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Shining light on the mechanisms of primate gastrulation: development of high-throughput, optogenetic tools for understanding embryonic development

Award Number: F32HD114539
ORGANIZATION: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS
PERIOD OF PERFORMANCE START DATE: 07/22/2024
PERIOD OF PERFORMANCE END DATE: 07/21/2027

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Shining light on the mechanisms of primate gastrulation: development of high-throughput, optogenetic tools for understanding embryonic development - ABSTRACT Gastrulation is a pivotal process in mammalian embryogenesis that is essential for the establishment of definitive germ layers and the formation of the body plan. Despite decades of research, the precise mechanisms that regulate cell differentiation, migration, and patterning during mammalian gastrulation remain poorly understood, particularly in primates. The study of primate gastrulation is especially challenging because this process occurs after the embryo has implanted into the uterus, making it difficult to directly observe. Recently, several three-dimensional (3D), stem cell-based models of primate embryos, termed “stembryos”, have been developed for in vitro studies of embryonic development. However, most approaches for generating stembryos exploit the inherent self-organizing capacity of stem cells to form 3D constructs and are unable to precisely control the number of cells and cell types per construct. A lack of tools for engineering stembryos has resulted in low yields and has limited the ability of stembryos to faithfully recapitulate primate gastrulation. We have recently developed a stembryo model using induced pluripotent stem cells (iPSCs) from chimpanzees along with an optofluidic (i.e., combination of optogenetic and microfluidic) device for performing Chip-based, High-throughput Investigations into Morphogenesis in Primates, called the OptoCHIMP platform. Our OptoCHIMP platform utilizes microfluidic encapsulation to place cells into hydrogel droplets. A downstream droplet sorter unit allows us to select droplets of interest with prescribed cell numbers of cell ratios. This enables us to fabricate stembryos in a precise and high-throughput manner. In this proposal, we will utilize the OptoCHIMP platform to define the roles that 3D structure, local tissue mechanics, and spatiotemporal signaling dynamics play in symmetry breaking during primate gastrulation. By utilizing optogenetic tools to stimulate signaling pathways in chimpanzee stembryos, we can recreate the complex interplay of signaling pathways between embryonic and extraembryonic tissues, induce symmetry breaking in our stembryo, and develop a model for understanding how morphogen signaling dynamics impact tissue patterning. This proposal will be the first time that microfluidic droplet encapsulation of stem cells, which offers precise control over the initial conditions, has been combined with optogenetic, providing a promising approach to uncovering the mechanisms that drive primate gastrulation.


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 = $78,040 )
2025	2025	UNIVERSITY OF CALIFORNIA, SANTA BARBARA	3227 CHEADLE HALL	SANTA BARBARA	CA	93106	SANTA BARBARA	USA	Child Health and Human Development Extramural Research	000	2	6/16/2025	NON-COMPETING CONTINUATION	$78,040
														Subtotal = $78,040

Issue Date FY: 2024 ( Subtotal = $74,284 )
2024	2024	UNIVERSITY OF CALIFORNIA, SANTA BARBARA	3227 CHEADLE HALL	SANTA BARBARA	CA	93106	SANTA BARBARA	USA	Child Health and Human Development Extramural Research	000	1	6/24/2024	NEW	$74,284
2024	2024	UNIVERSITY OF CALIFORNIA, SANTA BARBARA	3227 CHEADLE HALL	SANTA BARBARA	CA	93106	SANTA BARBARA	USA	Child Health and Human Development Extramural Research	001	1	6/26/2024	NEW	$0
														Subtotal = $74,284

Grand Total All Awards = $152,324

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Shining light on the mechanisms of primate gastrulation: development of high-throughput, optogenetic tools for understanding embryonic development

Award Number: F32HD114539

ORGANIZATION: NATIONAL INSTITUTE OF CHILD HEALTH AND HUMAN DEVELOPMENT

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS

PERIOD OF PERFORMANCE START DATE: 07/22/2024

PERIOD OF PERFORMANCE END DATE: 07/21/2027

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