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Regulation of protein dynamics in Drosophila embryos' ectoderm germ layer

Award Number: F30GM154411
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS
PERIOD OF PERFORMANCE START DATE: 06/01/2024
PERIOD OF PERFORMANCE END DATE: 05/31/2028

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Regulation of protein dynamics in Drosophila embryos' ectoderm germ layer - PROJECT SUMMARY/ ABSTRACT Gene expression is a complex process involving transcription, translation, and the turnover of mRNAs and proteins. Proper spatiotemporal regulation of gene expression and protein abundance plays a crucial role in guiding the differentiation of the fertilized egg into a complex animal. Understanding the regulation of protein dynamics can provide valuable insights into the differentiation of germ layers (endoderm, mesoderm, ectoderm), as well as the development of specialized cells and tissues. However, quantifying lineage-specific protein dynamics in developing embryos is challenging due to technical limitations in single-cell proteomics and the inherent heterogeneity of embryonic tissues. As a result, researchers have often used mRNA levels as a proxy for protein expression, but this approach has significant limitations due to the poor correlation between mRNA and protein levels. This discrepancy highlights the importance of posttranscriptional mechanisms, such as protein degradation and translation efficiencies, in determining cell fate. To address these issues and enhance our understanding of embryogenesis, it is crucial to measure proteome dynamics during cell differentiation directly. These measurements will provide a more comprehensive and accurate view of the intricate processes at play during embryonic development. This study proposes using genetically modified morphogen-flattened Drosophila embryos expressing four morphogens uniformly. These embryos contain a uniform cell type at gastrulation, different for each morphogen-flattened embryo. The focus will be on quantifying protein dynamics and posttranscriptional regulation within the ectoderm germ layer. My approach involves deep multiplexed proteomics and single-cell RNAseq to analyze proteome differentiation and stochastic cell fate diversification (Aim 1 & 2). Additionally, by achieving the desired outcomes of measuring posttranscriptional regulation (translation efficiency and protein turnover) in Aim 3, the proposed mathematical model will provide valuable insights into mRNA-protein dynamics during cell differentiation in developing embryos, which will address some of the discrepancies between the two. In conclusion, this proposal aims to decipher the mechanisms of embryonic cell-type differentiation by using morphogen-flattened embryos and employing an integrative analysis of genomics and proteomics measurements. This comprehensive approach will yield valuable insights into the intricate interplay of transcriptional and posttranscriptional regulation during development, enhancing our understanding of the fundamental principles in embryonic development and informing strategies for manipulating cell fate in regenerative medicine and tissue engineering.


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 = $34,114 )
2026	2026	THE TRUSTEES OF PRINCETON UNIVERSITY	1 NASSAU HALL	PRINCETON	NJ	08544	MERCER	USA	Biomedical Research and Research Training	000	3	5/11/2026	NON-COMPETING CONTINUATION	$34,114
														Subtotal = $34,114

Issue Date FY: 2025 ( Subtotal = $33,538 )
2025	2025	THE TRUSTEES OF PRINCETON UNIVERSITY	1 NASSAU HALL	PRINCETON	NJ	08544	MERCER	USA	Biomedical Research and Research Training	000	2	5/16/2025	NON-COMPETING CONTINUATION	$32,974
2025	2025	THE TRUSTEES OF PRINCETON UNIVERSITY	1 NASSAU HALL	PRINCETON	NJ	08544	MERCER	USA	Biomedical Research and Research Training	001	2	5/22/2025	NON-COMPETING CONTINUATION	$564
														Subtotal = $33,538

Issue Date FY: 2024 ( Subtotal = $32,974 )
2024	2024	THE TRUSTEES OF PRINCETON UNIVERSITY	1 NASSAU HALL	PRINCETON	NJ	08544	MERCER	USA	Biomedical Research and Research Training	002	1	6/11/2024	NEW	$1,280
2024	2024	THE TRUSTEES OF PRINCETON UNIVERSITY	1 NASSAU HALL	PRINCETON	NJ	08544	MERCER	USA	Biomedical Research and Research Training	001	1	5/1/2024	NEW	$0
2024	2024	THE TRUSTEES OF PRINCETON UNIVERSITY	1 NASSAU HALL	PRINCETON	NJ	08544	MERCER	USA	Biomedical Research and Research Training	000	1	5/1/2024	NEW	$31,694
														Subtotal = $32,974

Grand Total All Awards = $100,626

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Regulation of protein dynamics in Drosophila embryos' ectoderm germ layer

Award Number: F30GM154411

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS

PERIOD OF PERFORMANCE START DATE: 06/01/2024

PERIOD OF PERFORMANCE END DATE: 05/31/2028

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