Introduction
Familial Mediterranean Fever (FMF) is the most
common Mendelian autoinflammatory disorder, which is characterized
by recurrent attacks of fever with peritoneal, pleural or synovial
inflammation (1–4). Missense mutations in the Mediterranean
fever (MEFV) gene have been shown to be causative of the
disease (5). The 781-aa protein
product of MEFV, denoted pyrin (also known as marenostrin)
is produced in neutrophils, dendritic cells, eosinophils,
monocytes, and synovial fibroblasts (6–10).
Currently, >270 inherited variants and polymorphisms in
MEFV have been reported in the Infevers Database (http://fmf.igh.cnrs.fr/ISSAID/infevers).
Most of the inherited variants in MEFV are located in exon 2
and 10 of the transcript (11).
Pyrin contains several domains, including a pyrin domain (12–14),
which is involved in homotypic protein-protein interactions in
inflammatory and apoptotic signaling pathways (7,15).
Although the underlying mechanism is still being investigated,
pyrin potentially plays a role in the modulation of interleukin-1β
(IL-1β) and nuclear factor-κB (NF-κB) (16–20).
Any inherited variants in the MEFV gene may cause
inflammation and apoptosis due to the altered control of pyrin in
the activation of IL-1β and NF-κB (21,22).
Multiple myeloma (MM) is a neoplastic plasma-cell
disorder that is characterized by the aberrant expansion of
monotypic plasma cells within the bone marrow (23,24).
Perturbed signaling pathways that control normal physiological
processes, and mutations in several protooncogenes and tumor
suppressor genes, can lead to the development of MM (23). Although NF-κB functions in the
pathogenesis of MM (25,26), whether inherited variants in MEFV
can lead to constitutive NF-κB activation and cause a tendency for
MM remains to be determine. Accumulated evidence has shown that
there is a high frequency of inherited variants in MEFV in
patients with hematological malignancies as compared with the
general population (27–31). This association has been included in
the Genetic Association Database, Record 704091 (http://geneticassociationdb.nih.gov/cgi-bin/view.cgi?table=allview&id=704091).
In one of our previous studies, an increased frequency of inherited
variants in MEFV in patients with MM was observed (28). Since the sample size was small in
the present pilot study, the actual frequency of inherited variants
in MEFV in patients with MM was able to be investigated.
Materials and methods
Subjects
Twenty-eight (17 male and 11 female) patients with
MM and 65 healthy controls (40 male and 25 female) were included in
the study. FMF patients or subjects who had a family history of FMF
were excluded. The study protocol conformed to the ethical
guidelines of the Helsinki Declaration. Informed consent was
obtained from all patients and controls. The local Ethics Committee
and Institutional Review Board approved the study. All the patients
donated 2 ml of blood, collected in an ethylenediaminetetraacetic
acid tube. The eight inherited variants in the MEFV gene
(M694I, M694V, M680I (G/C-A), V726A, R761H, E148Q and P369S) were
detected using the Dr. Zeydanli® FMF Type I PCR system
(Ankara, Turkey) 5′ nuclease assay method using an ABI 7500
(Applied Biosystems, Foster City, CA, USA) quantitative polymerase
chain reaction system, as previously reported (27).
Statistical analysis
Data were analyzed using SPSS 17.0 (SPSS Inc.,
Chicago, IL, USA) statistical software. Differences between the
groups were analyzed using a χ2 test. P≤0.05 was
considered to indicate a statistically significant difference.
Results
The mean age of the patients with MM and healthy
controls was 59.38±22.88 years (age range, 32–84) and 30.25±10.62
years (age range, 20–45), respectively. Hematological
characteristics and identified MEFV gene variants in
patients with MM are shown in Table
I. Six heterozygous and one homozygous (E148Q/E148Q) variant in
patients with MM was identified. None of the subjects had a family
history compatible with FMF. In the healthy control group, 11
heterozygous variants were identified. M680I, M694I and R761H
inherited MEFV gene variants were not found in any of the
groups. The P369S variant was found in one healthy control.
 | Table IHematological characteristics and
distribution of inherited variants in the Mediterranean fever gene
in patients with multiple myeloma. |
Table I
Hematological characteristics and
distribution of inherited variants in the Mediterranean fever gene
in patients with multiple myeloma.
