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Calcium signaling controls Pseudomonas aeruginosa invasion and adaptation to the host intracellular environment

Award Number: R15GM124670
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 08/01/2017
PERIOD OF PERFORMANCE END DATE: 07/31/2025

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Calcium signaling controls Pseudomonas aeruginosa invasion and adaptation to the host intracellular environment - ABSTRACT The currently funded research 2R15GM124670-02 is focused on discovering the role of Ca2+ signaling in the interactions of P. aeruginosa with host cells. These studies include establishing the signaling patterns of Ca2+ homeostasis by monitoring the changes of Ca2+ levels in P. aeruginosa during interactions with host cells, including invasion, intracellular survival, and escape. To achieve these goals, a careful examination of host and pathogen Ca2+ homeostasis on one side and their cellular responses to spatial and temporal Ca2+ alterations on the other is required. To monitor the changes in spatial relationship between Pa and host cells, we plan to use a combination of fluorescent tagging (GFP, RFP, YFP) and fluorescent (DAPI, Alexa Fluor Phalloidin) or immunofluorescent staining. To monitor micro-spatial and temporal translocations of free Ca2+ within subcellular compartments where Pa interacts with host cells, we plan to use the Ca2+-sensitive fluorescent Genetic Encoded Calcium Indicators (GECIs) engineered to localize near the outer membrane facing the extracellular space, in the periplasm, near the cytoplasmic membrane facing the cytoplasm, and in the cytoplasm. All these approaches are based on the application of fluorescent molecules. Therefore, we are requesting to support a purchase of Inverted Fluorescence Phase Contrast microscope, Leica DMi8, the premium-class modular research microscope. This will be our first fluorescence microscope in the lab. The requested instrument will be placed in the PI’s lab and will be available for the use by all the students in the PI’s lab and the collaborating lab of Dr. Lutter. All the participating students will be trained to use the instrument and the accompanying software by the company representative. The anticipated research advances will significantly enhance our ability to study calcium signaling in P. aeruginosa interacting with its eukaryotic host. The obtained new knowledge will improve our understanding of this important human pathogen and inform the development of more efficient means for controlling or preventing its deadly infections.


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: 2024 ( Subtotal = $0 )
2024	2023	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	003	2	6/24/2024	SUPPLEMENT FOR EXPANSION	$0
2024	2022	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	002	2	6/24/2024	SUPPLEMENT FOR EXPANSION	$0
2024	2022	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	001	2	6/24/2024	SUPPLEMENT FOR EXPANSION	$0
2024	2021	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	000	2	6/24/2024	COMPETING CONTINUATION	$0
														Subtotal = $0

Issue Date FY: 2023 ( Subtotal = $88,140 )
2023	2023	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	000	2	6/28/2023	SUPPLEMENT FOR EXPANSION	$88,140
														Subtotal = $88,140

Issue Date FY: 2022 ( Subtotal = $138,043 )
2022	2022	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	003	2	9/12/2022	SUPPLEMENT FOR EXPANSION	$38,531
2022	2022	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	002	2	9/7/2022	SUPPLEMENT FOR EXPANSION	$99,512
2022	2021	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	001	2	3/2/2022	COMPETING CONTINUATION	$2,128
2022	2019	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	000	1	3/2/2022	SUPPLEMENT FOR EXPANSION	-$2,128
														Subtotal = $138,043

Issue Date FY: 2021 ( Subtotal = $441,822 )
2021	2021	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	000	2	7/20/2021	COMPETING CONTINUATION	$441,822
														Subtotal = $441,822

Issue Date FY: 2019 ( Subtotal = $29,001 )
2019	2019	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	000	1	2/5/2019	SUPPLEMENT FOR EXPANSION	$29,001
														Subtotal = $29,001

Issue Date FY: 2018 ( Subtotal = $0 )
2018	2017	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	001	1	11/29/2017	NEW	$437,357
2018	2017	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	000	1	11/28/2017	NEW	-$437,357
														Subtotal = $0

Issue Date FY: 2017 ( Subtotal = $437,357 )
2017	2017	OKLAHOMA STATE UNIVERSITY	401 WHITEHURST HALL	STILLWATER	OK	74078	PAYNE	USA	Biomedical Research and Research Training	000	1	7/31/2017	NEW	$437,357
														Subtotal = $437,357

Grand Total All Awards = $1,134,363

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Calcium signaling controls Pseudomonas aeruginosa invasion and adaptation to the host intracellular environment

Award Number: R15GM124670

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

PERIOD OF PERFORMANCE START DATE: 08/01/2017

PERIOD OF PERFORMANCE END DATE: 07/31/2025

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