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Published on: May 16, 2020
RNA-Seq analysis in an avian model of maternal phenylketonuria
Jamie N Watson1, Nikki J Seagraves1
1Department of Biology, University of Central Oklahoma, Edmond, OK, USA.
Insights
Maternal Phenylketonuria (PKU) causes congenital heart defects in infants. This study reveals that high phenylalanine exposure alters Retinoic Acid (RA) pathways, offering new insights into these cardiovascular malformations.
Area of Science:
- Developmental Biology
- Genetics
- Cardiovascular Science
Background:
- Congenital heart malformations (CVMs) are a major cause of infant mortality.
- Maternal Phenylketonuria (MPKU) significantly increases the risk of CVMs in developing fetuses due to in-utero phenylalanine exposure.
- The molecular mechanisms underlying MPKU-induced CVMs remain poorly understood.
Purpose of the Study:
- To investigate the molecular mechanisms of CVMs in an avian model of MPKU.
- To identify differentially expressed genes (DEGs) during early embryonic development (stages HH10, 12, and 14).
- To explore the role of the Retinoic Acid (RA) pathway in MPKU-associated cardiovascular defects.
Main Methods:
- RNA sequencing (RNA-Seq) was employed to analyze gene expression profiles in an avian model of MPKU.
- Differential gene expression analysis was performed across key early developmental stages.
- Quantitative reverse transcription PCR (qRTPCR) was used to validate findings, particularly for genes in the RA pathway.
Main Results:
- A total of 633 significantly differentially expressed genes were identified across the studied developmental stages.
- Functional annotation revealed enrichment of genes related to cardiac function, cell processes, and known MPKU clinical phenotypes.
- Significant alterations were observed in genes involved in Retinoic Acid (RA) metabolism and its downstream targets, with 42 RA pathway genes found to be differentially expressed.
Conclusions:
- This study provides the first elucidation of molecular mechanisms underlying cardiovascular malformations in MPKU at early developmental timepoints.
- Evidence suggests a direct link between phenylalanine (PHE) exposure and the dysregulation of the RA pathway.
- These findings offer novel insights into congenital heart defects associated with MPKU and highlight the RA pathway as a potential therapeutic target.
Abstract:
Cardiac malformations (CVMs) are a leading cause of infant morbidity and mortality. CVMs are particularly prevalent when the developing fetus is exposed to high levels of phenylalanine in-utero in mothers with Phenylketonuria. Yet, elucidating the underlying molecular mechanism leading to CVMs has proven difficult. In this study we used RNA-Seq to investigate an avian model of MPKU and establish differential gene expression (DEG) characteristics of the early developmental stages HH10, 12, and 14. In total, we identified 633 significantly differentially expressed genes across stages HH10, 12, and 14. As expected, functional annotation of significant DEGs identified associations seen in clinical phenotypes of MPKU including CVMs, congenital heart defects, craniofacial anomalies, central nervous system defects, and growth anomalies. Additionally, there was an overrepresentation of genes involved in cardiac muscle contraction, adrenergic signaling in cardiomyocytes, migration, proliferation, metabolism, and cell survival. Strikingly, we identified significant changes in expression with multiple genes involved in Retinoic Acid (RA) metabolism and downstream targets. Using qRTPCR, we validated these findings and identified a total of 42 genes within the RA pathway that are differentially expressed. Here, we report the first elucidation of the molecular mechanisms of cardiovascular malformations in MPKU conducted at early developmental timepoints. We provide evidence suggesting a link between PHE exposure and the alteration of RA pathway. These results are promising and offer novel findings associated with congenital heart defects in MPKU.
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