Trapping Cardiac Recessive Mutants via Expression-Based Insertional Mutagenesis Screening

Ding, Yonghe; Liu, Weibin; Deng, Yuan; Jomok, Beninio; Yang, Jingchun; Huang, Wei; Clark, Karl J.; Zhong, Tao; Lin, Xueying; Ekker, Stephen C.; Xu, Xiaolei

doi:10.1161/circresaha.112.300603

Cited by 47 publications

(71 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…By checking cardiac monomeric red fluorescent protein (mRFP) expression in 609 GBT lines, we were able to identify 44 Zebrafish Insertional Cardiac (ZIC) mutants (Supplemental Table 1; supplemental material available online with this article; doi:10.1172/jci. insight.88797DS1) (15). Having shown cardiac remodeling in adult zebrafish heart upon stress imposed by either anemia or doxorubicin (DOX) injection (16, 17), we applied DOX stress to these heterozygous ZIC mutants.…”

Section: Resultsmentioning

confidence: 99%

“…GBT0411 homozygous mutants demonstrated cardiomegaly at 1 year, whereas both heterozygous and homozygous GBT0411 mutants accelerated DOX-induced fish death at only 3 months of age (Figure 1A and Supplemental Figure 2A) (15). At the molecular level, GBT0411 specifically disrupted the long isoform of DnaJ (Hsp40) homolog, subfamily B, member 6b ( dnajb6b(L) ) (Figure 1, B and C).…”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

A modifier screen identifies DNAJB6 as a cardiomyopathy susceptibility gene

Ding¹,

Long²,

Bos³

et al. 2016

JCI Insight

Self Cite

View full text Add to dashboard Cite

Mutagenesis screening is a powerful forward genetic approach that has been successfully applied in lower-model organisms to discover genetic factors for biological processes. This phenotype-based approach has yet to be established in vertebrates for probing major human diseases, largely because of the complexity of colony management. Herein, we report a rapid strategy for identifying genetic modifiers of cardiomyopathy (CM). Based on the application of doxorubicin stress to zebrafish insertional cardiac (ZIC) mutants, we identified 4 candidate CM-modifying genes, of which 3 have been linked previously to CM. The long isoform of DnaJ (Hsp40) homolog, subfamily B, member 6b (dnajb6b(L)) was identified as a CM susceptibility gene, supported by identification of rare variants in its human ortholog DNAJB6 from CM patients. Mechanistic studies indicated that the deleterious, loss-of-function modifying effects of dnajb6b(L) can be ameliorated by inhibition of ER stress. In contrast, overexpression of dnajb6(L) exerts cardioprotective effects on both fish and mouse CM models. Together, our findings establish a mutagenesis screening strategy that is scalable for systematic identification of genetic modifiers of CM, feasible to suggest therapeutic targets, and expandable to other major human diseases.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

A modifier screen identifies DNAJB6 as a cardiomyopathy susceptibility gene

Ding¹,

Long²,

Bos³

et al. 2016

JCI Insight

Self Cite

View full text Add to dashboard Cite

show abstract

“…Recognizing this weakness, we scanned the literature for reported cardiac modifying genes in animal models. This led us to examine four genes ( SORBS2 , RXRA , DNAJB6 and ANO5 ) reported as cardiac modifiers of doxorubicin-induced cardiotoxicity in zebrafish and/or mouse models 46,47 . Suggesting that our data can be replicated we also identified variants in our study of N9831 in SORBS2 and RXRA and p<0.01 in DNAJB6 , p<0.001 (Supplementary Table 3).…”

Section: Discussionmentioning

confidence: 99%

Genome-wide association study of cardiotoxicity in the NCCTG N9831 (Alliance) adjuvant trastuzumab trial

