Single-cell whole exome and targeted sequencing in NPM1/FLT3 positive pediatric acute myeloid leukemia

Pediatric Blood & Cancer
Christiane WalterHelmut Hanenberg

Abstract

The small portion of leukemic stem cells (LSCs) in acute myeloid leukemia (AML) present in children and adolescents is often masked by the high background of AML blasts and normal hematopoietic cells. The aim of the current study was to establish a simple workflow for reliable genetic analysis of single LSC-enriched blasts from pediatric patients. For three AMLs with mutations in nucleophosmin 1 and/or fms-like tyrosine kinase 3, we performed whole genome amplification on sorted single-cell DNA followed by whole exome sequencing (WES). The corresponding bulk bone marrow DNAs were also analyzed by WES and by targeted sequencing (TS) that included 54 genes associated with myeloid malignancies. Analysis revealed that read coverage statistics were comparable between single-cell and bulk WES data, indicating high-quality whole genome amplification. From 102 single-cell variants, 72 single nucleotide variants and insertions or deletions (70%) were consistently found in the two bulk DNA analyses. Variants reliably detected in single cells were also present in TS. However, initial screening by WES with read counts between 50-72× failed to detect rare AML subclones in the bulk DNAs. In summary, our study demonstrated that single-cell WE...Continue Reading

References

Aug 15, 2002·Blood·D Gary Gilliland, James D Griffin
Jun 12, 2003·International Journal of Cancer. Journal International Du Cancer·Jochen GreinerMichael Schmitt
Mar 6, 2007·Best Practice & Research. Clinical Haematology·Craig T Jordan
May 3, 2008·The New England Journal of Medicine·Richard F SchlenkUNKNOWN German-Austrian Acute Myeloid Leukemia Study Group
Jun 10, 2008·Cancer Cell·Junping WeiJames C Mulloy
Aug 7, 2009·The New England Journal of Medicine·Elaine R MardisTimothy J Ley
Sep 2, 2010·Current Opinion in Oncology·Ulrike BacherTorsten Haferlach
Feb 22, 2012·Oncogene·M Jan, R Majeti
Jul 24, 2012·Cell·John S WelchRichard K Wilson
Aug 11, 2012·Blood·Ursula CreutzigUNKNOWN AML Committee of the International BFM Study Group
Aug 31, 2012·Science Translational Medicine·Max JanRavindra Majeti
Mar 13, 2014·Cell Stem Cell·Antonija Kreso, John E Dick
Aug 20, 2014·PloS One·Charles F A de BourcyStephen R Quake
Jan 1, 2015·Blood·R Coleman LindsleyBenjamin L Ebert
Jun 27, 2015·Seminars in Hematology·Andreas ReinischRavindra Majeti
Aug 26, 2015·Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology·C Michel ZwaanGertjan J L Kaspers
Jan 26, 2016·Nature Reviews. Genetics·Charles GawadStephen R Quake
Mar 10, 2016·Journal of Clinical Medicine·Jennifer N Saultz, Ramiro Garzon
May 11, 2016·Current Opinion in Genetics & Development·Sheng LiAri Melnick

❮ Previous
Next ❯

Citations

May 9, 2019·Physiological Reviews·Julhash U Kazi, Lars Rönnstrand
Oct 12, 2019·British Journal of Haematology·Marcus C HansenCharlotte G Nyvold

❮ Previous
Next ❯

Related Concepts

Related Feeds

AML: Role of LSD1 by CRISPR (Keystone)

Find the latest rersearrch on the ability of CRISPR-Cas9 mutagenesis to profile the interactions between lysine-specific histone demethylase 1 (LSD1) and chemical inhibitors in the context of acute myeloid leukemia (AML) here.

Acute Myeloid Leukaemia & RNA

Acute myeloid leukaemia (AML) is a common hematological type of cancer. As the population ages, there has been a rise in the frequency of AML. RNA expression has been used to see if there are different genetic profiles that exist within AML and whether these may underpin the variations in survival rates. Here is the latest research on AML and RNA.

Acute Myeloid Leukemia

Acute myeloid leukemia (AML) is a clinically and genetically heterogeneous disease with approximately 20,000 cases per year in the United States. AML also accounts for 15-20% of all childhood acute leukemias, while it is responsible for more than half of the leukemic deaths in these patients. Here is the latest research on this disease.

Blood And Marrow Transplantation

The use of hematopoietic stem cell transplantation or blood and marrow transplantation (bmt) is on the increase worldwide. BMT is used to replace damaged or destroyed bone marrow with healthy bone marrow stem cells. Here is the latest research on bone and marrow transplantation.