scholarly journals Establishment of a Dihydrofolate Reductase Gene Knock-In Zebrafish Strain to Aid Preliminary Analysis of Congenital Heart Disease Mechanisms

2021 ◽  
Vol 8 ◽  
Author(s):  
Ke Gong ◽  
Ting Xie ◽  
Yifeng Yang ◽  
Yong Luo ◽  
Yun Deng ◽  
...  
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
Western Blot ◽  
Dihydrofolate Reductase ◽  
Blot Analysis ◽  
Heart Development ◽  
Western Blot Analysis ◽  
Congenital Heart ◽  
Copy Number Variations ◽  
Hematoxylin And Eosin

Background: The dihydrofolate reductase (DHFR) gene is imperative in development, therefore it is essential to explore its effects on heart development. Thus, here a dhfr zebrafish knock-in (KI) strain was constructed.Methods: CRISPR/Cas9 technology was used to establish the dhfr KI zebrafish strain. This strain was hybridized with TgG fluorescent strain zebrafish to observe the phenotypes of heart shape, size, and circularization direction. Wild-type (WT) and KI zebrafish were then dissected and histologically stained to observe pathological changes. Western blot analysis was used to verify the increased expressions of zebrafish genes after KI. Hybridization experiments were used to confirm the presence of abnormal gonadal dysplasia.Results: The zebrafish dhfr KI strain was successfully constructed through CRISPR/Cas9 technology. At 6 days post fertilization (dpf), microscopic examinations of KI (homozygous) specimens revealed pericardial effusions, heart compressions, and curled tails. Compared with WT, the Hematoxylin and Eosin (H&E) tissue sections of KI-homozygous zebrafish showed defects such as reduced atria and ventricles. Western blot analysis indicated that the expression of the DHFR protein increased in both heterozygotes and homozygotes of dhfr KI zebrafish. Hybridization experiments revealed that dhfr KI may affect gonadal function.Conclusion: The DHFR gene plays an important regulatory role in the process of heart development, and copy number variations (CNVs) of this gene may constitute a new pathogenic mechanism of congenital heart disease (CHD).

scholarly journals Navigating the non-coding genome in heart development and Congenital Heart Disease

2019 ◽  
Vol 107 ◽  
pp. 11-23 ◽  
Author(s):  
Gulrez Chahal ◽  
Sonika Tyagi ◽  
Mirana Ramialison
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
Heart Development ◽  
Congenital Heart

scholarly journals A Non-coding HES1 Variant Predisposes Children to Congenital Heart Disease in Chinese Population

2021 ◽  
Vol 9 ◽  
Author(s):  
Yangliu Song ◽  
Weicheng Chen ◽  
Zitong Huang ◽  
Guixiang Tian ◽  
Mengru Li ◽  
...  
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
Association Analysis ◽  
Heart Development ◽  
Congenital Heart ◽  
Genetic Model ◽  
Gain Of Function ◽  
Coding Sequence ◽  
Homozygous Variant ◽  
Increased Risk

Background: As a key component in the NOTCH signaling pathway, HES1 plays an important role in vertebrate heart development. Variants in the HES1 coding sequence are known to be associated with congenital heart disease (CHD). However, little is known about HES1 non-coding sequence variants and their association with the risk of developing CHD.Method and Results: We initially analyzed the non-coding sequence of the HES1 gene in 12 unrelated CHD families by direct sequencing and identified a previously unreported promoter region variant (NM_005524.4: c.−1279−1278 insAC, rs148941464) in the HES1 gene in four CHD families. The homozygous variant in patients was inherited from carrier parents with normal phenotypes, indicating a likely recessive genetic model. Given that the HES1 gene is predicted to be likely to exhibit haploinsufficiency (%HI: 11.44), we hypothesized that the HES1 homozygous variant is a genetic risk factor underlying CHD. We then carried out sequencing of this HES1 variant in 629 sporadic non-syndromic CHD cases and 696 healthy controls and performed association analysis. Interestingly, we observed a significant association of the homozygous HES1 promoter variant with CHD (18.92% of cases vs. 9.91% of controls; OR: 2.291, 95% CI: 1.637-3.207, p = 9.72 × 10−7). No significant association with CHD was observed for the HES1 promoter heterozygous variant (p > 0.05). However, association analysis tests of the HES1 homozygous variant with each subtype of CHD revealed that this homozygous variant was strongly associated with transposition of the great arteries (TGA) (OR: 3.726, 95% CI: 1.745-7.956, p = 0.0003). Moreover, the prevalence of HES1 homozygous variants in CHD patients with TGA (27.66%) was significantly higher than that in patients with other CHD subtypes or controls. Similar results were observed in a replication group of TGA (n = 64). Functional studies demonstrated that the homozygous variant in the HES1 promoter can disrupt its ability to bind RXRA, an inhibitory transcription factor, which results in abnormally high expression of the HES1 gene, indicating that this variant harbors gain-of-function effects.Conclusions: Our findings reveal that the non-coding homozygous variant in the HES1 promoter has a gain-of-function effect and is associated with an increased risk of CHD development, especially the severe TGA subtype.

