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Investigating the spatial and temporal regulation of nkx2.5+ ALPM progenitors in outflow tract development

Award Number: F32HL186603
ORGANIZATION: NATIONAL HEART, LUNG, & BLOOD INSTITUTE
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS
PERIOD OF PERFORMANCE START DATE: 12/28/2026
PERIOD OF PERFORMANCE END DATE: 12/27/2027

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Investigating the spatial and temporal regulation of nkx2.5+ ALPM progenitors in outflow tract development - PROJECT SUMMARY Conotruncal defects are among the most severe and life-threatening forms of congenital heart disease (CHD), including conditions like Tetralogy of Fallot and Double Outlet Right Ventricle. Notably, anomalies in the formation or remodeling of the outflow tract (OFT), located within the conotruncus, are often associated with genetic syndromes such as 22q11 deletion syndrome. The OFT develops from a subset of multipotent cells in the anterior lateral plate mesoderm (ALPM), known as the second heart field (SHF), which expresses the cardiac transcription factor nkx2.5. Recently, we discovered that the heterogeneous nkx2.5+ ALPM is spatially and transcriptionally organized based on their specific cardiopharyngeal fates earlier in development than previously believed. However, the mechanisms orchestrating the migration and differentiation of OFT-fated progenitors from this heterogeneous pool into their respective final cell lineages remain unknown. In this study, we aim to (1) create a spatiotemporal map of nkx2.5+ progenitor development and (2) identify the gene regulatory networks that govern their migration and differentiation to the OFT. To address these aims, I will use the zebrafish model system, which offers unparalleled capabilities for tracking and analyzing progenitor cells throughout their external embryonic development. I will employ an integrated approach that includes Kaede-mediated photoconversion, whole-mount immunofluorescence, and in situ hybridization to determine when and where OFT-fated progenitors differentiate. Additionally, I will combine innovative spatial transcriptomics with loss-of-function studies to uncover the mechanistic signals that drive progenitor migration and differentiation within their pharyngeal microenvironment. Ultimately, understanding the developmental pathways and mechanisms underlying OFT formation will enhance our knowledge of CHD etiology. This work will be carried out in the ideal environment provided by the Department of Cardiology at Boston Children’s Hospital, under the mentorship of cardiovascular development expert Dr. Caroline Burns and bioinformatician expert Dr. Kaifu Chen. Additional support from my scientific advisory committee, along with resources from Harvard Medical School, will ensure the success of the proposed research and my growth as an independent scientist.


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 = $82,348 )
2026	2026	CHILDREN'S HOSPITAL CORPORATION, THE	300 LONGWOOD AVE	BOSTON	MA	02115	SUFFOLK	USA	Cardiovascular Diseases Research	000	1	7/1/2026	NEW	$82,348
														Subtotal = $82,348

Grand Total All Awards = $82,348

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Investigating the spatial and temporal regulation of nkx2.5+ ALPM progenitors in outflow tract development

Award Number: F32HL186603

ORGANIZATION: NATIONAL HEART, LUNG, & BLOOD INSTITUTE

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: FELLOWSHIP/SCHOLARSHIP/STUDENT LOANS

PERIOD OF PERFORMANCE START DATE: 12/28/2026

PERIOD OF PERFORMANCE END DATE: 12/27/2027

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