Establishment of a "multi-species high-throughput platform" to evaluate candidate congential heart disease genes

Theis JL, Vogler G, Missinato MA, Li X, Nielsen T, Zeng XI, Martinez- Fernandez A, Walls SM, Kervadec A, Kezos JN, Birker K, Evans JM, O'Byrne MM, Fogarty ZC, Terzic A, Grossfeld P, Ocorr K, Nelson TJ, Olson TM, Colas AR, Bodmer R. Patient-specific genomics and cross-species functional analysis implicate LRP2 in hypoplastic left heart syndrome. Elife. 2020 Oct 2;9:e59554.
doi: 10.7554/eLife.59554. Epub ahead of print. PMID: 33006316.

 

From the abstract:

"Congenital heart diseases (CHDs) ... are genetically complex and poorly understood. Here, a multi-disciplinary platform was established to functionally evaluate novel CHD gene candidates, based on whole genome and iPSC RNA sequencing of a HLHS family-trio. ... siRNA/RNAi-mediated knockdown in generic human iPSC-derived cardiomyocytes (hiPSC-CM) and in developing Drosophila and zebrafish hearts revealed that LDL receptor-related protein LRP2 is required for cardiomyocyte proliferation and differentiation. ... Collectively, we have established a multi-species high-throughput platform to rapidly evaluate candidate genes and their interactions during heart development, which are crucial first steps towards deciphering oligogenic underpinnings of CHDs, including maladaptive left hearts."

Protocol for testing of candidate heart-disease genes in Drosophila

Zhu JY, Fu Y, Richman A, Han Z. Validating Candidate Congenital Heart Disease Genes in Drosophila. Bio Protoc. 2017 Jun 20;7(12). pii: e2350. PMID: 29276722; PMCID: PMC5739056.

From the abstract: "... A high throughput in vivo model system to validate candidate gene association with pathology is ... useful. We present such a system employing Drosophila to validate candidate congenital heart disease (CHD) genes. The protocols exploit comprehensive libraries of UAS-GeneX-RNAi fly strains ... These protocols were recently used to evaluate more than 100 candidate CHD genes implicated by patient whole-exome sequencing (Zhu et al., 2017)."

Using flies to functionally validate candidate heart disease-related genes

Zhu JY, Fu Y, Nettleton M, Richman A, Han Z. High throughput in vivo functional validation of candidate congenital heart disease genes in Drosophila. Elife. 2017 Jan 13;6. pii: e22617. PMID: 28084990.

From the abstract: "... We developed a Drosophila-based functional system to screen candidate disease genes identified from Congenital Heart Disease (CHD) patients. 134 genes were tested in the Drosophila heart using RNAi-based gene silencing. Quantitative analyses of multiple cardiac phenotypes demonstrated essential structural, functional, and developmental roles for more than 70 genes ... We also demonstrated the use of Drosophila to evaluate cardiac phenotypes resulting from specific, patient-derived alleles of candidate disease genes. ... This approach has the potential to facilitate development of precision medicine approaches for CHD and other diseases associated with genetic factors."

Review highlights usefulness of Drosophila as a model for study of congenital heart disease

Vogler G, Bodmer R. Cellular Mechanisms of Drosophila Heart Morphogenesis. J Cardiovasc Dev Dis. 2015 Mar 1;2(1):2-16.  PMID: 26236710

From the abstract: "Many of the major discoveries in the fields of genetics and developmental biology have been made using the fruit fly, Drosophila melanogaster. With regard to heart development, the conserved network of core cardiac transcription factors that underlies cardiogenesis has been studied in great detail in the fly, and the importance of several signaling pathways that regulate heart morphogenesis, such as Slit/Robo, was first shown in the fly model. ... The specific limitations of the various cardiac model systems currently employed (mammalian and fish models) provide a niche for the fly model ... Here, we review recent advances made using the Drosophila embryo that identify factors relevant for heart formation. ..."