Monday, November 25, 2024 11/25/2024

Expanding the set of genetically encoded tools for compartment-specific manipulation of redox metabolism in living cells

Award Number: R35GM142495
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

Group Awards By:

View Award Abstract

Abstract One of the most important organizing principles in all life forms is the uninterrupted flow of electrons through reactions that involve reduction-oxidation (redox) changes. Not surprisingly, an imbalance in this fundamental cellular process, i.e. redox homeostasis, has been attributed to numerous diseases, including mitochondrial disorders, cancer, diabetes, neurodegeneration and the aging process itself. The redox cofactors, NADH and NADPH, and their oxidized forms are key contributors to the cellular redox environment, but it is unclear whether perturbations in their metabolism contribute directly to disease etiology or is simply a reflection of ongoing pathology. For most of these conditions, it is not known whether the observed redox imbalance is linked to altered bioenergetic efficiency or to a cellular process that is neither linked to ATP production nor to maintenance of the mitochondrial membrane potential. Another major challenge is that some of these redox reactions are redundant, i.e. have overlapping substrate dependency (towards NAD(P)H) or are found in more than one cellular compartment. To systematically address these pressing questions, methodology to modulate the steady-state concentrations of the NADH and NADPH cofactors is needed. Recently, we have developed genetically encoded tools to selectively decrease the NADH/NAD+ and NADPH/NADP+ ratios in live cells that are based on the heterologous expression of native or engineered versions of bacterial H 2O- forming NAD(P)H oxidases. In this proposal, we plan to expand our toolkit by developing a genetically encoded tool for the direct modulation of NADH reductive stress (i.e. increased NADH/NAD+ ratio) (Project 1). Preliminary screening of several bacterial enzymes has furnished promising candidates for driving NADH overproduction in different cellular compartments. The development of compartment-specific tools will enable studies to elucidate how overproduction of reducing equivalents in one cellular compartment is communicated to another and how NADH reductive stress remodels cellular metabolism (Project 2a). Multiple lines of evidence indicate that NAD(P)H-consuming redox cycling agents at low concentrations mildly exhaust antioxidant systems and that the resulting pro-oxidative shift promotes stress resistance and improves heathspan in several model organisms. We are using Drosophila as a model organism, to directly test whether redox modulation in either the oxidative or reductive direction are correlated with stress resistance, healthspan and lifespan (Project 2b). A third goal is to develop variants of our genetically encoded tools that are controlled by small molecules or by light to afford greater spatiotemporal control (Project 3). The latter is especially important as many redox processes crucial for redox signaling or energy metabolism and dysregulated in pathologies, occur rapidly (on an acute time scale). The successful completion of our studies will lead to enabling technologies for modulating the redox environment, which will be widely useful for metabolic studies on an acute or chronic time scale at the resolution of subcellular compartments.


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 = $480,000 )
2024	2024	THE SCINTILLON INSTITUTE	6868 NANCY RIDGE DR STE 200	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	000	4	6/13/2024	NON-COMPETING CONTINUATION	$480,000
														Subtotal = $480,000

Issue Date FY: 2023 ( Subtotal = $480,000 )
2023	2023	THE SCINTILLON INSTITUTE	6868 NANCY RIDGE DR	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	000	3	7/27/2023	NON-COMPETING CONTINUATION	$480,000
														Subtotal = $480,000

Issue Date FY: 2022 ( Subtotal = $726,546 )
2022	2022	SCINTILLON INSTITUTE,THE	6868 NANCY RIDGE DR	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	001	2	5/25/2022	SUPPLEMENT FOR EXPANSION	$246,546
2022	2022	SCINTILLON INSTITUTE,THE	6868 NANCY RIDGE DR	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	002	2	6/23/2022	NON-COMPETING CONTINUATION	$48,000
2022	2022	SCINTILLON INSTITUTE,THE	6868 NANCY RIDGE DR	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	000	2	4/5/2022	NON-COMPETING CONTINUATION	$432,000
														Subtotal = $726,546

Issue Date FY: 2021 ( Subtotal = $313,219 )
2021	2021	SCINTILLON INSTITUTE,THE	6868 NANCY RIDGE DR	SAN DIEGO	CA	92121	SAN DIEGO	USA	Biomedical Research and Research Training	000	1	6/23/2021	NEW	$313,219
														Subtotal = $313,219

Grand Total All Awards = $1,999,765

Top

All Categories

About

Search

Reports

Data Submission

Award Information

Expanding the set of genetically encoded tools for compartment-specific manipulation of redox metabolism in living cells

Award Number: R35GM142495

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

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

Federal Websites

Department of Health & Human Services

HHS Operating Divisions

HHS Staff Divisions

Download A Document Viewer