Tuesday, May 13, 2025 5/13/2025

Development and validation of novel optical methods for direct screening of taste receptor activation

Award Number: R21DC020041
ORGANIZATION: NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

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Development and validation of novel optical methods for direct screening of taste receptor activation - PROJECT SUMMARY Taste family 1 receptors (T1Rs, forming sweet and umami receptors) and taste family 2 receptors (T2Rs, forming bitter receptors) are important G-protein couple receptors (GPCRs) expressed in taste buds of the tongue. T1Rs and T2Rs also play important chemosensory roles in tissues all over the body, including the airway epithelium and in immune cells. We and other have characterized T1Rs and T2Rs as novel players in innate immunity. They detect bacterial metabolites to stimulate rapid innate immune responses. While current screening methods have revealed a lot about specific agonists that activate specific T1Rs and T2Rs, understanding T1R and T2R activation by complex mixtures (e.g., plant extracts or conditioned media from bacteria) is currently very difficult. Existing screening methods are focused on calcium signaling as a read-out. Many compounds activate calcium signals independently of taste receptors in cultured cells, and the HEK293 cells commonly used express endogenous T2Rs. Calcium-focused screening methods have also likely missed “biased agonists,” that is, agonists which do not activate T2R G protein signaling but activate arrestin signaling. This has been reported for many GPCRs, but no one has yet screened for biased agonists of taste receptors. This is a critical gap in the studies of T1R and T2R biology that will be addressed here. To overcome limitations of calcium-based assays and reveal new insights into T2R-arrestin signaling, we will adapt a fluorescence-based tripartite GFP-based assay (known as the TRIO assay) that directly visualizes heterologously-expressed receptor activation through arrestin binding to the activated receptor. This assay is much less limited by off-target effects of complex mixtures or single compounds compared with current methods. It is also adaptable to high throughput plate-reader instrumentation. This assay is faster (1-2 hrs) than other GPCR assays which rely on transcription as a readout (12-24 hrs). We have utilized this assay to study activation and inhibition of protease-activated and adrenergic receptors in prior studies. Preliminary data suggest this assay works well with T2Rs. We hypothesize that T1R and T2R TRIO assays will reveal novel ligand-T1R/T2R interactions and may be useful to de-orphanize the remaining orphan T2R isoforms. In this proposal, Aim 1 will focus on validation of this optical assay using known bitter and sweet compounds. Aim 2 will use this assay to screen taste receptors against common pathogenic bacteria and fungi to characterize which receptors are activated by which pathogens. New clinically relevant data will be revealed, and the expression constructs validated in this study will become useful tools for taste receptor research. All expression vectors will be made widely available via Addgene after initial publication. This research has important utility for both immune research as well as molecular sensory nutrition and taste research.


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: 2025 ( Subtotal = -$4,253 )
2025	2024	TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA, THE	3451 WALNUT ST STE 440A	PHILADELPHIA	PA	19104	PHILADELPHIA	USA	Research Related to Deafness and Communication Disorders	000	2	3/31/2025	NON-COMPETING CONTINUATION	-$4,253
														Subtotal = -$4,253

Issue Date FY: 2024 ( Subtotal = $203,125 )
2024	2024	TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA, THE	3451 WALNUT ST STE 440A	PHILADELPHIA	PA	19104	PHILADELPHIA	USA	Research Related to Deafness and Communication Disorders	000	2	11/28/2023	NON-COMPETING CONTINUATION	$182,813
2024	2024	TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA, THE	3451 WALNUT ST STE 440A	PHILADELPHIA	PA	19104	PHILADELPHIA	USA	Research Related to Deafness and Communication Disorders	001	2	5/3/2024	NON-COMPETING CONTINUATION	$20,312
														Subtotal = $203,125

Issue Date FY: 2023 ( Subtotal = $243,750 )
2023	2023	TRUSTEES OF THE UNIVERSITY OF PENNSYLVANIA, THE	3451 WALNUT ST STE 440A	PHILADELPHIA	PA	19104	PHILADELPHIA	USA	Research Related to Deafness and Communication Disorders	000	1	11/19/2022	NEW	$243,750
														Subtotal = $243,750

Grand Total All Awards = $442,622

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Development and validation of novel optical methods for direct screening of taste receptor activation

Award Number: R21DC020041

ORGANIZATION: NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

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