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L265P mutation of the MYD88 gene is frequent in Waldenström's macroglobulinemia and its absence in myeloma.

Mori N, Ohwashi M, Yoshinaga K, Mitsuhashi K, Tanaka N, Teramura M, Okada M, Shiseki M, Tanaka J, Motoji T - PLoS ONE (2013)

Bottom Line: We next tested for the mutation with BSiE1 digestion and allele-specific polymerase chain reaction in the 28 patients and 38 patients with myeloma.No siginificant difference was observed in the incidence of the L265P mutation between BSiE1 digestion and allele-specific polymerase chain reaction (p=0.32).BSiE1 digestion and allele-specific polymerase chain reaction may detect a small fraction of mutated cells in some cases.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Tokyo Women's Medical University, Tokyo, Japan.

ABSTRACT
L265P mutation in the MYD88 gene has recently been reported in Waldenström's macroglobulinemia; however the incidence has been different according to the methods used. To determine the relevance and compare the incidence by different methods, we analyzed the L265P mutation in bone marrow mononuclear cells from lymphoid neoplasms. We first performed cloning and sequencing in 10 patients: 8 Waldenström's macroglobulinemia; 1 non-IgM-secreting lymphoplasmacytic lymphoma; and 1 low grade B-cell lymphoma with monoclonal IgG protein. The L265P mutation was detected in only 1/8 Waldenström's macroglobulinemia patients (2 of 9 clones). To confirm these results, direct sequencing was performed in the 10 patients and an additional 17 Waldenström's macroglobulinemia patients and 1 lymphoplasmacytic lymphoma patient. Nine of 28 patients (7/25 Waldenström's macroglobulinemia, 1/2 lymphoplasmacytic lymphoma, and B-cell lymphoma) harbored the mutation. We next tested for the mutation with BSiE1 digestion and allele-specific polymerase chain reaction in the 28 patients and 38 patients with myeloma. Aberrant bands corresponding to the mutation were detected by BSiE1 digestion in 19/25 patients with Waldenström's macroglobulinemia (76%), 1/2 lymphoplasmacytic lymphoma and B-cell lymphoma, but not in the 38 myeloma patients. The L265P mutation was more frequent in patients with Waldenström's macroglobulinemia than in those with myeloma (p=1.3x10(-10)). The mutation was detected by allele-specific polymerase chain reaction in 18/25 Waldenström's macroglobulinemia patients (72%). In the 25 Waldenström's macroglobulinemia patients, the L265P was more frequently detected by BSiE1 digestion than by direct sequencing (p=5.3x10(-4)), and in males (15/16, 94%) than in females (4/9, 44%) (p=1.2x10(-2)). No siginificant difference was observed in the incidence of the L265P mutation between BSiE1 digestion and allele-specific polymerase chain reaction (p=0.32). These results suggest that the L265P mutation is involved in the majority of Waldenström's macroglobulinemia. BSiE1 digestion and allele-specific polymerase chain reaction may detect a small fraction of mutated cells in some cases.

