMAN	WA	99164	WHITMAN	USA	Allergy and Infectious Diseases Research	000	1	12/11/2023	NEW	$267,750
														Subtotal = $267,750

Grand Total All Awards = $779,124

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Tool development for Anaplasma phagocytophilum to understand determinants of infection

Award Number: R61AI179933

ORGANIZATION: NATIONAL INSTITUTE OF ALLERGY & INFECTIOUS DISEASES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 01/01/2024

PERIOD OF PERFORMANCE END DATE: 12/31/2026

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