| Patient no. | Age | Gender | Bone marrow
plasmacytosis (%) | WBC
(x109/l) | PLTS
(x109/l) | Hb (g/dl) | ESR (mm/h) | Inherited variants in
MEFV gene |
|---|
| 1 | 78 | M | 65 | 4,050 | 104 | 10 | 102 | E148Q/Unknowna |
| 2 | 54 | M | 90 | 93,700 | 2,28 | 8,7 | 91 | E148Q/Unknown |
| 3 | 70 | M | 2 | 5,980 | 240 | 12,4 | 46 | E148Q/Unknown |
| 4 | 65 | M | 35 | 31,200 | 156 | 9,4 | 92 | E148Q/E148Q |
| 5 | 74 | F | 40 | 29,300 | 111 | 13,9 | 91 | V726A/Unknown |
| 6 | 49 | F | 45 | 7,660 | 218 | 15 | 135 | M694V/Unknown |
| 7 | 68 | F | 40 | 4,440 | 58,200 | 9,3 | 60 | E148Q/Unknown |
Analytical data concerning the overall inherited
MEFV variant frequency between patients with MM and
comparisons with healthy controls are given in Table II. The difference in the overall
frequency of the inherited variants in the MEFV gene between
MM patients and the controls was statistically significant
(χ2=4.905; P=0.027). When the distribution was compared
between the patients and the controls, the frequency of the E148Q
variant was significantly higher in the patient group as compared
with the controls (χ2=7.438; P=0.006), while the M694V
was significantly higher in the control group than MM patients
(χ2=5.658; P=0.017).
| Table IIComparison of the inherited variant
frequency in the Mediterranean fever gene between patients with
multiple myeloma and normal controls. |
Table II
Comparison of the inherited variant
frequency in the Mediterranean fever gene between patients with
multiple myeloma and normal controls.
| | | Heterozygote variant
frequencies in MEFV gene |
|---|
| | |
|
|---|
| Variables | n | Overall inherited
variant frequency in MEFV gene | Homozygote
(E148Q/E148Q) | M694V/Unknowna | E148Q/Unknown | M680I/Unknown | V726A/Unknown | M694I/Unknown | R761H/Unknown | P369S/Unknown |
|---|
| Normal controls | 65 | 0.084 | 0 | 0.038 | 0.015 | 0 | 0.023 | 0 | 0 | 0.007 |
| Multiple myeloma | 28 | 0.143 | 1 | 0.017 | 0.107 | 0 | 0.017 | 0 | 0 | 0 |
| χ2 | | 4.905 | - | 5.658 | 7.438 | | 0.535 | - | - | 0.433 |
| P-value | | 0.027 | - | 0.017 | 0.006 | - | NS | - | - | NS |
Discussion
In the current study, a high frequency of inherited
variants in the MEFV gene was identified in patients with MM
as compared with the healthy controls. These results are in
concordance with our previous study (28). Of note, E148Q is the predominant
inherited MEFV variant in patients with MM. Pyrin, the
protein product of the MEFV gene, functions in the
modulation of IL-1β and NF-κB. Since IL-1β is important for the
anti-tumor immune response, it has been speculated that genetic
variations that modify the expression of IL-1β may influence the
risk of MM (32). NF-κB is another
important transcription factor for the expression of genes critical
for tumor promotion, cell proliferation, inflammation, metastasis,
angiogenesis, and suppression of apoptosis (33). The function of NF-κB in
lymphopoiesis is well recognized and it is an important factor for
the regulation of cellular homeostasis of T and B lymphocytes
(34–36). Altered NF-κB activation may cause an
increased production of cell cycle regulatory and antiapoptotic
proteins and may contribute to the abnormal proliferation and
survival of neoplastic cells (37–39).
It has been reported that the NF-κB signaling pathway is critical
in myeloma cell proliferation and the inhibition of apoptosis
(40). Furthermore, constitutive
nuclear NF-κB activity has been reported in numerous human MM cell
lines and primary myeloma cells (41,42).
Specifically, as a distinct mechanism, constitutive activation in
the NF-κB signaling pathway or blockade of IL-1β secretion due to
defective pyrin may be associated with an increased frequency of
inherited MEFV variants in patients with MM.
The present study has some limitations. Firstly,
only eight inherited variants in the MEFV gene were screened
in the patients. Rare or novel variants therefore have not been
identified. Secondly, the family members of the patients included
in this study were not screened for the inherited MEFV
variants. However, the individual and family history for FMF
manifestations was negative in these subjects.
In conclusion, a high frequency of inherited
MEFV gene variants was shown to be associated with MM. Based
on the current data, it may be hypothesized that the MEFV
gene is a cancer susceptibility gene. Additional evidence, such as
familial aggregation, monozygotic versus dizygotic twin concordance
and analysis of tumors in genetically-engineered model organisms,
is required in future studies.
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