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Single Cell Analysis of MAPK Signaling Dynamics in Multicellularity

Award Number: R35GM133499
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: 08/07/2019
PERIOD OF PERFORMANCE END DATE: 12/31/2029

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Single Cell Analysis of MAPK Signaling Dynamics in Multicellularity - Project Summary Cells have a remarkable ability to reproducibly respond to a changing environment even with promiscuous protein-protein interactions, stochastic gene expression, and macromolecular crowding. Our goal is to identify and characterize the mechanisms that robustly amplify signaling events over noise to govern cell biology and ultimately coordinate collective cell behavior. We use as a model the Mitogen Activated Protein Kinase (MAPK) pathway, a conserved signaling network involved in virtually all aspects of cell biology. Our focus on quantitatively understanding the timing of signaling events at the single cell and single molecule levels in the context of physiologically relevant multicellular systems allows us to dissect how signal and noise relate to functional outcome. We have developed biosensors and fully automated imaging platforms to simultaneously quantify multiple kinase activities in thousands of live single cells. More recently, we have generated transgenic mouse models to monitor signaling events in unperturbed mouse tissues and developing embryos. Using these approaches we have uncovered spatial and temporal patterns of signaling events that filter biological noise to effect deterministic outcomes. Our unique methodology features the high temporal resolution, sensor multiplexing capabilities, and single cell resolution essential for studying signaling network dynamics in single cells of a multicellular system. My research program is divided in two areas: (A) connecting single molecule behaviors to changes in cell state and (B) connecting single cell signaling activities to collective cell behaviors. This grant cycle, we propose to focus in four main questions: 1- What are the most upstream molecular events that engage the MAPK signaling network? 2- How do cells monitor and adjust the biophysical properties of the cytoplasm? 3- How are the collective properties of cytoskeleton driving cell-cell communication in tissues?, and 4- What are the mechanisms driving the unicellular to multicellular transition in mammalian embryos? Through these fundamental questions, our research program has expanded into several exciting new areas of modern biology including mechanobiology, phase transitions, and the regulation of macromolecular crowding. Using genetic and biochemical approaches coupled with powerful fluorescent biosensor and live cell imaging, we will define new biological mechanisms that control cell biology.


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 = $395,772 )
2026	2026	THE JOHNS HOPKINS UNIVERSITY	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	000	7	1/23/2026	NON-COMPETING CONTINUATION	$395,772
														Subtotal = $395,772

Issue Date FY: 2025 ( Subtotal = $479,038 )
2025	2025	THE JOHNS HOPKINS UNIVERSITY	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	001	6	1/23/2025	COMPETING CONTINUATION	$479,038
2025	2023	THE JOHNS HOPKINS UNIVERSITY	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	000	5	1/7/2025	NON-COMPETING CONTINUATION	$0
														Subtotal = $479,038

Issue Date FY: 2023 ( Subtotal = $409,375 )
2023	2023	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	000	5	7/7/2023	NON-COMPETING CONTINUATION	$409,375
														Subtotal = $409,375

Issue Date FY: 2022 ( Subtotal = $590,175 )
2022	2022	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	000	4	7/21/2022	NON-COMPETING CONTINUATION	$409,375
2022	2022	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	001	4	7/22/2022	SUPPLEMENT FOR EXPANSION	$180,800
														Subtotal = $590,175

Issue Date FY: 2021 ( Subtotal = $474,451 )
2021	2021	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	000	3	7/29/2021	NON-COMPETING CONTINUATION	$409,375
2021	2021	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	001	3	7/30/2021	SUPPLEMENT FOR EXPANSION	$65,076
														Subtotal = $474,451

Issue Date FY: 2020 ( Subtotal = $486,287 )
2020	2020	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	000	1	6/30/2020	SUPPLEMENT FOR EXPANSION	$5,918
2020	2020	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	001	2	7/29/2020	NON-COMPETING CONTINUATION	$480,369
														Subtotal = $486,287

Issue Date FY: 2019 ( Subtotal = $390,829 )
2019	2019	JOHNS HOPKINS UNIVERSITY, THE	3400 N CHARLES ST	BALTIMORE	MD	21218	BALTIMORE CITY	USA	Biomedical Research and Research Training	000	1	8/7/2019	NEW	$390,829
														Subtotal = $390,829

Grand Total All Awards = $3,225,927

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Single Cell Analysis of MAPK Signaling Dynamics in Multicellularity

Award Number: R35GM133499

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: 08/07/2019

PERIOD OF PERFORMANCE END DATE: 12/31/2029

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