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Allele-specific polymerase chain reaction (AS-PCR) of the MYD88 gene in Waldenström’s macroglobulinemia and lymphoma.Ten μl of PCR products was separated by electrophoresis through a 2% agarose gel, stained with ethidium bromide, and visualized by ultraviolet illumination. The size of the products is indicated on the left.(A) Sensitivity of AS-PCR. L265P-positive DNA (WM5) was diluted into wild-type DNA (WM2) before amplification. Mutations were detected in samples containing 0.1% or more of the L265P mutation. Lane 1, 100 bp ladder; lane 2, 0%; lane 3, 0.1%; lane 4, 0.5%; lane 5, 1%; lane 6, 0%; lane 7, 0.1%; lane 8, 0.5%; lane 9, 1%.(B) Aberrant bands were detected in 12 of 16 samples from WM patients (lanes 2, 5, 6, 8, 9, 10, 11, 12, 14, 15, 16, and 17). Lane 1, 100 bp ladder; lane 2, WM1; lane 3, WM2; lane 4, WM3; lane 5, WM4; lane 6, WM5; lane 7, WM6; lane 8, WM7; lane 9, WM8; lane 10, WM9; lane 11, WM10; lane 12, WM11; lane 13, WM12, lane 14, WM13; lane 15, WM14; lane 16, WM15; lane 17, WM16.(C) Aberrant bands were detected in 6 of 10 samples from WM patients (lanes 2, 3, 6, 7, 8, and 10) and 2 of 3 non-Hodgkin’s lymphoma patients (lanes 12 and 13). Lane 1, 100 bp ladder; lane 2, WM17; lane 3, WM18; lane 4, WM19; lane 5, WM20; lane 6, WM21; lane 7, WM22; lane 8, WM23; lane 9, WM24; lane 10, WM25; lane 11, NHL1; lane 12, NHL2; lane 13, NHL3; lane 14, normal lymphocyte; lane 15, water.
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pone-0080088-g003: Allele-specific polymerase chain reaction (AS-PCR) of the MYD88 gene in Waldenström’s macroglobulinemia and lymphoma.Ten μl of PCR products was separated by electrophoresis through a 2% agarose gel, stained with ethidium bromide, and visualized by ultraviolet illumination. The size of the products is indicated on the left.(A) Sensitivity of AS-PCR. L265P-positive DNA (WM5) was diluted into wild-type DNA (WM2) before amplification. Mutations were detected in samples containing 0.1% or more of the L265P mutation. Lane 1, 100 bp ladder; lane 2, 0%; lane 3, 0.1%; lane 4, 0.5%; lane 5, 1%; lane 6, 0%; lane 7, 0.1%; lane 8, 0.5%; lane 9, 1%.(B) Aberrant bands were detected in 12 of 16 samples from WM patients (lanes 2, 5, 6, 8, 9, 10, 11, 12, 14, 15, 16, and 17). Lane 1, 100 bp ladder; lane 2, WM1; lane 3, WM2; lane 4, WM3; lane 5, WM4; lane 6, WM5; lane 7, WM6; lane 8, WM7; lane 9, WM8; lane 10, WM9; lane 11, WM10; lane 12, WM11; lane 13, WM12, lane 14, WM13; lane 15, WM14; lane 16, WM15; lane 17, WM16.(C) Aberrant bands were detected in 6 of 10 samples from WM patients (lanes 2, 3, 6, 7, 8, and 10) and 2 of 3 non-Hodgkin’s lymphoma patients (lanes 12 and 13). Lane 1, 100 bp ladder; lane 2, WM17; lane 3, WM18; lane 4, WM19; lane 5, WM20; lane 6, WM21; lane 7, WM22; lane 8, WM23; lane 9, WM24; lane 10, WM25; lane 11, NHL1; lane 12, NHL2; lane 13, NHL3; lane 14, normal lymphocyte; lane 15, water.

Mentions: Sensitivity to the L265P mutation by AS-PCR was 0.1-0.5% in our study (Figure 3 A). The mutation was detected in 20 of the 28 patients: 18 of the 25 WM (72%), one of the 2 non-IgM-secreting LPL, and B-cell lymphoma with IgG M-protein (Table 2, Figure 3 B-C). All of the 18 patients, in whom the L265P mutation was detected by AS-PCR, were also shown to have the mutation by BSiE1 digestion (Table 2).


L265P mutation of the MYD88 gene is frequent in Waldenström's macroglobulinemia and its absence in myeloma.

Mori N, Ohwashi M, Yoshinaga K, Mitsuhashi K, Tanaka N, Teramura M, Okada M, Shiseki M, Tanaka J, Motoji T - PLoS ONE (2013)

Allele-specific polymerase chain reaction (AS-PCR) of the MYD88 gene in Waldenström’s macroglobulinemia and lymphoma.Ten μl of PCR products was separated by electrophoresis through a 2% agarose gel, stained with ethidium bromide, and visualized by ultraviolet illumination. The size of the products is indicated on the left.(A) Sensitivity of AS-PCR. L265P-positive DNA (WM5) was diluted into wild-type DNA (WM2) before amplification. Mutations were detected in samples containing 0.1% or more of the L265P mutation. Lane 1, 100 bp ladder; lane 2, 0%; lane 3, 0.1%; lane 4, 0.5%; lane 5, 1%; lane 6, 0%; lane 7, 0.1%; lane 8, 0.5%; lane 9, 1%.(B) Aberrant bands were detected in 12 of 16 samples from WM patients (lanes 2, 5, 6, 8, 9, 10, 11, 12, 14, 15, 16, and 17). Lane 1, 100 bp ladder; lane 2, WM1; lane 3, WM2; lane 4, WM3; lane 5, WM4; lane 6, WM5; lane 7, WM6; lane 8, WM7; lane 9, WM8; lane 10, WM9; lane 11, WM10; lane 12, WM11; lane 13, WM12, lane 14, WM13; lane 15, WM14; lane 16, WM15; lane 17, WM16.(C) Aberrant bands were detected in 6 of 10 samples from WM patients (lanes 2, 3, 6, 7, 8, and 10) and 2 of 3 non-Hodgkin’s lymphoma patients (lanes 12 and 13). Lane 1, 100 bp ladder; lane 2, WM17; lane 3, WM18; lane 4, WM19; lane 5, WM20; lane 6, WM21; lane 7, WM22; lane 8, WM23; lane 9, WM24; lane 10, WM25; lane 11, NHL1; lane 12, NHL2; lane 13, NHL3; lane 14, normal lymphocyte; lane 15, water.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3818242&req=5

