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HelpTest Code CXCFX MYD88 Reflex to CXCR4 Mutation Detection, Varies
Reporting Name
CXCR4, Gene Mutation, ReflexUseful For
The prognosis and clinical management of lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia
Method Name
Only orderable as a reflex. For more information see LPLFX / Lymphoplasmacytic Lymphoma/Waldenstrom Macroglobulinemia, MYD88 L265P with Reflex to CXCR4, Varies.
Bridged Nucleic Acids (BNA) Clamp Sanger Sequencing/Routine Sanger Sequencing
(BNAClamp is utilized pursuant to a license agreement with BNA Inc.)
Performing Laboratory
Mayo Clinic Laboratories in Rochester
Specimen Type
VariesSpecimen Required
No additional specimen is required.
Only orderable as a reflex. For more information see LPLFX / Lymphoplasmacytic Lymphoma/Waldenstrom Macroglobulinemia, MYD88 L265P with Reflex to CXCR4, Varies.
Specimen Minimum Volume
Blood, Bone Marrow: 1 mL
Extracted DNA: 20 mcL with a concentration of at least 10 nanograms per mcL
Specimen Stability Information
| Specimen Type | Temperature | Time | Special Container |
|---|---|---|---|
| Varies | Varies | 10 days |
Reject Due To
| Gross hemolysis | OK |
| B5-fixed tissues Bone marrow biopsies Frozen tissue Methanol acetic acid (MAA)-fixed pellets Moderately to severely clotted Paraffin shavings Slides |
Reject |
Reference Values
Only orderable as a reflex. For more information see LPLFX / Lymphoplasmacytic Lymphoma/Waldenstrom Macroglobulinemia, MYD88 L265P with Reflex to CXCR4, Varies.
An interpretive report will be provided
Day(s) Performed
Monday through Friday
CPT Code Information
81479-Unlisted molecular pathology procedure
Secondary ID
601508Clinical Information
Lymphoplasmacytic lymphoma/Waldenstrom macroglobulinemia (LPL/WM) is a B-cell lymphoma characterized by an aberrant accumulation of malignant lymphoplasmacytic cells in the bone marrow, lymph nodes, and spleen. It is a B-cell neoplasm that can exhibit excess production of serum IgM symptoms related to hyperviscosity, tissue filtration, and autoimmune-related pathology. CXCR4 mutations are identified in approximately 30% to 40% of patients with LPL/WM and are almost always in association with MYD88 L265P, which is highly prevalent in this neoplasm. The status of CXCR4 muations in the context of MYD88 L265P is clinically relevant as important determinants of clinical presentation, overall survival, and therapeutic response to ibrutinib. A MYD88-L265P/CXCR4-WHIM (C-terminus nonsense/frameshift mutations) molecular signature is associated with intermediate to high bone marrow disease burden and serum IgM levels, less adenopathy, and intermediate response to ibrutinib in previously treated patients. A MYD88-L265P/CXCR4-WT (wildtype) molecular signature is associated with intermediate bone marrow disease burden and serum IgM levels, more adenopathy, and highest response to ibrutinib in previously treated patients. The MYD88-WT/CXCR4-WT molecular signature is associated with inferior overall survival, lower response to ibrutinib therapy in previously treated patients, and lower bone marrow disease burden in comparison to those harboring a MYD88-L265 variant.
Interpretation
Mutation present or not detected; an interpretive report will be issued under LPLFX / Lymphoplasmacytic Lymphoma/Waldenstrom Macroglobulinemia, MYD88 L265P with Reflex to CXCR4, Varies.
Cautions
This test is a targeted assay for the C-terminus end of the CXCR4 gene only. It examines c.898-1059 of the CXCR4 gene (NCBI NM_003467.2 GRCh37) and does not detect variants outside this region. A 1% analytical sensitivity was established at 50-ng DNA input for the hotspot mutations c.1013C>G/A only, which uses bridged nucleic acids clamped Sanger sequencing, and DNA not meeting established criteria can lead to false-negative results. In the extremely rare event that a rare polymorphism or indel occurs at the Sanger sequencing primer binding sites, in cis with c.1013C>G/A, data can yield a failed result. Routine Sanger sequencing is used to interrogate other mutations in the tested region with a 15% to 20% analytical sensitivity. The analytical sensitivity of the assay can be affected by a variety of factors, including biologic availability (ie, tumor burden), fixation of paraffin-embedded specimens, rare polymorphisms or indels at the primer binding sites, or nonspecific polymerase chain reaction interferences.
Clinical Reference
1. Hunter Z, Xu L, Yang G, et al: The genomic landscape of Waldenstrom macroglobulinemia is characterized by highly recurring MYD88 and WHIM-like CXCR4 mutations, and small somatic deletions associated with B-cell lymphomagenesis. Blood. 2014 Mar 13;123(11):1637-1646. doi: 10.1182/blood-2013-09-525808
2. Landgren O, Tageja N: MYD88 and beyond: novel opportunities for diagnosis, prognosis and treatment in Waldenstrom's Macroglobulinemia. Leukemia. 2014 Sep;28(9):1799-1803. doi: 10.1038/leu.2014.88
3. Poulain S, Roumier C, Venet-Caillault A, et al: Genomic Landscape of CXCR4 Mutations in Waldenstrom Macroglobulinemia. Clin Cancer Res. 2016 Mar 15;22(6):1480-1488. doi: 10.1158/1078-0432.CCR-15-0646
4. Roccaro A, Sacco A, Jimenez C, et al: C1013G/CXCR4 acts as a driver mutation of tumor progression and modulator of drug resistance in lymphoplasmacytic lymphoma. Blood. 2014 Jun 26;123(26):4120-4131. doi: 10.1182/blood-2014-03-564583
5. Schmidt J, Federmann B, Schindler N, et al: MYD88 L265P and CXCR4 mutations in lymphoplasmacytic lymphoma identify cases with high disease activity. Br J Haematol. 2015 Jun;169(6):795-803. doi: 10.1111/bjh.13361
6. Treon SP, Cao Y, Xu L, Yang G, Liu X, Hunter ZR: Somatic mutations in MYD88 and CXCR4 are determinants of clinical presentation and overall survival in Waldenstrom macroglobulinemia. Blood. 2014 May 1;123(18):2791-2796. doi: 10.1182/blood-2014-01-550905
7. Treon SP, Tripsas CK, Meid K, et al: Ibrutinib in previously treated Waldenstrom's macroglobulinemia. N Engl J Med. 2015 Apr 9;372(15):1430-1440. doi: 10.1056/NEJMoa1501548
8. Xu L, Hunter ZR, Tsakmaklis N, et al: Clonal architecture of CXCR4 WHIM-like mutations in Waldenstrom Macroglobulinaemia. Br J Haematol. 2016 Mar;172(5):735-744. doi: 10.1111/bjh.13897
Method Description
The C-terminus end of CXCR4 (NM_003467.2, c.898-1059) is amplified from extracted genomic DNA by polymerase chain reaction, followed by Sanger sequencing and capillary electrophoresis analysis. Review of the sequence data is performed using a combination of automated calls and manual inspection.(Unpublished Mayo method)
The hotspot mutations c.1013C>G/A (p.S338X) are examined using bridged nucleic acids clamped Sanger sequencing with an analytic sensitivity of 1%. All other genetic mutations in the test region are examined by routine Sanger sequencing with an analytic sensitivity of 15% to 20%.(Unpublished Mayo method)