Applications of high-throughput DNA sequencing to benign hematology

Abstract

The development of novel technologies for high-throughput DNA sequencing is having a major impact on our ability to measure and define normal and pathologic variation in humans. This review discusses advances in DNA sequencing that have been applied to benign hematologic disorders, including those affecting the red blood cell, the neutrophil, and other white blood cell lineages. Relevant examples of how these approaches have been used for disease diagnosis, gene discovery, and studying complex traits are provided. High-throughput DNA sequencing technology holds significant promise for impacting clinical care. This includes development of improved disease detection and diagnosis, better understanding of disease progression and stratification of risk of disease-specific complications, and development of improved therapeutic strategies, particularly patient-specific pharmacogenomics-based therapy, with monitoring of therapy by genomic biomarkers.

Insight into GATA1 transcriptional activity through interrogation ofciselements disrupted in human erythroid disorders

“…In this study, we interrogated natural genetic variations occurring in MEDs to provide a window into the functions of erythroid gene regulatory networks (2). In primary HSPCs and an erythroid cell line, we used CRISPR/Cas9 genome editing to verify the regulatory effects of NC genetic variants associated with XLSA, CEP, and PKD, and were able to recapitulate the cellintrinsic phenotypes of the associated disorder (20)(21)(22)(23)43).…”

“…In hematology, WES has been extremely useful for identifying unknown genetic etiologies for various disorders, such as those affecting red blood cell (RBC) production, including Diamond-Blackfan anemia and congenital dyserythropoietic anemia (3)(4)(5), disorders of RBC structure and function (6,7), and disorders affecting other aspects of hematologic function (2,8). Despite this considerable success, however, more than 50% of cases of presumed monogenic diseases are refractory to current WES approaches (9).…”

“…W hole-exome sequencing (WES) and targeted sequencing approaches have greatly accelerated our ability to identify causal genetic lesions in both previously implicated and novel genes underlying monogenic disorders (1,2). In hematology, WES has been extremely useful for identifying unknown genetic etiologies for various disorders, such as those affecting red blood cell (RBC) production, including Diamond-Blackfan anemia and congenital dyserythropoietic anemia (3)(4)(5), disorders of RBC structure and function (6,7), and disorders affecting other aspects of hematologic function (2,8).…”

Prenatal Screening for Co-Inheritance of Sickle Cell Anemia and β-Thalassemia Traits

“…In case of real time PCR based single gene disorder assay for β-thalassemia screening, there is great risk of missing sickle cell anemia disease or trait, although, sickle cell mutation is present in the same β-globin gene. Nucleotide sequencing covers a larger target region and can detect unexpected and/ or novel variants which can be missed by target specific probe based real time PCR technologies [16]. Hence, nucleotide sequencing proves to be an advantageous method over real time PCR based detection especially in cases like this wherein the parents are asymptomatic.…”

Society for Pediatric Research 2015 Young Investigator Award: genetics of human hematopoiesis—what patients can teach us about blood cell production

“…Instead, disease gene identification can be accomplished by assessing multiple unrelated individuals who all have a particular disease after removal of variants found in healthy individuals in the general population (17). This has led to the widespread use of such approaches that enrich and sequence the 1-2% of the genome containing protein-coding genes, termed the exome, and has commonly been referred to as whole-exome sequencing (17)(18)(19). While using such broad-based sequencing approaches can lead to the identification of a number of rare variants, most of which will not be causal for a disease, specific approaches can enrich for true causal alleles, such as by looking at multiple unrelated probands and using segregation of mutations in affected individuals within a family (17).…”