scholarly journals Missense Mutation in the Transcription Factor NKX2–5: A Novel Molecular Event in the Pathogenesis of Thyroid Dysgenesis

2006 ◽  
Vol 91 (4) ◽  
pp. 1428-1433 ◽  
Author(s):  
Monica Dentice ◽  
Viviana Cordeddu ◽  
Annamaria Rosica ◽  
Alfonso Massimiliano Ferrara ◽  
Libero Santarpia ◽  
...  
Keyword(s):  
Congenital Heart Disease ◽  
Transcription Factor ◽  
Heart Disease ◽  
Heart Development ◽  
Congenital Heart ◽  
Functional Characterization ◽  
Dominant Negative Effect ◽  
Thyroid Dysgenesis ◽  
Thyroid Development

Context: Congenital hypothyroidism (CH) is a common endocrine disorder with an incidence of 1:3000–4000 at birth. In 80–85% of cases, CH is caused by defects in thyroid organogenesis, resulting in absent, ectopically located, and/or severely reduced gland [thyroid dysgenesis (TD)]. Mutations in genes controlling thyroid development have demonstrated that in a few cases, TD is a Mendelian trait. However, accumulating evidence supports the view that the genetics of TD are complex, possibly with a polygenic/multifactorial basis. A higher prevalence of congenital heart disease has been documented in children with CH than in the general population. Such an association suggests a possible pathogenic role of genes involved in both heart and thyroid development. NKX2–5 encodes a homeodomain-containing transcription factor with a major role in heart development, and mutations affecting this gene have been reported in individuals with congenital heart disease. Objective: In the present work we investigated the possible involvement of NKX2–5 mutations in TD. Results: Our results indicate that Nkx2–5−/− embryos exhibit thyroid bud hypoplasia, providing evidence that NKX2–5 plays a role in thyroid organogenesis and that NKX2–5 mutations contribute to TD. NKX2–5 mutational screening in 241 patients with TD allowed the identification of three heterozygous missense changes (R25C, A119S, and R161P) in four patients with TD. Functional characterization of the three mutations demonstrated reduced DNA binding and/or transactivation properties, with a dominant-negative effect on wild-type NKX2–5. Conclusion: Our results suggest a previously unknown role of NKX2–5 in the pathogenesis of TD.

Molecular insight into heart development and congenital heart disease: An update review from the Arab countries

2015 ◽  
Vol 25 (4) ◽  
pp. 291-301 ◽  
Author(s):  
Elhadi H. Aburawi ◽  
Hanan E. Aburawi ◽  
Keith M. Bagnall ◽  
Zahurul A. Bhuiyan
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
Heart Development ◽  
Congenital Heart ◽  
Arab Countries ◽  
Insight Into

scholarly journals Genetics of Congenital Heart Disease

Biomolecules ◽  
2019 ◽  
Vol 9 (12) ◽  
pp. 879 ◽  
Author(s):  
Kylia Williams ◽  
Jason Carson ◽  
Cecilia Lo
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
Cell Signaling ◽  
Heart Development ◽  
Congenital Heart ◽  
Genetic Etiology ◽  
Genes Encoding ◽  
Related Proteins ◽  
Improve Patient ◽  
Cell Signaling Pathways

Congenital heart disease (CHD) is one of the most common birth defects. Studies in animal models and humans have indicated a genetic etiology for CHD. About 400 genes have been implicated in CHD, encompassing transcription factors, cell signaling molecules, and structural proteins that are important for heart development. Recent studies have shown genes encoding chromatin modifiers, cilia related proteins, and cilia-transduced cell signaling pathways play important roles in CHD pathogenesis. Elucidating the genetic etiology of CHD will help improve diagnosis and the development of new therapies to improve patient outcomes.

scholarly journals Rare copy number variations in congenital heart disease patients identify unique genes in left-right patterning

2011 ◽  
Vol 108 (7) ◽  
pp. 2915-2920 ◽  
Author(s):  
K. A. Fakhro ◽  
M. Choi ◽  
S. M. Ware ◽  
J. W. Belmont ◽  
J. A. Towbin ◽  
...  
Keyword(s):  
Congenital Heart Disease ◽  
Heart Disease ◽  
Copy Number ◽  
Congenital Heart ◽  
Copy Number Variations ◽  
Unique Genes
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