Related Experiment Video
Updated: May 6, 2026
Engineering Oncogenic Heterozygous Gain-of-Function Mutations in Human Hematopoietic Stem and Progenitor Cells
Published on: March 10, 2023
Targeted cancer exome sequencing reveals recurrent mutations in myeloproliferative neoplasms
E Tenedini1, I Bernardis2, V Artusi2
11] Center for Genome Research, University of Modena and Reggio Emilia, Modena, Italy [2] Life Sciences Department, University of Modena and Reggio Emilia, Modena, Italy [3] Center for Regenerative Medicine 'Stefano Ferrari', University of Modena and Reggio Emilia, Modena, Italy.
Abstract:
With the intent of dissecting the molecular complexity of Philadelphia-negative myeloproliferative neoplasms (MPN), we designed a target enrichment panel to explore, using next-generation sequencing (NGS), the mutational status of an extensive list of 2000 cancer-associated genes and microRNAs. The genomic DNA of granulocytes and in vitro-expanded CD3+T-lymphocytes, as a germline control, was target-enriched and sequenced in a learning cohort of 20 MPN patients using Roche 454 technology. We identified 141 genuine somatic mutations, most of which were not previously described. To test the frequency of the identified variants, a larger validation cohort of 189 MPN patients was additionally screened for these mutations using Ion Torrent AmpliSeq NGS. Excluding the genes already described in MPN, for 8 genes (SCRIB, MIR662, BARD1, TCF12, FAT4, DAP3, POLG and NRAS), we demonstrated a mutation frequency between 3 and 8%. We also found that mutations at codon 12 of NRAS (NRASG12V and NRASG12D) were significantly associated, for primary myelofibrosis (PMF), with highest dynamic international prognostic scoring system (DIPSS)-plus score categories. This association was then confirmed in 66 additional PMF patients composing a final dataset of 168 PMF showing a NRAS mutation frequency of 4.7%, which was associated with a worse outcome, as defined by the DIPSS plus score.
Insights
Researchers explored gene mutations in Philadelphia-negative myeloproliferative neoplasms (MPN) using next-generation sequencing. They identified new mutations in genes like NRAS, linking NRAS mutations to worse outcomes in primary myelofibrosis (PMF).
Area of Science:
- Genetics
- Oncology
- Molecular Biology
Background:
- Philadelphia-negative myeloproliferative neoplasms (MPN) are complex blood cancers.
- Understanding their molecular landscape is crucial for diagnosis and treatment.
- Previous studies have identified key mutations, but a comprehensive analysis is needed.
Purpose of the Study:
- To dissect the molecular complexity of Philadelphia-negative MPN.
- To identify novel somatic mutations and microRNAs associated with MPN.
- To investigate the prognostic significance of identified mutations, particularly in primary myelofibrosis (PMF).
Main Methods:
- Utilized next-generation sequencing (NGS) with a target enrichment panel of 2000 cancer-associated genes and microRNAs.
- Sequenced DNA from granulocytes and T-lymphocytes (as germline control) in learning and validation cohorts of MPN patients.
- Employed Roche 454 and Ion Torrent AmpliSeq NGS technologies for mutation identification and frequency testing.
Main Results:
- Identified 141 novel somatic mutations in MPN patients.
- Found mutation frequencies between 3-8% for 8 genes (SCRIB, MIR662, BARD1, TCF12, FAT4, DAP3, POLG, NRAS).
- NRAS mutations (NRASG12V, NRASG12D) were significantly associated with higher DIPSS-plus scores in PMF, indicating a worse prognosis.
Conclusions:
- The study identified novel genetic alterations in Philadelphia-negative MPN.
- NRAS mutations are potential prognostic markers for primary myelofibrosis.
- These findings contribute to a deeper understanding of MPN molecular pathogenesis and clinical outcomes.
Related Concept Videos
02:57Targeted Cancer Therapies
There are several types of targeted therapies against...
02:21Cancers Originate from Somatic Mutations in a Single Cell
02:23Abnormal Proliferation