pone-0080088-g003: Allele-specific polymerase chain reaction (AS-PCR) of the MYD88 gene in Waldenström’s macroglobulinemia and lymphoma.Ten μl of PCR products was separated by electrophoresis through a 2% agarose gel, stained with ethidium bromide, and visualized by ultraviolet illumination. The size of the products is indicated on the left.(A) Sensitivity of AS-PCR. L265P-positive DNA (WM5) was diluted into wild-type DNA (WM2) before amplification. Mutations were detected in samples containing 0.1% or more of the L265P mutation. Lane 1, 100 bp ladder; lane 2, 0%; lane 3, 0.1%; lane 4, 0.5%; lane 5, 1%; lane 6, 0%; lane 7, 0.1%; lane 8, 0.5%; lane 9, 1%.(B) Aberrant bands were detected in 12 of 16 samples from WM patients (lanes 2, 5, 6, 8, 9, 10, 11, 12, 14, 15, 16, and 17). Lane 1, 100 bp ladder; lane 2, WM1; lane 3, WM2; lane 4, WM3; lane 5, WM4; lane 6, WM5; lane 7, WM6; lane 8, WM7; lane 9, WM8; lane 10, WM9; lane 11, WM10; lane 12, WM11; lane 13, WM12, lane 14, WM13; lane 15, WM14; lane 16, WM15; lane 17, WM16.(C) Aberrant bands were detected in 6 of 10 samples from WM patients (lanes 2, 3, 6, 7, 8, and 10) and 2 of 3 non-Hodgkin’s lymphoma patients (lanes 12 and 13). Lane 1, 100 bp ladder; lane 2, WM17; lane 3, WM18; lane 4, WM19; lane 5, WM20; lane 6, WM21; lane 7, WM22; lane 8, WM23; lane 9, WM24; lane 10, WM25; lane 11, NHL1; lane 12, NHL2; lane 13, NHL3; lane 14, normal lymphocyte; lane 15, water.
Mentions: Sensitivity to the L265P mutation by AS-PCR was 0.1-0.5% in our study (Figure 3 A). The mutation was detected in 20 of the 28 patients: 18 of the 25 WM (72%), one of the 2 non-IgM-secreting LPL, and B-cell lymphoma with IgG M-protein (Table 2, Figure 3 B-C). All of the 18 patients, in whom the L265P mutation was detected by AS-PCR, were also shown to have the mutation by BSiE1 digestion (Table 2).

Bottom Line: We next tested for the mutation with BSiE1 digestion and allele-specific polymerase chain reaction in the 28 patients and 38 patients with myeloma.No siginificant difference was observed in the incidence of the L265P mutation between BSiE1 digestion and allele-specific polymerase chain reaction (p=0.32).BSiE1 digestion and allele-specific polymerase chain reaction may detect a small fraction of mutated cells in some cases.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, Tokyo Women's Medical University, Tokyo, Japan.

ABSTRACT
L265P mutation in the MYD88 gene has recently been reported in Waldenström's macroglobulinemia; however the incidence has been different according to the methods used. To determine the relevance and compare the incidence by different methods, we analyzed the L265P mutation in bone marrow mononuclear cells from lymphoid neoplasms. We first performed cloning and sequencing in 10 patients: 8 Waldenström's macroglobulinemia; 1 non-IgM-secreting lymphoplasmacytic lymphoma; and 1 low grade B-cell lymphoma with monoclonal IgG protein. The L265P mutation was detected in only 1/8 Waldenström's macroglobulinemia patients (2 of 9 clones). To confirm these results, direct sequencing was performed in the 10 patients and an additional 17 Waldenström's macroglobulinemia patients and 1 lymphoplasmacytic lymphoma patient. Nine of 28 patients (7/25 Waldenström's macroglobulinemia, 1/2 lymphoplasmacytic lymphoma, and B-cell lymphoma) harbored the mutation. We next tested for the mutation with BSiE1 digestion and allele-specific polymerase chain reaction in the 28 patients and 38 patients with myeloma. Aberrant bands corresponding to the mutation were detected by BSiE1 digestion in 19/25 patients with Waldenström's macroglobulinemia (76%), 1/2 lymphoplasmacytic lymphoma and B-cell lymphoma, but not in the 38 myeloma patients. The L265P mutation was more frequent in patients with Waldenström's macroglobulinemia than in those with myeloma (p=1.3x10(-10)). The mutation was detected by allele-specific polymerase chain reaction in 18/25 Waldenström's macroglobulinemia patients (72%). In the 25 Waldenström's macroglobulinemia patients, the L265P was more frequently detected by BSiE1 digestion than by direct sequencing (p=5.3x10(-4)), and in males (15/16, 94%) than in females (4/9, 44%) (p=1.2x10(-2)). No siginificant difference was observed in the incidence of the L265P mutation between BSiE1 digestion and allele-specific polymerase chain reaction (p=0.32). These results suggest that the L265P mutation is involved in the majority of Waldenström's macroglobulinemia. BSiE1 digestion and allele-specific polymerase chain reaction may detect a small fraction of mutated cells in some cases.

Show MeSH
Related in: MedlinePlus