Genomics & Metabolomics

Background

Genomics and metabolomics are crosscutting technologies that enables molecular dissection of disease subtypes and mechanisms. Our core is equipped with 3 next-generation sequencers (NGS) as well as supported by multiple targeted gas and liquid chromatography mass spectrometers from the Duke-NUS metabolomics core facility (led by Asst Prof Jean Paul Kovalik, www.duke-nus.edu.sg/research/research-facilities/metabolomics-facility) that was relocated to NHCS in Year 2018 to expand research capability for metabolomics analyses. These state-of-the-art equipment and skillsets enable basic science and translational research into the genome, transcriptome, proteome and metabolome of cardiometabolic diseases. This ‘one-stop’ resource allows scientists to incorporate new genomics and metabolomics data into their studies. Together with NHCS Biobank and other NHRIS core facilities, the core is aimed to build phenotype-genotype-metabolomics profiles in the Asian context, bridging the gaps in national (SG100k) and global precision medicine initiatives.

We support multiple NGS analyses based on Illumina platform such as gene panel design, targeted DNA-seq, bulk RNA-seq, miRNA-seq and single cell RNA-seq. This knowledge enable scientists to translate basic research into meaningful insight concerning challenges by many diseases.

Prof Stuart Alexander Cook NHCS Edmund Pua NHCS Researcher NHRIS

Prof Stuart Alexander Cook Dr Edmund Pua Chee Jian

Contacts

For further information please contact our Genomics & Metabolomics core facility from Monday to Friday (except Public Holidays), 8.30am to 5pm, at:

Email: [email protected]
Tel: (65) 6704 2199

Prof Stuart Cook | Distinguished Clinician Scientist, NHCS; Core Lead, NHRIS
Dr Edmund Pua Chee Jian | Senior Research Associate, NHRIS
Ms Lim Shiqi | Research Associate, NHRIS

Expertise

Genomics Metabolomics NHCS NHRIS

Genomics:

(1) Clinical grade (ISO15189:2012) NGS genetic tests for inherited cardiac conditions; (2) Targeted re-sequencing of panels; (3) Whole Exome Sequencing (WES); (4) Sanger sequencing (5) mRNASeq and ChiPSeq; (6) Single-cell RNA-seq (7) Sanger sequencing; (8) Bioinformatics analyses and data storage; (9) Gene differential expression analysis (bulk/single-cell); (10) Custom gene panel design

Metabolomics analyses:

(1) Amino Acids; (2) Acyl Carnitines; (3) Free/Total Carnitines; (4) Organic Acids (TCA cycle intermediates); (5) Sphingolipids & phosphatidylcholines; (6) 1-Methylhistidine; (7) Tryptophan metabolism pathway.

Methodology

Equipment:

Illumina MiniSeq

Medium throughput next generation sequencer
Read types: paired-end; single-end
Read cycles: 50, 150, 300

Illumina MiSeq

Medium throughput next generation sequencer with longer read cycles
Read types: paired-end; single-end
Read cycles: 50, 150, 300, 600

Illumina NextSeq 500

High throughput next generation sequencer
Read types: paired-end; single-end
Read cycles: 75, 150, 300

Applied Biosystems 3500 Series Genetic Analyzer

Sanger sequencer

Perkin Elmer Caliper Sciclone G3 and LabChip GXii Touch System

Automated NGS library preparation and quality check
Up to 96 samples

Services:

Custom gene panel design and bioinformatics support

Common and rare diseases

Targeted re-sequencing

Illumina TruSight panels

Whole Exome Sequencing

Illumina Nextera Rapid Capture exome kit
Illumina Nextera Rapid Capture expanded exome kit

Bulk RNA sequencing

TruSeq stranded mRNA kit
TruSeq total RNA kit
TruSeq small RNA kit

Single cell RNA sequencing

Single cell gene expression kit

Sanger sequencing

8 capillaries

Research Area

Inherited Cardiac Conditions (ICC) are a diverse group of heart conditions including cardiomyopathy, arrhythmic disorders, aortopathies and lipid disorders such as falimial hypercholesterolemia. They can affect people of any age, and can be life-threatening so improving diagnosis is important for effective treatment. We use the commercialized TruSight Cardio panel and in-house bioinformatics pipeline to study Inherited Cardiac Condition (ICC) genes among patient/research samples to define novel pathological mutations that are unique to Asian including Singaporean. These mutations will be used to stratify sub-disease and we hope to use this information to pin down the underlying cause.

We are also interested to investigate the fundamental cause and therapeutic targets of cardiometabolic diseases using genomics and metabolomics technologies. We provide genotyping, DNA sequencing, bulk and single cell RNA sequencing services for research projects involving heart, kidney, lung and liver in cardiometabolic diseases.

Achievements

We have developed jointly with Royal Brompton Hospital, a genetic test kit that uses high-throughput DNA sequencing technology to look for variation in 174 genes that can cause inherited cardiac conditions (ICC). The test panel was commercialized by Illumina as the TruSight Cardio Sequencing Kit and available globally for clinical diagnostics of inherited cardiac conditions and case-control research study. Using local case-control cohort that we have recruited over the past 8 years, we identified two common variants in TNNI3 and TNNT2 in the Chinese hypertrophic cardiomyopathy (HCM) patients. The re-classification of both troponin variants is expected to affect nearly 200,000 HCM cases worldwide in the clinical care settings. At NHCS, we also use NGS technology to sequence cases of sudden unexplained death for the Health Services Authority (HSA) and up to 30% cases have likely pathogenic variants to explain the cause of death.

Awards

The President’s Technology Award 2018 was awarded to Prof Cook for his research in human genetics.

The lab is accredited by the Singapore Accreditation Council to ISO 15189:2012 (Medical laboratories) for clinical grade ICC genetic test

Publications

Pua, C. J., Tham, N., Chin, C. W. L., Walsh, R., Khor, C. C., Toepfer, C. N., . . . Cook, S. A. (2020). Genetic Studies of Hypertrophic Cardiomyopathy in Singaporeans Identify Variants in TNNI3 and TNNT2 That Are Common in Chinese Patients. Circ Genom Precis Med, 13(5), 424-434. doi:10.1161/circgen.119.002823　
Mazzarotto, F., Tayal, U., Buchan, R. J., Midwinter, W., Wilk, A., Whiffin, N., . . . Walsh, R. (2020). Reevaluating the Genetic Contribution of Monogenic Dilated Cardiomyopathy. Circulation, 141(5), 387-398. doi:10.1161/circulationaha.119.037661　
Schafer, S., de Marvao, A., Adami, E., Fiedler, L. R., Ng, B., Khin, E., . . . Cook, S. A. (2017). Titin-truncating variants affect heart function in disease cohorts and the general population. Nat Genet, 49(1), 46-53. doi:10.1038/ng.3719　
Walsh, R., Buchan, R., Wilk, A., John, S., Felkin, L. E., Thomson, K. L., . . . Cook, S. A. (2017). Defining the genetic architecture of hypertrophic cardiomyopathy: re-evaluating the role of non-sarcomeric genes. Eur Heart J, 38(46), 3461-3468. doi:10.1093/eurheartj/ehw603　
Pua, C. J., Bhalshankar, J., Miao, K., Walsh, R., John, S., Lim, S. Q., . . . Cook, S. A. (2016). Development of a Comprehensive Sequencing Assay for Inherited Cardiac Condition Genes. J Cardiovasc Transl Res, 9(1), 3-11. doi:10.1007/s12265-016-9673-5