Serie

Crook²,

Necela

et al. 2017

Pharmacogenetics and Genomics

View full text Add to dashboard Cite

Objectives The major clinical side-effect of the ERBB2 targeted breast cancer therapy, trastuzumab, is a decline in left ventricular ejection fraction (LVEF). Improved markers are needed to better identify patients susceptible to cardiotoxicity. Methods The NCCTG N9831 trial compared adjuvant doxorubicin and cyclophosphamide followed by either weekly paclitaxel (Arm A); paclitaxel then trastuzumab (Arm B); or concurrent paclitaxel and trastuzumab (Arm C) in patients with HER2-positive breast cancer. A GWAS was performed on all patients with available DNA (N=1,446). We used linear regression to identify SNPs associated with decline in LVEF, adjusting for age, baseline LVEF, anti-hypertensive medications and the first two principle components. Results 618,863 SNPs passed quality control (QC) and DNA from 1,191 patients passed genotyping QC and were identified as whites of non-Hispanic origin. SNPs at six loci were associated with a decline in LVEF (p=7.73×10−6 to 8.93×10−8), LDB2, BRINP1, chr6 intergenic, RAB22A, TRPC6 and LINC01060, in patients who received chemotherapy plus trastuzumab (Arms BC, N=800). None of these loci were significant in patients who received chemotherapy only (Arm A, N=391) and did not increase in significance in analysis of all patients combined. We did not observe association, p<0.05 with SNPs previously associated with trastuzumab induced cardiotoxicity at ERBB2, I655V and P1170A. We replicated association, p<0.05, with SNPs previously associated with anthracycline cardiotoxicity at CBR3 and ABCB1. Conclusions Our study identified six putative novel cardiotoxicity loci in patients treated with combination chemotherapy and trastuzumab that require further investigation and confirmed known associations of anthracycline-induced cardiotoxicity.

show abstract

“…The value of these advanced gene-trap models has clearly been demonstrated for analyses of spatiotemporal dynamics of protein expression, cellular behaviors and mutant phenotypic traits [67]. Although conditional gene-trap models have yet to be established for thyroid-specific genes, the rapidly expanding collection of conditional mutant lines (http://zfishbook.org/, http://www.fliptrap.org) will hopefully become an important resource also for zebrafish thyroid research [66,68]. In summary, a series of technological advancements in the recent past have made zebrafish an ideal model organism to study the basic mechanisms underlying organ development, and application of these novel techniques and methods in the context of thyroid morphogenesis is expected to provide for new and innovative research strategies to enhance the understanding of vertebrate thyroid development.…”

Section: The Zebrafish Genetic Toolkit On Thyroid Organogenesismentioning

confidence: 99%

New Model Systems to Illuminate Thyroid Organogenesis. Part I: An Update on the Zebrafish Toolbox

2013

View full text Add to dashboard Cite

Thyroid dysgenesis (TD) resulting from defects during embryonic thyroid development represents a major cause of congenital hypothyroidism. The pathogenetic mechanisms of TD in human newborns, however, are still poorly understood and disease-causing genetic variants have been identified in only a small percentage of TD cases. This limited understanding of the pathogenesis of TD is partly due to a lack of knowledge on how intrinsic factors and extrinsic signalling cues orchestrate the differentiation of thyroid follicular cells and the morphogenesis of thyroid tissue. Recently, embryonic stem cells and zebrafish embryos emerged as novel model systems that allow for innovative experimental approaches in order to decipher cellular and molecular mechanisms of thyroid development and to unravel pathogenic mechanisms of TD. Zebrafish embryos offer several salient properties for studies on thyroid organogenesis including rapid and external development, optical transparency, ease of breeding, relative short generation time and amenability for genome editing. In this review, we will highlight recent advances in the zebrafish toolkit to visualize cellular dynamics of organ development and discuss specific prospects of the zebrafish model for studies on vertebrate thyroid development and human congenital thyroid diseases.

show abstract

Trapping Cardiac Recessive Mutants via Expression-Based Insertional Mutagenesis Screening

Cited by 47 publications

References 33 publications

A modifier screen identifies DNAJB6 as a cardiomyopathy susceptibility gene

A modifier screen identifies DNAJB6 as a cardiomyopathy susceptibility gene

Genome-wide association study of cardiotoxicity in the NCCTG N9831 (Alliance) adjuvant trastuzumab trial

New Model Systems to Illuminate Thyroid Organogenesis. Part I: An Update on the Zebrafish Toolbox

Contact Info

Product

Resources

About